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AR226-1523 FINAL REPORT RECEIVED OPPT NClC 03 SEP 30 PM1:OO PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 FINAL REPORT DATE: 3 1 JULY 2003 PROTOCOL 418-028 - ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTIONLDEVELOPMENTAL TOXICITY SCREENINGTEST SPONSOR'S STUDY NUMBER: T-7706.1 TABLE OF CONTENTS SUBJECT 1. SUMMARY AND CONCLUSION 1.1. Methods 1.2. Results 1.3. Conclusion 2. DESCRIPTION OF TEST PROCEDURES 2.1. Conduct of Study 2.2. Test Substance Information 2.3. Vehicle Information 2.4. Test Substance Preparation and Storage Conditions 2.5. Test System 2.6. Husbandry 2.7. Methods 3. RESULTS - Male Rats 3.1. Mortality, Clinical and Necropsy Observations 3.2. Terminal Body Weights and Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight and Brain Weight PAGE 1-1 1-1 1-3 1-6 2-1 2-1 2-4 2-4 2-5 2-6 2-7 2-10 3-1 3-1 3-2 .. 11 SUBJECT PAGE 3.3. Hematology and Clinical Chemistry 3-2 3.4. Body Weights and Body Weight Changes 3-3 3.5. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values 3-3 3.6. Mating and Fertility 3-3 3.7. Functional Observational Battery 3-3 3.8. Motor Activity 3-4 3.9. Sperm 3-4 4. RESULTS - Female Rats 4- 1 4.1. Mortality, Clinical and Necropsy Observations 4- 1 4.2. Terminal Body Weights, Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight and Brain Weight and Primordial Follicle Counts 4-2 4.3. Hematology and Clinical Chemistry 4-2 4.4. Body Weights and Body Weight Changes 4-3 4.5 Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values 4-3 4.6. Estrous Cycling, Mating and Fertility 4-4 4.7. Functional Observational Battery 4-4 4.8. Motor Activity 4-4 4.9. Natural Delivery and Litter Observations 4-5 4.10. Pup Clinical and Necropsy Observations 4-5 4.11. Pup Liver Weight and Ratio of Liver Weight to Terminal Body Weight 4-5 REFERENCES 4-6 APPENDIX A - REPORT FIGURES Figure 1. Fo Generation Male Rats A- 1 ... 111 SUBJECT PAGE Figure 2. Fo Generation Female Rats A-2 Figure 3. Motor Activity - Number of Movements - Fo Generation Male Rats A-3 Figure 4. Motor Activity - Time Spent in Movement - Fo Generation Male Rats A-4 Figure 5. Motor Activity - Number of Movements - Fo Generation Female Rats A-5 Figure 6. Motor Activity - Time Spent in Movement - Fo Generation Female Rats A-6 APPENDIX B - REPORT TABLES - FOGENERATION MALE RATS Table B1. Clinical Observations - Summary - Fo Generation Male Rats B-1 Table B2. Necropsy Observations - Summary - Fo Generation Male Rats B-2 Table B3. Terminal Body Weights and Organ Weights - Summary - Fo Generation Male Rats B-3 Table B4. Ratios (%) of Organ Weight to Terminal Body Weight - Summary Fo Generation Male Rats B-5 Table B5. Ratios (%) of Organ Weight to Brain Weight - Summary - Fo Generation Male Rats B-7 Table B6. Hematology - Summary - Fo Generation Male Rats B-9 Table B7. Clinical Chemistry - Summary - Fo Generation Male Rats B-12 Table B8. Body Weights - Summary - Fo Generation Male Rats B-15 Table B9. Body Weight Changes - Summary - Fo Generation Male Rats B-16 Table B 10. Absolute Feed Consumption Values (g/day) - Summary - Fo Generation Male Rats B-17 Table B1 1. Relative Feed Consumption Values (g/kg/day) - Summary - Fo Generation Male Rats B-18 Table B12. Mating and Fertility - Summary - Fo Generation Male Rats B-19 Table B 13. Functional Observational Battery Observations - Summary - Fo Generation Male Rats B-20 iv SUBJECT PAGE Table B14. Motor Activity - Summary - Fo Generation Male Rats B-26 Table B 15. Sperm Motility, Count and Density - Summary - Fo Generation Male Rats B-28 Table B 16. Sperm Morphology - Summary - Fo Generation Male Rats B-29 Table B17. Clinical Observations - Individual Data - Fo Generation Male Rats B-30 Table B 18. Necropsy Observations - Individual Data - Fo Generation Male Rats B-33 Table B 19. Terminal Body Weights and Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight - Individual Data - Fo Generation Male Rats B-38 Table B20. Organ Weights and Ratios (%) Of Organ Weight to Brain Weight - Individual Data - Fo Generation Male Rats B-48 Table B21. Body Weights - Individual Data - Fo Generation Male Rats B-58 Table B22. Feed Consumption Values - Individual Data - Fo Generation Male Rats B-68 Table B23. Mating and Fertility - Individual Data - Fo Generation Male Rats B-71 Table B24. Functional Observational Battery Observations - Individual Data - Fo Generation Male Rats B-74 Table B25. Motor Activity - Individual Data - Fo Generation Male Rats B-79 Table B26. Sperm Motility, Count and Density - Individual Data - Fo Generation Male Rats B-89 Table B27. Sperm Morphology - Individual Data - Fo Generation Male Rats B-92 APPENDIX C - REPORT TABLES - FOGENERATION FEMALE RATS Table C 1. Clinical Observations - Summary - Fo Generation Female Rats c-1 Table C2. Necropsy Observations - Summary - Fo Generation Female Rats c-4 Table C3. Terminal Body Weights and Organ Weights and Primordial Follicle Count - Summarv - Fo Generation Female Rats c-5 V SUBJECT PAGE Table C4. Ratios (%) of Organ Weight to Terminal Body Weight - Summary - Fo Generation Female Rats c-7 Table C5. Ratios (%) of Organ Weight to Brain Weight - Summary - Fo Generation Female Rats C-8 Table C6. Hematology - S u m a r y - Fo Generation Female Rats c-9 Table C7. Clinical Chemistry - Summary - Fo Generation Female Rats c-12 Table C8. Body Weights - Precohabitation - Summary - Fo Generation Female Rats C-15 Table C9. Body Weight Changes - Precohabitation - S u m a r y - Fo Generation Female Rats C-16 Table C10. Maternal Body Weights - Gestation - Summary - Fo Generation Female Rats C-17 Table C l 1. Maternal Body Weight Changes - Gestation - Summary - Fo Generation Female Rats c-19 Table C12. Maternal Body Weights - Lactation - Summary - Fo Generation Female Rats c-20 Table C13. Maternal Body Weight Changes - Lactation - S u m a r y - Fo Generation Female Rats c-22 Table C14. Absolute Feed Consumption Values (g/day) - Precohabitation - Summary - Fo Generation Female Rats C-23 Table C15. Relative Feed Consumption Values (g/kg/day) - Precohabitation - Summary - Fo Generation Female Rats C-24 Table C16. Maternal Absolute Feed Consumption Values (g/day) - Gestation Summary - Fo Generation Female Rats C-25 Table C 17. Maternal Relative Feed Consumption Values (g/kg/day) - Gestation - Summary - Fo Generation Female Rats C-26 Table C18. Maternal Absolute Feed Consumption Values @/day)- Lactation - Summary - Fo Generation Female Rats C-27 vi SUBJECT Table C19. Table C20. Table C21. Table C22. Table C23. Table C24. Table C25. Table C26. Table C27. Table C28. Table C29. Table C30. Table C31. Table C32. Table C33. PAGE Maternal Relative Feed Consumption Values (g/kg/day) - Lactation - Summary - Fo Generation Female Rats C-28 Mating and Fertility, Estrous Cycling and Days in Cohabitation - Summary - Fo Generation Female Rats C-29 Functional Observational Battery - Summary Fo Generation Female Rats C-3 1 Motor Activity - Summary - Fo Generation Female Rats c-37 Natural Delivery Observations - Summary - Fo Generation Female Rats c-39 Litter Observations (Naturally Delivered Pups) - Summary F1 Generation Litters C-40 Clinical Observations from Birth to Day 22 Postpartum - Summary - F1 Generation Pups c-43 Necropsy Observations - Summary - F1 Generation Pups c-44 Pup Liver Weights - Summary - F1 Generation Pups c-45 Clinical Observations - Individual Data - Fo Generation Female Rats C-46 Necropsy Observations - Individual Data Fo Generation Female Rats C-5 1 Terminal Body Weights and Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight - Individual Data - Fo Generation Female Rats c-55 Organ Weights and Ratios (%) Of Organ Weight to Brain Weight - Individual Data - Fo Generation Female Rats C-60 Primordial Follicle Count - Individual Data - Fo Generation Female Rats C-65 Body Weights - Precohabitation - Individual Data - Fo Generation Female Rats C-67 vii SUBJECT PAGE Table C34. Maternal Body Weights - Presumed Gestation - Individual Data Fo Generation Female Rats C-70 Table C35. Maternal Body Weights - Lactation - Individual Data - Fo Generation Female Rats c-75 Table C36. Feed Consumption Values - Precohabitation - Individual Data - Fo Generation Female Rats c-80 Table C37. Maternal Feed Consumption Values - Presumed Gestation - Individual Data - Fo Generation Female Rats c-83 Table C38. Maternal Feed Consumption Values - Lactation - Individual Data Fo Generation Female Rats C-86 Table C39. Mating and Fertility, Estrous Cycling and Days in Cohabitation Individual Data - Fo Generations Female Rats C-89 Table C40. Functional Observational Battery - Individual Data - Fo Generation Female Rats C-92 Table C41. Motor Activity - Individual Data - Fo Generation Female Rats c-97 Table C42. Natural Delivery, Implantation Sites, and Pup Viability and Sex Individual Data - Fo Generation Female RatsP1 Generation Litters C-107 Table C43. Pup Body Weight Litter Averages from Birth to Day 22 Postpartum - Individual Data - F1 Generation Litters c-110 Table C44. Pup Body Weights from Birth to Day 22 Postpartum Individual Data - Fl Generation Pups C-113 Table C45. Pup Vital Status and Sex from Birth to Day 22 Postpartum Individual Data - F1 Generation Pups C-125 Table C46. Clinical Observations from Birth to Day 22 Postpartum Individual Data - F1 Generation Pups c-128 Table C47. Necropsy Observations - Individual Data - F l Generation Pups C-130 Table C48. Pup Liver Weights - Individual Date - F1 Generation Pups C-136 APPENDIX D - PROTOCOL AND AMENDMENTS D-1 to D-52 ... Vlll SUBJECT PAGE APPENDIX E - DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY E-1 to E-3 APPENDIX F - CERTIFICATE OF ANALYSIS F-1 to F-3 APPENDIX G - ANALYTICAL AND BIOANALYTICAL REPORT G-1 to G-153 APPENDIX H - TEMPERATUREAND RELATIVE HUMIDITY REPORT H- 1 APPENDIX I - POSITIVE CONTROL DATA 1-1 to 1-4 APPENDIX J - HISTOPATHOLOGY REPORT J-1 to J-105 APPENIDX K - HEMATOLOGY AND CLINICAL CHEMISTRY REPORTS K-1 to K-150 APPENDIX L - STATEMENTOF THE STUDY DIRECTOR L- 1 APPENDIX M - QUALITY ASSURANCE STATEMENT M-1 to M-2 ix 418-028:PAGE 1-1 TITLE: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTIONDEVELOPMENTALTOXICITY SCREENING TEST ARGUS RESEARCH PROTOCOL NUMBER: 418-028 SPONSOR'S STUDY NUMBER: T-7706.1 1. SUMMARY AND CONCLUSION 1.1. Methodsa Seventy-five Crl:CDB(SD)IGS VAFPlusB rats per sex were assigned to five dosage groups (Groups I through V), 15 rats per sex per group. An additional three rats per sex per group were assigned to Groups I through V for toxicokinetic sample collection. The test substance, T-7706 [Perfluorohexane Sulfonate Potassium Salt (PFHS)],or vehicle, aqueous 0.5% carboxymethylcellulose (CMC), was administered via gavage to male rats once daily beginning 14 days before cohabitation and continuing through the day before sacrifice, after completion of the cohabitation period, after a minimum of 42 days of administration, and to female rats once daily beginning 14 days before cohabitation and continuing through the day before sacrifice, day 21 of lactation (DL 21) or day 25 of presumed gestation (DG 25, rats that did not deliver a litter). Dosages were 0,0.3, 1, 3 and 10 mg/kg/day. The dosage volume, 10 d k g , was adjusted daily on the basis of the individual body weights recorded before intubation. F1 generation pups were not directly administered the test substance or vehicle. Within each dosage group, rats were assigned to cohabitation, one male rat per female rat. Rats were observed for viability at least twice each day of the study. Observations for clinical signs of effects of the test substance and deaths were made on the first day of dosage at approximately hourly intervals for the first four hours and at the end of the normal working day. Observations for clinical signs of effects of the test substance and deaths were made on subsequent days daily before dosage and approximately 60 k 10 minutes after dosage administration and on the day of sacrifice. Once before the first dosage and at least once weekly thereafter, detailed clinical observations were conducted for all male and female rats assigned to the main study. a. Detailed descriptions of all procedures used in the conduct of this study are provided in the appropriate sections of this report and in APPENDIX D (PROTOCOL AND AMENDMENTS). 418-028:PAGE 1-2 Body weights were recorded daily during the dosage period and at sacrifice. Feed consumption values for male rats assigned to the main study were recorded weekly during the dosage period. Feed consumption values for female rats assigned to the main study were recorded weekly to cohabitation, on DGs 0,7, 10, 12, 15, 18,20 and 25 (if necessary) and on DLs 1 , 5 , 8 and 15. During cohabitation, individual values were not recorded or tabulated. Estrous cycling was evaluated in rats assigned to the main study by examination of vaginal cytology beginning with the day after the first administration and then until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ during the cohabitation period. Estrous cycling was evaluated in rats assigned to the toxicokinetic study during the cohabitation period until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ. Female rats were evaluated for adverse clinical signs observed during parturition, duration of gestation, litter sizes and pup viability at birth. Maternal behavior was evaluated on DLs 1,5,8,15 and 22. Shortly before scheduled sacrifice, a functional observational battery (FOB) was conducted and motor activity was evaluated on 10 male and 10 female rats per group. On days 14 and 42 of study, blood samples were collected from each male rat assigned to the toxicokinetic sample collection portion of the study and on day 14 of study and DG 21, blood samples were collected from each female rat assigned to the toxicokinetic sample collection portion of the study. Each litter was evaluated for viability at least twice daily. The pups in each litter were counted once daily. Clinical observations were recorded once daily. Pup body weights were recorded on DLs 1,8, 15 and 22. Male and female rats assigned to the toxicokinetic study were sacrificed on day 42 of study and DG 2 1, respectively. Liver weights were recorded. The median liver lobe was shipped for analysis. Blood samples were collected from each fetus and pooled by litter and serum was shipped for analysis. The liver from each fetus was collected, pooled per litter and shipped for analysis. The number of implantation sites was recorded. Male and female rats assigned to the main study were sacrificed after a minimum of 42 days of dosage and on DL 22, respectively. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The number of implantation sites were recorded. Gross lesions were examined histologically. Ten rats per sex per group assigned to functional observational battery and motor activity tests were assigned to histological evaluations. The following organs were individually weighed: liver, kidneys, adrenals, thymus, testes, right epididymis, left epididymis (corpus and caput), seminal vesicles (with and without fluid), prostate, spleen, brain, heart, ovaries and uterus (with cervix). The following tissues or representative samples 4 18-028:PAGE 1-3 were retained: brain, small and large intestines, lungs, lymph nodes, peripheral nerve, stomach, kidneys, spleen, thymus, trachea, urinary bladder, spinal cord, liver, adrenals, heart, thyroid/parathyroid, bone marrow, testes, prostate, seminal vesicles, ovaries, uterus, vagina, mammary gland (female rats only) and gross lesions. Histological examination of retained tissues, including reproductive organs, was conducted for the assigned ten rats per sex from the control and high dosage groups. A quantitative evaluation of primordial follicles was conducted for Fo generation female rats. Sperm evaluations (concentration, motility and morphology) were performed for 10 male rats in each dosage group. At scheduled sacrifice, blood samples were collected from the 10 male and 10 female rats per group assigned to hematology and clinical chemistry sample collection. The following hematologic parameters were evaluated: erythrocyte count, hematocrit, hemoglobin, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, mean corpuscular volume, total leukocyte count, differential leukocyte count, platelet count, mean platelet volume and cell morphology. Two blood smear slides were prepared for measurements of differential leukocyte count. Plasma samples evaluated for prothrombin time and activated partial thromboplastin time. Sera samples were evaluated for total protein, triglycerides, albumin, globulin, albumirdglobulin ratio, glucose, cholesterol, total bilirubin, urea nitrogen, creatinine, creatinine kinase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, calcium, phosphorus, sodium, potassium and chloride. On DL 22, pups were sacrificed and examined for gross lesions. Necropsy included a single cross-section of the head at the level of the frontal-parietal suture and examination of the cross-sectioned brain for apparent hydrocephaly. Blood samples were collected from five pups per sex per litter from the 10 female rats per group selected for FOB and motor activity assessment, blood sample collection for hematology and clinical chemistry and histological evaluations. Sera was shipped for analysis. The liver from each selected pup was weighed. 1.2. Results 1.2.1. Male Rats All male rats survived to scheduled sacrifice and all clinical and necropsy observations were considered unrelated to the test substance. Body weight gains were significantly reduced in the 0.3,3 and 10mg/kg/day dosage groups on study days (DSs) 29 to 36. Significantlyreduced body weight gain occurred in the 0.3, 1 , 3 and 10 mg/kg/day dosage groups on study days DS 29 to termination. Body weight gain was also significantly reduced for the 10 mg/kg/day dosage group for the entire dosage period. Absolute and relative feed consumption values for the male rats were unaffected by dosages of the test substance as high as 10 mg/kg/day. 418-028:PAGE 1-4 Terminal body weights of the male rats were slightly reduced in the 10 mg/kg/day dosage group. The absolute weights of the liver, the ratios of the liver weights to terminal body weights and ratios of the liver weights to brain weight were significantly increased in the 3 and 10 mg/kg/day dosage groups. The ratio of the heart weights to brain weight was significantly decreased in the 10 mg/kg/day dosage group. Hemoglobin concentrations were significantly decreased in the 1 , 3 and 10 mg/kg/day dosage groups and average values for red blood cells and hematocrit were significantly decreased in the 3 and 10 mg/kg/day dosage groups. Prothrombin time was significantly increased in the 0.3,3 and 10 mg/kg/day dosage groups. Average values for cholesterol were significantly decreased in the 0.3, 1 , 3 and 10mg/kgday dosage groups and the average values for triglycerides was significantly decreased in the 10 mg/kg/day dosage group. Albumin, blood urea nitrogen, alkaline phosphatase, calcium and albumin/globulin ratio levels were significantly increased in the 10 mg/kg/day dosage group. Treatment-related microscopic changes were observed in the liver and thyroid gland of male rats in the 3 and 10 mg/kg/day dosage groups. The treatment-related microscopic changes in the liver consisted of minimal to moderate enlargement of centrilobular hepatocytes and in the thyroid gland as an increased incidence of hypertrophy of follicular cells and hyperplasia. These microscopic changes are consistent with the known effects of compounds that cause microsomal enzyme induction where the hepatocellular hypertrophy results in a compensatory hypertrophy and hyperplasia of the thyroid due to increased plasma turnover of thyroxine and associated stimulation of thyroid-stimulating hormone in rats? There were no treatment-related microscopic changes observed in the liver of the F1 generation pups from the dams given up to 10 mg/kg/day of the test substance. All mating and fertility parameters were unaffected by dosages of the test substance as high as 10 mg/kg/day. There were no other statistically significant or biologically important differences among the four dosage groups in the measures of the functional observational battery (FOB) or motor activity on DSs 36 through 39. Sperm motility was unaffected by dosages of the test substance as high as 10 mg/kg/day. The sperm count and sperm density were comparable among the five dosage groups. 1.2.2. Female Rats All female rats survived to scheduled sacrifice and all clinical and necropsy observations were considered unrelated to the test substance. Body weights, body weight gains, and absolute and relative feed consumption values were comparable and did not differ significantly during the precohabitation, gestation or lactation periods at dosages of the test substance up to 10 mg/kg/day. Terminal body weights, absolute and relative weights of the reproductive organs, brain, liver, left and right kidneys and adrenals, spleen, thymus and heart of the female rats were 418-028:PAGE 1-5 comparable among dosage groups. Average primordial follicle counts for the 10 mg/kg/day dosage group were comparable to the control group. No treatment-related microscopic changes were observed in any of the female rats administered up to 10 mg/kg/day of the test substance. There were no treatment-related microscopic changes observed in the liver of the Fl generation pups from the dams given up to 10 mg/kg/day of the test substance. Dosages as high as 10 mg/kg/day did not affect any hematology or clinical chemistry values evaluated. The average numbers of estrous stages per 13 days were comparable among the five dosage groups. All mating and fertility parameters, including the gestation index, viability index and lactation indices were unaffected by dosages of the test substance as high as 10 mg/kg/day. There were no statistically significant or biologically important differences among the five dosage groups in the measures of the functional observational battery (FOB) or motor activity on DL 17. All pregnant dams delivered a litter of one or more live pups. Values for the numbers of dams delivering litters, the duration of gestation, averages for implantation sites per delivered litter, the numbers of dams with stillborn pups, the numbers of dams with no liveborn pups, dams with all pups dying, were comparable among the five dosage groups. The number of pups surviving per litter, pup sex ratios, litter size and pup body weights per litter were comparable among the five dosage groups. No clinical or necropsy observations in the F1 generation pups were attributable to dosages of the test substance as high as 10 mg/kg/day. F1 generation male and female pup terminal body weights, absolute liver weight and ratio of liver weight to terminal body weight were comparable across all five dosage groups. 418-028:PAGE 1-6 1.3. Conclusion On the basis of these data, t,ie maternal no-observable-adverse-effect-leve(lNOAEL) for T-7706 [Perfluorohexane Sulfonate Potassium Salt] is greater than 10 mg/kg/day (the 10 mg/kg/day dosage caused no deaths, adverse clinical or necropsy/pathology observations, changes in body weight, feed consumption, or hematology or clinical chemistry values throughout precohabitation, gestation or lactation. The paternal NOAEL is less than 0.3 mg/kg/day (the 0.3 and 1 mg/kg/day dosages caused significant differences in body weight gain, hematology and clinical chemistry values and the 3 and 10 mg/kg/day dosages caused significant changes in absolute and relative organ weights and microscopic changes in the liver and thyroid gland). The reproductive NOAEL is greater than 10 mgkg/day (the 10 mg/kg/day dosage had no effect on the durations of gestation and parturition or any mating and fertility parameters. There was no effect on the sperm parameters in the male rats). The NOAEL for viability and growth in the offspring is also greater than 10 mg/kg/day (dosages of 10 mg/kg/day had no effect on perinatal mortality, clinical or necropsy observations or body or liver weights in the Fl generation offspring). Alan M. Hoberman, Ph.D., DABT Date Director of Research Study Director Date Study Director 4 18-028:PAGE 2- 1 2. DESCRIPTION OF TEST PROCEDURES 2.1. Conduct of Study 2.1.1. Sponsor 3M Corporate Toxicology, 3M Center, Building 220-2E-02, St. Paul, Minnesota 55144-1000 2.1.2. Testing Facility Argus Research, 905 Sheehy Drive, Building A, Horsham, Pennsylvania 19044-1297 2.1.3. Study Number 418-028 2.1.4. Sponsor's Study Number T-7706.1 2.1.5. Purpose of the Study The purpose of this study was to provide information on the possible health hazards that may result from repeated exposure of Crl:CDB(SD)IGS BR VAFPlusB male and female rats to a test substance beginning before cohabitation, through mating and continuing for at least 42 days (male rats) or through parturition until day 21 of lactation (female rats). This repeated dose study incorporated a reproductioddevelopmental toxicity screening test that can be used to provide initial infomation on possible effects on male and female reproductive performance (e.g., gonadal function, mating behavior, conception, development of the conceptus and parturition). The study also placed emphasis on neurological effects as a specific endpoint and should identify the neurotoxic potential of a test substance, which may warrant further in-depth investigation. Because of the selectivity of the endpoints and the short duration of the study, the screening test did not provide evidence for definitive claims of no reproductive/ developmental effects. In particular, it offered only limited means of detecting postnatal manifestations of prenatal exposure or effects that may be induced during postnatal exposure. 2.1.6. Study Design . The requirements of the Organisation for Economic Co-operation and Development (OECD)'2' were used as the basis for study design. 2.1.7. Regulatory Compliance This study was conducted in compliance with Good Laboratory Practice (GLP) regulations of the OECD'3',U.S. Food and Drug Administration (FDA)'4', the Japanese 4 18-028:PAGE 2-2 Ministry of Health and Welfare (MHW)? There were no deviations from the GLP regulations that affected the quality or integrity of the study. Quality Assurance Unit findings derived from the inspections during the conduct of this study are documented and have been provided to the Study Director and the Testing Facility Management. 2.1.8. Ownership of the Study The Sponsor owns the study. All raw data, analyses, reports and preserved tissues are the property of the Sponsor. 2.1.9. Study Monitor John Butenhoff, Ph.D., CIH, DABT 2.1.10. Study Director Raymond G. York, Ph.D., DABT (Associate Director of Research) Address as cited previously for Testing Facility. 2.1.11. Technical Performance John F. Barnett, B.S. (Director of Laboratory Operations) Joseph W. Lech, B.S. (Senior Research Associate) Mary P. Howard, B.S. (Team Leader - General Laboratory) Jaclyn S. Fox, B.S. (Laboratory Technician) Josette M. Provost, B.S. (Necropsy Laboratory Technician/Fixed Tissue Coordinator) Christopher K. Ruppert, B.S.(Formulation Laboratory Technician) 2.1.12. Report Preparation Raymond G. York, Ph.D., DABT Jo Ann Frazee, M.S. (Study Coordinator) JoAnne M. Conklin, B.S. (Data Management Specialist) Cristina Petrescu (Report Administrator) 2.1.13. Report Review Valerie A. Sharper, M.S. (Director of Study Management) 2.1.14. Date Protocol Signed 26 March 2002 2.1.15. Dates of Technical Performance 2.1.15.1. Male Rats Rat Arrival Dosage Period (14 days before cohabitation and through a 14-day cohabitation period until sacrifice after at least 42 days of dosage) FOB and Motor Activity Evaluation Toxicokinetic Sample Collections DSa 14 DS 42 Scheduled Sacrifice - Toxicokinetic Study Scheduled Sacrifice - Main Study 2.1.15.2. Female Rats Rat Arrival Dosage Period (14 days before cohabitation through DLb 21) Cohabitation Period Male 1 Male 2 Toxicokinetic Sample Collections DS 14 DGC21 DG 0 DG 25 Sacrifice (Rats that did not deliver a litter) Delivery Periodd(DL 1) FOB and Motor Activity Evaluation DL 22 Sacrifice Female Rats and Pups 418-028:PAGE 2-3 26 MAR 02 01 APR 02 - 16 MAY 02 06 MAY 02 - 09 MAY 02 14 APR 02 12 MAY 02 12 MAY 02 13MAY 02 - 17 MAY 02 26 MAR 02 01 APR 02 - 09 JUN 02 14 APR 02 PM - 21 APR 02 AM 21 APR 02 PM - 28 APR 02 AM 14 APR 02 06 MAY 02 - 19 MAY 02 15 APR 02 - 28 APR 02 10MAY 02 - 23 MAY 02 07 MAY 02 - 20 MAY 02 23 MAY 02 - 26 MAY 02 28 MAY 02 - 10 JUN 02 a. DS is an abbreviation used for day of study. b. DL is an abbreviation used for day of lactatiodpostpartum. c. DG is an abbreviation used for day of (presumed) gestation. d. The day of birth is designated lactation day 0 (postpartum day 0) in the Health Effects Test Guidelines - Reproduction and Fertility Effects (Office of Prevention, Pesticides and Toxic Substances 870.3800, August, 1998) and in the OECD Guideline for the Testing of Chemicals - Combined Repeated Dose Toxicity Study with the ReproductionDeveloprnentalToxicity Screening Test (Section 4, No. 422,22 March 1966). This same day is designated day 1postpartum (day 1 of lactation) in the Standard Operating Procedures of the Testing Facility. Throughout this study, the day of birth was designated as day 1 postpartum (day 1 of lactation) and all subsequent ages of the F1 generation rats and days of the lactation period were determined and cited accordingly. 418-028:PAGE 2-4 .2.1 16. Records Maintained The original report, raw data and reserve samples of each lot of bulk test substance and bulk vehicle components are retained in the archives of Argus Research. Any preserved tissues are retained in the archives of the Testing Facility for one year after the mailing of the draft final report, after which time the Sponsor will decide their final disposition. All unused prepared formulations were discarded at the Testing Facility. All remaining bulk test substance was returned to the Sponsor. 2.2. Test Substance Information 2.2.1. Description T-7706 [Perfluorohexane Sulfonate Potassium Salt (PFHS)] - a white powder 2.2.2. Date Received and Storage Conditions The test substance was received on 11 March 2002 and stored at room temperature. 2.2.3. Special Handling Instructions Standard safety precautions (use of protective clothing, gloves, dust-mist/HEPA-filtered mask, safety goggles or safety glasses and a face-shield) were taken during formulation preparation and dosage. A half-face respirator was worn during formulation preparation. 2.2.4. Analysis of Purity Information to document or certify the identity, composition, method of synthesis, strength and purity of the test substance was provided by the Sponsor to the Testing Facility. A Certificate of Analysis is available in APPENDIX F. 2.3. Vehicle Information 2.3.1. Description Aqueous 0.5% carboxymethylcellulose (CMC) (sodium salt; medium viscosity) prepared using carboxymethylcellulose (sodium salt; medium viscosity), an off-white powder, and reverse osmosis membrane processed deionized water (R.O. deionized water). 2.3.2. Lot Number 120K0252 2.3.3. Date Received and Storage Conditions The carboxymethylcellulose was received from Sigma Chemical Co., St. Louis, Missouri, on 11 September 2001 and stored at room temperature. R.O. deionized water is available from a continuous source at the Testing Facility and is maintained at room temperature. 4 18-028:PAGE 2-5 2.3.4. Special Handling Instructions Standard safety precautions (use of protective clothing, gloves, dust-mist/HEPA-filtered mask, safety goggles or safety glasses and a face-shield) were taken when handling the vehicle components and prepared vehicle. 2.3.5. Analysis of Purity Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the vehicle that would have interfered with the results of this study. The expiration date for the carboxymethylcelluloseis September 2005. 2.4. Test Substance Preparation and Storage Conditions Formulations were prepared weekly at the Testing Facility. Prepared test substance and vehicle formulations were stored refrigerated (2C to 8C). 2.4.1. Sample Information 1 1 Date Storage Conditions Shipped To/ Shipping Conditions 1 Date Shimed (all levels) Room temperature Refrigerated Exygenb/ Ambient conditions Sponsor/ Refrigerated 10 JUN 02 01 APR02 Concentrationd Bulk Test Substance Reserve Refrigerated Exygenb/ 28 MAY 02 Refrigerated 10 JUN 02 Refrigerated Sponsor/ 01 APR 02 Refrigerated l g 10JUN02 Room temperature Testing Facility 10JUN 02 Archives Vehicle Components Room temperature Testing Facility Reserve Archives Carboxymethylcellulose 1 g 10 JUN 02 R.O. deionized water 5 m L 10JUN02 10 JUN 02 I 10 JUNO2 a. A sample of the test article was retained on th :last day of treatmen :and shipped for analysis. b. Exygen Research, State College, Pennsylvania. C. Quadruplicatesamples were taken from the top, middle and bottom of each concentration on the first day of preparation. Two samples from each quadruplicate set were shipped for analysis. The remaining samples were retained at the Testing Facility as backup samples. d. Quadruplicate samples were taken from each concentration on the last day of preparation. Two samples from each quadruplicate set were shipped for analysis. The remaining samples were retained at the Testing Facility as backup samples. e. Samples for 0.03,O.l and 1mg/mL concentrationsonly. f. Samples for 0 and 0.3 mg/mL concentrations only. g. Two sets of duplicate samples from each concentration were taken on the first day of preparation. One sample of each duplicate set was shipped on the day of preparation. These samples were analyzed as soon as possible after preparation and ten days after the first analysis. The remaining samples were retained at the Testing Facility as backup samples. 418-028:PAGE 2-6 2.4.2. Analytical Results Results of the analytical analyses are available in APPENDIX G. 2.5. Test System 2.5.1. Species Rat 2.5.2. Strain Crl:CD8(SD)IGS VAF/Plus8 2.5.3. Supplier (Source) Charles River Laboratories, Inc. Male Rats - St. Constant, Quebec, CANADA Female Rats - Raleigh, North Carolina 2.5.4. Sex Male and Female 2.5.5. Rationale for Test System The Crl:CDB(SD)IGS BR VAF/PlusB rat was selected as the Test System because: 1) it is one mammalian species accepted for use in toxicity studies and it has been widely used throughout industry; 2) this strain of rat has been demonstrated to be sensitive to reproductive and developmental toxins; and 3) historical data and experience exist at the Testing 2.5.6. Test System Data 2.5.6.1. Male Rats Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) the Day after Arrival Weight (g) at Study Assignment 100 20 JAN 02 66 days 277 - 318 305 - 348 2.5.6.2. Female Rats Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) the Day after Arrival Weight (g) at Study Assignment 100 21 JAN02 65 days 195 - 227 207 - 230 418-028:PAGE 2-7 2.5.7. Method of Randomization Upon arrival, the male and female rats were assigned to individual housing on the basis of computer-generated random units. During an acclimation period of at least five days, male and female rats were selected for study on the basis of physical appearance and body weights recorded during acclimation. The rats were assigned to five dosage groups (Groups I through V), 15 rats per sex per group, using a computer-generated (weightordered) randomization procedure. An additional three rats per sex per group were assigned to Groups I through V for toxicokinetic sample collection. At study initiation, the weight variation of the rats did not exceed & 20% of the mean weight of each sex. Litters were not culled during the lactation period because random selection of pups for culling could have resulted in potential biases in pup viabilities and body weight gains over this period. Within each dosage group, consecutive order was used to assign the first 10 male and the first 10 female Fo generation rats to a functional observational battery (FOB) and motor activity assessment, blood sample collection for clinical chemistry and hematology and histological evaluations. On DL 22, a table of random units was used to select five male and five female pups per litter for blood sample and liver collection. These pups were only selected from the 10 dams selected for FOB, motor activity, clinical chemistry and hematology and histological evaluation. 2.5.8. System of Identification Male and female rats were assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study before administration of the first dosage. Rats were permanently identified using a MonelB self-piercing ear tag (Gey Band and Tag Co., Inc., No. MSPT 20101). Cage tags were marked with the study number, permanent rat number, sex, test substance identification, generation, dosage group and dosage level. Pups were not individually identified during lactation; all parameters were evaluated in terms of the litter. 2.6. Husbandry 2.6.1. Research Facility Registration USDA Registration No. 14-R-0144under the Animal Welfare Act, 7 U.S.C. 2131 et seq. 2.6.2. Study Room The study room was maintained under conditions of positive airflow relative to a hallway and independently supplied with a minimum of ten changes per hour of 100% fresh air that had been passed through 99.97% HEPA filters. Room temperature and humidity 4 18-028:PAGE 2-8 were monitored constantly throughout the study. Room temperature was targeted at 64F to 79F (18C to 26C); relative humidity was targeted at 30% to 70%". 2.6.3. Housing Rats were individually housed in stainless steel, wire-bottomed cages except during cohabitation and postpartum periods. During cohabitation, each pair of male and female rats was housed in the male rat's cage. Beginning no later than DG 20, Fo generation female rats were individually housed in nesting boxes until they either naturally delivered litters or were sacrificed (DG 25). Each dam and delivered litter was housed in a common nesting box during the postpartum period. All cage sizes and housing conditions were in compliance with the Guidefor the Care and Use of Laboratory animal^'^'. Argus Research is an AAALAC-accredited facility. 2.6.4. Lighting An automatically-controlledfluorescent light cycle was maintained at 12-hourslight: 12-hoursdark, with each dark period beginning at 1900hours EST. 2.6.5. Sanitization Cage pan liners were changed at least three times weekly. Cages were changed approximately every other week. Bedding was changed as often as necessary to keep the rats dry and clean. 2.6.6. Feed Rats were given ad libitum access to Certified Rodent Diet8#5002 (PMI Nutrition International, Inc., St. Louis, Missouri) in individual feeders. Feed was removed the evening prior to the scheduled sacrificeb. 2.6.7. Feed Analysis Analyses were routinely performed by the feed supplier. No contaminants at levels exceeding the maximum concentration for certified feed or deviations from expected nutritional requirements were detected by these analyses. Copies of the results of the feed analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the feed at concentrations that would have interfered with the results of this study. a. See APPENDIX H (TEMPERATURE AND RELATIVE HUMIDITY REPORT). b. See APPENDIX E (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY), item 1. 418-028:PAGE 2-9 2.6.8. Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad Libitum from individual water bottles attached to the cages and/or from an automatic watering access system. Chlorine was added to the processed water as a bacteriostat. 2.6.9. Water Analysis The processed water is analyzed twice annually for possible chemical contamination (Lancaster Laboratories, Lancaster, Pennsylvania) and monthly for possible bacterial contamination (Analytical Laboratories, Inc., Chalfont, Pennsylvania). Copies of the results of the water analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the water that would have interfered with the results of this study. 2.6.10. Bedding Material Bed-o'cobs8 bedding (The Andersons Industrial Products Group, Maumee, Ohio) was used as the nesting material. 2.6.11. Bedding Analysis Analyses for possible contamination are conducted semi-annually. Copies of the restults of the bedding analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the bedding that would have interfered with the results of this study. 418-028:PAGE 2-10 2.7. Methods 2.7.1. Dosage Administration Dosage Dosagea :oncentratio Dosage Number Volume of Rats Assigned Numbers Group I mg/kgfday 0 (mg/d) 0 ( d k ) per Sex 10 15 + 3b Male Rats Main Study: Female Rats ~~ Main Study: 19109,19115,19116,19119, 19010,19012,19019,19021, 19122,19125,19131,19132, 19023,19041,19042,19044, 19134,19135,19143,19151, 19050,19053,19065,19068, 19157,19161,19167 19072,19074,19075 Toxicokinetic Study: Toxicokinetic Study: 19176 - 19178 I1 0.3 0.03 10 15 + 3b Main Study: 19076 - 19078 Main Study: 19102,19106,19108,19110, 19004,19009,19016,19018, 19113,19120,19129,19136, 19026,19036,19037,19043, 19138,19139,19147,19153, 19047,19048,19052,19055, 19164,19165,19171 19061,19067,19071 Toxicokinetic Study: Toxicokinetic Study: 19179 - 19181 19079 - 19081 I11 0.1 10 15 + 3b Main Study: Main Study: 19101,19105,19107,19112, 19003,19007,19008,19013, 19114,19121,19123,19130, 19014,19015,19017,19024, 19137,19146,19155,19159, 19029,19034,19038,19056, 19172,19173,19174 19057,19060,19064 Toxicokinetic Study: Toxicokinetic Study: 19182 - 19184 19082 - 19084 IV 0.3 10 15 + 3b Main Study: Main Study: 19100,19103,19104,19118, 19002,19005,19035,19039, 19133,19141,19142,19144, 19040,19045,19046,19051, 19148,19150,19156,19160, 19054,19058,19062,19063, 19162,19163,19166 19066,19069,19073 V 10 Toxicokinetic Study: Toxicokinetic Study: 19185 - 19187 19085 - 19087 1 10 15 + 3b Main Study: Main Study: 19111,19117,19124,19126, 19001,19006,19011,19020, 19127,19128,19140,19145, 19022,19025,19027,19028, 19149,19152,19154,19158, 19030,19031,19032,19033, 19168,19169,19170 19049,19059,19070 Toxicokinetic Study: Toxicokinetic Study: 19188 - 19190 19088 - 19090 a. e test substance was coni xed 100' pure for i e purpose of dosage calculations. b. Three additional rats per sex were assigned to toxicokinetic sample collection. 2.7.2. Rationale for Dosage Selection Dosages were selected by the Sponsor based on previous studies conducted with the test substance, taking into account possible differences in sensitivity between pregnant and nonpregnant rats. The highest dosage was expected to cause toxic effects but not mortality or obvious suffering. The descending sequence of the lower dosage levels was selected for the purpose of demonstrating any dosage-related response, with no adverse effects expected at the lowest level. 418-028:PAGE 2-11 2.7.3. Route and Rationale for Route of Administration The oral (gavage) route was selected for use because: 1) in comparison with the dietary route, the exact dosage can be accurately administered; and 2) it is one of the possible routes for environmental exposure. 2.7.4. Frequency of Administration 2.7.4.1. Fo Generation Rats Male rats were administered the test substance or the vehicle once daily beginning 14 days before cohabitation (maximum 14 days) and continuing through the day before sacrifice, after completion of the cohabitation period, after a minimum of 42 days of administration. Female rats were administered the test substance or the vehicle once daily beginning 14 days before cohabitation (maximum 14 days) and continuing through the day before sacrifice, DL 21 or DG 24 (rats that did not deliver a litter). The dosage volume was adjusted daily on the basis of the individual body weights recorded before intubation. The rats were intubated once daily at approximately the same time each day. The first day of dosage was designated as DS 1. Dams in the process of delivering pups were not intubated until completion of parturition, in order to preclude possible disruption of maternal behavior and/or cannibalization of pups. Consequently, some dams were not administered one daily dosage during the delivery period. No dam missed more than one daily dosage. 2.7.4.2. F1 Generation Pups F1 generation pups were not directly administered the test substance or vehicle, but may have been possibly exposed to the test substance or vehicle during maternal gestation (in utero exposure) or via maternal milk during the lactation period. 2.7.5. Method of Study Performance 2.7.5.1. Fo Generation Rats Within each dosage group, consecutive order was used to assign rats to cohabitation, one male rat per female rat. The cohabitation period consisted of a maximum of 14 days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug in situ were considered to be DG 0 and assigned to individual housing. Female rats not mated with a male rat within the first seven days of cohabitation were assigned an alternate male rat that had mated (same dosage group) and remained in cohabitation for a maximum of seven additional days. Rats were observed for viability at least twice each day of the study. Rats were examined for clinical observations and general appearance weekly during the acclimation period. Observations for clinical signs of effects of the test substance and deaths were made on the first day of dosage at approximately hourly intervals for the first four hours and at the end of the normal working day. Observations for clinical signs of effects of the test 418-028:PAGE 2-12 substance and deaths were made on subsequent days daily before dosage and approximately 60 f 10 minutes after dosage administration and on the day of sacrificea. Once before the first dosage and at least once weekly thereafter, detailed clinical observations were conducted for all male and female rats assigned to the main studyb. These observations were made outside the cage in a standard arena at the same time each day of conduct. Effort was made to ensure that variations in the test conditions were minimal and that observations were conducted by observers unaware of treatment groups. The rats were observed for (but observations were not limited to) the following signs: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g., lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypic behavior (e.g., excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behavior (e.g., self-mutilation, walking backwards) were also recorded. Body weights were recorded weekly during the acclimation period, daily during the dosage period and at sacrifice. Feed consumption values for male rats assigned to the main study were recorded weekly during the dosage period. Feed consumption values for female rats assigned to the main study were recorded weekly to cohabitation, on DGs 0, 7,10,12,15,18,20 and 25 (if necessary) and on DLs 1 , 5 , 8 and 15'. Because pups begin to consume maternal feed on or about DL 15, feed consumption values were not tabulated after DL 15. During cohabitation, when two rats occupied the same cage with one feed jar, replenishment of the feed jars was documented but individual values were not recorded or tabulated. Estrous cycling was evaluated in rats assigned to the main study by examination of vaginal cytology beginning with the day after the first administration and then until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ during the cohabitation period. Estrous cycling was evaluated in rats assigned to the toxicokinetic study during the cohabitation period until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ. Female rats were evaluated for adverse clinical signs observed during parturition, duration of gestation (DG 0 to the day the first pup was observed), litter sizes (all pups delivered) and pup viability at birth. Maternal behavior was evaluated on DLs 1,5,8, 15 and 22. Variations from expected maternal behavior were recorded, if and when present, on all other days of the postpartum period. On one occasion during the course of the study, a functional observational battery (FOB)(10-13w)as conducted on 10 male and 10female rats per group. For male rats, this assessment was conducted shortly before scheduled sacrifice, but prior to blood sample a. See APPENDIX E, items 2 and 3. b. See APPENDIX E, item 4. C. See APPENDIX E, item 5. 418-028:PAGE 2-13 collection for hematology and clinical chemistry evaluations. Female rats were tested during the lactation period, shortly before scheduled sacrifice. The FOB evaluation was conducted by an observer unaware of the group assignment of the rat. The following parameters were assessed: 1. Lacrimation, salivation, palpebral closure, prominence of the eye, pupillary reaction to light, piloerection, respiration, and urination and defecation (autonomic functions). 2. Sensorimotor responses to visual, auditory, tactile and painful stimuli (reactivity and sensitivity). 3. Reactions to handling and behavior in the open field (excitability). 4. Gait pattern in the open field, severity of gait abnormalities, air righting reaction and landing foot splay (gait and sensorimotor coordination). 5. Forelimb and hindlimb grip strength. 6. Abnormal clinical signs including but not limited to convulsions, tremors and other unusual behavior, hypotonia or hypertonia, emaciation, dehydration, unkempt appearance and deposits around the eyes, nose or mouth. The ability of this battery to detect the effects of positive control substances has been established (Testing Facility Positive Control Data) and is available in APPENDIX I. Motor activity was evaluated on 10 male and 10 female rats per group once, shortly before scheduled sacrifice, prior to blood sample collection. The movements of each rat were monitored by a passive infrared sensor mounted outside a stainless steel, wirebottomed cage (40.6 x 25.4 x 17.8 cm). Each test session was 1.5 hours in duration with the number of movements and time spent in movement tabulated at each five-minute interval. The apparatus monitored a rack of up to 32 cages and sensors during each session, with each rat tested in the same location on the rack across test sessions. Groups were counterbalanced across testing sessions and cages. Data demonstrating that the test system is capable of detecting increases in activity produced by positive control substances (Testing Facility Positive Control Data) is available in APPENDIX I. On DSs 14 and 42, blood samples (approximately 1 mL each) were collected from each male rat assigned to the toxicokinetic sample collection portion of the study (three rats per group) and on DS 14 and DG 21, blood samples (approximately 1 mL each) were collected from each female rat assigned to the toxicokinetic sample collection portion of the study (three rats per group). Samples were collected prior to dosage on DS 14. The time of each blood collection was recorded in the raw data. Blood was collected from the orbital sinus on DS 14 of the study and from the vena cava on DS 42 of study (male rats) and DG 21 (female rats). The samples were transferred into EDTA-coated (purple top) tubes and spun in a centrifuge. The resulting serum was transferred into polypropylene 418-028:PAGE 2-14 tubes labeled with the protocol number, Sponsor study number, rat number, sex, group number, dosage level, day of study, collection interval, date of collection, species, generation and storage conditions. All samples were immediately frozen on dry ice and maintained frozen (5-70C) until shipment to Exygen Research, State College, Pennsylvania, for analysis. Results of these analyses are available in APPENDIX G. 2.7.5.2. F1 Generation Pups Day 1 of lactation (postpartum) was defined as the day of birth and was also the first day on which all pups in a litter were individually weighed (pup body weights were recorded after all pups in a litter were delivered and groomed by the dam). Each litter was evaluated for viability at least twice daily. The pups in each litter were counted once daily. Clinical observations were recorded once dailya. Pups were observed if they were warm and clean, for evidence of a nest and if pups were grouped together and nursing or had milk in stomach. Each pup was examined for general shape of the head, trunk, limbs, tail and presence of anus. Pup body weights were recorded on DLs 1, 8, 15 and 22 (terminal weight). 2.7.6. Gross Necropsy 2.7.6.1. Fo Generation Rats Male and female rats assigned to the toxicokinetic study were sacrificed by carbon dioxide asphyxiation on DS 42 and DG 21, respectively. Blood samples were collected from the rats as previously described. After sacrifice, the liver of each rat was excised and the liver weight was recorded. The median liver lobe was frozen and stored (I-20C) until shipment for analysisb. The fetuses were removed from the uterus and blood samples were collected from each fetus via decapitation. Blood was placed into tubes, pooled per litter, allowed to clot and spun in a centrifuge. The resulting serum was transferred into labeled polypropylene tubes. All samples were immediately frozen on dry ice and maintained frozen (I-70C)until shipment for analysis. The liver from each fetus was collected, pooled per litter and placed into labeled tubes. The samples were frozen and stored (G20"C) until shipment for analysis. The number of implantation sites was recorded. Carcasses were discarded without further evaluation. The livers were shipped to Exygen Research, State College, Pennsylvania, for analysis. Results of the analyses are available in APPENDIX G. Male and female rats assigned to the main study were sacrificed by carbon dioxide asphyxiation following the last dosage administration, after a minimum of 42 days of dosage and on DL 22, respectively. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Gross necropsy included an initial physical examination of external surfaces and all orifices, as well as the cranial, thoracic and abdominal cavities and their contents. Special attention was paid to the organs of the reproductive system. The ovaries and the uterus with cervix of each female rat were weighed, and the a. See APPENDIX E, items 6 and 7. b. See APPENDIX E, item 8. 418-028:PAGE 2-15 ovaries, uterus, vagina and a mammary gland were retained in neutral buffered 10%formalin. The number of implantation sites were recorded. Uteri of apparently nonpregnant rats were examined after being pressed between glass plates to confirm the absence of implantation sites and were retained in neutral buffered 10%formalin. Gross lesions were retained in neutral buffered 10%formalin and examined histologically. Representative photographs of gross lesions are available in the raw data. Tissue trimming and histopathology were performed under the supervision of or by a Board Certified Veterinary Pathologist. Ten rats per sex per group assigned to functional observational battery and motor activity tests were assigned to histological evaluations. The following organs were excised, trimmed and individually weighed as soon as possible after excision to avoid drying: liver, kidneys, adrenals, thymus, testes, right epididymis, left epididymis (corpus and caput), seminal vesicles (with and without fluid), prostate, spleen, brain, heart, ovaries and uterus (with cervix). The following tissues or representative samples were retained in neutral buffered 10% formalin: brain (representative regions including cerebrum, cerebellum, pons), small and large intestines (including Peyer's patches), lungs (perfused with neutral buffered 10% formalin), lymph nodes (submandibular and mediastinal), peripheral nerve (sciatic or tibial), stomach, kidneys, spleen, thymus, trachea, urinary bladder, spinal cord (cervical, thoracic and lumbar), liver, adrenals, heart, thyroid/parathyroid, bone marrow (sternum), testes (fixed in Bouin's solution for 48 to 96 hours before being retained in neutral buffered 10% formalin), prostate, seminal vesicles (with coagulating gland), the remaining portion of the left epididymis (corpus and caput), the right epididymis, ovaries, uterus, vagina, mammary gland (female rats only) and gross lesions. Histological examination of retained tissues, including reproductive organs, was conducted for the assigned ten rats per sex from the control and high dosage groups. A quantitative evaluation of primordial follicles was conducted for Fo generation female rats. Examination included enumeration of number of primordial follicles, which were combined with small growing follicles, for comparison of ovaries of rats assigned to treated and control groups. Tissues that were examined histologically were shipped to Research Pathology Services, Inc., New Britain, Pennsylvania, for evaluation. Results of the histological evaluation are available in APPENDIX J. To assess the potential toxicity of the test substance on the male reproductive system, sperrn evaluations were performed for 10 male rats in each dosage group. Sperm concentration and motility were evaluated using computer-assisted sperm analysis (CASA). Motility was evaluated by the Hamilton Thorne NOS by collection of a sample from the left vas deferens. A homogenate was prepared from the left cauda epididymis for evaluation by the Hamilton Thorne NOS to determine sperm concentration (sperm per gram of tissue weight). The remaining portion of the left cauda epididymis was used to manually evaluate sperm morphology. Sperm morphology evaluations included the following: 1) determination of the percentage of normal sperm in a sample of at least 200; and 2) qualitative evaluation of abnormal sperm, including such categories as abnormal head, abnormal tail, and abnormal head and tail. 418-028:PAGE 2-16 At scheduled sacrifice, the 10 male and 10 female rats per group assigned to hematology and clinical chemistry sample collection were exsanguinated from the inferior vena cava following sacrifice. Rats were fasted overnight before sacrificea. Approximately 5 mL of blood was collected. The tubes containing the samples were labeled with the protocol number, Sponsor study number, rat number, sex, group number, dosage level, day of study, collection interval, date of collection, species, generation and storage conditions. Approximately 1 mL of blood was collected into EDTA-coated tubes and maintained on wet ice or refrigerated until shipment for analysis of the following hematologic parameters: erythrocyte count (RBC), hematocrit (HCT), hemoglobin (HGB), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), total leukocyte count (WBC), differential leukocyte count, platelet count (PLAT), mean platelet volume (MPV) and cell morphology. Two blood smear slides were prepared at the Testing Facility for each sample for measurements of differential leukocyte count. Approximately 1.8 mL of blood was added to a tube containing 0.2 mL of sodium citrate (0.129 M). The contents were mixed and maintained on wet ice until the tubes were centrifuged (within 30 minutes of the collection time). The resulting plasma was transferred to 2.0 mL polypropylene tubes and immediately frozen. Plasma samples were maintained on dry ice or in a freezer (S7O"C) until shipment for measurement of prothrombin time (PT) and activated partial thromboplastin time (APTT). Approximately 2 mL of blood was collected into serum separator tubes and centrifuged. The resulting sera samples were immediately frozen on dry ice and maintained frozen (I-70C) until shipment for analysis of the following parameters: total protein (TP), triglycerides (TRI), albumin (A), globulin (G), albumidglobulin Ratio (NG),glucose (GLU), cholesterol (CHOL), total bilirubin (TBILI), urea nitrogen (BUN), creatinine (CREAT), creatinine kinase (CK), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALK), calcium (CA), phosphorus (PHOS), sodium (NA), potassium (K) and chloride (CL). Samples for hematology and clinical chemistry analyses were shipped to Redfield Laboratories, A Division of CRL-DDS, Redfield, Arkansas. Samples were shipped on dry ice and slides were shipped at ambient conditions. Results of these analyses are available in APPENDIX K. Female rats that did not deliver a litter were sacrificed on DG 25. Gross necropsy, examination and tissue retention were conducted as described above for rats at scheduled sacrifice. Gross lesions were preserved in neutral buffered 10%formalin for possible future evaluation. Representative photographs of gross lesions are available in the raw data. a. See APPENDIX E, item 1. 418-028:PAGE 2- 17 2.7.6.2. F1 Generation Pups On DL 22, pups were sacrificed by carbon dioxide asphyxiation and examined for gross lesions. Necropsy included a single cross-section of the head at the level of the frontalparietal suture and examination of the cross-sectioned brain for apparent hydrocephaly. Gross lesions were preserved in neutral buffered 10%formalin for possible future evaluation. Representative photographs of gross lesions are available in the raw data. Blood samples were collected from each selected pup (five per sex per litter from the 10 female rats per group selected for FOB and motor activity assessment, blood sample collection for hematology and clinical chemistry and histological evaluations) from the vena cava. The blood was placed into tubes, pooled per litter, allowed to clot and spun in a centrifuge; the resulting serum was transferred into labeled polypropylene tubes. All samples were immediately frozen on dry ice and maintained frozen (I-70C) until shipment for analysis. The liver from each selected pup was excised and the organ weight recordeda. The median lobe was frozen and stored (I-20C) until shipment for possible analysis. The remaining portion of each liver was retained in neutral buffered 10% formalin for possible histological evaluation. The livers were processed and evaluated histologically as described for the Fo generation rats. Pups that died before initial examination of the litter for pup viability were evaluated for vital status at birth. The lungs were removed and immersed in water. Pups with lungs that sink were identified as stillborn; pups with lungs that float were identified as liveborn, and to have died shortly after birth. Pups found dead were examined for gross lesions and for the cause of death. 2.7.7. Data Collection and Statistical Analyses Data generated during the course of this study were recorded either by hand or using the Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, the Coulbourn Instruments Passive Infrared Motor Activity System, the Coulbourn Instruments Auditory Startle System, the CoulbournInstruments Spatial Delayed Alternation System, and/or the passive avoidance software. All data were tabulated, summarized and/or statistically analyzed using the Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, Microsoft Excel [part of Microsoft Office 97 (version SR-2)] and/or The SAS System (version 6.12). a. See APPENDIX E, item 9. 4 18-028:PAGE 2- 18 Averages and percentages were calculated. Litter values were used where appropriate. The following schematic represents the statistical analyses of the data: I. Parametric Tvpe of Testa II. Nonparametricb A. Bartlett's Test S ignificant at ps0.0 0 1 Nonparametric Not Significant Analysis of Variance A. Kruskal-Wallis Test ( 2 7 5 % ties at any concentration) & Significant at ps0.05 Not Significant I Dunn's Test I Significant at ps0.05 I Dunnett's Test I Not Significant B. Fisher's Exact Test on Proportion of Ties (>75% ties at any concentration) + B. Analysis of Variance with Repeated Measures Significant at ps0.0 5 Not Significant I (Dosage) Dunnett's Test I (Dosage x Block Interaction) One-way ANOVA for each block S ignificant at ps0.05 I Dunnett's Test Not Significant Ill. Test for Proportion Data Variance Test for Homogeneity of the Binomial Distribution ................................. a. Statistically significant probabilities are reported as either ~10.0o5r ~ 5 0 .10. b. Proportion data are not included in this category. c. Test for homogeneity of variance. 418-028:PAGE 2-19 Adult data was evaluated with the individual rat as the unit measured. Litter values were used in evaluation of pup data, as appropriate. Variables with interval or ratio scales of measurement, such as body weights, feed consumption values, latency and errors per trial scores in behavioral tests and percent mortality per litter were analyzed as described under the Parametric heading of the schematic. Bartlett's Test of Homogeneity of Variances(14)was used to estimate the probability that the dosage groups have different variances. A non-significant result (pO.001) indicated that an assumption of homogeneity of variance was not inappropriate, and the data was compared using the Analysis of Variance? If that test was significant (pSO.OS), the groups given the test substance were compared with the control group using Dunnett's Test(? If Bartlett's Test was significant (p10.001),the Analysis of Variance Test was not appropriate, and the data was analyzed as described under the Nonparametric heading of the schematic. When 75% or fewer of the scores in all the groups were tied, the Kruskal-Wallis Test(17)was used to analyze the data, and in the event of a significant result (pS0.05),Dunn's Test(18)was used to compare the groups given the test substance with the control group. When more than 75% of the scores in any dosage group were tied, Fisher's Exact Test(lg)was used to compare the proportion of ties in the groups. Data from the motor activity test, with measurements recorded at intervals (Blocks) . throughout each test session, were analyzed using an Analysis of Variance with Repeated Measures(20)a, s described under that heading in the schematic. A significant result (p10.05) in that test could have appeared as effect of Concentration (differences among dosage groups in the totals of all measurements in a session) or as an interaction between Concentration and Block (differences in the patterns of dosage group values across the measurement periods). If the Concentration effect was significant, the totals for the control group and the groups given the test substance were compared using Dunnett's Test`? If the Concentration x Block interaction was significant, an Analysis of Variance(") was used to evaluate the data at each measurement period, and a significant result (pS0.05)was followed by a comparison of the dosage groups using Dunnett's Test(? Variables that had graded or count scores, such as litter size, the number of trials to a criterion in a behavioral test or the day a developmental landmark appeared, were analyzed using the procedures described under the Nonparametric heading of the schematic. Clinical observation incidence data, as well as the descriptive and quanta1 data from the FOB, were analyzed as contingency tables using the Variance Test for Homogeneity of the Binomial Distributiod2`I. Sperm motility data were expressed as percentages and analyzed, as indicated above, by parametric methods. 418-028:PAGE 3-1 3. 3.1. 3.1.1. RESULTS - Male Rats - Mortality, Clinical and Necropsy Observations (Summaries Tables B l - and B2; Individual Data Tables B17 and B18) Mortality All male rats survived to scheduled sacrifice. 3.1.2. Clinical Observations All clinical observations were considered unrelated to the test substance because: 1) the incidences were not dosage-dependent; and/or 2) the observation occurred in only one or two male rats in any dosage group. These observations included chromorhinorrea, missinghroken or misaligned incisors, chromodacryorrhea, lacrimation, localized alopecia on the head, underside or limbs, dry, brown perioral substance, soft or liquid feces, scab on head and dehydration. 3.1.3. Necropsy Observations All necropsy observations were considered unrelated to the test substance because the incidences were not dosage-dependent; and/or 2) the observation occurred in only one or two male rats in any dosage group. These observations included discolored/mottled kidneys, slight pelvic dilation of the left kidney, small epididymides, small testes and diverticulum jejunum. 3.1.4. Histopathology (APPENDIX J) No treatment-related microscopic changes were observed in any of the male rats administered 0.3 or 1 mg/kg/day of the test substance. Treatment-related microscopic changes were observed in the liver and thyroid gland of male rats in the 3 and 10 mg/kg/day dosage groups. The treatment-related microscopic changes in the liver consisted of minimal to moderate enlargement (hypertrophy) of centrilobular hepatocytes. The affected hepatocytes were enlarged with an increased amount of dense eosinophilic granular cytoplasm. The treatment-related microscopic changes in the thyroid gland consisted of an increased incidence of the male rats in the 3 and 10 mg/kg/day dosage groups with hypertrophy (enlargement) of follicular cells and hyperplasia (increased follicular cells and small follicles). Although the incidence in the 10 mg/kg/day dosage group was minimally increased over the control group values, the hypertrophy and hyperplasia could have been associated with the liver-cell changes. 418-028:PAGE 3-2 3.2. Terminal Body Weights and Organ Weights and Ratios (%) of Organ - Weight to Terminal Body Weight and Brain Weight (Summaries - Tables B3 through B5; Individual Data Tables B19 and B20) Terminal body weights of the male rats were slightly reduced (approximately6%),albeit not significantly, in the 10 mgkg/day dosage group, as compared with control group values. The absolute weights of the liver, the ratios of the liver weights to terminal body weights and ratios of the liver weights to brain weight were significantly increased (p10.05and/or ~10.01i)n the 3 and 10mgkgday dosage groups, as compared with the control group value. The ratio of the heart weights to brain weight was significantly decreased (p10.05) in the 10mgkgday dosage group, as compared with the control group value. The absolute and relative (terminal body and brain) weights of the male reproductive organs (left and right epididymes, left cauda epididymes, left and right testes, seminal vesicles with and without fluid and prostate), brain, left and right kidneys and adrenals, spleen, thymus and heart (except relative heart weight as described above) were comparable across dosage groups and did not differ significantly. - 3.3. Hematology and Clinical Chemistry (Summaries Tables B6 and B7; - Individual Data APPENDIX K) Hemoglobin concentrations (HGB) were significantly decreased (1310.05or p10.01)in the 1 , 3 and 10mgkg/day dosage groups and average values for red blood cells (RBC) and hematocrit (HCT) were significantly decreased (pI0.05or ~10.01i)n the 3 and 10 mgkg/day dosage groups, as compared with the control group values. Prothrombin time (PT) was significantly increased (p10.05or ~ 1 0 . 0 1i)n the 0.3,3 and 10 mgkg/day dosage groups. Dosages of the test substance as high as 10 mg/kg/day did not affect mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets (PLAT), mean platelet volume (MPV), volume, white blood counts (WBC), activated partial thromboplastin time (APTT), nucleated red blood cell count (NRBC), lymphocytes, segmented neutrophils, bands, monocytes, eosinophils, basophils and abnormal lymphocytes in male rats. These values were comparable among the four dosage groups and did not differ significantly. Average values for cholesterol (CHOL) were significantly decreased (p10.05or ~10.10) in the 0.3, 1 , 3 and 10 mgkgday dosage groups and the average values for triglycerides (TRIG) was significantly decreased (p10.01) in the 10 mgkg/day dosage group. Albumin (A), blood urea nitrogen (BUN), alkaline phosphatase (ALK), calcium (CA) and albumirdglobulinratio (A/G) levels were significantly increased (p10.01) in the 10mgkg/day dosage group. Clinical chemistry values for total protein (TP), glucose (GLU), total bilirubin (TBILI), creatinine (CREAT), creatinine kinase (CK), alanine aminotransferase (ALT), aspartate 418-028:PAGE 3-3 aminotransferase (AST), phosphorus (PHOS), sodium (NA), potassium (K), chloride (CL) and globulin (G) in male rats were comparable among the four dosage groups and did not differ significantly. 3.4. - Body Weights and Body Weight Changes (Figure 1; Summaries Tables B8 and B9; Individual Data - Table B21) Body weight gains were significantly reduced (p10.05 orplO.O1) in the 0.3,3 and 10 mg/kg/day dosage groups on study days (DSs) 29 to 36. As a result of these reductions, significantly reduced (p10.05 or p10.01)body weight gain occurred in the 0.3, 1 , 3 and 10 mg/kg/day dosage groups on study days DS 29 to termination. Body weight gain was also significantly reduced (p10.01) for the 10 mg/kg/day dosage group for the entire dosage period (calculated as DS 1 to termination). 3s. - - Absolute (g/day)and Relative (g/kg/day) Feed Consumption Values (Summaries Tables B10 and B l l ; Individual Data Table B22) Absolute (g/day) and relative (g/kg/day) feed consumption values for the male rats were unaffected by dosages of the test substance as high as 10 mg/kg/day. - - 3.6, Mating and Fertility (Summary Table B12; Individual Data Table B23) All mating and fertility parameters (numbers of days in cohabitation, rats that mated, fertility index, rats with confirmed mating dates during the first and second week of cohabitation and the number of pregnant rats per number of rats in cohabitation) were unaffected by dosages of the test substance as high as 10 mg/kg/day. - - 3.7, Functional 0bservational Battery (Summary Table B13; Individual Data Table B24) Home cage behavior of sleeping and imrnobile/awake were significantly different (p10.01) in the 3 mg/kg/day dosage group, compared to the control group. These increases were not considered treatment-related because they were not dosage-dependent. There were no other statistically significant or biologically important differences among the four dosage groups in the measures of the functional observational battery (FOB). There were no alterations in autonomic functions (lacrimation, salivation, palpebral closure, prominence of the eye, pupillary reaction to light, piloerection, respiration, defecation and urination), sensorimotor functions [responses to visual, auditory, tactile and painful stimuli (reactivity and sensitivity)], excitability (reactions to handling and behavior in the open field), gait and sensorimotor coordination (gait pattern in the open field, severity of gait abnormalities, air righting reaction and landing foot splay) and forelimb and hindlimb grip strength and abnormal clinical observations including but not limited to: convulsions, tremors, unusual behavior, hypotonia or hypertonia, emaciation, dehydration, unkempt appearance and deposits around the eyes, nose or mouth. 418-028:PAGE 3-4 Body weights recorded during the functional operational battery for the treated groups were not significantly different than the control group for the male rats. - 3.8. Motor Activity (Figures 3 and 4; Summary Table B14; - Individual Data Table B25) There were no statistically significant or biologically important differences among the five dosage groups in the number of movements or time (in seconds) spent in movement measures of motor activity on DSs 36 through 39. 3.9. Sperm (Summary - Tables B15 and B16; Individual Data - Tables B26 and B27) 3.9.1. Sperm Motility Sperm motility was unaffected by dosages of the test substance as high as 10 mg/kg/day. Group mean values were comparable among the five dosage groups and ranged from 85.5% to 93.2% motile sperm. 3.9.2. Sperm Count and Sperm Density The sperm count and sperm density were comparable arnong the five dosage groups. No treatment-related differences were observed. 3.9.3. Sperm Morphology A low incidence of head and/or tail abnormalities was observed for the male rats in all dosage groups. No treatment-related differences were observed. 4 18-028 PAGE 4- 1 4. RESULTS - Female Rats 4.1. - Mortality, Clinical and Necropsy (Summaries Tables C1 and C2; Observations Individual Data - Tables C28 and C29) 4.1.1. Mortality All female rats survived to scheduled sacrifice. 4.1.2. Clinical Observations All clinical observations were considered unrelated to the test substance because: 1) the incidences were not dosage-dependent; and/or 2) the observation occurred in only one female rat in any dosage group except for localized alopecia which is a common finding in rats that are allowed to deliver. These observations included brown period substance, rales, localized alopecia on the limbs, back or head, urine-stained abdominal fur, miosis, abdominal distention, emaciation, cold to touch, swollen nose, scab or ulceration on back or head, dehydration and tip of tail missing. 4.1.3. Necropsy Observations All necropsy observations were considered unrelated to the test substance because: 1) the incidences were not dosage-dependent; and/or 2 ) the observation occurred in only one female rat in each of the 1 , 3 and 10mg/kg/day dosage groups. These observations included the cortex of the right kidney adhered to the right lateral liver lobe in a 1mg/kg/day dosage group female rat (19907), small left ovaries (0.003g) in one 3 mg/kg/day dosage group female rat (19051) that was not pregnant, outer capsule of spleen adhered to the omentum in another 3 mg/kg/day dosage group female rat (19063) and the large and small intestines distented with gas, small spleen and thymus, large adrenals, and approximately 12 black areas (pinpoint to 0.3 cm) on the fundic mucosal surface of the stomach in a 10mgkgday dosage group female rat (19020). 4.1.4. Histopathology (APPENDIX J) No treatment-related microscopic changes were observed in any of the female rats administered up to 10 mg/kg/day of the test substance. There were no treatment-related microscopic changes observed in the liver of the F1 generation pups from the dams given up to 10 mg/kg/day of the test article. There were a few microscopic changes observed in the various organs and tissues which were considered to have occurred spontaneously and not to be treatment-related. 418-028:PAGE 4-2 4.2. Terminal Body Weights, Organ Weights and Ratios (%) of Organ Weight Follicle - to Terminal Body Weight and Brain Weight and Primordial Counts (Summaries Tables C3 through C5; Individual Data - Tables C30 through C32) Terminal body weights of the female rats were comparable among dosage groups and did not differ significantly. The absolute and relative weights (to terminal body and to brain weight) of the female reproductive organs (left and right ovaries and uterus with cervix), brain, liver, left and right kidneys and adrenals, spleen, thymus and heart were comparable across dosage groups and did not differ significantly (except on one occasion). The ratio of the left kidney to the brain weight of the female rats in the 3 mg/kg/day dosage group was significantly increased (p10.05)over the corresponding control group value. This significant finding was not considered treatment-related because: 1) the incidence was not dosage-dependent; and/or 2) the observation was a single occurrence. Average primordial follicle counts for the 10 mg/kg/day dosage group were comparable to the control group and did not differ significantly. - 4.3. Hematology and Clinical Chemistry (Summaries Tables C6 and C7; - Individual Data APPENDIX K) Dosages as high as 10 mg/kg/day did not affect any hematology or clinical chemistry values evaluated. Average values for red blood cells (RBC), white blood cells (WBC), hemoglobin concentration (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets (PLAT), mean platelet volume (MPV), prothrombin time (PT), activated partial thromboplastin time (APTT), nucleated red blood cell count (NRBC), lymphocytes, segmented neutrophils, bands, monocytes, eosinophils, basophils, abnormal lymphocytes in female rats were comparable among the five dosage groups and did not differ significantly (except on one occasion). The concentration of WBCs (lymphocytes) for of the female rats in the 0.3 mg/kg/day dosage group was significantly increased (p10.05)over the corresponding control group value. This significant finding was not considered treatment-related because: 1) the incidence was not dosage-dependent; and/or 2) the observation was a single occurrence. Average values for total protein (TP), albumin (A), glucose (GLU), cholesterol (CHOL), total bilirubin (TBILI), blood urea nitrogen (BUN), creatinine (CMAT), creatinine kinase (CK), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALK), calcium (CA), phosphorus (PHOS), triglycerides (TRIG), sodium (NA), potassium (K), chloride (CL), globulin (G) and albumin/globulin ratio (A/G) in female rats were comparable among the five dosage groups and did not differ significantly (except on two occasions). The concentration of G was significantly decreased (p10.01) and the concentration of A/G was significantly increased (p10.01) for of the female rats in the 0.3 mg/kg/day dosage group over the corresponding control group values. These significant findings were not considered treatment-related because: 4 18-028:PAGE 4-3 1) the incidences were not dosage-dependent; and/or 2 ) the observations were a single occurrence for both parameters. - 4.4. Body Weights and Body Weight Changes (Figure 2; Summaries - Tables CS through C13; Individual Data Tables C33 through C35) 4.4.1. Precohabitation Body weights and body weight gains were comparable and did not differ significantly during the precohabitation period at dosages of the test substance up to 10 mg/kg/day. 4.4.2. Gestation Body weights and body weight gains were comparable and did not differ significantly during the gestation period at dosages of the test substance up to 10 mg/kg/day. 4.4.3. Lactation Body weight gains were not significantly affected by dosages of the test substance up to 10 mg/kg/day during lactation. Body weights were significantly reduced (p10.05or p10.01) on days of lactation (DLs) 4, 6 through 8, 11 and 13 in the 0.3 rng/kg/day dosage group, DLs 7 and 8 in the 3 mg/kg/day dosage group, DLs 4 , 6 through 9,11,13 and 14 in the 10 mg/kg/day dosage group. These significant findings were not considered treatment-related because they were not dosage-dependent or they did not persist. 4.5. 4.5.1. - - Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values (Summaries Tables C14 through C19; Individual Data Tables C36 through C38) Precohabitation Absolute (g/day) and relative (g/kg/day) feed consumption values were not significantly affected by dosages of the test substance up to 10 mg/kg/day during the precohabitation period. 4.5.2. Gestation Absolute (g/day) and relative (g/kg/day) feed consumption values were not significantly affected by dosages of the test substance up to 10 mg/kg/day during the gestation period. 4.583. Lactation Absolute and relative feed consumption values were not affected by dosages of the test substance up to 10 mg/kg/day during lactation. 418-028:PAGE 4-4 Absolute feed consumption values were significantly reduced (pI0.05 or p10.01) on DLs 1 to 5 and 8 to 15 in the 0.3 mg/kg/day dosage group, DLs 1 to 5 and 8 to 15 in the 3 mg/kg/day dosage group and DLs 8 to 15 in the 10 mg/kg/day dosage group. Relative feed consumption values were significantly reduced (p10.01) on DLs 1 to 5 in the 0.3 mg/kg/day and DLs 5 to 8 in the 3 mg/kg/day dosage group. These significant findings were not considered treatment-related because they were not dosage-dependent and they did not persist. - 4.6. Estrous Cycling, Mating and Fertility (Summary Table C20; - Individual Data Table C39) The average numbers of estrous stages per 13 days were comparable among the five dosage groups and did not significantly differ. The number of rats with six or more consecutive days of diestrus or estrus did not differ significantly. All mating and fertility parameters (numbers of days in cohabitation, rats that mated, fertility index, rats with confirmed mating dates during the first and second week of cohabitation and the number of pregnant rats per number of rats in cohabitation) were unaffected by dosages of the test substance as high as 10 mg/kg/day. - 4.7. Functional Observational Battery (Summary Table C21; Individual - Data Table C40) There were no statistically significant or biologically important differences among the five dosage groups in the measures of the functional observational battery (FOB). There were no alterations in home cage behavior, autonomic functions (lacrimation, salivation, palpebral closure, prominence of the eye, pupillary reaction to light, piloerection, respiration, defecation and urination), sensorimotor functions [responses to visual, auditory, tactile and painful stimuli (reactivity and sensitivity)], excitability (reactions to handling and behavior in the open field), gait and sensorimotor coordination (gait pattern in the open field, severity of gait abnormalities, air righting reaction and landing foot splay) and forelimb and hindlimb grip strength and abnormal clinical observations including but not limited to: convulsions, tremors, unusual behavior, hypotonia or hypertonia, emaciation, dehydration, unkempt appearance and deposits around the eyes, nose or mouth. Body weights recorded during the functional operational battery for the treated groups were not significantly different than the control group for the female rats. - 4.8. Motor Activity (Figures 5 and 6; Summary Table C22; - Individual Data Table C41) There were no statistically significant or biologically important differences among the five dosage groups in the measures of number of movements or time (in seconds) spent in movement motor activity on DL 17. 418-028:PAGE 4-5 4.9. Natural Delivery and C24; Individual Data -Litter Observations (Summary Tables C42 through C45) - Tables C23 and Pregnancy occurred in all 15 (100%)rats assigned to the 0 (Vehicle), 0.3 and 10 mg/kg/day dosage groups and 13 (88.7%) of the rats assigned to the 1 and 3 mg/kg/day dosage groups. All pregnant dams delivered a litter of one or more liveborn pups. Values for the numbers of dams that delivered litters, the duration of gestation (calculated in days), averages for implantation sites per delivered litter, the numbers of dams with stillborn pups, the numbers of dams with no liveborn pups, dams with all pups dying days 1 to 4 and 5 to 22, were comparable among the five dosage groups and did not significantly differ. The number of liveborn pups, the number of pups found dead or presumed cannibalized on day 1, and days 2 to 8,9 to 15 and 16 to 22 postpartum were comparable among the five dosage groups and did not significantly differ. The gestation index (number of dams with one or more liveborn pups per number of pregnant rats), viability index (number of live pups on DL 8 per number of liveborn pups on DL 1) and lactation index (number of live pups on DL 22 per number of liveborn pups on DL 8) were compared to the control group value and did not differ significantly. The number of pups surviving per litter, pup sex ratios, litter size and pup body weights per litter on DLs 1, 8, 15 and 22 were comparable among the five dosage groups and did not significantly differ. 4.10. - Pup Clinical and Necropsy Observations (Summary Tables C25 and C26; Individual Data - Tables C46 and C47) No clinical or necropsy observations in the F1 generation pups were attributable to dosages of the test substance as high as 10 mg/kg/day because: 1) the incidences were not dosage-dependent; and 2) the observation occurred in only one to two litters. These clinical observations included: not nursing, emaciation, dehydration, whole body discolored purple, scab, cold to touch, left eye discolored purple, right eye enlarged, laceration on right hindlimb, bruise on back, not nesting, pale in appearance and corneal opacity of right eye. Necropsy observations of 228,203, 196, 172 and 211 pups in the five respective dosage groups on postpartum days 5 and 22 were limited to moderate dilation of the renal pelvis of one pup from a 0.3 mg/kg/day dosage group litter. 4.11. - - Pup Liver Weight and Ratio of Liver Weight to Terminal (Summary Table C27; Individual Data Table C48) Body Weight F1 generation male and female pup terminal body weights, absolute liver weight (by sex) and ratio of liver weight to terminal body weight (by sex) were comparable across all five dosage groups and did not differ significantly. 418-028:PAGE 4-6 REFERENCES 1. Sanders, J.E., Eigenberg, D.A., Bracht, L.J., Wang, W.R., and Zwieten, M.J. (1988): Thyroid and liver trophic changes in rats secondary to liver microsomal enzyme induction caused by an experimental leukotriene antagonist (L-649,923). Toxicology and Pharmacology, 95: 378-387. 2. Organisation for Economic Co-operation and Development (1996). OECD Guidelinefor Testing of Chemicals. Section 4, No. 422: Combined Repeated Dose Toxicity Study with the ReproductiodDevelopmentalToxicity Screening Test, adopted 22 March 1996. 3. Organisation for Economic Co-operation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Final]. 4. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. 5. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standardfor Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. 6. Christian, M.S. and Voytek, P.E. (1982). In VivoReproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield,VA 22161. 7. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone. (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 8. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River Crl:CDBBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 9. Institute of Laboratory Animal Resources (1996). Guidefor the Care and Use of LaboratoryAnimals. National Academy Press, Washington, D.C. 10. Haggerty, G.C. (1989). Development of Tier I neurobehavioral testing capabilities for incorporation into pivotal rodent safety assessment studies. J. Amer. Col. Toxicol. 853-70. 11. Irwin, S. (1968). Comprehensiveobservational assessment: Ia. A systemic quantitative procedure for assessing the behavioral and physiologic state of the mouse. Psychopharmacologia (Berlin) 13:222-257. 12. Moser, V.C. (1989). Screening approaches to neurotoxicity: A functional observational battery. J. Amer. Col. Toxicol. 8:85-94. 418-028:PAGE 4-7 13. O'Donoghue, J.L. (1989). Screening for neurotoxicity using a neurologically based examination and neuropathology. J. Amer. Col. Toxicol. 8:97-116. 14. Sokal, R.R. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances.Biornetry, W.H. Freeman and Co., San Francisco, pp. 370-371. 15. Snedecor, G.W. and Cochran, W.G. (1967). Analysis of Variance. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 258-275. 16. Dunnett, C.W. (1955). A multiple comparison procedure for comparing several treatments with a control. J. Amer. Stat. Assoc. 50:1096-1121. 17. Sokal, R.R. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometry, W.H. Freeman and Co., San Francisco, pp. 388-389. 18. Dunn, O.J. (1964). Multiple comparisons using rank sums. Technometrics 6(3):241-252. 19. Siegel, S. (1956). Nonparametric Statisticsfor the Behavioral Sciences. Fisher's Exact. McGraw-Hill Co., New York, pp. 96-105. 20. SAS Institute, Inc. (1988). Repeated measures analysis of variance. SAS/STATrM User's Guide, Release 6.03 Edition, Cary, NC, pp. 602-609. 21. Snedecor, G.W. and Cochran, W.G. (1967). Variance test for homogeneity of the binomial distribution. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 240-241. APPENDIX A REPORT FIGURES PROTOCOL 418-0281 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST (SPONSORS STUDY NUMBER: T-7706.1) BODY WEIGHTS Fo GENERATION MALE RATS Figure 1 500 + 480 0 MG/KG/DAY 460 440 n + 420 I 400 380 0.3 MG/KG/DAY & 1 MG/KG/DAY + 3 MG/KG/DAY +z- 10 MG/KG/DAY 360 340 320 I 1 a. Last value recorded before cohabitation. b. First value recorded after cohabitation. b c. Terminal body weight. h) I I I I I 7 8 14a 29b 36 43 C DAY OF STUDY 450 - 400 - s!3,50 - I- I- c3 300 - PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST (SPONSORS STUDY NUMBER: T-7706.1) BODY WEIGHTS Fo GENERATION FEMALE RATS Figure 2 A 0 MG/KG/DAY -0.3 MG/KG/DAY 1 MG/KG/DAY + 3 MG/KG/DAY & 10 MG/KG/DAY 250 a. Last value recorded before cohabitation. 1 *p<0.05 1 1 3 5 7 91113 1 3 5 7 9 11 13 15 17 19 2 4 6 8 10 12 14a 0 2 4 6 8 10 12 14 16 18 20 DAY OF STUDY DAY OF GESTATION Illllllllllllll II1111 I 2 4 6 8 10 12 14 16 18 20 22 1 3 5 7 9 11 13 15 17 19 21 DAY OF LACTATION 1 **p10.01 1 PROTOCOL 418-0281 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSORS STUDY NUMBER: T-7706.1) MOTOR ACTIVITY - NUMBER OF MOVEMENTS - FOGENERATION MALE RATS Figure 3 80 + 70 0 MG/KG/DAY 60 Yc/) I- zzW 50 0 840 OL W 30 z 3 20 0.3 MGIKGIDAY -c2+ 1 MG/KG/DAY -Ic- 3 MG/KG/DAY -2Ek 10 MG/KG/DAY 10 0 0 10 20 30 40 50 60 70 80 90 100 TIME (MINUTES) PROTOCOL 418-0281ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) MOTOR ACTIVITY - TIME SPENT IN MOVEMENT - FOGENERATION MALE RATS Figure 4 250 2 200 z z W W > 0 2 +- 150 z w a. v) G? tz2 100 8cW z W iZ 50 + 0 MG/KG/DAY 0.3 MG/KG/DAY * -1 MG/KG/DAY 3 MG/KG/DAY * 10 MG/KG/DAY P c-r 0 0 20 40 60 80 100 TIME (MINUTES) ? P PROTOCOL 418-0281 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTIONIDEVELOPMENTAL TOXICITY SCREENING TEST (SPONSORS STUDY NUMBER: T-7706.1) MOTOR ACTIVITY - NUMBER OF MOVEMENTS - FOGENERATION FEMALE RATS Figure 5 80 --%%- 70 0 MG/KG/DAY 60 zv) I- w 50 z 9 0 40 LL 0 2fY W 30 3 z 20 0.3 MG/KG/DAY + -1 MG/KG/DAY 3 MG/KG/DAY * 10 MG/KG/DAY 10 P C'L 0 0 10 20 30 40 50 60 70 80 90 100 TIME (MINUTES) PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSORS STUDY NUMBER: T-7706.1) MOTOR ACTIVITY - TIME SPENT IN MOVEMENT - FOGENERATION FEMALE RATS Figure 6 200 9cn I- $ 150 z 0 z z Iz- 100 cn 5n z 0 Ys. 50 F VJ? 0 MG/KG/DAY 0.3 MG/KG/DAY + -1 MG/KG/DAY 3 MG/KG/DAY * 10 MG/KG/DAY 0 0 20 40 60 80 100 TIME (MINUTES) APPENDIX B REPORT TABLES - FO GENERATION MALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B1 (PAGE 1): CLINICAL OBSERVATIONS - SUMMARY - FO GENERATION MALE RATS MORTALITY CHROMORHINORRHEA INCISORS: TOTAL MISSING/BROKEN MISALIGNED CHROMODACRYORRHEA 0 o/ 0 o/ 0 0 1/ 1 o/ 0 0 I/ 1 3/ 1 0 1/ 1 ' 13/ 1 6/ 1 13/ 1 9/ 1 0 2/ 2 13/ 1 13/ 1 o/ 0 2/ 1 LOCALIZED ALOPECIA: TOTAL HEAD UNDERSIDE LIMBS DRY, BROWN PERIORAL SUBSTANCE SOFT OR LIQUID FECES o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 27/ 2 16/ 1 o/ 0 11/ 1 I/ 1 1/ 1 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 DEHYDRATION 7/ 1 o/ 0 o/ 0 o/ 0 o/ 0 .................................................................................................................................... STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. 418-028:PAGE B-2 I I I I I I I 1 I I I I I I I I I I I I * m I I 130 I I rllrl 0 rl I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I * I I I >m I m 0 IH Id rl I I I I I I I I I I I I I I I I I 1 I I w I I 1 I X I I b I I I I I I I I I I I H r l I Ul 0 m IH Irl rl IH I I I I I I I I I I I I I w I I I I 2 I I z I I I I I I I I I I I I I I I I 0 N rl 0 I I Frc I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I w 1 I I HO I 0 I Irl rl I 1 I I I 1 I I a I I I 2 I I I I I k I I I I I I I I I I I I I I I I I I rl 03 N 0 I a, *rl I I I I I I I I I I 0 0 0 0 0 rl I I I I G Y I 8 I r n Ik IU I$ IU II c d I S 0 0 rl rl rl 0 I EI -rl IW Ig IU I I -;I IU 0 0 0 0 0 0 I Id Ild IE I Ik I0 IW I I- ~r Irl rl 0 0 0 0 rl 0 0 0 0 0 Irn I% a 22 z a I 9 l a , I I0 II U I I I. !Id I 4 18-028:PAGE B-3 E N u x H 0 E 0 w d n .i: 2 3 w w X E E X W 0 r P I E Frc 0 k h 6 rl 6 W 0 r P E I 6 W H x 0 E w tn 0 n m L2n tn E X H a E 0u V 2 E w twn 0a W E 4 W 3 wv 2 32U 0 tn rl 3 8 m 0u0 aw d E$ I I I I I I I I I I I I I I I I I ?0 I 0 I rl I rl I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I 1 I 0 I I rl I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I H rl I 0 I H I rl I H I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I Hm I 0 I H I rl I 0 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I H0 I 0 I I rl I I I I I I I I I I I I I I I I I I I I I I I I I I 2 I I I 1 I I I I I 1 I I I I I I I I I I I I I I I 1 I I I 1 I I I I I I I I I I I 1 I I I I I I I I w I I I W 1 I I 3 I I I 0 I n I I I I W cv m 6\ m P 0 d W cv d m m W rl 0 0 rl 0 0 0 +I +I +I +I co d rl m m d 03 0 rl 0 d dW mP d 0 0 rl m cv 0 0 cv m In m CO co 0 W m rl m rl 0 m m 0 0 0 0 +I +I +I +I +I m u, m W m rl rl rl P cv m m rl d P m cv W m d 0 0 I+ rl cv m m W cv 0 03 d rl In In cv W m m 0 0 rl cv 00 0 0 +I +I +I +I +I CO r cv P m P m rl CO m cv cv P m mW mW W d 0 0 rl rl CO P u, m cv m W m P P m 01 cv u) m 0 0 rl cv 00 0 0 +I +I +I +I +I 03 CO cv d m m co m m CO m rl rl d dW mP m d 0 0 rl rl u) W P dm mP 0 r rl m cv CO m m 0 0 cv 0 0 0 +I +I +I +I r u) CO m P 0 m m dm 0 u) u) m P d 0 0 rl :6' :6' 9 9 9 9 tn tn tn tn w w w E E E E ::E X E2! E wkl d 0 m w tw n d W w H E 9 w d d m H w k! 6w E E tn 5 m m W 0 rl rl 0 +I cv P P r 0 r 03 P rl d cv rl rl 0 0 +I +I 0 W P 0 W rl r W 0 0 03 m \D m 0 m rl 0 0 0 +I +I m d u) m m m r W 0 0 P: 1 m rl 0 rl d rl 0 0 0 +I +I a, W rl d rl m CO W 0 0 CO d m cv 0 d rl 0 0 0 +I +I 0 m d d CdO r W 0 0 6'n 9 tn w x E X W E 5 0 X P d d d d m rl cv 0 0 +I +I d CO 0 rl W m P rl rl rl m m m m m u) m rl 0 0 +I +I m d r m rl P Lo cv rl rl cv dm d W W rl cv 0 0 +I +I cv P cv W m cv W cv rl rl r P CO m cv cv rl rl 0 0 +I +I rl 0 CO 0 cv 0 P cv rl rl W CO m m u) m 0 cv 0 0 +I +I r cv r 0 P m W cv rl rl 6'9 9 m tn E $wE Etn H tEn I I 0 I rl I I 0 I 1 I +I I I 0 I d I I cv I I I I I I I I m I 0 I I 0 I 1 I +I I I co I m I I N I I I I I I I I m I 0 I I 0 I I I +I I I 03 I m I I cv I I I I I I I I rl I rl I I 0 I I I +I I I d I d I I cv I I I I I I I I rl I rl I I 0 I I I +I I I 0 I d I I cv I I I I I I I n I I I 6' w tn I I I I E I I I I I I I I I I I I I I I I I I I I I I I w I I d I 2 I I PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B3 (PAGE 2): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS - SUMMARY - Fo GENERATION MALE RATS FLATS TESTED N 10 10 lo TERMINAL BODY WEIGHT MEANfS.D . 469.7 k 31.6 448.8 f 37.8 455.8 f 55.2 LIVER MEANfS .D. 14.63 f 1.81 14.22 & 1.58 15.94 f 3.56 KIDNEY LEFT MEANfS .D . 1.98 f 0.25 1.86 f 0.12 1.96 f 0.26 KIDNEY RIGHT MEANfS.D . 2.02 f 0.26 1.90 f 0.15 2.00 f 0.24 ADRENAL LEFT MEANfS.D . 0.035 f 0.007 0.035 f 0.007 0.034 f 0.005 ADRENAL RIGHT MEAN+S .D . 0.034 f 0.006 0.034 f 0.006 0.030 f 0.007 SPLEEN MEANfS .D. 0.79 f 0.14 0.81 f 0.07 0.76 f 0.12 THYMUS MEANfS .D . 0.41 f 0.12 0.45 f 0.09 0.36 f 0.10 HEART MEANfS .D . 1.55 f 0.10 1.52 f 0.12 1.54 + 0.13 ............................................................................................ ALL WEIGHTS WERE RECORDED IN GRAMS (G). * Significantly different from the Group I value (~50.05). * * Significantly different from the Group I value (~50.01). 10 472.3 f 38.3 17.60 f 1.64" 2.04 f 0.17 2.13 f 0.18 0.032 f 0.006 0.032 f 0.007 0.88 f 0.19 0.42 f 0.09 1.60 f 0.13 .................... lo 440.8 f 12.6 22.83 f 2.73** 2.01 f 0.18 2.04 f 0.14 0.032 f 0.004 0.029 f 0.007 0.78 f 0.10 0.42 f 0.05 1.44 0.07 ---------------- PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B4 (PAGE 1): RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - SUMMARY - Fo GENERATION MALE RATS TERMINAL BODY WEIGHT EPIDIDYMIS LEFT MEANfS .D. MEANfS .D . 469.7 f 31.6 0.138 f 0.014 448.8 f 37.8 0.142 f 0.015 455.8 f 55.2 0.138 f 0.018 472.3 f 38.3 0.126 f 0.027 440.8 f 12.6 0.138 f 0.011 CAUDA EPIDIDYMIS LEFT TESTIS LEFT MEAN+S.D. MEANfS.D . 0.068 f 0.009 0.354 f: 0.024 0.071 f 0.012 0.385 f 0.047 0.071 f 0.007 0.370 f 0.041 0.063 f: 0.015 0.342 f 0.078 0.068 f 0.008 0.397 f 0.041 SEMINAL VESICLES WITH FLUID MEANfS .D . 0.366 f 0.064 0.348 f 0.073 0.359 f: 0.063 0.329 f 0.081 0.326 f 0.062 SEMINAL VESICLES WITHOUT FLUID MEANfS .D. 0.181 f 0.031 0.183 f: 0.038 0.176 f 0.022 0.163 f: 0.038 0.176 f 0.026 EPIDIDYMIS RIGHT MEANfS.D . 0.143 f: 0.016 0.147 f 0.018 0.145 f 0.016 0.127 f 0.028 0.141 f 0.012 TESTIS RIGHT MEAN+S .D . 0.357 f 0.030 0.386 f 0.049 0.374 f 0.042 0.344 f 0.078 0.400 f 0.035 PROSTATE MEANfS .D. 0.276 f 0.073 0.267 f 0.022 0.271 f 0.061 0.274 f 0.052 0.270 f 0.054 BRAIN MEANfS .D . 0.510 f 0.030 0.546 f 0.053 0.528 f 0.050 0.506 f 0.041 0.545 f 0.027 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). RATIOS ( % ) = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B4 (PAGE 2): RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - SUMMARY - FO GENERATION MALE RATS TERMINAL BODY WEIGHT MEANkS .D. 469.7 f 31.6 448.8 f 37.8 455.8 k 55.2 472.3 f 38.3 440.8 f 12.6 LIVER MEAN+S . D. 3.109 f 0.264 3.165 f 0.208 3.466 f 0.396 3.728 f 0.212** 5.190 f 0.702** KIDNEY LEFT MEANfS .D . 0.419 f 0.032 0.415 f 0.033 0.429 f 0.030 0.433 f 0.025 0.456 f 0.042 KIDNEY RIGHT MEANfS .D . 0.428 f 0.036 0.426 f 0.044 0.441 f 0.030 0.450 f 0.032 0.463 f 0.034 ADRENAL LEFT a MEANfS .D. 7.419 f 1.226 7.837 f 1.573 7.427 f 1.196 6.929 f 1.461 7.280 f 0.836 ADRENAL RIGHT a MEANfS .D . 7.202 k 1.181 7.587 & 1.353 6.603 f 1.636 6.788 f 1.503 6.680 f 1.514 SPLEEN MEANfS.D . 0.168 f 0.025 0.180 f 0.024 0.167 f 0.019 0.183 f 0.030 0.177 f 0.023 THYMUS MEANfS.D . 0.086 & 0.024 0.099 f 0.017 0.077 f 0.019 0.089 f 0.019 0.095 f 0.013 HEART MEANfS.D . 0.331 f 0.021 0.339 k 0.014 0.340 f 0.024 0.337 f 0.015 0.327 f 0.012 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). RATIOS (%I = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) x 100. a. Value was multiplied by 1000. * * Significantly different from the Group I value (p10.01). PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B5 (PAGE 1): RATIOS ( % ) OF ORGAN WEIGHT TO BRAIN WEIGHT - SUMMARY - FO GENERATION MALE RATS BRAIN WEIGHT EPIDIDYMIS LEFT MEANf:S.D . MEANkS.D . 2.40 f: 0.11 27.10 f 2.14 2.44 f 0.11 26.11 f 1.77 2.38 f 0.09 26.39 f: 1.99 2.38 f 0.09 24.77 f: 4.51 2.40 f 0.10 25.55 ?r 2.21 CAUDA EPIDIDYMIS LEFT TESTIS LEFT MEAN+S.D. MEANfS.D . 13.83 f 1.28 69.36 f 4.17 13.05 f 0.96 70.39 f 4.40 13.59 f: 1.18 70.28 f 6.50 12.20 f 2.80 67.41 f 13.64 12.66 f 1.32 72.79 f 6.71 SEMINAL VESICLES WITH FLUID MEANfS .D . 71.40 f: 11.06 63.64 f 12.45 68.84 f 12.04 64.77 f: 12.58 59.83 f 10.03 SEMINAL VESICLES WITHOUT FLUID MEANfS.D . 35.34 f 4.98 33.48 f 5.71 33.44 f 4.03 32.14 f 5.94 32.30 f 4.57 EPIDIDYMIS RIGHT MEANfS .D. 28.22 f 2.23 26.90 f 2.06 27.47 f 2.16 25.57 f 5.13 26.08 f 2.00 TESTIS RIGHT MEAN+S.D . 70.19 f 5.17 70.94 k 5.02 71.14 f 6.42 67.66 f 13.99 73.30 f 6.34 PROSTATE MEANkS .D . 54.35 f 14.08 49.41 f 6.63 51.57 f: 11.28 53.52 f 7.70 49.48 f: 9.23 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). RATIOS ( % ) = (ORGAN WEIGHT/BRAIN WEIGHT) X 100. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B5 (PAGE 2): RATIOS ( % ) OF ORGAN WEIGHT TO BRAIN WEIGHT - SUMMARY - Fo GENERATION MALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) RATS TESTED BRAIN WEIGHT LIVER KIDNEY LEFT KIDNEY RIGHT ADRENAL LEFT ADRENAL RIGHT SPLEEN THYMUS N MEANkS .D. MEAN+S.D. MEANkS .D. MEANfS .D . MEANfS .D. MEANfS .D . MEANfS.D . MEANfS .D . I 0 10 2.40 j: 0.11 611.89 & 79.55 82.68 f 9.11 84.22 & 9.20 1.46 k 0.27 1.41 f 0.25 32.90 f 5.93 17.22 f 5.29 I1 0.3 10 2.44 f 0.11 584.43 f 68.39 76.46 f 5.95 77.94 f 6.26 1.44 i: 0.31 1.40 & 0.26 33.34 f 3.91 18.37 f 4.02 I11 1 10 2.38 j: 0.09 667.08 f129.02 82.16 f 9.05 84.13 j: 7.94 1.42 f 0.24 1.26 f. 0.33 31.83 j: 4.39 14.93 f 4.04 IV 3 10 2.38 k 0.09 739.11 & 73.28** 85.78 & 7.04 89.26 f 6.52 1.36 & 0.25 1.34 j: 0.26 36.64 f 7.27 17.75 f 3.63 V 10 lo 2.40 & 0.10 948.61 f 95.12** 83.64 f 8.05 85.00 & 6.56 1.34 f 0.18 1.23 & 0.29 32.44 f 3.96 17.46 f 2.39 z b E H x 0 E-c w E X I? 3 W 0 IP I E-r E 0 n a, rl 0 br ELHI u H E-r x oa, E Cu 1 418-028:PAGE B-9 % r- cv c c W 0 cn 0 w w W rl W rl m In Iz cn m 0 rl rl 0 0 In rl 0 +I +I +I +I +I +I +I +I i+I I Iz cn cn w rl W * P m m m W 0 W cv W rl w In cv m rl * $: $: $: r In $: In In 0 w cn 0 P cn rl w W m W N rl m rl 0 0 m 0 0 cv +I +I +I +I U +I +I +I * cv m w cv cn cv w C u It m 0 W In 0 r- N W rl rl W rl w In cv m rl a: I I * W w rl w cn cv W In cn N I cvI PI w m cv P Iz P cn * f cv 0 0 cv rl 0 0 rl 0 *1I I +I +I +I +I +I +I +I +I +I I I a3 CJ m m rl r- r- 0 In N rl w yf w In In In I rl r- In rl cv P m rl rl WI I I I m 0 W cn m c*v Iz Iz In cv m In rl rl w: I n. 1I m 0 0 rl cv rl rl 0: I +I +I +I +I +I +I +I +I I cn m m w rl P cn N In cv rl w L- W P Iz rl r- In N m I . . rl m m1 rl rl Pi mW W W w rl 0 W W rl w rl m m r- m cn W W cn 0 rl 0 0 mrl +I +I +I +I +I +I +I +I m rl In In Irnl r- W d m 0 0 m Iz LPn cv W W rrll 9 9 9 9 9 9 n ul ul ul ul ul co ul +I +I +I +I +I +I +I h rl a3 (u 0 I W rl w e h O\O W v h O\O W v 2 u W E-r ? X X V X V x V x V x PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B6 (PAGE 2): HEMATOLOGY - SUMMARY - Fo GENERATION MALE RATS (See abbreviations on the last page of this table.) RATS TESTED N 10 10 PT (SECONDS) MEAN f S.D. 13.4 f 0.24 14.2 f 0.27** APTT (SECONDS) MEAN f S . D . 21.0 f 3.09 22.1 f 4.41 NRBC COUNT MEAN f S.D. 0 f 0.0 Lymphocytes (THSN/CU MM) MEAN f S.D. 11.9 -+ 1.72 0 f 0.0 11.0 f 2.68 Segmented (THSN/CU MM) MEAN f S.D. 2.2 f 0.73 2.9 f 1.18 Bands (THSN/CU MM) MEAN f S.D. 0 . 0 f 0 . 0 0 0.0 f 0.00 Monocytes (THSN/CUMM) MEAN f S.D. 0.8 f 0.56 0.7 f 0.50 Eosinophils (THSN/CU MM) MEAN f S.D. 0.3 f 0.20 0.3 f 0.21 Basophils (THSN/CU MM) Abnormal L (THSN/CU MM) MEAN f S.D. 0 . 0 MEAN f S.D. 0.1 f 0.00 -+ 0.20 0.0 f 0.00 0.1 f 0.14 0.0 * * Significantly different from the Group I value (pS0.01). 10 13.6 f 0.23 20.6 f 2.38 0 f 0.0 10.8 f 2.02 2.0 f 1.06 0 . 0 f 0.03 0.6 4 0.51 0.3 f 0.21 0.0 f 0.00 0.1 f 0.07 10 13.8 f 0.40* 21.2 f 2.24 0 f 0.0 12.1 f 3.57 2.8 f 0.73 0.0 f 0.03 0.8 f 0.67 0.3 f 0.26 0.0 f 0.00 0.2 f 0.22 10 14.0 21.3 f 0.47** -+ 1.43 0 f 0.0 12.4 f 2.75 1.6 f 0.78 0.0 f 0.00 0.5 f 0.33 0.2 f 0.15 0.0 -4. 0 . 0 0 0.1 f 0.21 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B6 (PAGE 3 ) : HEMATOLOGY - SUMMARY - FO GENERATION MALE RATS RBC HGB HCT MCV MCH MCHC PLAT MPV PT APTT NRBC Segmented Red Blood Cells (Erythrocytes) Hemoglobin Hematocrit (Packed Cell Volume) Mean Corpuscular Volume Mean Corpuscular Hemoglobin Mean Corpuscular Hemoglobin Concentration Platelet Count Mean Platelet Volume Prothrombin Time Activated Partial Thromboplastin Nucleated Red Blood Cell Count + P Segmented Neutrophils PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B7 (PAGE 1): CLINICAL CHEMISTRY - SUMMARY - FO GENERATION MALE RATS (See abbreviations on the last page of this table.) RATS TESTED TP (G/DL) A (G/DL) GLU (MG/DL) CHOL (MG/DL) TBILI (MG/DL) BUN (MG/DL) CREAT (MG/DL) CK (U/L) N MEAN f S.D. MEAN f S.D. MEAN f S.D. MEAN f S.D. MEAN f S.D. MEAN * S.D. MEAN f S.D. MEAN f S.D. 10 6.3 f 0.36 4.3 f 0.22 145 f 16.8 57 f 8.3 0.2 f 0 . 0 7 16 5 1.5 0.3 f 0.04 288 f 172.6 10 6.1 f 0.28 4.1 f 0.19 141 f 26.0 41 f 11.1** 0.1 f 0.05 16 f 0.8 0.3 f 0.05 246 f 111.5 10 6.3 f 0.33 4.3 f 0.15 182 f 58.2 46 * 11.6" 0.1 f 0.06 16 f 1.8 0.3 f 0.06 239 -4- 112.9 10 6.2 f 0.28 4.2 f 0.17 162 f 25.2 43 f 13.1** 0.1 f 0.05 17 f 1.6 0.3 f 0.03 196 f 53.5 10 6.3 f 0.28 4.5 f 0.20** 165 f 33.7 33 f 7.2** 0.2 f 0.06 21 f 2.4** 0.3 f 0.05 243 f 117.4 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B7 (PAGE 2): CLINICAL CHEMISTRY - SUMMARY - Fo GENERATION MALE RATS (See abbreviations on the last page of this table.) RATS TESTED AST (U/L) ALK (U/L) CA (MG/DL) PHOS (MG/DL) TRI (MG/DL) NA (MMOL/L) K (MMOL/L) CL (MMOL/L) G (G/DL) N MEAN f S.D. MEAN f S.D. MEAN f S.D. MEAN f S . D . MEAN f S.D. MEAN f S.D. MEAN f S.D. MEAN f S.D. MEAN f S.D. 10 96 f 22.4 105 -+ 14.3 10.9 f 0.48 9.5 f 1.40 52 f 21.8 146 f 1.4 6.6 f 2.10 98 -+ 2.7 2 .o f 0.19 10 121 f 92.6 111 f 36.9 10.7 f 0.44 9.4 f 1.11 47 f 17.2 146 f 1.3 6.1 f 0.89 99 f 1.5 2.0 k 0.16 10 117 f 40.9 100 f 12.4 11.1 f 0.44 9.9 * 2.09 36 f 13.8 147 f 1.6 6.5 -+ 1.27 100 f 2.3 2.0 -+ 0.24 10 198 f 248.7 115 f 25.1 11.1 f 0.34 9.7 f 1.43 36 f 27.9 147 f 1.0 6.0 --+ 0.60 100 f 2.7 2.0 f 0.16 10 97 f 15.6 144 f 37.5** 11.5 f 0.38** 10.2 f 1.33 17 f 8.0** 146 f 2.2 6.9 f 1.55 100 f 1.2 1.8 f 0.13 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B7 (PAGE 3 ) : CLINICAL CHEMISTRY - SUMMARY - FO GENERATION MALE RATS A GLU CHOL TBILI BUN CREAT ALT AST ALK CA PHOS TRI NA K CL G Albumin Glucose Cholesterol Total Bilirubin Blood Urea Nitrogen Creatinine Alanine Aminotransferase Aspartate Aminotransferase Alkaline Phosphatase Calcium Phosphorus Triglycerides Sodium Potassium Chloride Globulin I I I I I I I I 1 I * I I E I u I >o I I I I I I m x H I rllrl I I 0 I I I3 I I I I I I I I I I 1 I I I I I I I I I I I I I r dr rl m rl rl +I +I P P * W P P m m I I I I VlI .I WI rll I +I I I mI *I rll 91 *I I I I I 1 I I rll *I I I I I I I I I I I I I PI mi I +I I I mi .I 031 u)I *I I I I I I I I I I w I I I I X I I I3 I I E I I I 1 I H r l I Ul H IH Irl IH I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I m I I I HIT I IH * I d I 0 1 I I I I I I I I I I co W rn cv * * +I +I m u) * u) * * 0 m m rn rl +I rl W P m I cvl -1 031 *I I +I I I rll *I ml u)I *I I I I I I I I mI *I *I m II +I I I PI I I I I I I I I I I I I I I I I I I I 0 I I cy I I I H O I Ul I I Irl I I I I I I I I I I I I I I I I CJ * cv m cv cv +I +I P m cv P 03 m 0 * cv cv 03 r m m +I +I cv m 0 0 c*o r*n I I rnl .I mI ml I +I I I I I I I I I I I I I I I I I I2 I I I I 1 I I I I I I I I I I 6' 6' 9 ? rn rn w w E z 6' 6' ? 9 rn u1 2! E I I I I i 2 ; h I -I rl 03 0 cv w I 03 *rl .. v u h v *Id rl 03 rl W m m * co a n2 n2 I I 418-028:PAGE B-15 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B9 (PAGE 1): BODY WEIGHT CHANGES - SUMMARY - FO GENERATION MALE RATS BODY WEIGHT CHANGE ( G ) DAYS 1 - 8 DAYS 8 - 14a DAYS 1 - 14a DAYS 29b- 36 DAYS 36 - 43 MEANfS.D . MEANkS .D . MEANfS.D . MEANfS.D. MEAN&S .D. +39.9 f 12.3 +24.7 f 6.8 +64.6 f 16.5 +23.6 f 8.6 +10.3 f 10.8 +37.2 f 11.0 +24.6 f 5.0 +61.8 & 14.6 +16.2 f 7.8* +8.7 f 11.2 +38.5 f 10.0 +23.8 f 11.1 +62.3 f 18.3 +20.2 f 5.6 +5.7 f 14.4 +41.0 f 12.1 +26.3 f 8.2 +67.3 & 16.3 +12.6 f 5.4** +9.4 f 12.9 +33.4 f 5.6 +20.6 f 4.8 +54.0 f 7.0 +16.5 f 8.4* +7.9 & 13.7 DAYS 29bTERMINATION MEANfS.D . +15.9 f 11.7 +3.4 f 7.9** +6.5 f 10.1* -0.2 f 7.7** +3.8 & 11.8** DAYS 1 - TERMINATION MEANfS.D. +129.8 f 25.5 +114.8 f 27.5 +121.2 f 37.7 +122.7 f 30.0 +98.2 f 13.9** ________________________________________------------------------------------------------------------------------------------------ DAY (S) = DAY (S) OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. * Significantly different from the Group I value (~10.05). * * Significantly different from the Group I value (~10.01). I I I I I I I I I I I I I I I I ?:I2 +I I I 0-l I I I I u, I I (v I I I I I I I I I I I I I I I I I I I I I I I I I I I > m I In +I IH 114 I I W I I I I W I I (v 3cll I I I I I 1 I I I I I I w I I I I m I I I I m X I I B I I I I E X I H r l I Ln +I IH 11-4 IH I rl rn I I I I u, I I (v r 0 I I I I P 0 I I I k I I B I I I I I E I I 0 I I 0 I I I I cu I I I I I I +I I co I I I I P I I (v I I I I I I I I I 1 I I I I I I rl I I I I m I I I I I I I H O I Ul +I w I Irl I I I I I I a, I I (v I I I I I I I I I I I I I I I I I I I I I I I IZ 1 I I I I I I I I I I I h I I I I I I I I h I I rl I -I td W W (v 0 I I 03 rl rl w 0 rl a n w w 418-028:PAGE B-17 PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B11 (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - SUMMARY - Fo GENERATION MALE RATS FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8a MEANfS.D . 78.3 f 7.2 77.7 f 3.9 80.1 f 6.9 78.2 f 4.3 74.7 * 3.3 DAYS 29b- 36 MEANfS .D . 64.2 f 3.2 62.8 f 4.5 64.1 f 4.2 61.7 f 3.7 63.2 * 3.6 [ 14lc .................................................................................................................................... DAYS = DAYS OF STUDY [ ] = NUMBER OF VALUES AVERAGED a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Excludes values that were associated with spillage. w X E 5%N cEn w. 0 0 E I * I Ec-7 H h rl a, cv 0 I a, *rl I I I I I I - w I I I I I I I I rl In- n I I 0 rlo 0 -+e - I I I 3 0 I In +I 0 \O In 0 I rllrl 0 In0 rl 0 I I In rl rl I I Y I I cv In In I I rl rl I I I I I I I I I I I I I I I I I I 0 I I cv I I h r I I n 0 I I +I I0 0 ;?Im:: a, 0 I I rl rl I I Y Y 1 I m m m I I rl rl I I I I I I I I I I I I I I - I I I I cv I I I I rl I I 0 0 I I I H r l I In +I 0 IH Irl 0 IH I a, rl In 0 rl 0 rl 1 I Y Y I I cv In In I I rl rl I I I I I I I I I I I I I I I I I I cv I I I I rl n In- I I 0 rlo 0 I I I H m I Ul +I 0 \O In 0 IH *Id 0 In0 rl 0 I 0 1 cn rl rlrl rl I I Y Y Y I I rl In In I I rl rl I I I I I I I I I I I I I I * I I I I I I I I rl h w - A I I m rlo 0 I I cv I H O I In I Irl +I mcn \O wo I I * -+z! m I I v I I w 0 rl 0 rl v w I I rl rl I I I I I I I I I I I I I I I 12 I I I h z - O\O z z - z Y \ o\o I I I I I I I I I I I I u I I I I I I rd I I I I X 0 W I I I I I I In- I rlo I .I \o I In0 I rlrl I -1 I I I I I I I I I I I I In- I rlo I .I \o I ma, I rl-; I I I I I I I I I I I I Inn I rlr I *I \w I ma, I rlu; lm I& Ird Ik I 418-028:PAGE B-19 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B13 (PAGE 1): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - Fo GENERATION MALE RATS HOME CAGE BEHAVIOR 1: Sleeping 8** 2: Awake, Immobile 2** 3: Normal movement 0 4: Unusual posture 0 5: Unusual behavior 0 ALTERATIONS (HOME CAGE) 1: None N 9 10 10 10 10 2: Stereotyped behavior N la 0 0 0 0 3: Bizarre behavior N 0 0 0 0 0 4: Limb twitches/tremor N 0 0 0 0 0 5: Whole body tremor/spasm N 0 0 0 0 0 6: Unusual posture N 0 0 0 0 0 7: Tonic-clonic seizure N 0 0 0 0 0 REACTION TO REMOVAL (1) Sits quietly N 10 10 10 10 10 ( 21 Vocalization N 0 0 0 0 0 ( 3 ) Runs or freezes N 0 0 0 0 0 ( 4 ) Tail or throat rattles N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 REACTION TO HANDLING (1) No resistance N 10 10 10 10 10 (2) Vocalization N 0 0 0 0 0 (3) Tense N 0 0 0 0 0 ( 4 ) Squirming N 0 0 0 0 0 .....................................M.E.A..N..S.C..O.R.E............1...0......................1...0.......................1...0.......................1...0........................1....0......... n: = Category number for descriptive test item. (n) = Score assigned to graded test items; mean score was calculated by multiplying each score by the number of rats with that score and then dividing the sum of the products by the total number of rats a. Excludes a value for rat 19131, which appeared incorrectly recorded. * * Significantly different from the Group I value (~10.01). PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B13 (PAGE 2): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - Fo GENERATION MALE RATS REARS IN OPEN FIELD DEFECATION IN OPEN FIELD 1: None 2: Feces normal 3: Soft or liquid feces MEANfS.D . N N N URINATION IN OPEN FIELD (1) None (2) Normal urination ( 3 ) Excess urination N N N MEAN SCORE LEVEL OF AROUSAL (1) Stuporous (2) Sluggish (31 Apparently normal (4) Sudden darting (5) Freezing, vocalization N N N N N MEAN SCORE ALTERATIONS (OPEN FIELD) 1: None N 2: Stereotyped behavior N 3 : Bizarre behavior N 4: Limb twitches/tremor N 5 : Whole body tremor/spasm N 6: Unusual posture N 3.3 f 2.5 10 0 0 6 4 0 1.4 0 0 10 0 0 3.0 10 0 0 0 0 0 6.2 * 3.2 1 9 0 1.9 0 0 10 0 0 3.0 10 0 0 0 0 0 3.9 f 1.9 7 3 0 6 4 0 1.4 0 0 10 0 0 3.0 10 0 0 0 0 0 4.5 * 2.1 2 8 0 1.8 0 0 10 0 0 3.0 10 0 0 0 0 0 4.5 f 3.1 3 7 0 1.7 0 0 10 0 0 3.0 10 0 0 0 0 0 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B13 (PAGE 3): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - Fo GENERATION MALE RATS GAIT PATTERN 1: Apparently normal 2: Ataxic 3: Limbs splay or drag 4: Spastic, tip-toe 5: Duck-walk 6: Scissors gait 10 10 10 10 10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GAIT ABNORMALITY, SEVERITY (1) Normal gait N 10 10 10 10 10 (2) Slight N 0 0 0 0 0 (3) Moderate N 0 0 0 0 0 (4) Extreme N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 PALPEBRAL CLOSURE (1) Wide open N 10 10 10 10 10 (2) Slightly drooping N 0 0 0 0 0 (3) Half-closed N 0 0 0 0 0 (4) Completely shut N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 PROMINENCE OF THE EYE 1: Normal N 10 10 10 10 10 2: Exophthalmos N 0 0 0 0 0 (n) = Score assigned to graded test items; mean score was calculated by multiplying each score by the number of rats with that score and then dividing the sum of the products by the total number of rats PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) LACRIMATION (1) No excess N 10 10 10 10 10 ( 2 ) Excess at eyelid margin N 0 0 0 0 0 ( 3 ) Margin persistently damp N 0 0 0 0 0 (4) Extends beyond margin N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 SALIVATION (1) No excess N 10 10 10 10 10 ( 2 ) Margin of mouth wet N 0 0 0 0 0 ( 3 ) 1/4 to 1/2 submandibular N 0 0 0 0 0 (4) Entire submandibular N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 PILOERECTION N 0 0 0 0 0 ABNORMAL RESPIRATION N 0 0 0 0 0 APPEARANCE (1) Clean and groomed N 10 10 10 10 10 (2) Unkempt N 0 0 0 0 0 ( 3 ) Urine and/or fecal stain N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 VISUAL REACTION (1) None N 0 0 0 0 0 (2) Orienting N 10 10 10 10 10 (3) Startle N 0 0 0 0 0 (4) More energetic reaction N 0 0 0 0 0 (5) Attacks N 0 0 0 0 0 PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B13 (PAGE 5): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - Fo GENERATION MALE RATS TACTILE REACTION (1) None N 0 0 0 0 0 ( 2 ) Orienting N 10 10 10 10 10 ( 3 ) Startle N 0 0 0 0 0 ( 4 ) More energetic reaction N 0 0 0 0 0 ( 5 ) Attacks N 0 0 0 0 0 MEAN SCORE 2.0 2.0 2.0 2 .o 2.0 AUDITORY REACTION (1) None N 0 0 0 0 0 ( 2 ) Orienting N 0 0 0 0 0 ( 3 ) Startle N 10 10 10 10 10 ( 4 ) More energetic reaction N 0 0 0 0 0 ( 5 ) Intense vocalization N 0 0 0 0 0 MEAN SCORE 3.0 3.0 3.0 3 .O 3 -0 TAIL-PINCH REACTION (1) None N 0 0 0 0 0 ( 2 1 Orienting N 10 10 10 10 10 ( 3 ) Startle N 0 0 0 0 0 ( 4 ) More energetic reaction N 0 0 0 0 0 ( 5 ) Attacks N 0 0 0 0 0 .....................................M.E..A.N..S..C.O.R.E............2...0......................2...0.......................2...0.......................2... 0.........................2.. .0......... n: = Category number for descriptive test item. ( n ) = Score assigned to graded test items; mean score was calculated by multiplying each score by the number of rats with that score and then dividing the sum of the products by the total number of rats PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B13 (PAGE 6): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - FO GENERATION MALE RATS MALE RATS N 10 AIR RIGHTING RESPONSE (1) All feet land on ground N 10 (2) Lands on side N 0 (3) Lands on back N 0 MEAN SCORE 1.0 PUPIL RESPONSE TO LIGHT N 10 FORELIMB GRIP TEST Maximum (GI Average (GI MEANfS.D . MEANfS.D . 444.0 f 49.6 410.5 f 52.7 HINDLIMB GRIP TEST Maximum (G) Average (GI MEANfS.D . MEANfS.D . 386.0 f 128.8 360.9 f 121.8 LANDING FOOT SPLAY Average (CM) MEANfS.D - 7.5 * 1.0 10 10 10 10 10 10 0 0 0 0 0 0 1.0 1.0 1.0 10 10 10 451.5 f 57.5 400.6 f 62.7 456.0 f 111.1 409.4 f 93.7 418.5 f 367.5 2 125.8 109.0 456.0 f 400.8 a 109.4 101.1 457.0 i: 102.5 419.5 f 97.8 414.5 f 58.9 364.5 f 67.5 8.6 f 2.2 9.0 f 1.3 8.6 2 1.9 10 10 0 0 1.0 10 406.5 f 120.7 359.2 f 104.6 467.5 ? 113.4 439.4 f 95.8 8.0 f 2.1 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B14 (PAGE 1): MOTOR ACTIVITY - SUMMARY - Fo GENERATION MALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) I I1 0 0.3 NUMBER OF MALE RATS N 10 10 NUMBER OF MOVEMENTS BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN i: S . D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN S.D 64.8 k 6.6 66.9 k 9.4 69.1 f 6.8 72.3 f 10.5 60.1 k 12.0 48.3 f 25 - 4 39.4 f 21.0 37.9 f 34.6 27.6 k 25.2 25.0 f 30.2 24.2 f 27.1 37.8 i: 28.0 42.1 k 31.9 37.6 f 30.7 23.3 k 26.0 23.6 f 31.2 23.6 k 26.8 22.8 24.7 746.4 f 217.7 64.3 63.6 59.8 61.2 54.7 52.8 50.0 46.7 49.2 53.1 35.1 35.1 43.0 43.8 35.5 37.6 39.2 34.4 859.1 f 6.7 f 11.5 f 16.2 f 20.0 f 23.1 ~t: 26.8 f 25.8 f 24.8 f 22.8 f 20.5 f 28.3 f 25.7 f 34.6 f 29.4 f 22.3 f 28.5 f 30.2 f. 23.6 f 251.7 TOTAL = SUM OF BLOCKS; EACH BLOCK CONSISTS OF A 5 MINUTE PERIOD. I11 IV 1 3 10 10 63.5 f 8.6 64.6 f 9.0 65.1 k 8.3 68.2 f 12.1 65.6 f 5.4 64.2 f 12.8 59.8 f 14.7 54.0 f 19.9 49.7 f 23.4 47.0 f 28.9 51.2 f 25.5 45.5 f 25.9 44.0 f 35.4 36.5 f 26.3 39.8 f 36.2 31.2 f 31.0 29.1 f 25.9 35.9 f 30.2 914.9 f 212.7 62.1 f 10.0 73.5 f 4.4 71.6 f 11.7 65.4 f 11.9 63.1 f 16.5 59.9 f 14.9 60.6 f 8.2 57.0 f 19.4 55.3 f 20.4 50.0 f 23.1 51.7 f 26.7 49.1 f 26.0 50.4 f 25.6 59.2 f 19.0 51.3 f 21.1 50.9 f 22.7 53.5 f 24.3 42.4 f 27.0 1027.0 f 176.0 V 10 10 64.0 f 8.0 69.6 f 7.2 67.5 f 7.8 56.0 +- 11.7 56.9 f 15.6 57.4 f 9.9 56.8 f 11.5 49.7 f 22.2 52.2 f 16.9 49.6 f 15.6 41.5 f 21.2 * 47.6 f 13.0 43.0 22.3 43.8 f 29.6 43.0 f 25 - 4 42.5 f 29.4 37.3 f 29.6 34.2 f 23.1 912.6 -+ 139.0 PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T - 7 7 0 6 . 1 ) NUMBER OF MALE RATS N 10 TIME (SECONDS) SPENT IN MOVEMENT BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 1 0 BLOCK 11 BLOCK 1 2 BLOCK 1 3 BLOCK 1 4 BLOCK 1 5 BLOCK 1 6 BLOCK 1 7 BLOCK 1 8 TOTAL MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN k S.D MEAN f S.D MEAN k S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN f S.D MEAN i: S.D MEAN f S.D 201.1 187.4 155.5 141.4 100.7 91.2 70.2 61.3 44.0 48.9 40.2 58.1 66.9 65.6 39.1 31.9 32 .O 30.5 1466.0 f 24.1 f 22.6 f 26.2 f 33.3 f 28.5 f 54.3 f 44.2 f 60.8 f 46.2 f 68.7 f 59.5 f 46.2 f 56.7 f 58.7 f 49.7 f 45.1 f 38.5 39.6 f 453.9 10 201.8 174.4 140.3 138.3 131.1 89.7 90.2 83.7 80.9 86.6 71.4 67.1 86.9 75.7 53.1 55.5 62.5 47.4 1736.6 f 17.8 ? 32.9 f 63.3 f 53.0 f 71.4 *k 52.6 55.5 f 55.9 f 40.5 f 40.3 f 63.0 k 60.9 f 82.2 f 55.9 *f 4 6 . 5 47.5 f 56.4 f 46.1 f 607.9 10 214.8 195.8 172.5 142.4 150.3 118.4 110.2 109.9 93.6 77.4 86.0 91.7 52.1 54.1 68.0 60.3 46.9 56.3 1900.7 f 13.0 f 33.8 f 35.6 f 24.7 k 20.6 f 41.2 i: 4 6 . 9 f 52.7 f 59.3 i: 5 5 . 3 f 49.1 f 62.1 f 44.8 f 47.4 f 65.3 f 73.9 k 53.5 f 54.7 f 469.7 10 204.6 180.2 140.6 140.9 119.2 109.7 118.6 116.1 103.6 83.1 97.7 81.1 80.2 107.8 74.2 87.2 91.4 66.9 2003.1 rf: 2 3 . 7 f 20.9 f 27.5 +- 3 4 . 1 f 29.0 i: 4 8 . 8 f 45.9 f 50.9 ~t: 5 6 . 4 f 47.0 k 61.8 f 49.1 f 41.2 f 40.9 f 35.4 f 43.5 i: 4 9 . 7 f 43.5 f 453.4 10 194.8 170.1 148.7 130.9 113.8 102.7 107.9 72.0 93.6 76.8 67.2 72.9 70.5 73.3 64.2 77.1 51.0 54.1 1741.6 f 27.2 f 32.5 f 39.4 +_ 5 0 . 6 k 45.7 f 40.8 f 38.8 f 35.9 f 47.3 f 44.1 f 40.0 f 33.7 +- 4 8 . 9 f 59.7 f 36.9 f 68.2 f 44.8 f 41.8 f 463.4 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B15 (PAGE 1): SPERM MOTILITY, COUNT AND DENSITY - SUMMARY - Fo GENERATION MALE RATS NUMBER MOTILE MOTILE PERCENT STATIC COUNT (NONMOTILE) TOTAL COUNT a SPERM COUNT b MEANfS.D MEANfS .D . 289.1 f 51.3 92.1 f 3.1 MEANfS .D . MEANfS.D. MEANfS.D. 25.2 314.3 ~f: 11.3 f 57.7 170.1 f 48.8 307.4 f 77.3 93.3 f 2.8 23.5 f 14.4 330.9 89.7 158.2 f 55.2 312.0 f 80.2 92.5 f 3.4 23.1 rt 7.8 335.1 f 75.6 161.0 f 41.6 307.3 85.5 f 149.6 f 22.3 325.4 f 81.5 91.1 f 7.4 32.7 340.0 f 15.3 & 152.4 124.6 k 50.2 32.4 f 23.4 357.8 & 81.2 174.5 f 37.9 20 fields were evaluated. b. Sperm count used in the calculation of sperm density. Ten fields were evaluated. c. The sperm density was calculated by dividing the sperm count by ,thevolume in the image area (34.3 x mL), multiplying by 2 (dilution factor) and multiplying by to obtain the sperm concentration. The calculated sperm concentration value (rounded to 1 decimal place) was multiplied by 50 (volume) and divided by the weight of the left cauda epididymis (see Table B19 for the weight of the left cauda epididymis) to obtain the sperm density. The calculated value will vary by approximately 0.8% from the Computer Automated Sperm Analysis because the digital image evaluated is slightly smaller (4 pixels) than the actual field causing a slight underestimate of the actual volume and an overestimate of the concentration. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B16 (PAGE 1): SPERM MORPHOLOGY - SUMMARY - FO GENERATION MALE RATS NORMAL DETACHED HEAD NO HEAD BROKEN FLAGELLUM BENT FLAGELLUM MEANfS.D . MEANfS.D . MEANfS.D . MEAN+S .D . MEANfS.D . 192.3 ?I 3.1 3.7 f 2.4 3.4 f 1.7 0.4 f 0.5 0.2 f 0.4 193.3 f 3.0 3.6 f 2.2 2.1 f 1.2 0.8 f 0.8 0.2 f 0.4 193.3 f 3.5 3.1 f 2.0 2.6 f 1.4 0.7 f 0.7 0.3 +- 0.7 173.8 f 59.8 6.8 f 11.0 10.5 f 23.1 8.5 f 24.8 0.4 f 1.3 193.3 f 3.1 3.8 f 2.6 1.8 f 1.3 1.0 f 0.9 0.1 f 0.3 w PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B17 (PAGE 1): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION MALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B17 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION MALE RATS b 13 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B17 (PAGE 3 ) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION MALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B18 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION MALE RATS I 0 19109 DS 43 42 ALL TISSUES APPEARED NORMAL. 19115 DS 43 42 ALL TISSUES APPEARED NORMAL. 19116 DS 44 43 KIDNEYS: BILATERAL, MOTTLED TAN AND RED. ALL OTHER TISSUES APPEARED NORMAL. 19119 DS 44 43 ALL TISSUES APPEARED NORMAL. 19122 DS 44 43 ALL TISSUES APPEARED NORMAL. 19125 DS 45 44 ALL TISSUES APPEARED NORMAL. 19131 DS 45 44 ALL TISSUES APPEARED NORMAL. 19132 DS 45 44 ALL TISSUES APPEARED NORMAL. 19134 DS 46 45 ALL TISSUES APPEARED NORMAL. 19135 DS 46 45 ALL TISSUES APPEARED NORMAL. 19143 DS 47 46 ALL TISSUES APPEARED NORMAL. 19151 DS 47 46 ALL TISSUES APPEARED NORMAL. 19157 DS 47 46 ALL TISSUES APPEARED NORMAL. 19161 DS 47 46 ALL TISSUES APPEARED NORMAL. 19167 DS 47 46 ALL TISSUES APPEARED NORMAL. ________________________________________------------------------------------------------------ DS = DAY OF STUDY a. Refer to the individual clinical observations table (Table B17) for external observations confirmed at necropsy. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B18 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION MALE RATS 19106 DS 43 42 ALL TISSUES APPEARED NORMAL. 19108 DS 44 43 ALL TISSUES APPEARED NORMAL. 19110 DS 44 43 ALL TISSUES APPEARED NORMAL. 19113 DS 44 43 KIDNEYS: LEFT, PELVIS, SLIGHT DILATION. ALL OTHER TISSUES APPEARED NORMAL. 19120 DS 45 44 ALL TISSUES APPEARED NORMAL. 19129 DS 45 44 ALL TISSUES APPEARED NORMAL. 19136 DS 45 44 KIDNEYS: BILATERAL, MOTTLED. ALL OTHER TISSUES APPEARED NORMAL. 19138 DS 46 19139 DS 46 19147 DS 47 19153 DS 47 19164 DS 47 19165 DS 47 19171 DS 47 ____________-__---__----------------- DS = DAY OF STUDY a. Refer to the individual clinical observations 45 45 46 46 46 46 46 table ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. (Table B17) for external observations confirmed at necropsy . 13 W W P PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B18 (PAGE 3): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION MALE RATS .................................................................................................................................... DOSAGE GROUP RAT DAY OF DOSES DOSAGE (MG/KG/DAY) NUMBER NECROPSY ADMINISTERED OBSERVATIONS a ____________________--------------- I11 1 19101 DS 43 42 ALL TISSUES APPEARED NORMAL. 19105 DS 43 42 ALL TISSUES APPEARED NORMAL. 19107 DS 43 42 ALL TISSUES APPEARED NORMAL. 19112 DS 44 43 ALL TISSUES APPEARED NORMAL. 19114 DS 44 43 ALL TISSUES APPEARED NORMAL. 19121 DS 45 44 ALL TISSUES APPEARED NORMAL. 19123 DS 45 44 ALL TISSUES APPEARED NORMAL. 19130 DS 46 45 ALL TISSUES APPEARED NORMAL. 19137 DS 46 45 ALL TISSUES APPEARED NORMAL. 19146 DS 46 45 ALL TISSUES APPEARED NORMAL. 19155 DS 47 46 ALL TISSUES APPEARED NORMAL. 19159 DS 47 46 ALL TISSUES APPEARED NORMAL. 19172 DS 47 46 ALL TISSUES APPEARED NORMAL. 19173 DS 47 46 ALL TISSUES APPEARED NORMAL. 19174 DS 47 46 ALL TISSUES APPEARED NORMAL. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B18 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION MALE RATS 19142 DS 45 44 EPIDIDYMIDES: BILATERAL, SMALL; LEFT CAUDA, SMALL. TESTES: BILATERAL, SMALL. ALL OTHER TISSUES APPEARED NORMAL. 19144 DS 45 44 19148 DS 46 45 19150 DS 46 45 19156 DS 47 46 19160 DS 47 46 19162 DS 47 46 19163 DS 47 46 19166 DS 47 46 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - -. --- -- --- ----- --- ----- --- -- --- - DS = DAY OF STUDY a. Refer to the individual clinical observations table ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ........................................................................... (Table B17) for external observations confirmed at necropsy. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B18 (PAGE 5): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION MALE RATS 19117 DS 43 19124 DS 43 19126 DS 44 19127 DS 44 19128 DS 45 19140 DS 45 19145 DS 46 19149 DS 46 19152 DS 46 19154 DS 47 19158 DS 47 19168 DS 47 19169 DS 47 19170 DS 47 .................................................. DS = DAY OF STUDY a. Refer to the individual clinical observations 42 42 43 43 44 44 45 45 45 46 46 46 46 46 table ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. (Table B17) for external observations confirmed at necropsy. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B19 (PAGE 1): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - INDIVIDUAL DATA - Fo GENERATION MALE RATS 19109 438. 1.6529 0.38 1.1854 0.27 2.29 0.52 14.18 3.24 1.51 0.34 1.51 0.34 19115 466. 1.6847 0.36 1.4274 0.31 2.27 0.49 16.71 3.58 1.99 0.43 1.99 0.43 19116 490. 1.6820 0.34 1.5838 0.32 2.28 0.46 15.18 3.10 2.16 0.44 2.12 0.43 19119 505. 1.6364 0.32 0.7632 0.15 2.63 0.52 14.66 2.90 2.26 0.45 2.38 0.47 19122 451. 1.6956 0.38 1.0942 0.24 2.45 0.54 13.10 2.90 1.92 0.42 1.81 0.40 19125 508. 1.6796 0.33 1.1823 0.23 2.44 0.48 16.13 3.18 2.31 0.45 2.29 0.45 19131 499. 1.6534 0.33 1.2496 0.25 2.45 0.49 16.94 3.39 2.13 0.43 2.22 0.44 19132 439. 1.8532 0.42 1.8597 0.42 2.34 0.53 11.94 2.72 1.78 0.40 1.89 0.43 19134 419. 1.4818 0.35 1.3858 0.33 2.35 0.56 12.01 2.87 1.76 0.42 1.84 0.44 19135 482. 1.7570 0.36 1.1710 0.24 2.45 0.51 15.48 3.21 2.00 0.41 2.15 0.45 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - - - - - - - - - -.-.-.-.-.-.-.-.-.-.-.-.-.-.-. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) x 100. a. Damaged during processing (weight not affected). PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B19 (PAGE 2): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - INDIVIDUAL DATA - FO GENERATION MALE RATS 19109 438. 0.027 6.16 19115 466. 0.041 8.80 19116 490. 0.035 7.14 19119 505. 0.048 9.50 19122 451. 0.033 7.32 19125 508. 0.034 6.69 19131 499. 0.041 8.22 19132 439. 0.036 8.20 19134 419. 0.024 5.73 19135 482. 0.031 6.43 ........................................ ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value was multiplied by 1000. 0.027 6.16 0.56 0.13 0.37 0.08 1.41 0.32 0.040 8.58 0.77 0.16 0.41 0.09 1.62 0.35 0.035 7.14 0.95 0.19 0.49 0.10 1.51 0.31 0.046 9.11 0.79 0.16 0.30 0.06 1.62 0.32 0.031 6.87 0.79 0.18 0.33 0.07 1.68 0.37 0.031 6.10 0.76 0.15 0.70 0.14 1.54 0.30 0.036 7.21 1.03 0.21 0.47 0.09 1.67 0.33 0.038 8.66 0.73 0.17 0.27 0.06 1.40 0.32 0.025 5.97 0.60 0.14 0.44 0.10 1.46 0.35 0.030 6.22 0.90 0.19 0.33 0.07 1.63 0.34 - - - - - - - - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - - -. .- .-.-.-.-.-.-. -. -. .-. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ABS . WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) x 100. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B 1 9 (PAGE 3): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - FO GENERATION MALE RATS .................................................................................................................................... DOSAGE GROUP I1 0.3 MG/KG/DAY .................................................................................................................................... EPIDIDYMIS CAUDA EPIDIDYMIS TESTIS SEMINAL VESICLES SEMINAL VESICLES EPIDIDYMIS RAT TERMINAL BODY LEFT LEFT LEFT WITH FLUID WITHOUT FLUID RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. RFAL. ABS. REL. WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW .................................................................................................................................... 19102 428. 0.6197 0.14 0.2867 0.07 1.5498 0.36 1.7556 0.41 0.9426 0.22 0.6523 0.15 19106 476. 0.6623 0.14 0.2962 0.06 1.6495 0.35 1.0913 0.23 0.5991 0.12 0.6675 0.14 19108 474. 0.6465 0.14 0.3502 0.07 1.8847 0.40 1.2460 0.26 0.7062 0.15 0.6572 0.14 19110 453. 0.5411 0.12 0.2779 0.06 1.4915 0.33 1.8646 0.41 0.8697 0.19 0.5439 0.12 19113 444. 0.6310 0.14 0.3379 0.08 1.7844 0.40 1.7236 0.39 0.8319 0.19 0.6568 0.15 19120 486. 0.6180 0.13 0.3209 0.07 1.6588 0.34 1.6952 0.35 0.8335 0.17 0.6724 0.14 19129 435. 0.6644 0.15 0.3181 0.07 1.8000 0.41 1.7928 0.41 0.8254 0.19 0.6538 0.15 19136 495. 0.6785 0.14 0.3219 0.06 1.7629 0.36 1.2650 0.26 0.6950 0.14 0.6914 0.14 19138 365. 0.6686 0.18 0.3604 0.10 1.7956 0.49 1.5421 0.42 0.8728 0.24 0.6831 0.19 19139 432. 0.6280 0.14 0.3114 0.07 1.7773 0.41 1.4625 0.34 0.9423 0.22 0.6675 0.15 ________________________________________-------------------------------------------------------------------------------------------- TESTIS PROSTATE BRAIN LIVER KIDNEY KIDNEY RAT TERMINAL BODY RIGHT LEFT RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW .................................................................................................................................... 19102 428. 1.5930 0.37 1.2374 0.29 2.19 0.51 14.53 3.39 1.92 0.45 1.95 0.46 19106 476. 1.6779 0.35 1.0360 0.22 2.57 0.54 17.19 3.61 1.97 0.41 2.00 0.42 19108 474. 1.9171 0.40 1.1892 0.25 2.52 0.53 14.75 3.11 1.90 0.40 1.98 0.42 19110 453 * 1.4694 0.32 1.1812 0.26 2.42 0.53 13.95 3.08 1.96 0.43 2.00 0.44 19113 444. 1.7350 0.39 1.2130 0.27 2.58 0.58 13.08 2.94 1.96a 0.44 2.11 0.48 19 12 0 19 12 9 19136 19138 19139 486. 435. 495. 365. 432. 1.6765 1.8164 1.8039 1.7840 1.8078 0.34 0.42 0.36 0.49 0.42 1.4290 1.1232 1.3313 0.9939 1.2660 0.29 0.26 0.27 0.27 0.29 2.38 2.45 2.41 2 -46 2 -40 0.49 0.56 0.49 0.67 0.56 14.26 13.37 15.75 11.25 14.10 2.93 3.07 3.18 3.08 3.26 1.72 1.89 1.83 1.58 1.88 0.35 0.43 0.37 0.43 0.44 1.76 1.90 1.69 1.66 1.93 0.36 0.44 0.34 0.45 0.45 ______________________________^_________-------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. a. Rat 19113 had slight dilation of the pelvis of the left kidney. See the individual necropsy observations table (Table B 1 8 ) . 5 (?O 0 19 PP 8?td b 0 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B19 (PAGE 4): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION MALE RATS .................................................................................................................................... DOSAGE GROUP I1 0.3 MG/KG/DAY .................................................................................................................................... ADRENAL ADRENAL SPLEEN THYMUS HEART RAT TERMINAL BODY LEFT RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW a WT. % TBW a W T . % TBW WT. % TBW WT. % TBW .................................................................................................................................... 19102 428. 0.041 9.58 0.038 8.88 0.89 0.21 0.52 0.12 1.38 0.32 19106 476. 0.024 5.04 0.024 5.04 0.86 0.18 0.42 0.09 1.56 0.33 19108 474. 0.030 6.33 0.030 6.33 0.73 0.15 0.47 0.10 1.64 0.34 19110 453. 0.037 8.17 0.038 8.39 0.85 0.19 0.41 0.09 1.59 0.35 19113 444. 0.046 10.36 0.044 9.91 0.81 0.18 0.48 0.11 1.47 0.33 19120 486. 0.042 8.64 0.038 7.82 0.83 0.17 0.40 0.08 1.56 0.32 19129 19136 435. 495. 0.037 0.033 8.50 6.67 0.034 0.034 7.82 6.87 0.76 0.72 0.17 0.14 0.34 0.58 0.08 0.12 1.55 1.71 0.36 0.34 19138 365. 0.028 7.67 0.027 7.40 0.73 0.20 0.30 0.08 1.30 0.36 19139 432. 0.032 7.41 0.032 7.41 0.92 0.21 0.54 0.12 1.46 0.34 ________________________________________------------------------------------------------------------------------------------------ ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) x 100. a. Value was multiplied by 1000. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B19 (PAGE 5): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION MALE RATS DOSAGE GROUP I11 1 MG/KG/DAY .................................................................................................................................... EPIDIDYMIS CAUDA EPIDIDYMIS TESTIS SEMINAL VESICLES SEMINAL VESICLES EPIDIDYMIS RAT TERMINAL BODY LEFT LEFT LEFT WITH FLUID WITHOUT FLUID RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW WT. % TBW WT . % TBW WT. % TBW WT. % TBW WT. % TBW .................................................................................................................................... 19101 19105 353. 505. 0.6021 0.7012 0.17 0.14 0.2993 0.3377 0.08 0.07 1.6371 2.0077 0.46 0.40 1.1004 1.7433 0.31 0.34 0.5923 0.7941 0.17 0.16 0.6071 0.7481 0.17 0.15 19107 437. 0.5355a 0.12 0.2667 0.06 1.4816 0.34 2.2034 0.50 0.9613 0.22 0.5625 0.13 19112 452. 0.6158 0.14 0.3199 0.07 1.6088 0.36 1.6734 0.37 0.8909 0.20 0.6460 0.14 19114 430. 0.5678 0.13 0.2915 0.07 1.4232 0.33 1.6586 0.38 0.7345 0.17 0.6091 0.14 19121 567. 0.6466 0.11 0.3316 0.06 1.8123 0.32 1.6481 0.29 0.8330 0.15 0.6559 0.12 19123 464. 0.6675 0.14 0.3384 0.07 1.7221 0.37 1.7452 0.38 0.9021 0.19 0.6825 0.15 19130 434. 0.6775 0.16 0.3513 0.08 1.7366 0.40 1.7430 0.40 0.7931 0.18 0.7227 0.17 19137 443. 0.6420 0.14 0.3467 0.08 1.6457 0.37 1.3000 0.29 0.7387 0.17 0.6680 0.15 19146 473. 0.6207 0.13 0.3492 0.07 1.6423 0.35 1.5693 0.33 0.7248 0.15 0.6320 0.13 .................................................................................................................................... TESTIS PROSTATE BRAIN LIVER KIDNEY KIDNEY RAT TERMINAL BODY RIGHT LEFT RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW WT . % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW ________________________________________-------------------------------------------------------------------------------------------- 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 353. 505. 437. 452. 430. 567. 464. 434. 443. 473. 1.5806 2.0016 1.5096 1.6041 1.4506 1.7745 1.8031 1.8184 1.6928 1.6866 0.45 0.40 0.34 0.35 0.34 0.31 0.39 0.42 0.38 0.36 0.8460 1.5220 1.5754 1.3786 1.1497 1.0712 0.9914 1.6000 1.0622 1.0704 0.24 0.30 0.36 0.30 0.27 0.19 0.21 0.37 0.24 0.23 2.22 2.40 2.35 2.41 2.36 2.54 2.43 2.37 2.45 2.26 0.63 0.48 0.54 0.53 0.55 0.45 0.52 0.55 0.55 0.48 11.34 17.13 13.84 14.51 14.34 23.98 19.27 13.79 14.59 16.57 3.21 3.39 3.17 3.21 3.33 4.23 4.15 3.18 3 -29 3.50 1.50 2.37 1.71 2.09 1.84 2.20 2.17 1.88 1.88 1.94 0.42 0.47 0.39 0.46 0.43 0.39 0.47 0.43 0.42 0.41 1.56 2.29 1.71 1.97 1.92 2.25 2.22 1.98 2.14 2.01 0.44 0.45 0.39 0.44 0.45 0.40 0.48 0.46 0.48 0.42 _______________-________^_______________-------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. a. Damaged during processing (weight not affected). REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B19 (PAGE 6): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - INDIVIDUAL DATA - Fo GENERATION MALE RATS .................................................................................................................................... DOSAGE GROUP I11 I MG/KG/DAY .................................................................................................................................... RAT TERMINAL BODY ADRENAL LEFT ADRENAL RIGHT SPLEEN THYMUS HEART NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW a WT. % TBW a WT. % TBW WT. % TBW WT . % TBW ________________________________________-------------------------------------------------------------------------------------------- 19101 353. 0.028 7.93 0.026 7.36 0.55 0.16 0.23 0.06 1.33 0.38 19105 505. 0.034 6.73 0.027 5.35 0.74 0.15 0.47 0.09 1.66 0.33 19107 437 * 0.039 8.92 0.038 8.70 0.69 0.16 0.26 0.06 1.36 0.31 19112 452. 0.026 5.75 0.022 4.87 0.65 0.14 0.55 0.12 1.48 0.33 19114 430. 0.033 7.67 0.031 7.21 0.68 0.16 0.32 0.07 1.52 0.35 19121 567. 0.032 5.64 0.035 6.17 0.92 0.16 0.44 0.08 1.70 0.30 19123 464. 0.040 8.62 0.025 5.39 0.78 0.17 0.40 0.09 1.56 0.34 19130 434. 0.030 6.91 0.033 7.60 0.87 0.20 0.33 0.08 1.48 0.34 19137 443. 0.032 7.22 0.019 4.29 0.91 0.20 0.29 0.06 1.65 0.37 19146 473. 0.042 8.88 0.043 9.09 0.80 0.17 0.28 0.06 1.65 0.35 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) x 100. a. Value was multiplied by 1000. b E3 W b W PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B19 (PAGE 7): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - INDIVIDUAL DATA - Fo GENERATION MALE RATS .................................................................................................................................... DOSAGE GROUP IV 3 MG/KG/DAY .................................................................................................................................... EPIDIDYMIS CAUDA EPIDIDYMIS TESTIS SEMINAL VESICLES SEMINAL VESICLES EPIDIDYMIS RAT TERMINAL BODY LEFT LEFT LEFT WITH FLUID WITHOUT FLUID RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW WT. % TBW WT. % TBW WT . % TBW WT. % TBW WT. % TBW .................................................................................................................................... 19100 421. 0.6712 0.16 0.3355 0.08 1.7129 0.41 2.0456 0.48 1.0125 0.24 0.6970 0.16 19103 487. 0.6486 0.13 0.2909 0.06 1.6530 0.34 2.0607 0.42 1.0014 0.20 0.6457 0.13 19104 476. 0.6201 0.13 0.2961 0.06 1.7770 0.37 1.4443 0.30 0.7874 0.16 0.5846 0.12 19118 442. 0.5834 0.13 0.3022 0.07 1.5495 0.35 1.6316 0.37 0.7381 0.17 0.6169 0.14 19133 448. 0.5667 0.13 0.2961 0.07 1.7262 0.38 1.7327 0.39 0.8511 0.19 0.5492 0.12 19141 545. 0.6531 0.12 0.3098 0.06 1.7814 0.33 1.4610 0.27 0.7077 0.13 0.6801 0.12 19142 479. 0.3058a 0.06 0.1333a 0.03 0.6988a 0.14 1.1736 0.24 0.6729 0.14 0.3002a 0.06 19144 523. 0.5626 0.11 0.2529 0.05 1.6357 0.31 1.3813 0.26 0.6369 0.12 0.6547 0.12 19148 454. 0.6884 0.15 0.3851 0.08 1.8767 0.41 1.3721 0.30 0.6888 0.15 0.7389 0.16 19150 448. 0.6161 0.14 0.3111 0.07 1.7049 0.38 1.1701 0.26 0.5734 0.13 0.6388 0.14 .................................................................................................................................... TESTIS PROSTATE BRAIN LIVER KIDNEY KIDNEY RAT TERMINAL BODY RIGHT LEFT RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW ________________________________________-------------------------------------------------------------------------------------------- 19100 421. 1.7274 0.41 1.5079 0.36 2.40 0.57 15.67 3.72 1.82 0.43 1.91 0.45 19103 487. 1.7657 0.36 1.4347 0.29 2.39 0.49 19.28 3.96 2 -40 0.49 2.39 0.49 19104 476. 1.7025 0.36 1.2209 0.26 2.43 0.51 17.78 3.74 1.98 0.42 2.16 0.45 19118 442. 1.6129 0.36 1.3183 0.30 2.42 0.55 15.97 3.61 1.90 0.43 2.12 0.48 19133 448. 1.7045 0.38 1.2429 0.28 2.47 0.55 15.06 3.36 2.08 0.46 2.24 0.50 19141 545. 1.7406 0.32 1.0238 0.19 2.52 0.46 20.42 3.75 2.18 0.40 2.20 0.40 19142 479. 0.6700a 0.14 1.0863 0.23 2.23 0.46 17.61 3.68 2.00 0.42 2.07 0.43 19144 523. 1.6766 0.32 1.1080 0.21 2 -40 0.46 18.23 3.48 2.21 0.42 2.36 0.45 19148 454. 1.8681 0.41 1.4524 0.32 2.24 0.49 18.08 3.98 1.91 0.42 1.92 0.42 19150 448. 1.7068 0.38 1.3391 0.30 2.35 0.52 17.94 4.00 1.97 0.44 1.92 0.43 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). A B S . WT. = ORGAN WEIGHT. REL. % TBW = (ORGANWEIGHT/TERMINAL BODY WEIGHT) x 100. a. Rat 19142 had small testes, epididymides and left cauda epididymis. See the individual necropsy observations table (Table B 1 8 ) . I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I /I g : 8 I \ Il \a IE I 1I m I I I I I I I I I I I I I I I I I I I I I 1 1 I I I I I I I I I I I I I I I I I2I I :w IX I I I I I I I I I I I I : E I C O I I I I I I I I II W I I I I I I I I I I gI G I Iw I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I . . . . . . . . . . ~m I * N * N W * N m * W I -31 m m m m m m m m m m I I I W E ;0000000000 I I 5 0 I I I I I I !$ i . . . . . . . . . . I I . I Plw**o***mnl I *mw*WmmPmw I I rlrlrlrlrlrlrlrlrlrl I I I I I I I I I . . . . . . . . . . ~m I 1 I o ~ c o c o c o o o w c o mI - 3 I rloooorlrloorl I I I M E I 0000000000 I I Io\o I I I : A I I : z I ~ocom~~rlm~rmi - 5I NrlNrlrlNrlNrlrl I . . . . . . . . . J ~ I .I wi3looooooooool I : p: ,\o I I I I I I I ld: I I ZI I I EI I o\o I I I I I I I I I I q. I I I I I I I I I I I . . . . . . . . . . I I. lmrlomOo*PPwI- IWNmPw*coodNIg I ~wco~~mvmr~coi I 418-028:PAGE B-45 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B19 (PAGE 9): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( 0 ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION MALE RATS EPIDIDYMIS CAUDA EPIDIDYMIS TESTIS SEMINAL VESICLES SEMINAL VESICLES EPIDIDYMIS RAT TERMINAL BODY LEFT LEFT LEFT WITH FLUID WITHOUT FLUID RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW .................................................................................................................................... 19111 432. 0.5886 0.14 0.2814 0.06 1.7973 0.42 1.4923 0.34 0.7724 0.18 0.6626 0.15 19117 462. 0.5554a 0.12 0.2921 0.06 1.5902 0.34 1.4692 0.32 0.8619 0.19 0.5723 0.12 19124 426. 0.7041 0.16 0.3378 0.08 1.9441 0.46 1.6428 0.38 0.8490 0.20 0.6631 0.16 19126 426. 0.5648 0.13 0.2550 0.06 1.5950 0.37 0.9950 0.23 0.5749 0.13 0.5576 0.13 19127 434. 0.5950 0.14 0.3045 0.07 1.7552 0.40 1.6404 0.38 0.7509 0.17 0.5793 0.13 19128 445. 0.5787 0.13 0.2725 0.06 1.5509 0.35 1.1019 0.25 0.6009 0.14 0.6242 0.14 19140 442. 0.6181 0.14 0.2951 0.07 1.6009 0.36 1.5387 0.35 0.8782 0.20 0.6487 0.15 19145 441. 0.6705 0.15 0.3537 0.08 1.8102 0.41 1.8705 0.42 0.9316 0.21 0.6428 0.14 19149 439. 0.6384 0.14 0.3200 0.07 1.9607 0.45 1.4543 0.33 0.8120 0.18 0.6610 0.15 19152 461. 0.6200 0.13 0.3288 0.07 1.8784 0.41 1.2190 0.26 0.7403 0.16 0.6498 0.14 .................................................................................................................................... TESTIS PROSTATE BRAIN LIVER KIDNEY KIDNEY RAT TERMINAL BODY RIGHT LEFT RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT. % TBW WT . % TBW .................................................................................................................................... 19111 432. 1.7246 0.40 1.0515 0.24 2.48 0.57 24.36 5.64 2.00 0.46 2.13 0.49 19117 462. 1.6160 0.35 1.0954 0.24 2.43 0.52 24.11 5.22 2.22 0.48 2.26 0.49 19124 426. 1.8910 0.44 1.5146 0.36 2.38 0.56 25.72 6.04 2.21 0.52 2.18 0.51 19126 426. 1.6044 0.38 0.7260 0.17 2.28 0.54 22.94 5.38 2.04 0.48 2.04 0 -48 19127 434. 1.7997 0.41 1.4010 0.32 2.62 0.60 27.38 6.31 1.90 0.44 1.95 0.45 19128 445. 1.5952 0.36 0.9706 0.22 2.31 0.52 21.74 4.88 1.78 0.40 1.82 0.41 19140 442. 1.6407 0.37 1.2239 0.28 2.42 0.55 20.21 4.57 1.93 0.44 1.88 0 -42 + P cp 19145 441. 1.8340 0.42 1.3756 0.31 2.40 0.54 23.20 5.26 2.27 0.51 2.16 0 -49 19149 439. 1.9989 0.46 1.2522 0.28 2.38 0.54 20.06 4.57 1.76 0 -40 1.92 0.44 0 w 19152 461. 1.8997 0.41 1.3077 0.28 2.34 0.51 18.58 4.03 1.97 0.43 2.07 0.45 .................................................................................................................................... PP ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. F a. Damaged during processing (weight not affected). 8 W b m I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I 1 I I I I I 1 I I I I I I I I I 1 I I I I I I I $ I I 1 w I I II II X I I I I I I I I I I I I I I I I I I I I I I I : g I I 3 I I ; g I I I I I I I I I I I I I I I I I I I I I I I I E I I I I w I I I 11-11 I I& l m I I I I I I 1 I I I I I 1 I I I I I I I I I lrll I I I 1 I I I I I I I I I I I I I I I I I I I . . . . . . . . . . I INmmcvmcvedmcv ~mmmmmmmmmm I 10000000000 I I I I . . . . . . . . . . I IcnoommcvowwP Imw*m**mm*e I lddddrlddrldd I I I I I 1 2 I o\Q I I I. IJ I I. Ik I I , . . . . . . . . . . Id I ~ w m c n r e e m m m *I lmommdwm*mN I I ~ m m r w r r w r w c nI I I I I I I . . . . . . . . . . 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I0 I0 Irl I IX I- IE I% II H w I I II I 418-028:PAGE B-49 N 1 1 I I 03 I I N 0 I 03 rl w 0 N Fo I I I I I I I I I I I I I fI W Pomwrlrlwalmol NwmwmmmmNml 00000000001 . . . . . . . . . . wI 00000000001 I I I I I I 418-028:PAGE B-50 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I ; I I E I I I X I I I I I I I I I I I I I I E I w I I I J I I I I I I I I I I I I I I I I I I I I I I * * * * I * I I rl I 0m m cv cv 0 m W W W W cv I I I I I I I m m In cv In m co In cn I * I I cv m m m m cv m m I I I I I I I * I I W rl P m In 0 W m I I I cv m m rl m m In cv cv I I * * I I I m m m W m m cv m r m I W W W W W W I I I I I I 00 0000000 I I I I I I I I I I I I I I I I I * * I I WW I rl In rl m W m 0 W cv rl m W m-=r II I I I I I 0 cv P W rl m cv cv 0 I * I I W PW Prm W W I I I I I I I * * * I I W m W W CJ 03 0 d In I I I I Lo rl I m m W m cv In m cm v m W cv CJ W I I I I P 0 W r W P cv In I I I I I I rl rl rl rl rl rl rl rl rl I I I I I I I I I I I I I I I I I I I I IPwmmWorInmm I .3 I P r l P W r l P W r l c n O I I I . . . . . . . . . . I I Jp: I I I I w a ~ o ~ ~ r l m m m m cI v w I I I I m a.!. IPWPWWWPPPP I I I I HE I w I I E Ik m I I 03 In P W 0 m W m Ip: w * I I m I m m rl 03 0 cv In P w * I2 I I ;I * * I I E II In W W W m In 0 W m r W co r P r I d I I I I s I I I rl rl rl rl rl rl rl rl rl I I I I I I I I 1 I I I I I I I I I I 1 I 1 I I I I I I I I I I la, I I *rl I CON l 0 . Q I c v m I ord rle I 00 I I ..I x I 00 I I I nl I I I * * I I I cv W r m W 03 P m cv a, In rl I I P W cv rl r 0 P I r r W 03 r r W I I I I I 0 W 0 rl O m W I 0 m 0 Fi m W W I I cv rl cv cv rl rl rl I I I I I I * * I In 0 m r W cn cn rl m m m cv I * I W In 0 m r m I P P a, P r P W I I I I I P 0 W Wrd m m W I m m m m WWm I I rl rl rl rl rl rl rl I I I I I I I I I I P P cv In W W rl m cv 0 I * * I W m m rl r m m In I I I I m 03 In W W m m m r P I * I W W W P 0 m Lfl m W I * 1 W W In In m m m W m I I I I I * * * * I I 1 m cn m In W W r In In 0 I 1 In rl r m 0 cv m P cv rl I I I I P m m m m rl I I rl rl rl rl rl rl rl rl rl rl I I I I I I I I I * * I 0 rl m rl cv TY 0 In I * I m m rl W 0 0 03 cv r I I I I W 0 r W r 0 Lfl m 0 cv I * * * * I m CY W TY m m I I I I I * I 0 cv cv 0 0 cv m cn 0 I I r W m rl m m m rl m W I I m m W W rl cv cv m m W I * I m 0 rl rl (\I rl m m cv I I I I rl rl rl rl rl rl rl rl 0 rl I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I HI I HI I I I I I I I I I I I I I I I .......... wm~ncvwmrwrw cvcvcvcvcv~~cvcvcv rmmrloo*InU)o mcvWrlrlo3*www . . . . . . . . . . r l W * * m r l W P W c v WWWInWWWWWW 0000000000 I I I . I .......... cvcvcvcvcvcvcvcvcvcv ; c v W w o m o c n W w m ooorlrlcvcvmmm I rlrlrlrlrlrlrlrlrlrl mmmmmmcnmmm rlrlrlrlrlrlrlrlrlrl I I E iE I giI I I PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B20 (PAGE 4): ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO BRAIN WEIGHT - INDIVIDUAL DATA - Fo GENERATION MALE RATS PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B20 (PAGE 5): ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO BRAIN WEIGHT - INDIVIDUAL DATA - Fo GENERATION MALE RATS RAT NUMBER 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 EPIDIDYMIS BRAIN LEFT WEIGHT ABS. REL. WT. % BRW .-------------- .--------------- 2.22 0.6021 27.12 2.40 0.7012 29.22 2.35 2.41 2.36 0.5355a 22.79 0.6158 25.55 0.5678 24.06 2.54 0.6466 25.46 2.43 0.6675 27.47 2.37 0.6775 28.59 2.45 0.6420 26.20 2.26 0.6207 27.46 CAUDA EPIDIDYMIS LEFT ABS. REL. WT. % BRW ..................... 0.2993 13.48 0.3377 14.07 0.2667 11.35 0.3199 13.27 0.2915 12.35 0.3316 13.06 0.3384 13.92 0.3513 14.82 0.3467 14.15 0.3492 15.45 TESTIS SEMINAL VESICLES LEFT WITH FLUID ABS. REL. ABS. REL. WT. % BRW WT. --------------- % BRW 1.6371 73.74 1.1004 49.57 2.0077 83.65 1.7433 72.64 1.4816 63.05 2.2034 93.76 1.6088 66.76 1.6734 69.44 1.4232 60.30 1.6586 70.28 1.8123 71.35 1.6481 64.88 1.7221 70.87 1.7452 71.82 1.7366 73.27 1.7430 73.54 1.6457 67.17 1.3000 53.06 1.6423 72.67 1.5693 69.44 SEMINAL VESICLES WITHOUT FLUID ABS. REL. WT. % BRW ..................... 0.5923 26.68 0.7941 33.09 0.9613 40.91 0.8909 36.97 0.7345 31.12 0.8330 32.80 0.9021 37.12 0.7931 33.46 0.7387 30.15 0.7248 32.07 EPIDIDYMIS RIGHT ABS. REL. WT. % BRW .---------------- 0.6071 27.35 0.7481 31.17 0.5625 23.94 0.6460 26.80 0.6091 25.81 0.6559 25.82 0.6825 28.09 0.7227 30.49 0.6680 27.26 0.6320 27.96 19101 2.22 1.5806 71.20 0.8460 38.11 11.34 510.81 1.50 67.57 1.56 70.27 0.028 1.26 19105 2.40 2.0016 83.40 1.5220 63.42 17.13 713.75 2.37 98.75 2.29 95.42 0.034 1.42 19107 2.35 1.5096 64.24 1.5754 67.04 13.84 588.94 1.71 72.76 1.71 72.76 0.039 1.66 19112 2.41 1.6041 66.56 1.3786 57.20 14.51 602.07 2.09 86.72 1.97 81.74 0.026 1.08 19114 2.36 1.4506 61.47 1.1497 48.72 14.34 607.63 1.84 77.97 1.92 81.36 0.033 1.40 19121 2.54 1.7745 69.86 1.0712 42.17 23.98 944.09 2.20 86.61 2.25 88.58 0.032 1.26 19123 2.43 1.8031 74.20 0.9914 40.80 19.27 793.00 2.17 89.30 2.22 91.36 0.040 1.65 19130 2.37 1.8184 76.72 1.6000 67.51 13.79 581.86 1.88 79.32 1.98 83.54 0.030 1.26 b 19137 2.45 1.6928 69.09 1.0622 43.36 14.59 595.51 1.88 76.73 2.14 87.35 0.032 1.31 19146 2.26 1.6866 74.63 1.0704 47.36 16.57 733.18 1.94 85.84 2.01 88.94 0.042 1.86 ................................................................................................................................. h, ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/BRAIN WEIGHT) x 100. 00 ;b a. Damaged during processing (weight not affected). b PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B20 (PAGE 6): ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO BRAIN WEIGHT - INDIVIDUAL DATA - Fo GENERATION MALE RATS W ul w PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B20 (PAGE 7): ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO BRAIN WEIGHT - INDIVIDUAL DATA - Fo GENERATION MALE RATS ________________________________________-------------------------------------------------------------------------------------------- DOSAGE GROUP IV 3 MG/KG/DAY .................................................................................................................................... EPIDIDYMIS CAUDA EPIDIDYMIS TESTIS SEMINAL VESICLES SEMINAL VESICLES EPIDIDYMIS RAT BRAIN LEFT LEFT LEFT WITH FLUID WITHOUT FLUID RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % BRW WT. % BRW WT . % BRW WT. % BRW WT. % BRW WT. % BRW .................................................................................................................................... 19100 2.40 0.6712 27.97 0.3355 13.98 1.7129 71.37 2.0456 85.23 1.0125 42.19 0.6970 29.04 19103 2.39 0.6486 27.14 0.2909 12.17 1.6530 69.16 2.0607 86.22 1.0014 41.90 0.6457 27.02 19104 2.43 0.6201 25.52 0.2961 12.18 1.7770 73.13 1.4443 59.44 0.7874 32.40 0.5846 24.06 19118 2.42 0.5834 24.11 0.3022 12.49 1.5495 64.03 1.6316 67.42 0.7381 30.50 0.6169 25.49 19133 2.47 0.5667 22.94 0.2961 11.99 1.7262 69.89 1.7327 70.15 0.8511 34.46 0.5492 22.23 19141 2.52 0.6531 25.92 0.3098 12.29 1.7814 70.69 1.4610 57.98 0.7077 28.08 0.6801 26.99 19142 2.23 0.3058a 13.71 0.1333a 5.98 0.6988a 31.34 1.1736 52.63 0.6729 30.17 0.3002a 13.46 19144 2.40 0.5626 23.44 0.2529 10.54 1.6357 68.15 1.3813 57.55 0.6369 26.54 0.6547 27.28 19148 2.24 0.6884 30.73 0.3851 17.19 1.8767 83.78 1.3721 61.25 0.6888 30.75 0.7389 32.99 19150 2.35 0.6161 26.22 0.3111 13.24 1.7049 72.55 1.1701 49.79 0.5734 24.40 0.6388 27.18 .................................................................................................................................... TESTIS PROSTATE LIVER KIDNEY KIDNEY ADRENAL RAT BRAIN RIGHT LEFT RIGHT LEFT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % BRW WT. % BRW WT. % BRW WT. % BRW WT. % BRW WT. % BRW ________________________________________------------------------------------------------------------------------------------------- 19100 2.40 1.7274 71.98 1.5079 62.83 15.67 652.92 1.82 75.83 1.91 79.58 0.028 1.17 19103 2.39 1.7657 73.88 1.4347 60.03 19.28 806.69 2.40 100.42 2.39 100.00 0.040 1.67 19104 2.43 1.7025 70.06 1.2209 50.24 17.78 731.69 1.98 81.48 2.16 88.89 0.030 1.23 19118 2.42 1.6129 66.65 1.3183 54.48 15.97 659.92 1.90 78.51 2.12 87.60 0.030 1.24 19133 2.47 1.7045 69.01 1.2429 50.32 15.06 609.72 2.08 84.21 2.24 90.69 0.043 1.74 19141 2.52 1.7406 69.07 1.0238 40.63 20.42 810.32 2.18 86.51 2.20 87.30 0.024 0.95 19142 2.23 0.6700a 30.04 1.0863 48.71 17.61 789.69 2.00 89.69 2.07 92.82 0.028 1.26 19144 2.40 1.6766 69.86 1.1080 46.17 18.23 759.58 2.21 92.08 2.36 98.33 0.037 1.54 19148 2.24 1.8681 83.40 1.4524 64.84 18.08 807.14 1.91 85.27 1.92 85.71 0.028 1.25 b 19150 2.35 1.7068 72.63 1.3391 56.98 17.94 763.40 1.97 83.83 1.92 81.70 _________^______________________________-------------------------------------------------------------------------------------------- x ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/BRAIN WEIGHT) 0.037 100. 1.57 h) 00 a. Rat 19142 had small testes, epididymides and left cauda epididymis. See the individual necropsy observations table (Table B18). I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I ; 2 I i X: w I I I I s I I I I I I I I 1 I I I I I I w I I d I I I I I Im & I I 3 I I I E I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I m* I I .I col w1 I I I I NI wl .I dI I I I I I WI cvf .I *I cvI I I I I PI mI *I 0 1I I I I 418-028:PAGE B-55 1 1 I I I I I I I I I I I I I I I I I I I I I I I I PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B20 (PAGE 9): ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO BRAIN WEIGHT - INDIVIDUAL DATA - Fo GENERATION MALE RATS RAT NUMBER 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 EPIDIDYMIS BRAIN LEFT . WEIGHT ABS. REL. WT % BRW . - - - - - - - - - - - - -. - - - - - - - - - - - - - 2.48 0.5886 23.73 2.43 0.5554a 22.86 2.38 0.7041 29.58 2.28 0.5648 24.77 2.62 0.5950 22.71 2.31 0.5787 25.05 2.42 0.6181 25.54 2.40 0.6705 27.94 2.38 0.6384 26.82 2.34 0.6200 26.50 CAUDA EPIDIDYMIS LEFT ABS. REL. WT. % BRW ------------------ 0.2814 11.35 0.2921 12.02 0.3378 14.19 0.2550 11.18 0.3045 11.62 0.2725 11.80 0.2951 12.19 0.3537 14.74 0.3200 13.44 0.3288 14.05 TESTIS LEFT ABS. REL. WT. % BRW 1.7973 1.5902 1.9441 1.5950 1.7552 1.5509 1.6009 1.8102 1.9607 1.8784 72.47 65.44 81.68 69.96 66.99 67.14 66.15 75.42 82.38 80.27 SEMINAL VESICLES SEMINAL VESICLES WITH FLUID WITHOUT FLUID ABS. REL. ABS. REL. WT. % BRW WT. % BRW ------------------ .................... 1.4923 60.17 0.7724 31.14 1.4692 60.46 0.8619 35.47 1.6428 69.02 0.8490 35.67 0.9950 43.64 0.5749 25.21 1.6404 62.61 0.7509 28.66 1.1019 47.70 0.6009 26.01 1.5387 63.58 0.8782 36.29 1.8705 77.94 0.9316 38.82 1.4543 61.10 0.8120 34.12 1.2190 52.09 0.7403 31.64 EPIDIDYMIS RIGHT ABS. REL. WT . % BRW ----------------- 0.6626 26.72 0.5723 23.55 0.6631 27.86 0.5576 24.46 0.5793 22.11 0.6242 27.02 0.6487 26.80 0.6428 26.78 0.6610 27.77 0.6498 27.77 19111 2.48 1.7246 69.54 1.0515 42.40 24.36 982.26 2.00 80.64 2.13 85.89 0.025 1.01 19117 2.43 1.6160 66.50 1.0954 45.08 24.11 992.18 2.22 91.36 2.26 93.00 0.030 1.23 19124 2.38 1.8910 79.45 1.5146 63.64 25.72 1080.67 2.21 92.86 2.18 91.60 0.035 1.47 19126 2.28 1.6044 70.37 0.7260 31.84 22.94 1006.14 2.04 89.47 2.04 89.47 0.029 1.27 19127 2.62 1.7997 68.69 1.4010 53.47 27.38 1045.04 1.90 72.52 1.95 74.43 0.031 1.18 19128 2.31 1.5952 69.06 0.9706 42.02 21.74 941.12 1.78 77.06 1.82 78.79 0.036 1.56 19140 2.42 1.6407 67.80 1.2239 50.57 20.21 835.12 1.93 79.75 1.88 77.68 0.036 1.49 19145 2.40 1.8340 76.42 1.3756 57.32 23.20 966.67 2.27 94.58 2.16 90.00 0.034 1.42 b 19149 2.38 1.9989 83.99 1.2522 52.61 20.06 842.86 1.76 73.95 1.92 80.67 0.029 1.22 19152 2.34 1.8997 81.18 1.3077 55.88 18.58 794.02 1.97 84.19 2.07 88.46 0.036 1.54 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - --------- - - - - - - - - - - t3 ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGANWEIGHT/BRAIN WEIGHT) x 1 0 0 . a. Damaged during processing (weight not affected). I I I I I I I I8 I I I I I 1 I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I 1 I I I I I I I 1 I I I 1 I I I I I I I I I I I I I I I I I I I 1 I I I I I ; $ ; I I I I I I ; I wa I I I~ ~ m m a ~ r c o r w m i wcocomm*mwmcoI ..........I I mmcoO*ddwrlml mwmwmwwmwwl p: o\o I I :E I I I I I I I 3 I . Icooocom~oww~~ I 1x mw*m**mm**l . . . . . . . . . . I I I !!g I I rldrlrlrldrldrlrll I I I I I I I I I I I I I I I . . . . . . . . . . w a I I I I I I I mrlmd*mcomPmI I I - 3I c n c o d w m m r l c o o r i I I I I !J& I I I* * m o ~ ~ c o m c o d i 1 I rlrldmrlrlrlrldml I I m &do: I I I I ? 1 I I I I I : z I . I rwwPPoqcomdi I mmm****m*ml I E . . . . . . . . . . I I ! ! g ; I 00000000001 I I I I I I I I I I I I I I I I I I I I I I 418-028:PAGE B-57 I I I E I I I I I I I a I I I I I I I I I I I PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B21 (PAGE 1): BODY WEIGHTS - INDIVIDUAL DATA - FO GENERATION MALE RATS .................................................................................................................................... RAT # DOSAGE GROUP I 0 MG/KG/DAY ________________________________________-------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 14a 15 16 ________________________________________-------------------------------------------------------------------------------------------- 19109 322. 326. 334. 342. 344. 352. 357. 359. 355. 363. 370. 374. 380. 380. 376. 384. 19115 357. 361. 370. 374. 377. 388. 385. 393. 402. 408. 412. 417. 423. 427. 422. 430. 19116 347. 348. 360. 363. 372. 378. 385. 388. 389. 394. 402. 402. 408. 403. 410. 416. 19119 350. 356. 370. 378. 388. 394. 400. 414. 420. 424. 431. 446. 450. 454. 449. 452. 19122 328. 329. 320. 312. 336. 346. 350. 354. 358. 366. 372. 379. 380. 382. 389. 396. 19125 359. 364. 372. 382. 384. 392. 400. 409. 414. 420. 428. 431. 436. 440. 435. 435. 19131 341. 350. 357. 362. 375. 384. 388. 388. 403. 400. 407. 407. 419. 417. 418. 418. 19132 338. 339. 343. 347. 348. 356. 361. 359. 353. 363. 363. 369. 370. 376. 376. 376. 19134 332. 330. 342. 345. 350. 356. 358. 358. 366. 367. 370. 373. 373. 380. 357. 378. 19135 358. 359. 372. 380. 381. 391. 393. 398. 395. 398. 408. 403. 416. 417. 416. 424. 19143 338. 334. 344. 347. 357. 358. 363. 366. 365. 374. 381. 381. 387. 390. 387. 388. 19151 346. 350. 362. 366. 370. 378. 380. 387. 389. 392. 400. 404. 407. 406. 404. 408. 19157 331. 337. 344. 350. 355. 362. 363. 370. 369. 377. 378. 384. 394. 393. 384. 392. 19161 362. 373. 385. 392. 399. 411. 417. 424. 425. 416. 436. 433. 436. 446. 449. 457. 19167 332. 340. 347. 351. 359. 365. 372. 373. 382. 382. 388. 391. 391. 399. 396. 401. ________________________________________-------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29b 30 31 32 .................................................................................................................................... 19109 390. 391. 392. 398. 400. 398. 396. 403. 406. 409. 420. 420. 421. 427. 430. 435. 19115 429. 442. 443. 451. 449. 455. 458. 460. 465. 463. 462. 453. 472. 469. 479. 472. 19116 413. 422. 420. 425. 431. 432. 442. 448. 448. 452. 461. 469. 462. 466. 476. 479. 19119 455. 463. 470. 476. 478. 485. 483. 492. 494. 504. 513. 511. 504. 510. 518. 516. 19122 396. 399. 399. 405. 406. 409. 413. 420. 419. 421. 426. 432. 432. 434. 440. 441. 19125 435. 434. 442. 448. 452. 452. 465. 471. 478. 482. 486. 489. 491. 493. 500. 498. 19131 417. 432. 425. 438. 435. 440. 447. 450. 456. 458. 470. 470. 479. 479. 484. 494. 19132 384. 392. 393. 410. 415. 406. 407. 406. 418. 420. 424. 426. 428. 428. 431. 438. P 19134 371. 377. 377. 380. 385. 381. 381. 387. 388. 394. 396. 404. 404. 406. 415. 419. c-r 19135 426. 432. 430. 440. 437. 440. 442. 446. 449. 453. 464. 465. 468. 469. 470. 477. 19143 392. 393. 400. 406. 408. 407. 413. 414. 417. 426. 432. 440. 435. 440. 444. 453. 19151 411. 413. 408. 423. 424. 428. 430. 434. 446. 445. 450. 452. 463. 461. 457. 464. 19157 394. 397. 396. 410. 405. 409. 411. 420. 424. 429. 432. 409. 438. 434. 439. 441. 19161 463. 468. 474. 483. 488. 499. 502. 508. 517. 518. 532. 539. 546. 544. 548. 555. 19167 410. 409. 419. 425. 422. 378. 368. 363. 373. 389. 396. 405. 406. 414. 398. 421. ________________________________________-------------------------------------------------------------------------------------------- . DAY = DAY OF STUDY W ALL WEIGHTS WERE RECORDED IN GRAMS (G) VI a. Last value recorded before cohabitation. 00 b. First value recorded after cohabitation. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B21 (PAGE 2): BODY WEIGHTS - INDIVIDUAL DATA - FO GENERATION MALE RATS 19109 442. 446. 452. 443. 19115 439. 482. 485. 476. 19116 477. 485. 490. 491. 19119 521. 532. 538. 533. 19122 437. 451. 449. 455. 19125 503. 509. 517. 517. 19131 492. 497. 501. 513. 19132 444. 446. 446. 448. 19134 421. 424. 426. 433. 19135 485. 486. 493. 490. 19143 458. 456. 466. 468. 19151 473. 466. 461. 474. 19157 441. 443. 447. 455. 19161 557. 561. 560. 566. 19167 424. 426. 438. 441. ........................................... DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 451. 490. 490. 523. 458. 527. 512. 450. 438. 504. 472. 481. 458. 567. 445. 447. 487. 492. 526. 467. 517. 506. 439 * 431. 494. 470. 474. 459. 571. 446. ----- 450. 491. 494. 527. 467. 528. 514. 454. 434. 496. 477. 481. 460. 564. 445. ----- 455. 463. 466. 438. 490. 498. 499. 466. 501. 501. 503. 509. 529. 526. 537. 538. 468. 468. 475. 474. 528. 528. 534. 534. 516. 515. 520. 528. 458. 465. 466. 463. 436. 441. 444. 436. 502. 502. 504. 504. 478. 476. 486. 491. 482. 482. 484. 482. 463. 470. 473. 478. 571. 562. 556. 559. 451. 451. 456. 458. .......................... 490. 505. 451. 543. 530. 474. 446. 511. 493. 490. 483. 569. 458. ---- 508. 499. 439. 445. 509. 495. 500. 491. 575. 462. 419. 482. 499. 503. 486. 583. 472. I i3 418-028:PAGE B-60 I I I I I I I I 1 I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I 1 I I III III III . . . . . . . . . . . . . . . III III . . . . . . . . . . . . . . .III I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I . . . . . . . . . . . . . . . I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II II . . . . . . . . . . . . . . . III I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I *I I II II C V II II I I I I I I I I I I I I I I I I I I mI 1 II 11 m II II 1 I I I I 1 I I I I I I PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B21 (PAGE 4): BODY WEIGHTS - INDIVIDUAL DATA - FO GENERATION MALE RATS 19102 431. 436. 440. 443. 455. 19106 489. 488. 488. 486. 499. 19108 483. 484. 490. 486. 489. 19110 465. 464. 464. 461. 462. 19113 451. 453. 453. 464. 458. 19120 491. 496. 497. 503. 509. 19129 448. 451. 451. 456. 450. 19136 503. 506. 508. 511. 513. 19138 375. 380. 379. 383. 389. 19139 433. 442. 451. 450. 451. 19147 466. 474. 471. 475. 478. 19153 482. 486. 490. 494. 495. 19164 487. 490. 497. 489. 501. 19165 470. 466. 464. 468. 470. 19171 426. 420. 425. 428. 428. ........................................... -------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 450. 501. 479. 471. 456. 493. 440. 504. 382. 451. 469. 498. 504. 473. 431. ---- 455. 508. 485. 473. 460. 506. 455. 524. 382. 447 * 478. 498. 493. 478. 424. 461. 502. 485. 474. 463. 508. 460. 523. 387. 461. 480. 500. 495. 467. 424. 453. 501. 488. 474. 466. 509. 461. 522. 386. 450. 484. 505. 500. 476. 427. 456. 506. 495. 481. 464. 505. 463. 528. 390. 462. 497. 502. 509. 480. 435. ----- 428. 476. 498. 474. 480. 453. 468. 444. 517. 524. 486. 459. 466. 435. 534. 537. 495. 388. 389. 396. 365. 458. 462. 464. 432. 481. 496. 499. 498. 470. 504. 511, 513. 506. 487. 516. 512. 517. 521. 484. 486. 490. 489. 492. 455. 434. 443. 444. 453. 421. ....................................... 418-028:PAGE B-62 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I 1 I . . . . . . . . . . . . . . . I I I I I I I I I I I I I I I I I I I I I mcv*om(vrlcvrlcvwmoom I I I I I I mmmowmrlmwrlwocvrlw I I I I I I m * m ~ m * * m m * m - ~ - ~ * mI I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I . . . . . . . . . . . . . . . I I I I I I I I I I I I I I I I 1 I I I IwcvowmwwmooommdP I I 1 I I I d c v m w m c v w w ~ o ~ w r l mI r I I 1 I I m*mmm*mmm*mm*mm I I I I I 1 I I I I I I 1 I I I I I I I I I 1 I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I . . . . . . . . . . . . . . . I I I I I I W I *mPrlmww*r**wcvmm I I I I I (vmwwmowPwmPmomw I I I I\ I I mmmmm*mmmmmm*mm I I I I I I I I x I I 1 I I I\ I I I I I tu I I I I I IP I I I I := I I I I I Irl I I I I I I I I I I I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I I I I I I I I I I I I I I I I I I I I W I o * o * w m d m m ~ P m w m oI I I I I I cvowP*oPwmwwwwww I I I I I I m*mmm*mmmmmmmmm I I I I I I I I I . . . . . . . . . . . . . . . I I I I I I I I I I I I I I 1 I I I I I II m I Pmwdmmmmmmwwwwm I dmwP*owwmPmPPPm 1 1 I I I I I I I m m m m m ~ m m m m m m m m mI I I I 1 I I I I . . . . . . . . . . . . . . . I I I I I I I I I I I I I I I I I PwPmwmwwm**wmmw I I I I * I dwwwmwmm*PmwPP* I 1 I I I mmmmmmmmmmmmmmm I I I I I I I . . . . . . . . . . . . . . . I I I I I I I I 1 I I I I I I I I m I wmwo*do*cvPPP(vmm I I IH I I dwmwmwmm*w*www* I I IH I I mmmmmmmmmmmmmmm I I IH I I I I I I I I I Ipc I I I I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I I I I Ig 1 0 1 I w c v o m r l w m w o P d d o m * I I Ip: I I o ~ m m m w m ~ m m ~ m mI m mI I I mmmmmmmmmmmmmmm I I fa I 1 I I IW I I tu I I I I I I . . . . . . . . . . . . . . . I I I I ; $ I d I m w ~ ~ m m m r m m * ~ w oI d oI 1 0I I ow**cvmm*(vmm*mmm 1 I ln I I mmmmmmmmmmmmmmm I I I I I I I I I I I I I I I 1 I I I ; % I I I I f 0 I I I I I I I I I I I I I I I I# I I I I 3 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I .. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B21 (PAGE 6): BODY WEIGHTS - INDIVIDUAL DATA - FO GENERATION MALE RATS ________________________________________-------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I11 1 MG/KG/DAY ________________________________________-------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 ________________________________________-------------------------------------------------------------------------------------------- 19101 369. 370. 374. 369. 373. 373. 374. 379. 375. 379. 353. 19105 515. 522. 519. 528. 538. 537. 536. 535. 533. 532. 505. 19107 454. 455. 461. 457. 458. 449. 449. 453. 456. 460. 437. 19112 468. 463. 471. 470. 474. 473. 480. 480. 480. 478. 482. 452. 19114 428. 430. 435. 436. 442. 442. 446. 445. 450. 451. 454. 430. 19121 573. 576. 579. 584. 598. 590. 602. 600. 606. 606. 603. 616. 567. 19123 472. 474. 481. 483. 478. 469. 482. 483. 489. 491. 497. 494. 464. 19130 449. 447. 450. 449. 452. 454. 448. 452. 464. 466. 466. 461. 470. 434. 19137 453. 457. 456. 462. 469. 465. 461. 474. 474. 471. 473. 473. 468. 443. 19146 478. 478. 479. 481. 481. 483. 475. 480. 493. 493. 495. 496. 499. 473. 19155 467. 474. 475. 479. 477. 481. 481. 486. 492. 494. 493. 502. 500. 504. 481. 19159 470. 466. 476. 480. 480. 482. 485. 483. 484. 494. 494. 484. 498. 506. 475. 19172 514. 508. 520. 528. 534. 531. 534. 531. 532. 538. 541. 547. 549. 556. 526. 19173 467. 471. 476. 486. 485. 483. 482. 466. 477. 483. 483. 485. 483. 486. 459. 19174 449. 453. 457. 460. 457. 462. 461. 457. 461. 461. 461. 466. 471. 473. 448. ________________________________________------------------------------------------------------------------------------------------ DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B21 (PAGE 7): BODY WEIGHTS - INDIVIDUAL DATA - Fo GENERATION MALE RATS ________________________________________-------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP IV 3 MG/KG/DAY .................................................................................................................................... DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 14a 15 16 .................................................................................................................................... 19100 332. 332. 341. 344. 348. 356. 363. 363. 366. 366. 378. 377. 376. 381. 375. 382. 19103 363. 363. 379. 386. 393. 403. 415. 417. 428. 425. 434. 436. 439. 446. 435. 441. 19104 19118 351. 342. 360. 352. 366. 358. 375. 362. 383. 363. 389. 370. 398. 374. 401. 376. 405. 380. 400. 383. 408. 389. 412. 394. 412. 397. 423. 399. 426. 399. 432. 394. 19133 338. 338. 339. 347. 354. 357. 362. 364. 368. 372. 384. 393. 393. 388. 386. 395. 19141 363. 374. 382. 391. 396. 406. 414. 422. 426. 431. 438. 451. 450. 457. 452. 466. 19142 334. 338. 345. 354. 363. 368. 374. 379. 389. 387. 391. 398. 400. 410. 407. 415. 19144 348. 357. 368. 378. 386. 393. 405. 410. 418. 422. 434. 436. 430. 436. 434. 437. 19148 336. 343. 342. 355. 360. 366. 376. 378. 388. 378. 388. 389. 393. 398. 395. 403. 19150 356. 360. 372. 377. 384. 390. 391. 396. 399. 395. 404. 407. 410. 411. 406. 419. 19156 333. 330. 333. 337. 346. 358. 361. 360. 368. 366. 384. 386. 382. 394. 385. 395. 19160 351. 351. 364. 375. 376. 388. 395. 392. 392. 396. 404. 401. 412. 417. 396. 417. 19162 331. 333. 334. 338. 341. 344. 347. 351. 352. 354. 358. 369. 366. 369. 365. 372. 19163 350. 356. 364. 368. 379. 387. 393. 394. 401. 405. 418. 412. 410. 422. 412. 419. 19166 356. 359. 373. 385. 388. 398. 404. 396. 415. 416. 430. 430. 439. 443. 437. 447. _______^________________________________-------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29b 30 31 32 ________________________________________-------------------------------------------------------------------------------------------- 19100 390. 391. 396. 400. 403. 405. 404. 408. 411. 414. 418. 425. 428. 428. 431. 432. 19103 451. 458. 457. 470. 468. 470. 479. 489. 483. 486. 488. 490. 492. 498. 497. 499. 19104 438. 439. 436. 452. 454. 454. 458. 461. 467. 464. 471. 478. 479. 481. 481. 481. 19118 398. 406. 409. 419. 425. 426. 430. 429. 433. 434. 442. 444. 446. 449. 447. 451. 19133 395. 404. 410. 415. 421. 421. 432. 427. 439. 440. 441. 445. 449. 446. 443. 451. 19141 462. 473. 476. 476. 486. 490. 499. 502. 505. 512. 519. 518. 531. 526. 533. 539. 19142 420. 425. 431. 435. 441. 438. 448. 450. 455. 463. 469. 471. 472. 475. 485. 490. 19144 435. 451. 444. 456. 459. 462. 471. 474. 480. 494. 497. 497. 509. 518. 519. 517. 19148 405. 412. 417. 421. 421. 424. 431. 434. 437. 438. 450. 452. 455. 451. 457. 465. 19150 418. 427. 432. 441. 440. 436. 448. 442. 454. 450. 462. 462. 464. 458. 467. 474. 19156 396. 392. 402. 411. 411. 409. 416. 417. 419. 423. 422. 430. 436. 436. 435. 438. 19160 421. 424. 427. 434. 437. 431. 446. 442. 449. 450. 461. 470. 464. 468. 475. 473. 19162 377. 376. 381. 382. 390. 392. 397. 402. 400. 408. 409. 410. 415. 414. 415. 417. 19163 429. 434. 438. 453. 455. 448. 462. 463. 459. 463. 468. 478. 484. 476. 481. 479. 19166 456. 460. 465. 473. 480. 480. 480. 487. 488. 493. 500. 502. 503. 505. 505. 505. ________________________________________------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. First value recorded after cohabitation. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B21 (PAGE 8): BODY WEIGHTS - INDIVIDUAL DATA - FO GENERATION MALE RATS .................................................................................................................................... RAT # DOSAGE GROUP IV 3 MG/KG/DAY .................................................................................................................................... DAY 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 .................................................................................................................................... 19100 435. 438. 440. 441. 445. 447. 448. 455. 449. 454. 421. 19103 497. 502. 503. 499. 508. 513. 516. 509. 511. 516. 487. 19104 484. 488. 484. 486. 497. 499. 506. 506. 502. 506. 509. 476. 19118 448. 452. 459. 458. 457. 459. 464. 467. 464. 469. 470. 442. 19133 452. 453. 454. 461. 457. 464. 470. 474. 470. 475. 484. 448. 19141 537. 537. 543. 545. 556. 557. 566. 570. 567. 564. 574. 585. 545. 19142 494. 489. 491. 496. 510. 497. 501. 504. 500. 504. 505. 515. 479. 19144 524. 524. 528. 531. 539. 533. 546. 536. 547. 552. 547. 553. 523. 19148 460. 461. 469. 473. 482. 482. 481. 489. 485. 486. 482. 495. 495. 454. 19150 468. 463. 466. 470. 470. 472. 472. 463. 478. 481. 476. 472. 479. 448. 19156 438. 442. 441. 448. 446. 450. 447. 452. 452. 457. 453. 447. 455. 460. 433. 19160 19162 473. 419. 475. 418. 479. 426. 479. 424. 485. 429. 488. 427. 486. 431. 480. 433. 483. 435. 481. 434. 478. 435. 496. 441. 493. 449. 491. 445. 461. 415. 19163 491. 483. 490. 492. 494. 492. 497. 492. 500. 503. 508. 516. 513. 512. 486. 19166 511. 511. 517. 513. 519. 525. 525. 520. 522. 523. 528. 535. 531. 537. 506. ________________________________________------------------------------------------------------------------------------------------ DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B21 (PAGE 9): BODY WEIGHTS - INDIVIDUAL DATA - FO GENERATION MALE RATS .................................................................................................................................... RAT # DOSAGE GROUP V 10 MG/KG/DAY .................................................................................................................................... DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 14a 15 16 .................................................................................................................................... 19111 326. 334. 339. 346. 351. 360. 370. 370. 374. 382. 390. 389. 391. 393. 389. 396. 19117 363. 365. 375. 381. 382. 387. 393. 393. 390. 392. 404. 401. 408. 416. 405. 416. 19124 345. 346. 354. 360. 365. 372. 372. 379. 382. 389. 389. 379. 389. 393. 385. 397. 19126 344. 349. 356. 360. 365. 370. 371. 371. 377. 377. 384. 383. 391. 389. 386. 390. 19127 329. 335. 342. 351. 357. 366. 365. 370. 376. 378. 379. 383. 387. 395. 362. 394. . 19128 358. 358. 367. 372. 376. 382. 382. 385. 389. 394. 390. 406. 409. 408. 411. 415. 19140 331. 328. 347. 346. 353. 359. 362. 366. 373. 379. 380. 387. 388. 393. 386. 396. 19145 348. 349. 358. 366. 375. 375. 382. 386. 388. 393. 395. 398. 406. 406. 387. 394. 19149 325. 324. 331. 339. 339. 346. 352. 354. 352. 359. 368. 367. 367. 369. 368. 375. 19152 347. 360. 358. 367. 371. 378. 384. 385. 382. 388. 394. 396. 401. 405. 399. 407. 19154 346. 346. 354. 355. 363. 369. 376. 371. 378. 381. 388. 385. 394. 401. 397. 409. 19158 350. 351. 362. 368. 373. 379. 384. 387. 389. 385. 398. 395. 394. 401. 398. 408. 19168 329. 329. 338. 341. 342. 352. 356. 359. 360. 369. 376. 378. 380. 378. 374. 385. 19169 358. 360. 365. 372. 375. 379. 381. 388. 390. 391. 403. 403. 409. 410. 411. 424. 19170 350. 353. 359. 365. 368. 382. 382. 386. 386. 390. 396. 395. 402. 402. 400. 396. .................................................................................................................................... DAY 17 18 19 20 21 22 23 24 25 26 27 28 29b 30 31 32 ________________________________________------------------------------------------------------------------------------------------ 19111 401. 403. 407. 410. 412. 414. 426. 424. 427. 429. 437. 436. 438. 440. 440. 442. 19117 410. 424. 423. 427. 428. 430. 438. 446. 444. 445. 456. 456. 463. 460. 464. 466. 19124 403. 397. 402. 409. 409. 410. 415. 410. 413. 416. 419. 425. 430. 429. 428. 433. 19126 394. 393. 394. 400. 402. 400. 400. 400. 406. 407. 415. 420. 418. 422. 420. 423. 19127 388. 382. 384. 398. 402. 405. 464.c 417. 416. 421. 432. 436. 436. 435. 440. 446. 19128 414. 425. 429. 429. 430. 434. 436. 433. 430. 440. 439. 443. 444. 444. 451. 449. 19140 401. 407. 415. 417. 424. 417. 420. 428. 430. 436. 434. 442. 440. 446. 450. 448. 19145 399. 401. 407. 414. 418. 414. 421. 424. 424. 430. 435. 437. 440. 440. 447. 453. P 19149 383. 376. 383. 386. 396. 399. 402. 405. 406. 407. 417. 417. 422. 422. 433. 434. c-r 19152 412. 415. 416. 418. 426. 399. 406. 410. 417. 424. 430. 432. 430. 440. 445. 449. 19154 408. 409. 411. 412. 417. 416. 414. 415. 419. 427. 437. 437. 443. 448. 452. 463. 19158 412. 412. 414. 422. 427. 428. 432. 431. 432. 439. 440. 447. 447. 443. 446. 450. 19168 390. 394. 388. 391. 394. 390. 387. 392. 390. 396. 403. 404. 405. 404. 409. 414. 19169 424. 429. 431. 438. 438. ,436. 446. 447. 444. 447. 454. 456. 463. 466. 461. 468. _ _ _ _1_91_7_0 _ _ _ _ _ _4_07_._ _ _41_2_._ _ _41_0_. _ _4_1_8._ _ _4_21_._ _ _4_20_._ - -4-2-4-.- - - -4- 2- 8- .- - - - 4- 2- 8- .- - - - -4-3-3-.- - - -4-4-3-.- - - -4-4-3-.- - - -4-4-6-.- - - -4--4--8--.-- -----4--5--2--.--------4--5- 5- .- - - - DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Value appeared incorrectly recorded and was excluded from group averages and statistical analyses. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B21 (PAGE 10): BODY WEIGHTS - INDIVIDUAL DATA - FO GENERATION MALE RATS ________________________________________-------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP V 10 MG/KG/DAY ________________________________________-------------------------------------------------------------------------------------------- _ _ _ _ _ _ _ _ _D_AY_ _ _ _33_ _ _ _ _34_ _ _ _ 3_5_ _ _ _3_6_ _ _ _3_7_ _ _ _3_8_ - - - -3-9- - - - - -4-0- - - - - -4- 1- - - - - - 4- -2- - - - - -4-3- - - - - -4-4- - - - - -4-5- - - - - -4-6- - - - - - 4- -7- - - - - - - - - - - - - - - - - - - - - - - - - - - - - 19111 446. 456. 449. 449. 457. 456. 455. 461. 468. 466. 432. 19117 465. 467. 473. 467. 477. 481. 484. 480. 486. 483. 462. 19124 434. 437. 443. 438. 448. 445. 449. 445. 448. 450. 426. 19126 430. 436. 431. 435. 429. 438. 437. 445. 446. 452. 452. 426. 19127 448. 456. 448. 454. 444. 454. 453. 458. 455. 470. 467. 434. 19128 458. 457. 459. 461. 473. 463. 473. 469. 472. 470. 481. 486. 445. 19140 452. 449. 456. 453. 464. 454. 466. 465. 466. 469. 473. 486. 442. 19145 454. 451. 455. 455. 460. 463. 459. 465. 464. 471. 473. 477. 466. 441. 19149 443. 442. 444. 451. 453. 460. 454. 463. 460. 459. 462. 466. 468. 439. 19152 450. 455. 458. 461. 464. 463. 464. 476. 479. 483. 485. 498. 491. 461. 19154 458. 466. 468. 472. 479. 477. 480. 491. 488. 496. 490. 496. 498. 498. 466. 19158 449. 448. 450. 453. 461. 458. 456. 460. 462. 469. 474. 461. 478. 471. 453. 19168 412. 414. 418. 414. 417. 423. 423. 420. 412. 416. 416. 421. 421. 425. 402. 19169 471. 469. 472. 484. 485. 485. 479. 471. 474. 486. 479. 477. 482. 480. 448. 19170 456. 460. 459. 465. 460. 462. 466. 457. 452. 457. 459. 463. 467. 471. 445. ________________________________________-------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B22 (PAGE 1): FEED CONSUMPTION VALUES - INDIVIDUAL DATA - FO GENERATION MALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B22 (PAGE 2): FEED CONSUMPTION VALUES - INDIVIDUAL DATA - Fo GENERATION MALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B22 (PAGE 3): FEED CONSUMPTION VALUES - INDIVIDUAL DATA - Fo GENERATION MALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY JWMBER: T-7706.1) TABLE B23 (PAGE 1): MATING AND FERTILITY - INDIVIDUAL DATA - Fo GENERATION MALE RATS 19102 4 M 19106 1 M 19108 1 M 19110 2 M 19113 4 M 19120 1 M 19129 2 M 19136 1 M 19138 1 M 19139 4 M 19147 1 M 19153 1 M 19164 2 M 19165 3 M 19171 1 M ............................................. M = MATED ( ) = FEMALE RAT NUMBER C = CONFIRMED P = PREGNANT NP = NOT PREGNANT a. Result of first cohabitation period. b. Result of second cohabitation period. P(19004) P (19009) P (19016) P (19018) P (19026) P(19036) P(19037) P(19043) P(19047) P(19048) P( 19052) P (19055) P (19061) P (19067) P( 19071) PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B23 (PAGE 2): MATING AND FERTILITY - INDIVIDUAL DATA - Fo GENERATION MALE RATS 19101 3 M C P (19003) 19105 3 M C P(19007) 19107 4 M C P(19008) 19112 2 M C P(19013) 19114 1 M C NP (19014) 19121 4 M C P (19015) 19123 1 M C P (19017) 19130 4 M C P (19024) 19137 2 M C P (19029) 19146 3 M C P(19034) 19155 1 M C NP(19038) 19159 4 M C P (19056) 19172 4 M C P (19057) 19173 2 M C P (19060) 19174 4 M C P (19064) .................................................................................................................................... DOSAGE GROUP IV 3 MG/KG/DAY .................................................................................................................................... 19100 7 19103 4 19104 4 19118 2 DID NOT MATE M C M C M C - (19002) P (19005) P (19035) P(19039) 19133 4 M C P (19040) 19141 1 M C P (19045) 19142 1 M C NP (19046) P 19144 7 DID NOT MATE - (19051) C'L 19148 3 M C P (19054) 19150 1 M 19156a 4 M 19156b 6 M 19160 2 M C P(19058) C P (19062) C P (19002) C P (19063) 19162 4 M C P (19066) 19163a 2 1916333 7 M C DID NOT MATE P(19069) - (19051) 19166 1 M C P (19073) .................................................................................................................................... M = MATED ( ) = FEMALE RAT NUMBER C = CONFIRMED 'P = PREGNANT NP = NOT PREGNANT a. Result of first cohabitation period. b. Result of second cohabitation period. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B23 (PAGE 3): MATING AND FERTILITY - INDIVIDUAL DATA - FO GENERATION MALE RATS 19111 1 M 19117 3 M 19124 4 M 19126 4 M 19127 4 M 19128 3 M 19140 4 M 19145 1 M 19149 4 M 19152 1 M 19154 4 M 19158 1 M 19168 1 M 19169 1 M 19170 1 M --------------------________________^__ M = MATED ( ) = FEMALE RAT NUMBER C = CONFIRMED P = PREGNANT NP = NOT PREGNANT C P (19001) C P (19006) C P(19011) C P(19020) C P (19022) C P(19025) C P(19027) C P (19028) C P (19030) C P (19031) C P (19032) C P (19033) C P (19049) C P (19059) C P (19070) HOME CAGE BEHAVIOR 2 2 2 2 2 2 1 2 2 1 ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 2a 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 1 1 REARS IN OPEN FIELD 2 5 3 4 3 6 1 1 8 0 DEFECATION IN OPEN FIELD 1 1 1 1 1 1 1 1 1 1 URINATION IN OPEN FIELD 2 1 1 2 1 2 1 2 1 1 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 1 1 1 1 1 1 1 1 1 1 GAIT ABNORMALITY, SEVERITY 1 1 1 1 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE 1 1 1 1 1 1 1 1 1 1 LACRIMATION 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION 0 0 0 0 0 0 0 0 0 0 ABNORMAL RESPIRATION 0 0 0 0 0 0 0 0 0 0 APPEARANCE 1 1 1 1 lb 1 1 1 1 1 VISUAL REACTION 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION 2 2 2 2 2 2 2 2 2 2 AUDITORY REACTION 3 3 3 3 3 3 3 3 3 3 TAIL-PINCH REACTION 2 2 2 2 2 2 2 2 2 2 AIR RIGHTING RESPONSE 1 1 1 1 1 1 1 1 1 1 PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 355 460 500 360 340 470 3 15 4 15 2 95 315 FORELIMB GRIP TEST #2 (G) 460 510 420 3 95 375 350 465 465 4.30 3 70 HINDLIMB GRIP TEST #1 (G) 450 320 620 315 315 490 255 300 280 345 HINDLIMB GRIP TEST #2 (G) 4 15 380 690 220 330 390 255 325 250 275 LANDING FOOT SPLAY #1 (CM) 6.7 9.8 8.8 7.9 4.5 8.5 7.8 8.0 7.7 8 -4 LANDING FOOT SPLAY #2 (CM) 6.8 7.6 7.7 7.6 5.7 7.1 7.2 6.8 7.5 8.4 ._B_OD_Y_W_E-I-G-H-T__(._GI.___--------------4--5-2--------4-8-7--------4--9-5--------5-2-7--------4-6--4--------5-2-9--------5-0--7--------4-4-5--------4-3-3---------5-0-3------------------------------------ VALUES ARE THE QUANTITY, CATEGORY NUMBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. a. Value appeared incorrectly recorded; value excluded from group averages and statistical analyses. b. Chromorhinorrhea. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B24 (PAGE 2): FUNCTIONAL OBSERVATIONAL BATTERY - INDIVIDUAL DATA - FO GENERATION MALE RATS HOME CAGE BEHAVIOR 2 2 2 2 2 2 2 1 1 1 ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 1 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 1 1 REARS IN OPEN FIELD 11 8 3 11 9 4 4 3 4 5 DEFECATION IN OPEN FIELD 1 1 1 1 2 2 1 1 1 1 URINATION IN OPEN FIELD 2 1 2 2 2 2 2 2 2 2 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 1 1 1 1 1 1 1 1 1 1 GAIT ABNORMALITY, SEVERITY 1 1 1 1 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE 1 1 1 1 1 1 1 1 1 1 LACRIMATION 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION 0 0 0 0 0 0 0 0 0 0 ABNORMAL RESPIRATION 0 0 0 0 0 0 0 0 0 0 APPEARANCE 1 1 1 1 1 1 1 1 1 1 VISUAL REACTION 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION 2 2 2 2 2 2 2 2 2 2 AUDITORY REACTION 3 3 3 3 3 3 3 3 3 3 TAIL-PINCH REACTION 2 2 2 2 2 2 2 2 2 2 AIR RIGHTING RESPONSE 1 1 1 1 1 1 1 1 1 1 PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 3 55 375 3 95 415 455 245 250a 505 2 15 435 FORELIMB GRIP TEST #2 (G) 400 490 360 470 465 4 10 365 3 90 465 550 HINDLIMB GRIP TEST #1 (G) 345 435 610 365 315 250 3 15 155 435 465 HINDLIMB GRIP TEST #2 (G) 460 505 575 450 520 550 34533 220 365 335 LANDING FOOT SPLAY #1 (CM) 11.6 9.5 7.6 8.0 6.8 6.2 12.5 7.6 8.0 9.9 LANDING FOOT SPLAY #2 (CM) 9.7 8.4 6.5 7.8 6 -5 4.8 12.7 7.8 9.2 11.5 ._B-O-D_Y_W_E_IG_H_T _(_G_) ____--_--------4-4-4--------4-8-9---------4-9-0--------4-6-7--------4--6-4--------4-9-6--------4-4--3--------5-1-7--------3--8-5--------4-6-0-----------------------------------~~ VALUES ARE THE QUANTITY, CATEGORY NUMBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. a. Vocalization and aggression observed during forelimb grip test. b. Soft or liquid feces observed during hindlimb grip test. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B24 (PAGE 3): FUNCTIONAL OBSERVATIONAL BATTERY - INDIVIDUAL DATA - FO GENERATION MALE RATS HOME CAGE BEHAVIOR 2 2 2 2 2 2 2 1 1 1 ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 1 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 1 1 REARS IN OPEN FIELD 3 3 7 2 2 3 2 5 6 6 DEFECATION IN OPEN FIELD 2 1 2 1 1 1 2 1 1 1 URINATION IN OPEN F1,ELD 2 2 2 1 2 1 1 1 1 1 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 1 1 1 1 1 1 1 1 1 1 GAIT ABNORMALITY, SEVERITY 1 1 1 1 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE 1 1 1 1 1 1 1 1 1 1 LACRIMATION 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION 0 0 0 0 0 0 0 0 0 0 ABNORMAL RESPIRATION 0 0 0 0 0 0 0 0 0 0 APPEARANCE 1 1 la 1 1 1 1 1 1 1 VISUAL REACTION 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION 2 2 2 2 2 2 2 2 2 2 AUDITORY REACTION 3 3 3 3 3 3 3 3 3 3 TAIL-PINCH REACTION 2 2 2 2 2 2 2b 2 2 2 AIR RIGHTING RESPONSE 1 1 1 1 1 1 1 1 1 lc PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 2 15 500 480 490 270 550 405 3 95 340 570 FORELIMB GRIP TEST #2 (G) 175 430 545 400 430 3 95 3 95 515 330 360 HINDLIMB GRIP TEST #1 (G) 520 480 540 415 3 95 465 375 280 305 220 HINDLIMB GRIP TEST #2 (G) 54 5 580 415 525 395 4 75 4 90 435 255 280 LANDING FOOT SPLAY #1 (CM) 8.2 8.7 9.7 6.8 9.0 11.8 8.5 9.7 11.1 8.7 LANDING FOOT SPLAY #2 (CM) 6.7 7.5 10.3 7.5 8.8 10.2 9.5 7.8 10.7 7.6 .-B-O--D-Y--W-E--I-G-H-T---(.-G-.)--------------------3-7-5---------5-3-2--------4--5-9--------4--8-2--------4-3-9---------6-0-1---------4-7-6--------4--5-7--------4-6--7--------4-8--3------------------------------ VALUES ARE THE QUANTITY, CATEGORY NUMBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. a. Localized alopecia: lower midline - right side (4.0 cm x 1.5 cm). b. Soft or liquid feces observed during tail-pinch reaction. c. Vocalization to touch and aggressive behavior observed during air righting response. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B24 (PAGE 4): FUNCTIONAL OBSERVATIONAL BATTERY - INDIVIDUAL DATA - Fo GENERATION MALE RATS HOME CAGE BEHAVIOR 2 2 2 2 2 2 1 2 2 1 ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 1 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 1 1 REARS IN OPEN FIELD 7 6 1 4 4 2 6 7 3 5 DEFECATION IN OPEN FIELD 1 2 1 1 1 1 1 1 1 1 URINATION IN OPEN FIELD 2 2 1 2 2 1 2 2 2 2 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 1 1 1 1 1 1 1 1 1 1 GAIT ABNORMALITY, SEVERITY 1 1 1 1 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE 1 1 1 1 1 1 1 1 1 1 LACRIMATION 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION 0 0 0 0 0 0 0 0 0 0 ABNORMAL RESPIRATION 0 0 0 0 0 0 0 0 0 0 APPEARANCE 1 1 1 1 1 1 1 1 1 la VISUAL REACTION 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION 2 2 2 2 2 2 2 2 2 2 AUDITORY REACTION 3 3 3 3 3 3 3 3 3 3 TAIL-PINCH REACTION 2 2 2 2 2 2 2 2 2 2 AIR RIGHTING RESPONSE 1 1 1 1 1 1 lb 1 1 1 PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 450 355 330 270 335 120 3 90 355 465 260 FORELIMB GRIP TEST #2 (G) 54 0 315 465 4 75 365 220 365 255 675 345 HINDLIMB GRIP TEST #1 (G) 425 520 450 380 350 270 215 200 4 05 270 HINDLIMB GRIP TEST #2 (G) 385 420 335 4 15 265 420 365 340 485 3 75 LANDING FOOT SPLAY #1 (CM) 7.6 10.8 6.6 9.0 6.6 6.8 9.7 8.5 11.9 9.4 LANDING FOOT SPLAY #2 (CM) 6.9 9.7 7.8 8.5 4.5 6.7 9.6 9.2 12.2 9.6 ._B_OD_Y_W_E-I_G_HT__(._G.I___---------------4-4-1---------5-0-5--------4--9-4--------4--7-. --2--------4-5-9---------2E-Ut-.J3--------5--0-6--------5-3--8--------4-8--1--------4-7-3-------------------------------------VALUES ARE THE QUANTITY, CATEGORY NUMBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. a. Chromodacryorrhea - right eye and incisors misaligned - upper bilateral. b. Vocalization and aggression observed during air righting response. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B24 (PAGE 5): FUNCTIONAL OBSERVATIONAL BATTERY - INDIVIDUAL DATA - Fo GENERATION MALE RATS HOME CAGE BEHAVIOR ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 1 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 1 1 REARS IN OPEN FIELD 6 6 2 8 2 0 6 5 9 1 DEFECATION IN OPEN FIELD 1 1 1 1 2 1 1 1 1 1 URINATION IN OPEN FIELD 2 2 1 2 2 1 2 2 2 1 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 1 1 1 1 1 1 1 1 1 1 GAIT ABNORMALITY, SEVER1TY 1 1 1 1 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE 1 1 1 1 1 1 1 1 1 1 LACRIMATION 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION 0 0 0 0 0 0 0 0 0 0 ABNORMAL RESPIRATION 0 0 0 0 0 0 0 0 0 0 APPEARANCE 1 1 1 1 1 1 1 1 1 1 VISUAL REACTION 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION 2 2 2 2 2 2 2 2 2 2 AUDITORY REACTION 3 3 3 3 3 3 3 3 3 3 TAIL-PINCH REACTION 2 2 2 2 2 2 2 2 2 2 AIR RIGHTING RESPONSE 1 1 1 1 1 1 1 1 1 1 PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 3 55 335 320 580 4 15 300 2 85 165 165 510 FORELIMB GRIP TEST #2 (G) 365 450 350 470 470 450 450 155 275 320 HINDLIMB GRIP TEST #1 (G) 500 420 510 485 440 430 430 500 250 305 HINDLIMB GRIP TEST #2 (G) 485 660 555 400 450 505 440 480 2 15 330 LANDING FOOT SPLAY #1 (CM) 8.1 7.5 8.1 8.8 9.1 5.3 13.0 8.1 5.8 8.5 LANDING FOOT SPLAY #2 (CM) 7.6 6.8 8.3 7.6 9.7 5.0 12.7 6.5 6.2 7.6 ._B_OD_Y_W_E-I~ -G_H~~_T -_I.G-_), ___-------------4--5-8--------4-7-5---------4-4-0--------4-3-9--------4-6--3--------4-7-0--------4--6-2--------4-7-3--------4-6--6--------4-7-3------------------------------------- VALUES ARE THE QUANTITY, CATEGORY NUMBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B25 (PAGE 1): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION MALE RATS DOSAGE GROUP I o MG/KG/DAY RAT NUMBER 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 71 60 64 68 67 72 58 62 73 53 78 74 56 64 56 61 83 67 71 59 59 66 58 72 76 66 76 76 73 69 68 80 71 66 70 55 83 70 93 67 61 46 56 71 72 75 55 47 73 45 64 0 55 62 55 61 78 52 5 51 29 9 69 45 50 29 56 50 4' 53 50 0 76 9 7 97 24 62 0 54 3 1 33 1 44 70 0 40 30 54 1 1 12 1 5 59 0 31 73 67 0 0 2 36 7 71 0 47 20 59 33 55 0 0 71 72 3 46 43 55 6 58 61 0 59 82 20 73 0 62 0 72 15 0 56 44 68 50 0 71 0 7 0 0 37 61 61 12 5 50 0 0 6 1 56 83 60 25 0 5 5 0 0 1 47 49 72 44 16 2 0 0 7 36 35 71 14 53 8 4 528 52 9 641 533 870 1178 811 907 587 880 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B25 (PAGE 2): MOTOR ACTIVITY - INDIVIDUAL DATA - FO GENERATION MALE RATS DOSAGE GROUP I o MG/KG/DAY RAT NUMBER 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 185 217 193 169 161 194 229 219 224 220 182 182 133 186 191 201 188 184 2 17 210 161 115 110 181 14 9 162 183 178 173 143 144 127 106 166 190 88 13 9 129 134 191 106 54 113 132 12 9 140 92 69 94 78 91 0 125 111 71 158 133 135 1 87 70 6 14 0 82 125 48 65 80 2 84 99 0 87 8 10 124 18 165 0 102 0 0 31 0 113 118 0 53 50 75 0 0 11 1 4 122 0 43 122 186 0 0 0 36 6 169 0 58 12 121 38 91 0 0 84 114 72 62 119 9 90 60 0 95 116 21 126 0 152 0 67 9 0 151 127 104 90 0 108 0 5 0 0 105 118 99 10 3 51 0 0 5 0 79 102 106 24 0 3 4 0 0 0 100 57 72 70 16 1 0 0 6 46 44 125 23 56 2 3 1089 954 1129 1118 1807 2283 1473 1761 1112 1934 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B25 (PAGE 3): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION MALE RATS DOSAGE GROUP I1 0.3 MG/KG/DAY RAT NUMBER 19102 19106 19108 19110 19113 19120 19129 19136 19138 19139 NUMBER OF MOVEMENTS BLOCK 1 64 55 64 64 71 69 52 64 74 66 BLOCK 2 69 55 61 60 56 94 58 57 61 65 BLOCK 3 68 59 47 66 24 82 67 72 50 63 BLOCK 4 63 68 65 73 15 93 67 47 60 61 BLOCK 5 44 60 61 59 0 93 61 55 51 63 BLOCK 6 32 24 51 70 0 86 77 70 55 63 BLOCK 7 5 54 71 65 0 59 70 62 63 51 BLOCK 8 3 45 60 57 2 74 59 67 48 52 BLOCK 9 34 77 37 64 0 70 48 45 70 47 BLOCK 10 6 75 60 37 51 65 56 48 56 77 BLOCK 11 0 53 3 0 16 64 72 63 40 40 BLOCK 12 1 64 0 51 1 45 66 51 35 37 BLOCK 13 5 58 65 0 12 0 74 54 78 84 BLOCK 14 14 61 64 22 0 8 59 61 81 68 BLOCK 15 51 53 39 20 0 17 27 78 25 45 BLOCK 16 36 81 51 65 0 2 2 57 33 49 BLOCK 17 64 78 52 46 0 0 3 63 22 64 BLOCK 18 63 58 41 40 0 38 6 53 1 44 P C'L TOTAL 622 1078 8 92 859 248 959 924 1067 903 1039 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B25 (PAGE 4): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION MALE RATS DOSAGE GROUP I1 0.3 MG/KG/DAY RAT NUMBER 19102 19106 19108 19110 19113 19120 19129 19136 19138 19139 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 2 15 201 192 192 176 197 240 2 16 191 198 133 164 152 182 138 160 238 209 170 198 150 126 71 150 24 211 223 2 03 144 101 116 140 162 201 16 144 209 129 143 123 35 147 166 116 0 171 2 14 197 83 182 40 31 103 142 0 135 157 125 76 88 1 125 136 98 0 148 153 102 71 68 1 77 81 55 0 162 117 146 69 129 33 105 99 85 0 117 116 63 125 66 4 114 78 42 94 94 137 95 75 133 0 117 4 0 49 135 177 123 58 51 0 72 0 60 0 90 146 181 62 60 1 88 86 0 7 0 190 135 207 155 26 90 120 29 0 2 114 152 122 102 113 59 60 17 0 26 28 146 15 67 57 125 63 84 0 0 1 121 28 76 93 151 66 56 0 0 0 117 21 121 111 54 37 41 0 43 2 139 1 46 1129 1986 1676 1550 504 1835 2462 2599 1661 1964 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B25 (PAGE 5): MOTOR ACTIVITY - INDIVIDUAL DATA - FO GENERATION MALE RATS IXlSAGE GROUP I11 1 MG/KG/DAY RAT NUMBER 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 67 57 63 79 60 74 60 51 56 68 55 57 63 76 57 67 69 55 81 66 76 72 52 71 56 70 59 57 71 67 80 68 63 73 61 71 79 43 84 60 62 63 66 61 66 75 60 71 72 60 72 57 52 60 47 78 64 52 74 86 74 57 52 47 65 71 69 28 76 59 57 68 51 6 66 78 49 43 68 54 59 86 64 22 31 69 22 31 74 39 56 73 66 14 35 75 17 47 85 2 21 44 52 2 69 74 84 73 50 43 71 8 37 14 12 79 60 60 59 55 109 5 54 49 8 70 22 37 81 5 29 17 81 0 29 55 0 53 57 44 27 69 71 1 2 85 0 65 75 3 50 72 5 0 0 66 3 47 64 5 72 55 1 18 0 54 5 13 27 46 73 65 0 55 5 57 1 42 2 59 1110 993 893 648 669 1268 723 868 1156 821 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B25 (PAGE 6): MOTOR ACTIVITY - INDIVIDUAL DATA - FO GENERATION MALE RATS DOSAGE GROUP I11 1 MG/KG/DAY RAT NUMBER 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 2 12 219 145 127 148 65 164 103 116 98 36 165 103 18 47 52 90 100 2008 226 224 156 125 191 110 79 139 154 120 76 8 1 20 135 100 81 156 2101 218 129 92 109 147 87 104 151 102 97 77 37 87 104 79 11 0 0 1631 207 144 160 138 134 93 60 2 18 6 0 11 54 0 0 0 16 76 1119 194 197 177 169 160 91 175 160 39 59 134 48 5 30 1 0 0 5 1644 201 198 197 138 157 164 112 161 208 153 170 180 113 95 139 152 162 85 2785 228 220 203 14 1 115 139 146 73 25 11 125 138 34 0 0 5 3 0 1606 236 230 187 167 151 99 29 51 82 99 99 109 24 136 140 71 18 43 1971 219 2 02 202 188 166 201 141 132 116 131 81 111 98 65 138 211 23 1 2426 207 195 206 122 134 135 92 127 76 0 62 110 2 73 1 1 76 97 1716 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE B25 (PAGE 7): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION MALE RATS DOSAGE GROUP IV 3 MG/KG/DAY RAT NUMBER 19100 19103 19104 19118 19133 19141 19142 19144 19148 19150 BLOCK 1 80 55 61 54 47 63 73 66 54 68 BLOCK 2 84 74 72 73 69 69 76 75 70 73 BLOCK 3 54 69 87 87 81 69 67 54 74 74 BLOCK 4 57 57 75 86 78 71 55 48 63 64 BLOCK 5 67 43 85 71 37 74 77 71 42 64 BLOCK 6 29 67 62 79 41 66 59 64 74 58 BLOCK 7 47 60 66 62 71 56 49 70 66 59 BLOCK 8 62 43 64 44 12 63 71 80 69 62 BLOCK 9 55 53 69 27 14 69 83 60 63 60 BLOCK 10 32 69 65 85 0 51 50 49 58 41 BLOCK 11 1 50 76 74 8 58 65 70 68 47 BLOCK 12 1 46 89 60 18 54 66 57 69 31 BLOCK 13 3 14 77 77 54 73 56 62 53 35 BLOCK 14 58 69 63 10 71 69 80 53 61 58 BLOCK 15 75 63 71 3 40 54 54 49 67 37 BLOCK 16 54 74 61 70 0 57 56 67 45 25 BLOCK 17 72 62 63 72 20 48 66 65 67 0 BLOCK 18 67 16 57 61 5 47 34 74 63 0 P U TOTAL 898 984 1263 1095 666 1111 1137 1134 1126 856 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B25 (PAGE 8): MOTOR ACTIVITY - INDIVIDUAL DATA - FO GENERATION MALE RATS DOSAGE GROUP IV 3 MG/KG/DAY RAT NUMBER 19100 19103 19104 19118 19133 19141 19142 19144 19148 19150 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 198 136 83 120 120 30 59 87 82 30 0 0 0 106 113 58 146 108 1476 244 202 105 193 90 12 9 186 223 196 155 83 114 18 136 111 134 127 36 2482 235 171 159 156 131 109 187 108 114 94 155 136 100 102 97 100 95 88 2337 219 163 138 139 137 128 87 59 27 120 92 101 102 14 1 61 91 78 1757 171 200 138 181 55 57 141 43 9 0 3 14 87 107 43 0 13 2 1264 200 206 143 114 145 154 142 131 93 126 143 71 105 106 60 114 109 65 2227 206 184 153 86 120 65 65 120 152 55 158 84 118 163 69 112 111 58 2079 182 182 160 125 144 146 122 123 151 107 143 125 124 96 107 134 143 115 2429 211 183 175 174 145 187 113 161 109 77 14 8 128 76 153 82 110 79 119 2430 180 175 152 121 105 92 84 106 103 67 52 38 72 95 59 49 0 0 1550 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B25 (PAGE 9): MOTOR ACTIVITY - INDIVIDUAL DATA - FO GENERATION MALE RATS DOSAGE GROUP V 10 MG/KG/DAY RAT NUMBER 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 62 69 57 61 72 68 64 49 77 61 68 66 67 72 63 75 87 63 65 70 67 56 63 76 61 78 72 60 77 65 58 50 73 63 68 37 41 48 59 63 62 49 65 42 25 51 62 76 76 61 39 50 59 51 67 56 55 66 57 74 59 63 62 62 30 49 54 52 68 69 55 74 24 61 12 54 78 44 67 28 46 37 42 35 75 41 87 56 47 56 73 40 47 51 64 23 47 67 52 32 44 0 45 50 33 23 82 54 43 41 28 39 57 37 67 39 43 64 44 58 44 31 64 30 86 42 21 62 39 11 77 0 29 64 66 62 16 56 68 0 75 0 39 62 57 41 61 39 56 0 43 0 66 61 3 49 69 64 68 2 71 0 72 49 56 34 6 68 14 3 65 1 58 31 63 38 1 31 35 19 1036 625 989 958 968 860 946 1019 1012 713 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B25 (PAGE 10): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION MALE RATS DOSAGE GROUP V 10 MG/KG/DAY RAT NUMBER 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 206 170 14 1 175 153 64 98 74 51 14 0 87 37 75 150 103 54 93 110 1981 161 174 110 169 75 67 81 101 63 64 .o 94 54 0 0 0 0 0 1213 173 143 118 118 124 97 112 36 47 80 98 59 107 32 60 77 98 91 1670 169 116 14 1 115 111 96 102 64 47 47 102 65 40 66 74 95 86 21 1557 212 14 9 109 103 30 87 39 7 107 67 33 105 163 106 68 0 70 106 1561 192 205 128 42 65 134 104 116 81 27 17 37 48 90 66 132 42 69 1595 181 176 197 86 108 64 107 90 125 49 126 42 29 14 100 142 5 0 1641 243 233 231 220 158 196 195 100 196 165 80 138 135 155 95 203 108 69 2920 230 158 146 126 173 94 121 98 129 87 60 57 38 120 76 68 6 55 1842 181 177 166 155 14 1 128 120 34 90 42 69 95 16 0 0 0 2 20 1436 418-028:PAGE B-89 . . . m a , r r w ( u * m m rl * m m w rl m rl m rl cn 0 m rl rl rl r cn (u rl m m 0 r rl (u rl rl * 0 m a, m w rl P m m (u 0 h I E c, rn 4 r (u m N r r m cn r ( u o r l r l * a, r a, cn c n c n c n c n m a, cn m m m r0 cna,cncn m B u w a, cn v rl w * r m (u (u m (u 0 m m (u * ( u r m r mwmm m (ummcu m * cn m w c n ( u m r l (u 0 rl r l r l ( u c v m m m rl rl r l r l r l r l r l rl rl cn cn c n c n c n c n c n cn cn rl rl r l r l r l r l r l rl rl (uwcoo m o o o r l rl r l r l r l r l rl c n c n c n m cn r l r l r l r l rl * E V H x 0 E 2I4 w 5 0 I4 W 2 n \ E E V 5E0 w p! w X E X i? 8 W 0 r P I E ih 0 *k E . mW 0 *P EP HI VE H x 0 ** E pw! wm ."Kn s$ 2 a wm p!- n po! Wrn ma $2 k- mE ww WE C!JH E%- 2 W Glw om 03 cv 0 I 03 *rl EI4 0 U 0 E 0 418-028:PAGE B-90 * * * m rl cn m * * m cv 0 m rl cn rl W cn W 0 W m m Ln cv rl m * rl rl rl rl cv d rl * . . (v ocv rl W 0 0 m cv PW cncv W W rl 0 P rl W mo rlcv P 0 rl * * W rl m 0 03 0 W cv P m W cn rl rl rl rl cv rl rl rl W m P m 03 P rl W *rl rlcv m 03 * m W 0 rl rl cv cv 0 OI W cn m P m m cv m m cv m cv * m * cv 0 3 0 0 0 m cnm Lnm W m m W rl W cv W m m m rl rl rl cv cv 0 W W rl WP *rl c*n cv m P a O m m cn P cn W cn 0 cn m cn 2 : I I I I * * m L n rl Ln m w c n WW P rl*o (vcv m mcvm m P(?O(vW cn c n c v c n c n a l W 03 cv m P cv 0 P 03 0 0 rl m m rl r l r l rl rl rl rl c n c n cn o\ cn cn r l r l rl rl rl rl PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE B26 (PAGE 3): SPERM MOTILITY, COUNT AND DENSITY - INDIVIDUAL DATA - FO GENERATION MALE RATS 19126 2 64 93 12 276 91 1032.3 19127 339 72 26 365 190 1805.0 19128 231 95 92 323 163 1730.3 19140 464 95 27 4 91 161 1578.2 19145 360 96 30 3 90 146 1194.1 19149 289 87 39 328 198 1789.9 ----__1_91_5_2_____--____2-5-6---------------------9-8----------------------6----------------------2-6-2--------------------2-0-4-------------------1-7--9-4-.-7----------------------------- a. Sum of number motile and static count. Groups of five fields were evaluated until a sperm count of at least 200 was achieved or 20 fields were evaluated. b. Sperm count used in the calculation of sperm density. Ten fields were evaluated. c. The sperm density was calculated by dividing the sperm count by the volume in the image area (34.3 x mL) , multiplying by 2 (dilution factor) and multiplying by to obtain the sperm concentration. The calculated sperm concentration value (rounded to 1 decimal place) was multiplied by 50 (volume) and divided by the weight of the left cauda epididymis (see Table B19 for the weight of the left cauda epididymis) to obtain the sperm density. The calculated value will vary by approximately 0.8% from the Computer Automated Sperm Analysis because the digital image evaluated is slightly smaller (4 pixels) than the actual field causing a slight underestimate of the actual volume and an overestimate of the concentration. 418-028:PAGE B-92 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I30 I I I I I I I I I I I I I I I I I I I I I I I I I I . . . . . . . I I 0 0m o o m o l n o 0 I *. I I r m mm*mmmCJ m 1 . . . . . . I I I m o m000momln I I *. I I m m CJ * m m m m m m I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I H I I I I I I I I I I I I I I I I I I I I I I I I I I 3 w I I X I I I E m I I w I 3w 0 I r I r I I E; 3 k 0 I I I 0 0oooorlrlo 0 I I I I I I I o r l rl 0 0 0 0 0 0 0 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I d o o o o r l o o r l rl I I I ; C J r l rl m o o rl o r l o I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I 0 I I I I I I Fr, I I I I I I I I I I I I I I I I I I I I I I I I I I ;g3; I I r* I I I m I I I * I I *rl m m r l m I I rlrlm I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I E I IW I I I I I I I I I I wCJ r c o m m m m r l m II 0. I I0 rl m m c o u, m m m I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I w3 I I I I I mPI I I I I I I I I I I I I h rl co I w CJ I 0 r3 I I I 2 03 rl v * r CJ a I I u,* Ia m I rlrl I I I I fH I I I I I I I I mw con lcnn m m w m o m I I r l r l rl r l r l r l I I I IH IH I I I m m u,mCJml-lm* m I orl rlrl(\lmmmm m I r l r l r l r l r l r l r l r l r l rl I m m mmmmcnmm m I r l r l r l r l r l r l r l r l r l rl I wmm cncnm rlrlrl I I I I I I 418-028:PAGE B-93 I I I I I I I I I 1 I I I I 1 I I I I 1 I I I 1 I I I I I I I I I I I I I I I I I 1 l l I I I I I I I I I I I 1 I x I I I I I I 0 I I I E I I I I I I I 1 I I I 1 I I I I I I I I I J l I I I I I I I I I I I I I I I . . . . . . . . . I I lommmmooooml 1 0 1 lrlrlmrl*m*u7*ml I I I 1 100000000001 I *.........I Im*rlu7**cocomrll I cn I I I I I I I I I 1 I I I I I 1 I I I I I I I I I I I I I I I 1 I I I I I I I I I I I 1 I f I I I I I I I I I I I I I I I I I 1 I I 5 ; I I I I I t I I I I 1 1 I wX I hl I I I I E 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orlrlrloorlmrlol I I I I lrloorlmrlcnorlol I P I I I I I I I I I I I I I I I I I I I I I I I t I I I I I 1 I I I I I I I I I I I I I O I h I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I 1 1 I I I :o$: F E II I I I Irlrlmrlmmm**ml I I I I I I I I Im*oPmmu7wmmI I P I I I I I I I I I I t I I I I I I I I2I I I I I f I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I Id l I I I I I I 0 1 I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I Y I P m a IHI IHI Id I51 m 3 w 2 mE w. 0 *P 0 F4 I 4 3 E H I n m a m w 0 I -3 a2 rl v P d m 0 u a 0 W s ii g I3 I I I I I I I I I I I I I I I I I k i iI u o I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I 0 2 I I 2 E I I I I I I I I I I I n I I I I 8 2 4 w I I I E X n I I I I I I I I I I I I I I I I I I I I I I I I I I I I 3 i I !% I W I I I I I I I I I I I I I I I I I I I I I I I I I I I I t I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I \ !2I I I I 0 I rl I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I W I W I 3 I I 0 n I I I I II I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I . . . . . . . . . . I I lolnoolnlnlnoom I 1 I lrirlvmln*vm*v I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 100000000rl0 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I orlrlommorlrlrl I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I rlrlrlmmlnrlrlmrl I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 418-028:PAGE B-94 APPENDIX C REPORT TABLES - FO GENERATION FEMALE RATS PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C1 (PAGE 1): CLINICAL OBSERVATIONS - SUMMARY - FO GENERATION FEMALE RATS MORTALITY BROWN PERIORAL SUBSTANCE LOCALIZED ALOPECIA: TOTAL HEAD LIMBS URINE-STAINED ABDOMINAL FUR ABDOMINAL DISTENTION 0 o/ 0 o/ 0 o/ 0 0 o/ 0 o/ 0 o/ 0 0 o/ 0 0 o/ 0 0 71 1 o/ 0 o/ 0 o/ 0 o/ 0 3/ 1 2/ 1 bh) PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C1 (PAGE 2): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS MAXIMUM POSSIBLE INCIDENCE MORTALITY LOCALIZED ALOPECIA: TOTAL LIMBS HEAD 330/ 15 0 329/ 15 0 338/ 15 0 312/ 14 0 30/ 2 30/ 2 o/ 0 330/ 15 0 21/ 1 14/ 1 7/ 1 BACK: SCAB 13/ 1 o/ 0 o/ 0 o/ 0 o/ 0 . . . B. A. C. K. .:. . .U.L.C.E.R.A.T.I.O.N. . . . . . . . . . . . . . . . . . . . . . .1./. . .1. . . . . . . . . . . . . . .o./. . .0. . . . . . . . . . . . . . . .o./ . . .0. . . . . . . . . . . . . .o./. . .0. . . . . . . . . . . . . o. ./ . . .0. . . . . . . . . . STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Restricted to rats with a confirmed mating date. bh) 418-028:PAGE C-3 I I I I I I I I I I I I I I I I I I I I I I I I I I I ILn 1 trl I I I Im I lm I I I I I I I I I I I I I I I I I I I I I t>m IH I I\ IW I la, I lcv I I I I I I I I w I I I I 3: I I I3 I I I I I I I I I Im I lrl I 10 I I I I I I I I I I I I I I I I I I I I 1 I 1 1 t tm I Irl 1 I IHm I\ IH - 1 0 I Olm I Im 0 I I h I I I I I I I I I I I I I I I I I I I I I I I I I I I 1U-l Irl I 1HO I I I I I I\ I0 ~m Irn I I I I I I I I I I I I I I I I I I I I I 1 I ; E l I I IZ I : , w I 1 li-i I IU I n m a, (\1 0 I CO rl Tr v ru l I I I I I I I I I I I I I c v r l r l o rl rl rl rl rl 0 1 I I 0 \ \ \ \ \ \I W ul r W rl 01 rl I I I mcvorl 0 0 0 0 0 0 \ \ \ \ \ \I& 0 0 0 0 0 WIW lvl I0 cvrlcvo 0 0 0 0 0 0 \ \ \ \ \ 0 0 0 0 0 rld00 0 0 0 N 0 0 \\\\ \ \ \ \ \ 0000 0 0 0 m 0 cvcv cv mcvrlo 0 0 0 m 0 0 \\\\ \ \ \ \ \ uloulo 0 00 m 0 mm rl I m -.< rsz 0 vl x w n PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C2 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS MORTALITY N 0 0 0 0 0 APPEARED NORMAL N 15 15 14 13 14 KIDNEYS : RIGHT, CORTEX ADHERED TO RIGHT LATERAL LIVER LOBE N 0 0 1 0 0 OVARIES : LEFT, SMALL N 0 0 1 0 SPLEEN : OMENTUM ADHERED TO THE LENGTH OF THE OUTER CAPSULE N 0 LARGE AND SMALL INTESTINES: DISTENDED WITH GAS N 0 THYMUS : SMALL N 0 SPLEEN : SMALL N 0 ADRENALS : BILATERAL, LARGE N 0 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C3 (PAGE 1): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND PRIMORDIAL FOLLICLE COUNT - SUMMARY - FO GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) I I1 I11 IV V 0 0.3 1 3 10 RATS TESTED PREGNANT TERMINAL BODY WEIGHT BRAIN LIVER KIDNEY LEFT KIDNEY RIGHT ADRENAL LEFT ADRENAL RIGHT SPLEEN THYMUS OVARY LEFT OVARY RIGHT UTERUS WITH CERVIX N N MEANfS .D. MEANfS.D . MEANfS .D . MEANfS.D . MEANfS .D . MEANfS.D . MEANfS.D . MEANfS.D . MEANfS .D . MEAN+S .D . MEANfS .D. MEANfS .D . 15 15 306.3 f 26.0 2.16 f 0.08 [ 10la 14.38 f 2.24 10la 1.39 k 0.14 [ 10la 1.47 f 0.12 10la 0.052 f 0.009 [ 10la 0.043 f 0.007 [ 10la 0.63 f 0.08 [ 10la 0.21 f 0.07 [ 10la 0.055 f 0.014 0.062 f 0.015 0.53 f 0.09 15 15 324.3 f 27.1 2.11 f 0.09 E 10la 15.98 f 2.71 [ 101a 1.47 f 0.14 [ 10la 1.52 f 0.16 10la 0.051 f 0.009 [ 10la 0.041 f 0.010 [ 10la 0.65 f 0.09 [ 10la 0.21 f 0.06 [ 10la 0.056 f 0.020 0.060 f 0.019 0.56 f 0.12 15 13 311.2 f 35.0 2.10 f 0.09 [ 10la 14.70 f 2.92 [ 10la 1.38 f 0.14 [ 10la 1.45 f 0.13 10la 0.051 f 0.009 [ 10la 0.050 f 0.011 [ 10la 0.60 f 0.11 10la 0.18 f 0.04 [ 10la 0.053 f 0.014 0.060 f 0.021 0.50 f 0.13 15 13 319.6 f 40.4 2.08 f 0.18 10la 15.11 f 3.28 [ 10la 1.49 f 0.19 [ 10la 1.52 f 0.13 [ 10la 0.053 f 0.012 [ 10la 0.049 f 0.006 10la 0.75 f 0.31 [ 10la 0.20 f 0.10 [ 10la 0.053 f 0.012 0.054 f 0.012 0.62 f 0.15 15 15 303.5 f 37.9 2.12 f 0.10 [ 10la 13.64 k 3.18 [ 10la 1.39 f 0.10 10la 1.41 f 0.15 [ 10la 0.053 f 0.009 [ 10la 0.046 f 0.008 [ 10la 0.54 f 0.10 [ 10la 0.18 f 0.04 [ 10la 0.054 f 0.015 0.054 f 0.018 0.56 f 0.16 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C3 (PAGE 2): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND PRIMORDIAL FOLLICLE COUNT - SUMMARY - Fo GENERATION FEMALE RATS RATS TESTED N 15 15 PREGNANT N 15 15 b E3 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C4 (PAGE 1): RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - SUMMARY - FO GENERATION FEMALE RATS PREGNANT TERMINAL BODY WEIGHT BRAIN LIVER KIDNEY LEFT KIDNEY RIGHT ADRENAL LEFT b ADRENAL RIGHT b SPLEEN THYMUS OVARY LEFT b N MEANfS .D. MEANfS .D . MEANfS .D . MEANfS .D . MEANfS .D . MEANfS.D. MEANfS.D. MEANfS .D . MEANfS .D . MEANfS.D. 15 306.3 f 26.0 0.711 f 0.062 [ 10la 4.703 & 0.450 [ 10la 0.459 f 0.049 [ 10la 0.482 f 0.043 [ 10la 17.095 & 2.132 [ 10la 13.984 f 1.612 [ 10la 0.208 f 0.024 [ 10la 0.070 & 0.022 [ 10la 18.005 & 4.064 OVARY RIGHT b UTERUS WITH CERVIX HEART MEAN+S.D. MEANfS .D . MEANfS .D. 20.307 f 4.390 0.175 & 0.031 0.459 f 0.067 [ 10la ALL WEIGHTS WERE RECORDED IN GRAMS (G). [ ] = NUMBER OF VALUES AVERAGED a. Only ten rats were tested. b. Value was multiplied by 1 0 0 0 . 15 324.3 f 27.1 0.659 f 0.055 [ 10la 4.947 f 0.703 10la 0.460 & 0.053 10la 0.472 & 0.052 10la 15.856 f 2.913 10la 12.842 & 3.379 [ 10la 0.205 f 0.028 E 10la 0.065 f 0.018 [ 10la 17.632 f 6.782 18.514 & 5.844 0.175 f 0.042 0.392 f 0.057 10la 13 13 15 311.2 f 35.0 319.6 f 40.4 303.5 f 37.9 0.684 f 0.082 [ 10la 4.697 f 0.512 [ 10la 0.446 & 0.054 [ 10la 0.467 f 0.042 [ 10la 16.799 f 4.091 [ 10la 16.479 k 4.381 [ 10la 0.194 f 0.035 [ 10la 0.059 & 0.016 [ 10la 17.053 f 3.559 0.675 f 0.099 [ 10la 4.795 f 0.517 [ 10la 0.479 f 0.070 [ 10la 0.492 & 0.058 [ 10la 16.867 f 3.532 10la 15.743 f 2.727 1Ola 0.238 f 0.076 [ 10la 0.065 f 0.037 [ 10la 16.883 f 4.629 0.732 f 0.102 [ 10la 4.582 f 0.541 10la 0.477 f 0.040 [ 10la 0.484 f 0 . 0 4 3 10la 18.402 f 4.737 [ 1Ola 15.959 f 4.763 [ 10la 0.181 f 0 . 0 2 6 [ 10la 0.059 f 0.016 10la 17.611 f 3.620 19.152 f 6.400 17.025 f 3.632 17.529 f 5.247 0.162 f 0.046 0.198 f 0.063 0.185 f 0.055 0.436 f 0.054 [ 10la 0.457 f 0.073 10la 0.453 & 0.056 [ 10la RATIOS ( % ) = (ORGANWEIGHT/TEFWNAL BODY WEIGHT) x 1 0 0 . PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C5 (PAGE 1): RATIOS ( % ) OF ORGAN WEIGHT TO BRAIN WEIGHT - SUMMARY - FO GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) ____________________----- RATS TESTED N BRAIN WEIGHT MEANfS .D . LIVER MEANfS.D . KIDNEY LEFT MEANfS .D . KIDNEY RIGHT ADRENAL LEFT MEANfS .D. . MEANkS D . ADRENAL RIGHT MEAN+S .D . SPLEEN THYMUS OVARY LEFT MEANfS .D . . MEANfS D . MEANfS .D. OVARY RIGHT MEANfS.D . UTERUS WITH CERVIX MEANfS.D. HEART MEANfS.D . 2.16 f 0.08 I1 I11 IV V 0.3 1 3 10 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ -------------- - - - - - - - - - - - 10 10 lo lo 2.11 f 0.09 2.10 f 0.09 2.08 f 0.18 2.12 * 0.10 664.58 f 96.72 757.16 f127.78 701.88 k155.49 729.66 f165.57 641.86 k141.49 64.31 f 5.18 69.72 f 4.89 65.73 f 6.72 71.48 f 6.75" 65.57 f 3.88 67.75 f 4.49 71.72 f 6.50 68.95 k 7.09 73.35 f 5.96 66.53 k 5.96 2.42 f 0.42 2.40 f 0.35 2.44 f 0.39 2.54 f 0.58 2.49 f 0.40 1.98 f 0.31 29.20 f 3.42 1.95 k 0.46 30.95 f 4.47 2.39 f 28.56 + 0.48 5.09 2.36 k 0.40 36.15 f 15.02 2.15 f 0.40 25.16 f 4.18 9.72 f 3.12 9.96 f 2.95 8.43 k 1.96 9.68 f 4.96 8.35 f 2.13 2.68 f 0.75 2.85 f 0.84 2.47 k 0.43 2.55 f 0.58 2.58 f 0.78 2.83 k 0.72 3.09 f 0.93 2.82 f 0.98 2.55 f 0.57 2.37 f 0.89 26.20 f 4.17 28.44 f 4.18 24.14 f 6.72 31.96 f 7.43 27.71 k 6.59 64.52 k 7.00 59.50 k 6.15 64.03 f 5.41 67.44 f 6.68 62.58 f 10.73 P c. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C6 (PAGE 1): HEMATOLOGY - SUMMARY - FO GENERATION FEMALE RATS (See abbreviations on the last page of this table.) RATS TESTED WBC (THSN/CU MM) RBC (MILL/CU MM) HGB (GRAMS/DL) HCT ( % ) MCV (CU MICRONS) MCH (PIC0 GRAMS) MCHC ( % ) PLAT (THSN/CU MM) MPV (CU MICRONS) N 10 MEAN f S.D. 9.0 f 3.09 MEAN f S.D. 6.51 f 0.461 MEAN f S.D. 15.5 f 0.95 MEAN f S.D. 40.5 k 2.93 MEAN f S.D. 62.2 f 1.19 MEAN f S.D. 23.9 f 0.55 MEAN f S.D. 38.4 f 0.76 MEAN f S.D. 1419 f 153.0 MEAN f S.D. 8.3 f 0.49 10 13.8 f 5.73* 6.22 f 0.423 15.0 f 0.77 39.3 f 2.72 63.3 f 2.01 24.2 f 0.76 38.3 f 0.94 1240 f 208.1 8.0 0.84 10 11.1 f 2.98 6.40 f 0.554 15.2 f 0.91 39.5 f 3.24 61.8 f 1.50 23.9 f 0.88 38.7 k 1.29 1274 f 226.3 8.3 0.82 10 10.8 f 3.14 6.39 f 0.532 15.2 f 0.90 40.1 f 3.02 62.9 f 2.04 23.8 f 1.25 37.8 f 1.22 1305 f 207.2 8.0 0.63 10 8.9 f 3.48 6.42 -+ 0.538 15.3 f 0.77 40.4 f 3.15 63.0 f 1.60 23.8 f 1.05 37.8 k 1.37 1355 f 176.8 7.7 0.74 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C6 (PAGE 2): HEMATOLOGY - SUMMARY - FO GENERATION FEMALE RATS (See abbreviations on the last page of this table.) RATS TESTED N 10 PT (SECONDS) MEAN f S.D. 13.2 f 0.41 APTT (SECONDS) NRBC COUNT MEAN f S.D. 21.7 f MEAN f S.D. 0 f 2.36 0 .o Lymphocytes (THSN/CU MM) MEAN f S.D. 6.6 f 2.44 Segmented (THSN/CU MM) MEAN f S.D. 2.2 f 0.90 Bands (THSN/CU MM) MEAN f S.D. 0 . 0 f 0 . 0 0 Monocytes (THSN/CU MM) MEAN f S.D. 0.1 f 0.15 Eosinophils (THSN/CU MM) MEAN f S.D. 0.1 f 0 -10 Basophils (THSN/CU MM) Abnormal L (THSN/CU MM) MEAN f S.D. 0 . 0 MEAN f S . D . 0 .o f 0.00 f 0.05 10 13.4 f 0.29 23.9 f 3.19 0 f 0 .o 9.8 f 3.44" 3.6 f 3.69 0.0 f 0.00 0.3 f 0.49 0.1 f 0.13 0.0 f 0.00 0.0 f 0 .oo 10 13.2 f 0.37 23.5 f 0 f 2.35 0 .o 8.4 f 2 -60 2.4 f 0.93 0.0 f 0.00 0.2 f 0.13 0.1 f 0.13 0.0 f 0.00 0.1 f 0 . 0 7 10 13.5 f 0.42 25.2 f 2.45 0 f 0.0 8.2 f 2.52 2.4 f 0.96 0 . 0 f 0.09 0.2 f 0.11 0.1 f 0.10 0.0 f 0.00 0 .o f 0.10 10 13.8 f 1.12 25.2 f 3.81 0 f 0.6 6.0 f 2.77 2 - 6 f 0.82 0.0 f 0- 2 -+ 0.00 0.14 0.1 f 0.15 0.0 f 0.00 0 . 0 f 0.05 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C6 (PAGE 3 ) : HEMATOLOGY - SUMMARY - FO GENERATION FEMALE RATS RBC HGB HCT MCV MCH MCHC PLAT MPV PT APTT NRBC Segmented Abnormal L Red Blood Cells (Erythrocytes) Hemoglobin Hematocrit (Packed Cell Volume) Mean Corpuscular Volume Mean Corpuscular Hemoglobin Mean Corpuscular Hemoglobin Concentration Platelet Count Mean Platelet Volume Prothrombin Time Activated Partial Thromboplastin Nucleated Red Blood Cell Count Segmented Neutrophils 0 Abnormal Lymphocytes h) ?? TP (G/DL) A (G/DL) GLU (MG/DL) CHOL (MG/DL) TBILI (MG/DL) BUN (MG/DL) CREAT (MG/DL) CK (U/L) MEAN f S.D. 6.1 f 0.52 5.5 MEAN f S.D. 4.1 f 0.42 4.0 MEAN f S . D . 152 f 19.8 162 MEAN f S.D. 77 f 27.8 72 MEAN f S.D. 0.2 f 0.07 0.2 MEAN f S.D. 28 f 4.4 24 MEAN f S.D. 0.4 f 0.05 0.4 MEAN f S.D. 202 f 54.1 250 0.50 0.34 20.0 14.0 0.03 4.7 0.05 85.6 5.9 f 0.41 4.1 f 0.24 155 f 18.7 71 f 20.0 0.2 f 0.05 28 k 6.7 0.3 f 0.05 192 f 21.7 10 5.8 4 0.32 4.0 f 0.29 146 f 16.2 62 f 14.2 0.1 f 0.05 25 f 4.4 0.4 +- 0.08 2 13 f 80.4 10 5.8 f 0.29 4.0 f 0.31 140 f 35.5 61 f 19.8 0.1 f 0.05 30 f 15.4 0.3 f 0.06 252 f 101.6 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C7 (PAGE 2): CLINICAL CHEMISTRY - SUMMARY - FO GENERATION FEMALE RATS (See abbreviations on the last page of this table.) RATS TESTED AST (U/L) ALK (U/L) CA (MG/DL) PHOS (MG/DL) TRI (MG/DL) NA (MMOL/L) K (MMOL/L) CL (MMOL/L) G (G/DL) N MEAN f S . D . MEAN f S.D. MEAN f S.D. MEAN f S.D. MEAN -I S.D. MEAN f S.D. MEAN f S.D. MEAN f S.D. MEAN f S . D . 10 142 f 22.8 145 f 88.7 10.6 k 0.72 8.8 f 1.33 54 f 19.9 143 f 6.3 5.8 f 0.67 96 f 3.4 1.9 f 0.22 10 135 f 21.6 159 f 81.7 11.2 f 0.63 7.9 f 1.94 50 f 10.2 143 f 2.4 5.6 f 0.58 94 f 2.7 1.6 -+ 0.26** I LO 137 5 11.8 136 f 107.3 10.4 f 0.87 9.0 f 2.80 49 f 29.4 142 f 2.5 5.6 f 0.32 95 f 3.8 1.8 f 0.21 10 124 f 12.1 178 f 131.0 10.4 f 0.46 8.7 f 2.88 45 -+ 19.5 142 f 2.6 6.1 f 0.70 95 f 3.4 1.7 f 0.13 10 156 f 40.7 128 f 44.7 10.4 -+ 0.82 10.1 f 1.64 44 -+ 18.3 142 f 2.6 6.2 f 0.90 94 f 3.1 1.8 f 0.21 b t3 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY' STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C7 (PAGE 3 ) : CLINICAL CHEMISTRY - SUMMARY - FO GENERATION FEMALE RATS A A1bumin GLU Glucose CHOL Cholesterol TBILI Total Bilirubin BUN Blood Urea Nitrogen CREAT Creatinine ALT Alanine Aminotransferase AST Aspartate Aminotransferase ALK Alkaline Phosphatase CA Calcium PHOS Phosphorus TRI Triglycerides * P NA Sodium F K Potassium 0 w 00 CL Chloride G Globulin PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C8 (PAGE 1): BODY WEIGHTS - PRECOHABITATION - SUMMARY - FO GENERATION FEMALE RATS BODY WEIGHT (G) DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 DAY 8 DAY 9 DAY 10 DAY 11 DAY 12 DAY 13 MEAN+S.D . . MEANfS D . MEANfS .D . MEANfS.D . MEANfS.D . MEANfS.D . MEANfS.D . MEANfS.D . MEANfS .D . MEANfS.D . MEANfS .D . MEANfS.D . MEANfS .D . 228.6 f 8.0 228.2 f 8.7 233.5 f 7.2 236.6 f 7.7 239.0 f 10.0 241.2 f 10.8 245.3 f 10.4 250.5 f 11.5 250.4 f 12.1 251.6 f 14.3 256.4 f 14.6 256.9 f 14.0 256.3 f 16.3 224.7 f 8.1 226.2 f 9.1 228.5 f 9.5 233.8 f 9.4 234.6 f 11.0 235.4 f 11.2 238.5 f 10.7 243.4 10.6 244.1 f 10.8 243.4 f 12.6 246.1 f 12.2 250.8 f 12.4 250.4 f 12.8 226.3 f 7.7 227.3 f 9.2 231.1 f 10.1 234.2 f 9.8 236.9 f 9.8 241.0 f 10.8 244.3 f 10.8 246.5 f 11.6 248.3 f 11.8 250.0 f 12.8 254.9 & 13.3 254.8 f 12.5 255.0 f 13.2 226.8 f 7.9 227.3 f 7.6 231.5 f 8.0 234.5 f 8.3 235.3 f 9.3 237.7 f 9.1 241.0 f 9.4 245.8 f 9.6 245.8 f 10.5 246.3 f 12.5 248.6 f 11.5 252.1 f 11.3 252.9 f 12.8 226.9 f 8.0 228.3 f 10.0 231.5 f 10.0 235.7 f 10.5 235.1 f 12.3 236.3 f 14.9 238.4 f 15.2 243.5 f 18.4 242.3 f 20.8 243.3 f 20.9 248.3 f 19.3 252.0 f 18.4 251.9 f 17.9 I I I I I I I I I I I I I I I I I I :Y151 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I > m I In IH Irl I I I I I I I I I I I I I I I I I I I I I I w I I I I X I I b I I I I I I I Hrl IH IH I Kl trl I I I v, I I 0 I I r I I r I I I I I E I I I I lit 0 I I 0 lk I I I I I I I I I I I I I I I I H W IKl IH *I d I 01 I I I I I I I I 1 I I I I I I I I I I I I I I I I 1 I I I I I I I I H O I Kl I Id I I I I I I I I I I I I I I I I I I I I I I I I I 5 I I W H cn -1 a3 *I rll I +I +I I I Kl CVI + -1 r *I CVI +I I I I I I I I * W W* II * u) a 3 1 I I +I +I +I I 0 CV C *V II cn r c o l + rl CVI + +I I I I I I r rl *I .I Kl co C O I I I +I +I +I I I 0 W u)l .I + + 0 rl rll CV rl w t +I I I I I I W C *V II co PI I I +I +I +I I r W I CnI .I a3 r LnI + rl CVI + +I I I I I I I * I W PI *I a3 r WI rll I +I +I +I I cn cn rl C+V r + n rl a3 CV 0 I a3 *rl *rd a3 rl I I rl a3 418-028:PAGE C-16 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C10 (PAGE 1): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS PREGNANT N MATERNAL BODY WEIGHT (G) DAY 0 MEANfS.D . DAY 1 DAY 2 MEANfS .D. MEANfS .D . DAY 3 MEANfS.D . DAY 4 MEANfS.D . DAY 5 DAY 6 DAY 7 DAY 8 DAY 9 MEANkS .D . MEANfS.D . MEANfS.D . MEANfS .D. MEANfS .D . 15 270.1 f 14.3 273.7 f 19.9 280.5 f 20.2 283.4 f 20.6 286.5 k 22.2 291.3 f 20.3 294.1 f 18.7 298.0 f 16.1 303.3 f 18.0 307.9 f 16.8 15 259.4 f 12.0 264.7 f 11.5 269.9 f 12.8 274.3 f 12.4 278.9 k 12.9 283.1 f 13.9 287.3 f 14.2 290.4 f 14.9 294.6 f 13.2 299.3 f 15.3 13 270.9 f 15.6 274.6 f 13.4 278.2 f 14.7 284.5 f 14.0 285.7 f 16.6 290.2 f 17.3 292.2 f 18.0 295.2 f 18.4 300.5 f 18.6 305.3 f 20.1 13 265.6 f 15.4 270.9 k 14.5 275.4 f 15.6 278.8 f 16.0 282.7 f 17.7 286.3 f 16.8 288.5 f 18.1 291.3 f 19.1 296.2 f 22.1 301.0 f 22.3 15 260.9 f 18.2 266.9 f 19.2 269.1 f 18.1 275.7 f 18.0 277.7 f 20.8 280.9 f 19.4 283.7 f 19.8 287.7 f 20.6 292.2 f 21.0 295.9 f 20.6 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C10 (PAGE 2): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - FO GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/ DAY) I I1 I11 IV V 0 0.3 1 3 10 PREGNANT N MATERNAL BODY WEIGHT (G) ' DAY 11 MEAN+S . D . DAY 12 MEANfS.D . DAY 13 MEANfS.D . DAY 14 MEANfS.D . DAY 15 MEANfS.D . DAY 16 MEANfS.D . DAY 17 MEANfS.D . DAY 18 MEANfS.D . DAY 19 MEANkS .D . 15 320.7 f 19.6 325.6 f 19.8 331.1 f 19.0 338.8 f 17.9 345.7 f 18.4 353.1 f 23.8 365.3 f 26.7 383.5 f 27.1 398.1 f 31.0 15 311.7 f 16.5 318.3 f 16.8 321.3 f 16.9 328.5 f 16.8 336.1 f 17.7 345.1 f 17.8 357.7 f 19.8 370.3 f 21.5 380.3 f 27.4 13 318.5 f 20.6 325.1 f 19.8 328.3 f 21.6 334.2 f 20.7 342.5 & 20.6 350.1 f 20.8 364.8 f 19.4 380.4 f 20.8 393.0 & 22.7 13 313.6 f 22.0 319.6 f 26.0 323.2 f 25.8 329.2 f 26.1 335.2 f 26.3 342.8 f 29.0 356.0 f 30.6 365.2 f 25.2 380.4 f 24.7 15 308.6 f 23.3 315.0 f 23.9 319.3 f 23.3 324.1 f 23.1 331.2 f 24.1 337.7 f 24.6 353.3 f 24.6 365.0 f 25.9 378.0 f 27.8 I I I I I I I I I I I I I I I I I I I30 I m m I rllrl rl I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I m I I I >m I m IH lrl rl I I I I I I I I I I I 1 I I I I I I I I I I I I I I I 1 I I I I I I I H r l I Ln m IH I d rl IH I I I I I I I I I El, 0 I I bl I I I I I I I bl I I 0 k I I I I I I I I I I I I I I I H m I v) In I I H * I d rl I 01 z I I 0 I I H I I E I I I I 2 I I vl I I w I I w I I I I I I I I I I I I I I I In rl a, P cv rl P cn * Ln m co *I In co c o l rll I +I +I +I +I +I +I +I I cvl * cn cn cv 02 rl .I * w cv w m *I 0 mI + + cv rl rl rl m m r l l + + + +I I I I I I I rl 03 m I cv c n l w m Ln -1 cn r l l NI +I +I +I I +I +I I 0 m In + + + In m Ln rl rl rl +w c *v lI mI m mI m rll +I I I I I I I m m co In 0 cv I P rll * m co . I I +I +I +I +I +I +I +I I * * m In (UI cn P .I rll w co m r P rl *I cv rl rl rl m m r l l + + + + + + +I I I I I I I ** I m cn * . II P m m r PI (UI +I +I +I +I +I I +I I 0 co rl P N + * *+ + + rl m P m rl rl rl m 01 .I WI mI rll +I I I I I I I P cn m w * w +I +I +I +I cn rl In rl I 01 .I cnl Pal I +I I cn~ .I + + + P 10 rl 0 cv rl rl cv rll *I rll +I I I I I I 9 9 n 8 8 vl vl w E x .I 8 8 : 9 9 1 vl r n l w x co cv 0 I m rl v w P 0 cv Ln co 0 rl rl rl rl cv I I I I I I 0 r 0 c\1 In 03 rl rl rl rl 418-028:PAGE C-19 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C12 (PAGE 1): MATERNAL BODY WEIGHTS - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS PREGNANT N 15 15 13 13 15 DELIVERED A LITTER N 15 15 13 13 15 MATERNAL BODY WEIGHT (G) DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 MEANfS.D . MEANfS.D . MEANfS .D . MEANfS.D . MEANfiS.D . MEANfS.D . MEANfiS.D . DAY 8 DAY 9 MEANfS.D . MEANfS.D. DAY 10 MEANfiS.D . DAY 11 MEANfS.D . 313.1 f 22.6 313.1 f 24.6 316.1 f 22.6 321.5 f 22.7 326.0 fi 20.5 332.7 f 26.1 340.4 f 20.8 342.3 f 19.8 349.9 f 25.6 14la 353.6 fi 17.9 [ 13la 354.4 f 18.5 [ 14la 302.2 f 16.8 300.1 f 17.2 300.2 f 18.6 13la 303.4 f 17.2* [ 13la 307.3 f 14.2 312.9 f 11.9* 321.4 f 12.6* 320.7 f 12.6* 331.7 f 11.9 14la 334.9 f 17.5 331.9 f 25.7* 309.5 f 20.6 305.4 f 18.9 310.4 f 18.1 [ llla 314.2 f 19.8 [ 12la 314.9 f 21.1 [ 12la 323.9 f 18.2 [ 12la 331.2 f 18.3 333.6 f 19.6 12la 337.4 fi 17.3 345.5 fi 18.2 349.2 f 20.3 C 12la 304.7 f 23.2 304.2 f 22.2 305.0 f 23.1 [ 12la 310.2 f 22.5 12la 311.5 f 23.0 [ 121a 316.9 f 16.1 [ llla 317.7 f 21.6** 12la 322.2 f 21.5* 331.8 f 21.2 333.5 f 23.0 E 12la 336.4 f 24.6 [ 12la 296.3 f 19.9 298.9 f 19.8 300.3 & 20.0 [ 14la 299.3 f 19.7" [ 13la 308.4 f 15.7 [ 13la 306.0 f 26.9** 315.2 f 25.4** 318.1 f 26.0** 324.7 f 29.7** 327.4 f 38.3 330.2 f 30.8" DAY = DAY OF LACTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes values that appeared incorrectly recorded, as well as those associated with interrupted water access. * Significantly different from the Group I value ( ~ 5 0 . 0 5 ) . * * Significantly different from the Group I value ( ~ ~ 0 . 0 1 ) . PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C12 (PAGE 2): MATERNAL BODY WEIGHTS - LACTATION - SUMMARY - FO GENERATION FEMALE RATS PREGNANT N 15 15 13 13 15 DELIVERED A LITTER N 15 15 13 13 15 MATERNAL BODY WEIGHT (G) DAY 12 DAY 13 DAY 14 DAY 15 DAY 16 DAY 17 DAY 18 MEANfS .D . MEANfS .D . MEANfS .D . MEANfS .D . MEANfS .D . MEANfS .D. MEANfS .D . 355.7 f 20.2 364.1 18.5 14la 359.8 f 21.5 355.3 f 31.2 360.7 f 22.0 351.7 f 21.4 360.3 f 21.5 344.2 f 13.6 [ 14la 342.9 f 17.2* I: 14la 342.7 f 14.5 1 14la 342.4 f 10.4 [ 14la 345.7 f 12.9 341.1 f 16.9 346.4 f 15.6 352.6 f 20.1 12la 353.5 f 20.2 [ 121a 353.1 f 17.3 343.2 & 17.2 C 121a 353.2 f 18.0 346.5 f 17.8 E 12la 352.2 f 21.5 347.5 f 24.4 [ 12la 352.0 f 21.8 12la 345.1 f 19.5 348.7 f 21.4 [ 12la 348.4 f 21.3 339.3 f 26.4 352.2 f 21.8 334.2 f 32.5 [ 14la 340.1 f 30.8* 335.2 f 32.2** [ 13la 333.4 f 33.6 14la 337.8 f 34.3 14la 333.0 f 33.4 340.6 f 34.3 DAY 19 DAY 20 MEANfS.D . MEANfS .D . 360.1 f 21.4 359.6 f 19.0 347.9 f 15.1 344.6 f 16.1 355.0 f 19.6 348.8 f 20.6 357.5 f 25.1 352.9 f 24.0 339.5 f 32.3 14la 338.9 f 34.9 DAY 21 MEANfS .D . 350.1 f 26.9 335.7 f 18.7 343.4 f 19.8 344.4 f 25.8 334.4 f 35.9 DAY 22 MEANfS .D . [ 141a 306.3 f 26.0 14la 324.3 f 27.1 311.2 f 35.0 319.6 f 40.4 303.5 f 37.9 ________________________________________------------------------------------------------------------------------------------------ DAY = DAY OF LACTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes values that appeared incorrectly recorded, as well as those associated with interrupted water access. * Significantly different from the Group I value (~50.05). * * Significantly different from the Group I value (p~O.01). PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C13 (PAGE 1): MATERNAL BODY WEIGHT CHANGES - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS PREGNANT N 15 15 13 13 15 DELIVERED A LITTER N 15 15 13 13 15 MATERNAL BODY WEIGHT CHANGE (G) DAYS 1 - 5 DAYS 5 - 8 DAYS 8 - 11 DAYS 11 - 15 DAYS 15 - 18 DAYS 18 - 22 MEANfS .D . MEANfS .D . MEANfS .D . MEANfS .D . MEANfS .D . MEANfS .D. +12.9 f 10.2 +16.3 f 9.6 +13.9 f 7.1 [ 14la -0.7 f 22.4 141a +4.9 f 21.8 -54.0 f 25.3 +5.1 f 10.7 +13.5 f 7.4 +11.1 f 19.2 +12.8 19.9 14la +2.2 f 11.9 [ 14la -22.1 f 26.7 +7.2 f 12.7 12la +17.3 f 7.6 [ Ills +14.4 f 5.6 C 11la -3.7 f 13.2 [ llla +7.9 f 18.0 [ 12la -41.1 f 33.6 +7.7 f 14.8 12la +11.0 f 11.0 [ 12la +13.6 f 10.6 [ 12la +13.8 f 9.7 111a +4.1 f 6.3 [ 12la -32.5 f 28.2 +7.8 f 15.1 [ 13la +16.5 f 10.1 E 13la +12.1 f 10.8 +3.7 f 11.2 14la +7.4 f 11.4 [ 14la -37.1 & 29.8 DAYS 1 - 22 MEANfS .D . -6.9 f 28.4 +22.1 f 26.0 +1.7 f 28.0 +14.9 f 31.8 +7.3 f 29.3 .................................................................................................................................... DAYS = DAYS OF LACTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes values that appeared incorrectly recorded, as well as those associated with interrupted water access. ? w w I I I I I I I 0 I I I I I I I I m I I I I I I I 13Z2 x I I m 0 I I E I I I I I I 3 I I I I w I I I I I I w4 I I I I lk I I I I I +I I 1 CD I I ln I rl I I I I I I I I co I I rl I I I I 1 I I >m IH I I I I I I I UI Irl I I I I I I +I I I m I I ln rl I I I I 0 I I lk 1 I I I I I I I I 0 w I I I I X I I E I I I I (Y I I I H r l I UI IH Irl IH I +I a, r I tn I I rl I I I I I I I I I I 4 0 rl cv I -rl I I I E I I I 1 h I I 0 I I I I I I I I I I I I I H C ) I UI IH - I d I I 01 I I I I I I I 1 2 I I \ I I I I u v I I I I I I I I I I I I I I I a CD rl U u-rl +I I In rn I rn ln rl I a, I 5 I m co rl I I I I I HO I I I I I I I I Irl I I I I I I +I * I 0 (Y I a I I I I I I I I I I n I I w I I w I I h I I I I I U I k I I I I IZ I I I I I I I I I I 22 I I I I a I I I I I I I I I -I I ; 8* I : co E I d 8m m 2 w U II w2 n w 3 E w h 418-028:PAGE C-23 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C15 (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PRECOHABITATION - SUMMARY - Fo GENERATION FEMALE RATS FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8 MEAN&S .D . 86.1 f 5.5 83.4 k 5.6 87.9 k 7.0 83.0 & 5.2 83.3 f 10.0 [ 13la ________________________________________-------------------------------------------------------------------------------------------- DAYS = DAYS OF STUDY [ ] = NUMBER OF VALUES AVERAGED a. Excludes values that were not recorded, as well as those associated with spillage. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C16 (PAGE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS PREGNANT N 15 15 13 13 15 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 - 7 MEANfS .D . 25.4 f 3.3 25.0 f 2.5 24.3 ?r 2.7 25.2 f 3.6 24.0 f 2.7 DAYS 7 - 10 MEANfS .D . 27.7 f 2.8 26.8 f 2.8 26.3 f 3.4 26.3 f 5.0 25.8 f 2.5 DAYS 10 - 12 DAYS 12 - 15 MEANfS .D . MEANfS .D . 32.0 f 10.5 [ 14la 28.4 f 2.8 27.0 f 27.5 f 3.2 2.4 28.0 f 27.5 & 3.2 2.1 26.6 & 26.7 f 3.9 3.9 27.0 f 27.5 f 2.8 2.6 DAYS 15 - 18 MEANfS.D . 25.4 f 4.0 26.7 & 3.1 25.9 & 2.1 25.3 f 3.6 26.0 f 3.3 DAYS 18 - 20 MEANfS .D . 26.6 f 5.7 22.8 f 6.2 25.8 f 3.2 25.0 f 3.7 26.1 f 4.8 DAYS 0 - 20 MEANfS .D . 26.9 f 3.2 25.9 f 2.4 25.8 f 2.0 25.7 f 3.0 25.6 f 2.3 .................................................................................................................................... DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes values that were associated with spillage. I I I I I I I I I I * I m I E 3 ; Y ; : xU H I I 0 I E I I I I I I Ln rl I I I I I I I I I I I I I I I I I I I I I I I I I I m I I I >m I m IH Id rl I I 0 I I Fr, I I I I I I I I I w I I E I I I I I H r l I Kl m IH Irl rl IH I I I w I I I I I I m rl I I I I I I I I I I I I I I I I I I I I I I m I I I H O I Ln I Id rl I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I z 1 I I 1 I I I I I I I I I I I I I I I I I h I I rl I -I ; 8 ; I *I co cv 0 I \I I a2 rl dl P rl U 418-028:PAGE C-26 I ...... I m~rmcvm~oi .I mm*PmoTrr rl 1 I I +I +I +I +I +I +I +I I ...... I WwmwrlPmI .I wPPLnma3NI c o c o c o c o P w c o 1I I I I I I ...... I mPomdlcoTrl .I mcvrw~rlmi rl rl I I +I +I +I +I +I I +I +I I . . . . . . c o P c o m mP*rlc cocococ I cvrlwl .I vwrll oPwcoI I I I I I I ...... I cn~rrlmcumrli .I WPwdlwPml I I I +I +I +I +I +I +I +I I ...... I w ~ m r m ~ m i .I mwPcvcvm01 cocoocoPwcoI I I I I I I . . . . . . .I rloOcom*wl I PPcomWmmI rl I I I . . . . . . . +I +I +I +I +I +I +I I Trom*WmwI I OOWTFmmcvl mma3coPmcoI I I * Tr Tr P m m I l a, WI bl . . -1 co P C O P co cv W I rl cv rl I rl I -rl I T 4 +I +I +I +I +I +I +I I rl * co m o m 0 . - . co co 0 m mm cd ol 0 P W W B I I I I U .. 9 9 99 P 0 cvm rl r l r l I I I I 0P o m rlrl PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C18 (PAGE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) RATS TESTED N PREGNANT N DELIVERED A LITTER N MATERNAL FEED CONSUMPTION (G/DAY) DAYS 1 - 5 DAYS 5 - 8 DAYS 8 - 15 MEANfS .D. MEANfS.D . MEAN+S .D . I 0 15 15 15 38.5 & 7.1 61.2 f 6.4 71.6 f 8.6 I1 I11 0.3 1 15 15 15 13 15 13 27.6 f 8.7** [ 14la 53.7 f 6.6 68.3 f 8.3* 33.2 f 4.0 [ 12la 55.9 & 4.4 69.2 & 4.0 IV V 3 10 15 15 13 15 13 15 31.4 & 5.5" 49.4 f 10.2 69.6 f 9.5** 34.4 f 7.9 [ 14la 52.3 f 9.0 68.2 & 9.4** PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE ci9 (PAGE 1): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - LACTATION - SUMMARY - FO GENERATION FEMALE RATS PREGNANT N 15 15 13 13 15 DELIVERED A LITTER N 15 15 13 13 15 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 5 MEANkS.D . 121.1 f 21.3 91.8 f 27.8** 107.6 f 15.0 102.7 f 21.2 118.7 f 22.1 DAYS 5 - 8 DAYS 8 - 11 DAYS 11 - 15 DAYS 1 - 15 MEANfS.D . MEANfS .D . MEANfS.D . MEANkS .D . 183.0 f 20.6 176.1 _+ 134.6 [ 14la 222.6 f 97.5 [ 14la 176.7 f 19.5 [ 14la 170.4 f 23.5 176.1 f 109.9 225.3 f 64.3 [ 14la 166.9 f 22.6 [ 121a 170.6 f 12.2 [ Ills 101.9 k 97.9 [ Ills 276.0 k 76.3 E Ills 172.4 f 11.5 [ 12la 155.2 f 32.4** [ 12la 155.4 f 134.0 [ 12la 245.4 f 89.7 Ills 166.6 f 20.2 12la 168.2 f 20.0 [ 12la 138.4 f 127.0 248.7 f 81.6 14la 175.1 k 14.9 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1.4. l. a. . . . . . . . . . . . . . .[ . 1. 2. l. a. . . . . . . . . . . . . . .[. .1.2.l.a. . . . . . . . . . . . . . [. .1.4.l.a. . . . . DAYS = DAYS OF LACTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes values that appeared incorrectly recorded, as well as those associated with interrupted water access or spillage. * * Significantly different from the Group I value ( ~ ~ 0 . 0 1 ) . I I I I 1 I I I I I N I I I I rl I I E I I I Y ; xU Ln +I H rl rl 0 I I E N CO 0 +I 0-l 01 I cv c4l I I I I I I I I w X E I I I E X I I 1 Hrl I IH I I H I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 HC) I - I H I I 0 I I I 2 I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I HO I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I h I i 2 rl I - CO N 0 I \ I CO rl v Tr 0 N U u) 0 Ln +I rl m m 0-l 0 Ln +I rl CO N 0-l 0 Ln +I rl N z I 1 I I I I I I I I I rl 0 1 I I I I I I I I I I I I I I I I I 0 rll I I I I I I I I I I I I I I I I I I 0 0 I I I I I I I I I I I I I I I I I I I 0 0 I I I I I I I I I I I I I I I I I I I rl 0 I I I I I 1 I I I 1 I I z I 2 1 I I I I I I I I I I I I I I ZII p: c n I O u)!25I ZHFil I Q8 Z E g cnwcn ;I I I I 418-028:PAGE C-29 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C20 (PAGE 2): MATING AND FERTILITY, ESTROUS CYCLING AND DAYS IN COHABITATION - SUMMARY - Fo GENERATION FEMALE RATS DAYS IN COHABITATION a MEANfS.D. 3.7 & 3.0 1.9 k 1.2 2.8 f 1.2 4.0 & 4.0 2.5 f 1.4 RATS THAT MATED N(%) 15 (100.0) 15(100.0) 15 (100.0) 14( 93.3) 15 (100.0) FERTILITY INDEX b N/N 15/ 15 (%I (100.0) 15/ 15 (100.0) 131 15 ( 86.7) 13/ 14 ( 92.8) 151 15 (100.0) RATS WITH CONFIRMED MATING DATES N 15 15 15 14 15 MATED BY FIRST MALE c DAYS 1-7 N(%) 14( 93.3) 15(100.0) 15 (100.0) 13( 92.8) 15 (100.0) MATED BY SECOND MALE c DAYS 8-14 N(%) 1( 6.7) O ( 0.0) O ( 0.0) 1( 7.2) O ( 0.0) RATS PREGNANT/MTS IN COHABITATION N/N 15/ 15 151 15 13/ 15 13/ 15 15/ 15 (%I (100.0) (100.0) ( 86.7) ( 86.7) (100.0) .................................................................................................................................... a. Restricted to rats with a confirmed mating date and rats that did not mate. b. Number of pregnancies/number of rats that mated. c. Restricted to rats with a confirmed mating date. PROTOCOL 418-028; ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C21 (PAGE 1): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - FO GENERATION FEMALE RATS HOME CAGE BEHAVIOR 1: Sleeping 2: Awake, Immobile 3: Normal movement 4: Unusual posture 5: Unusual behavior ALTERATIONS (HOME CAGE) 1: None N 10 10 10 10 10 2: Stereotyped behavior N 0 0 0 0 0 3: Bizarre behavior N 0 0 0 0 0 4: Limb twitches/tremor N 0 0 0 0 0 5: Whole body tremor/spasm N 0 0 0 0 0 6: Unusual posture N 0 0 0 0 0 7: Tonic-clonic seizure N 0 0 0 0 0 REACTION TO REMOVAL (1) Sits quietly N 10 10 10 10 10 (2) Vocalization N 0 0 0 0 0 ( 3 ) Runs or freezes N 0 0 0 0 0 (4) Tail or throat rattles N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 REACTION TO HANDLING (1) No resistance N 10 10 10 10 10 ( 2 ) Vocalization N 0 0 0 0 0 ( 3 ) Tense N 0 0 0 0 0 (41 Squirming N 0 0 0 0 0 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN FATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C21 (PAGE 2): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - Fo GENERATION FEMALE RATS REARS IN OPEN FIELD DEFECATION IN OPEN FIELD 1: None 2: Feces normal 3: Soft or liquid feces MEAN?S.D. 12.0 k 3.6 N 8 N 2 N 0 13.0 & 2.7 10 0 0 11.6 k 3.5 10.7 & 2.3. 10.9 & 2.6 URINATION IN OPEN FIELD (1) None N 6 7 7 10 8 (2) Normal urination N 4 3 3 0 2 (3) Excess urination N 0 0 0 0 0 MEAN SCORE 1.4 1.3 1.3 1.0 1.2 LEVEL OF AROUSAL ( 1) Stuporous N 0 0 0 0 0 (2) Sluggish N 0 0 0 0 0 (31 Apparently normal N 10 10 10 10 10 (4) Sudden darting N 0 0 0 0 0 ( 5 ) Freezing, vocalization N 0 0 0 0 0 MEAN SCORE 3.0 3.0 3.0 3.0 3.0 ALTERATIONS (OPEN FIELD) * P 1: None N 10 10 10 10 10 2: Stereotyped behavior N 0 0 0 0 0 3: Bizarre behavior N 0 0 0 0 0 4: Limb twitches/tremor N 0 0 0 0 0 5: Whole body tremor/spasm N 0 0 0 0 0 6: Unusual posture N 0 0 0 0 0 (n) = Score assigned to graded test items; mean score was calculated by multiplying each score by the number of rats with that score and then dividing the sum of the products by the total number of rats PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C21 (PAGE 3): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - Fo GENERATION FEMALE RATS GAIT PATTERN 1: Apparently normal 2: Ataxic 3: Limbs splay or drag 4: Spastic, tip-toe 5: Duck-walk 6: Scissors gait 10 10 10 10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GAIT ABNORMALITY, SEVERITY (1) Normal gait N 10 10 10 10 8 (2) Slight N 0 0 0 0 2 ( 3 ) Moderate N 0 0 0 0 0 (4) Extreme N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.2 PALPEBRAL CLOSURE (1) Wide open N 10 10 10 10 10 (2) Slightly drooping N 0 0 0 0 0 ( 3 ) Half-closed N 0 0 0 0 0 (4) Completely shut N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 * P b PROMINENCE OF THE EYE 00 1: Normal N 10 10 10 10 10 h) 2: Exophthalmos N 0 0 0 0 0 ? w w PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C21 (PAGE 4): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - Fo GENERATION FEMALE RATS LACRIMATION (1) No excess N 10 10 10 10 10 (2) Excess at eyelid margin N 0 0 0 0 0 ( 3 ) Margin persistently damp N 0 0 0 0 0 ( 4 ) Extends beyond margin N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 SALIVATION (1) No excess N 10 10 10 10 10 (2) Margin of mouth wet N 0 0 0 0 0 ( 3 ) 1/4 to 1/2 submandibular N 0 0 0 0 0 ( 4 ) Entire submandibular N 0 0 0 0 0 MEAN SCORE 1.0 1.0 1.0 1.0 1.0 PILOERECTION N 0 0 0 0 0 ABNORMAL RESPIRATION N 0 0 0 0 1 APPEARANCE (1) Clean and groomed N 10 10 9 9 10 ( 2 ) Unkempt N 0 0 1 0 0 (3) Urine and/or fecal stain N 0 0 0 I 0 MEAN SCORE 1.0 1.0 1.1 1.2 1.0 VISUAL REACTION (1) None N 0 0 0 0 0 (2) Orienting N 10 10 10 10 10 ( 3 ) Startle N 0 0 0 0 0 (4) More energetic reaction N 0 0 0 0 0 ( 5 ) Attacks N 0 0 0 0 0 PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C 2 1 (PAGE 5): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - Fo GENERATION FEMALE RATS TACTILE REACTION (1) None N 0 0 0 0 0 ( 2 ) Orienting N 10 10 10 10 10 ( 3 ) Startle N 0 0 0 0 0 ( 4 ) More energetic reaction N 0 0 0 0 0 (5) Attacks N 0 0 0 0 0 MEAN SCORE 2.0 2 .o 2.0 2.0 2.0 AUDITORY REACTION (1) None N 0 0 0 0 0 ( 2 Orienting N 0 0 0 1 1 ( 3 ) Startle N 10 10 10 9 9 ( 4 ) More energetic reaction N 0 0 0 0 0 (5) Intense vocalization N 0 0 0 0 0 MEAN SCORE 3.0 3.0 3.0 2.9 2.9 TAIL-PINCH REACTION (1) None N 0 ( 2 ) Orienting N 8 ( 3 ) Startle N 2 ( 4 ) More energetic reaction N 0 ( 5 ) Attacks N 0 0 10 0 0 0 P, 00 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C 2 1 (PAGE 6): FUNCTIONAL OBSERVATIONAL BATTERY - SUMMARY - Fo GENERATION FEMALE RATS AIR RIGHTING RESPONSE (1) All feet land on ground N 10 (2) Lands on side N 0 (3) Lands on back N 0 MEAN SCORE 1.0 PUPIL RESPONSE TO LIGHT N 10 FORELIMB GRIP TEST Maximum (G) Average (G) MEAN+S.D. MEANfS.D. 509.5 f 161.7 455.9 f 136.7 HINDLIMB GRIP TEST Maximum (G) Average (G) MEANkS.D. MEAN+S.D. 462.0 f 111.3 423.0 f 113.7 LANDING FOOT SPLAY Average (CM) MEANfS.D . 7.7 j, 1.8 10 10 0 0 0 0 1.0 1.0 10 10 396.5 rt 125.7 319.8 f 100.6 492.5 f 257.7 423.9 f 207.3 412.5 f 104.7 366.5 f 77.3 455.5 f 87.7 418.3 79.4 6.7 f 1.0 7.7 f 1.5 10 0 0 1.0 10 493.0 f 112.3 452.5 f 103.4 397.0 f 67.5 362.8 j, 79.8 7.1 f 1.4 10 0 0 1.0 10 518.5 f 161.9 460.3 f 166.4 485.5 427.3 f 71.5 j, 52.2 8.1 f 1.7 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) NUMBER OF FEMALE RATS NUMBER OF MOVEMENTS BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL N MEAN f MEAN f * MEAN f MEAN MEAN f MEAN k MEAN f MEAN f MEAN f. MEAN f MEAN f MEAN MEAN k MEAN & * MEAN f. MEAN MEAN f * MEAN f MEAN 10 72.0 k 6.9 53.8 f 25.7 50.4 f 26.5 28.6 f 34.7 27.5 f 24.0 23.9 f 26.5 11.6 f 16.0 19.3 f 18.1 21.4 f 21.0 19.6 f 22.1 9.2 f 10.6 18.1 f 25.5 12.1 f 18.3 16.0 f 19.0 22.2 f 30.6 23.2 f 27.2 32.2 f 33.8 25.9 f 28.5 487.0 f 244.9 10 10 10 10 67.1 f 7.6 * 53.6 f 20.8 41.2 34.2 30.7 -+ 25.5 21.2 f 23.6 19.4 f 20.7 17.8 f 23.2 13.2 f 18.6 10.5 f 24.3 6.5 f 7.2 7.0 f 16.6 5.9 f 5.3 15.0 f 22.1 19.0 f 19.6 13.0 f 15.7 15.0 f 16.7 11.7 f 18.6 * 10.8 f 14.7 378.6 202.4 * 68.3 f 6.8 66.6 12.4 50.2 f 17.0 43.9 -+ 26.8 43.6 f 26.4 42.6 f 29.5 44.5 f 21.3 38.0 f 20.5 28.1 f 25.4 29.4 f 29.3 26.9 f 28.2 21.1 f 26.2 24.3 f 26.3 24.4 f. 30.8 * 23.6 25.9 * 24.5 28.2 24.7 f 27.9 17.9 f 18.9 642.6 f 295.6 65.7 f 9.7 57.1 f 18.2 49.3 f 21.4 39.0 f 25.1 * 33.9 f 24.2 35.0 26.8 38.3 k 29.4 44.5 18.0 34.0 f 19.0 25.9 f 23.5 23.5 f 22.7 21.8 f 20.4 19.8 f 25.0 * 24.4 -+ 26.2 19.8 24.9 27.2 f 29.0 21.9 f 23.2 19.0 f 25.3 600.1 f 292.0 68.4 42.5 31.1 20.5 18.6 19.7 14.0 32.6 27.9 15.5 19.7 29.7 22.1 16.8 15.1 19.2 18.3 22.6 454.3 f 9.8 f 19.9 f 24.6 f 22.9 f. 22.4 -i- 18.0 f 17.3 & 26.2 f 24.6 f 16.5 f 22.6 -+ 21.4 f 25.5 f 18.3 f 20.2 *f 18.9 25.6 -+ 20.9 f 261.4 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C22 (PAGE 2): MOTOR ACTIVITY - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/ KG/ DAY) NUMBER OF FEMALE RATS N TIME (SECONDS) SPENT IN MOVEMENT BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL MEAN f MEAN f MEAN f MEAN f MEAN f MEAN f MEAN i MEAN f MEAN f MEAN f MEAN f MEAN f MEAN f MEAN f MEAN f MEAN f MEAN f MEAN f MEAN f I 0 10 154.8 f 95.3 f 65.7 f 30.6 f 33.6 f 27.5 f 11.7 f 25.4 f 25.5 f 25.2 f 11.8 f 22.9 f 14. O f 14.8 f 30.1 f 27.0 f 40.7 & * 38.2 f 694.8 27.4 50.4 36.2 41.4 36.5 37.9 18.4 29.0 28.6 30.0 14.7 35.7 23.3 20.2 46.0 33.9 46.3 45.8 331.8 I1 I11 0.3 1 10 10 IV V 3 10 10 10 141.5 f 46.3 94. O f 54.4 154.4 f 32.2 114.4 -+ 24.2 60.1 f 48.0 48.0 f 48.9 27.5 f 30.6 88.5 f 70.2 -+ 63- 4 --+ 56.9 52.2 42.7 31.0 f 41.8 56.1 f 41.1 21.4 f 31.9 69.0 k 46.5 15.5 f 26.5 60.7 f 36.2 13.8 k 37.1 37.9 f 41.2 6.2 f 9.6 38.7 f 44.1 8.3 f 21.8 39.7 f 44.2 4.2 f 3.7 34.4 f 48.6 14.7 f 21.0 38.4 f 52.5 24.1 f 27.9 34.5 f 48.1 15.5 f 23.7 36.4 f 44.5 17.6 f 24.1 36.1 f 42.1 13.7 f 27.6 37.8 f 48.8 9.6 f 14.8 18.1 f 23.2 566.7 f 350.6 1028.7 f 477.6 181.0 & 36.4 119.5 f 56.0 94.4 f 55.9 73.0 f 61.1 52.0 f 47.4 * 56.7 f 47.1 65.3 59.7 70.1 k 26.6 66.6 k 58.8 41.6 f 49.9 37.2 f 46.8 26.6 f 25.8 33.4 f 49.6 33.1 f 42.5 33.0 f 54.3 38.6 f 40.7 34.9 f 46.8 29.3 f 48.7 1086.3 f 711.3 146.2 f 26.1 96.6 f 63.6 52.3 f 51.5 34.6 f 45.9 27.6 f 35.5 31.0 f 30.6 16.5 f 29.0 41.7 f 38.3 40.1 f 38.6 14.1 f 16.3 24.6 f 33 .O 38.7 f 34.5 32.7 f 43.3 25.6 f 30.6 17.3 f 25.7 18.5 f 23.1 21.7 f 33.6 27.6 f 30.1 707.4 f 442.6 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C23 (PAGE 1): NATURAL DELIVERY OBSERVATIONS - SUMMARY - FO GENERATION FEMALE RATS PREGNANT N(%) DELIVERED LITTERS N(%) DURATION OF GESTATION a MEAN&S.D. IMPLANTATION SITES PER DELIVERED LITTER N MEAN+S.D. DAMS WITH STILLBORN PUPS N(%) DAMS WITH NO LIVEBORN PUPS N GESTATION INDEX b % N/N DAMS WITH ALL PUPS DYING DAYS 1-4 POSTPARTUM N(%) 15(100.0) 15 (100.0) 22.5 f 0.5 251 16.7 & 1.3 O ( 0.0) O ( 0.0) 100.0 15/ 15 O ( 0.0) 15(100.0) 15 (100.0) 22.7 -L 0.4 230 15.3 f 2.9 O ( 0.0) O ( 0.0) 100.0 15/ 15 O ( 0.0) 13( 88.7) 13 (100.0) 22.7 f 0.5 206 15.8 & 1.3 1( 7.7) O ( 0.0) 100.0 13/ 13 O ( 0.0) 13 ( 88.7) 13 (100.0) 22.8 f 0.4 192 14.8 k 2.8 3( 23.1) O ( 0.0) 100.0 13/ 13 O ( 0.0) 15(100.0) 15(100.0) 22.6 f 0.5 231 15.4 f 2.3 2( 13.3) O ( 0.0) 100.0 15/ 15 O ( 0.0) I I I I I I I I I I I I I I I I I I I30 I I rll I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I >m I IH I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I t Hrl I IH I IH I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I Hm I IH - 1 I 0 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I HO I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I -I I $; I I In 9 9 m m 0 h h rl m m 0 +I rl hl mr +I m m Ln +I 0 rl . - . w * *a, o m rl r l r l m rl a * m m 0 h h co W . -m m I+ +I +I r +I rl rl a) rlm W m .w m m r om rl rlr rl - 9 W m rl rl 0 h m In . - 4 - m rl +I rl 0 mm +I m m +I 0 rl m m o orl rl r c o m m 01 0 m m 0 h h 0 0 m 9 +I rl rl 1 e cum +I 0 0 +I 0 0 .w * * * rl rlrl 0 0 m r r 0 rl rl 0 h h 0 0 m Ln +I rl m mr +I 0 r e0 +I 0 0 .w Ln InLn 00 rl r l m m 2 $2 9 9 9 z m m - mi?- : m EE E h 2 0 H Y n w 3z 2 3E 2 0a a GI Iwn4 w?I HI4 m I4 PI . . . . h h h h *ern0 rlrloo wvvv cornhlrl rlrldrl mmmm \\\\ mmrco . . . . h h h h rlrlW0 rlrloo \\\\ mmrlo . . . . o m m o - A h - orloo vvvw OOPW oomm mmrlrl \\\\ omrlo . . . . - A h - mcooo omrlo WVVY 9mw* rlrloo mmcvcv \\\\ mWm0 . . . . h h h h *WOO om00 VVYY In*coco mmmm cvmmm \\\\ rlW00 zzzz \ z \ z \ z \ z 418-028:PAGE C-40 I W 1 wo .. I N I I588 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C24 (PAGE 2): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - F1 GENERATION LITTERS SURVIVING PUPS/LITTER a DAY 1 DAY 8 DAY 15 DAY 22 MEANfS .D. MEANfS .D . MEANfS .D . MEANfS .D . 15.7 f 1.7 15.2 f 1.8 15.2 f 1.8 15.2 f 1.8 14.3 f 3.2 13.7 f 2.9 13.6 f 2.8 13.6 f 2.8 15.4 f 1.6 15.2 f 1.5 15.1 f 1.3 15.1 f 1.3 13.6 f 3.2 13.3 f 3.3 13.2 f 3.2 13.2 f 3.2 14.5 f 2.4 14.1 f 2.1 14.1 f 2.2 14.1 2.2 PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY 1 MEANfS .D . 45.2 f 12.4 51.9 f 13.1 51.6 f 6.8 43.6 f 13.4 51.4 f 13.4 DAY 8 DAY 15 MEANfS . D. MEANkS .D . 45.2 f 13.0 45.2 f 13.0 52.7 f 12.0 53.1 f 11.8 51.8 f 6.3 52.0 f 6.2 42.9 f 13.1 43.1 f 13.0 51.4 f 13.9 51.8 f 14.2 DAY 22 MEANfS.D . 45.2 f 13.0 53.1 k 11.8 52.0 f 6.2 43.1 f 13.0 51.8 f 14.2 .................................................................................................................................... DAY = DAY POSTPARTUM a. Average number of live pups per litter, including litters with no surviving pups. b. Includes pups born alive, found dead day 1 postpartum. P PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C24 (PAGE 3): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - F1 GENERATION LITTERS LIVE LITTER SIZE AT WEIGHING DAY 1 DAY 8 MEANkS .D . MEANfS.D . 15.6 f 1.8 15.2 f 1.8 14.1 f 3.1 13.7 f 2.9 15.4 f 1.6 15.2 f 1.5 13.5 f 3.3 13.3 f 3.3 14.3 k 2.2 14.1 f 2.1 DAY 15 DAY 22 MEANfS .D. MEANkS .D . 15.2 f 1.8 15.2 k 1.8 13.6 f 2.8 13.6 k 2.8 15.1 k 1.3 15.1 f 1.3 13.2 f 3.2 13.2 k 3.2 14.1 f 2.2 14.1 f 2.2 PUP WEIGHT/LITTER (GRAMS) DAY 1 MEANkS .D . 6.3 k 0.3 6.3 f 0.5 6.4 f 0.3 6.5 k 0.7 6.3 f 0.5 DAY 8 MEANfS .D . 14.3 k 1.6 13.3 f 1.7 13.8 f 1.0 14.1 f 3.1 13.1 k 1.5 DAY 15 MEANfS.D . 26.2 f 2.8 26.3 f 3.9 25.9 k 2.6 27.4 k 5.7 25.1 k 3.3 DAY 22 MEANfS.D . 37.4 f 4.7 39.3 f 6.9 38.0 k 5.7 40.0 f 7.9 36.2 f 7.7 .................................................................................................................................... bAY = DAY POSTPARTUM PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C25 (PAGE 1): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 22 POSTPARTUM - SUMMARY - F1 GENERATION PUPS TRANSIENT CLINICAL OBSERVATIONS: a TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS NOT NURSING EMACIATION DEHYDRATION N/N O/O N/N o/o N/N o/o 1/1 o/o o/o o/o 4/1 2/2 o/o 13/1 o/o 13/1 o/o 13/1 WHOLE BODY, EXCLUDING HEAD, DISCOLORATION, PURPLE SCAB b COLD TO TOUCH LEFT EYE, PERIORBITAL DISCOLORATION, PURPLE RIGHT EYE, ENLARGED RIGHT HINDLIMB, LACERATION BACK, BRUISE NOT NESTING PALE IN APPEARANCE C PI RIGHT EYE, CORNEAL OPACITY I I I I I I I I I I I I I I I I I I I I > o I ul I rllrl I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I >m IH I I I m Irl I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I Hrl IH IH I I I m Irl I I I I I I I I I I 2 I 1 -m I I PI I I I I I I I I 32 I I 0 I I H I I I I I I I H m I ul IH -Id I 0 1 w I I u I 1 I I rl I I h I I I I I I I I I 2 I I I I I I I I I I 1 I Cll I I I I I 1 H O I ul I -m I 2 I 0 I H I I 2 I I I 2w I Irl I I I I I I I I I aCI] I I I I 0 I I I I I I CI I I PI 1 I 0 I -I %UI E rl 2 u w v w cu U w ib $ 4 18-028:PAGE C-44 ulrl rlrl cu 0. 0. 00 00 rlrl ww ulrl rlrl cu 00 a . 00 --I 00 I I I I I I u I I & I 1.d I& 13 I E: I-rl A- 00 .. 00 --I 00 I8 I Ba I E Ik I0 IW Ik a, I I mu) 4m rl 0. 0. 00 00 rlrl vv mw rlm rl --I a, 00 I d I la,. m dLi M I I. Itd PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C27 (PAGE 1): PUP LIVER WEIGHTS - SUMMARY - F1 GENERATION PUPS LITTERS EXAMINED TERMINAL BODY WEIGHT WEIGHT/LITTER MALE PUPS FEMALE PUPS N MEANf.S.D . MEANkS .D . MEANkS .D . 10 37.42 f. 4.53 38.17 5 4.72 36.67 f. 4.44 lo 40.00 5 8.08 41.57 5 8.53 38.35 f. 7.94 10 38.56 5 4.78 40.27 5 4.60 36.86 f. 5.30 10 40.11 2 8.35 41.04 5 8.18 39.22 5 8.57 10 34.70 2 8.26 35.33 5 8.27 34.07 5 8.30 LIVER WEIGHT/LITTER MALE PUPS FEMALE PUPS MEAN2S.D . MEANkS.D . MEANkS .D . 1.54 5 0.24 1.48 2 0.26 1.59 5 0.24 1.65 f. 0.41 1.64 2 0.43 1.65 2 0.40 1.60 5 0.28 1.62 2 0.31 1.58 5 0.31 1.70 2 0.38 1.65 2 0.40 1.75 5 0.39 1.40 2 0.40 1.37 2 0.40 1.44 f. 0.40 RATIOS ( % ) OF LIVER TO TERMINAL BODY WEIGHT WEIGHT/LITTER MEANf.S.D . 4.10 2 0.24 4.1 -+ .2 4.14 f. .3 4.23 f. 0. 7 -+ .34 MALE PUPS MEAN5S.D . 3.86 f. 0.27 3.91 f. 0.33 4.02 f. 0.44 4.01 2 0.47 3.79 2 0.37 FEMALE PUPS MEANkS .D . 4.33 5 0.24 4.28 f. 0.26 4.26 5 0.34 4.46 f. 0.35 4.17 f. 0.36 ________________________________________-------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). RATIOS ( % ) = (LIVER WEIGHT/TERMINAL BODY WEIGHT) X 100. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C28 (PAGE 1): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C28 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C28 (PAGE 3): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .................................................................................................................................... DOSAGE GROUP I11 1 MG/KG/DAY .................................................................................................................................... RAT # DESCRIPTION .................................................................................................................................... 19003 19007 19008 19013 19014 19015 19017 19024 19029 19034 19038 19056 19057 19060 19064 DL( 17- 22) DG( 20- 21) DL( 1- 22) DL( 22 ) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS HEAD a LIMBS LIMBS a HEAD b ______________-^-_------------------------------------------------------------------------------------------------------------------ DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. b. First observed at necropsy. DL = DAY OF LACTATION PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C28 (PAGE 4): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # DESCRIPTION ------------- .................................................................................................................... 19002 19005 19035 19039 19040 19045 19046 19051 19054 19058 DL( 19- 22) DG( 14 ) DG( 18- 21) DL( 1- 22) DS( 11- 14) DG( 0- 25) DL( 17- 22) LOCALIZED ALOPECIA: NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NOSE: SWOLLEN LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: NO ADVERSE FINDINGS LOCALIZED ALOPECIA: NO ADVERSE FINDINGS BACK a LIMBS LIMBS a LIMBS LIMBS a LIMBS a 19062 DL( 17- 22) TIP OF TAIL MISSING a 19063 NO ADVERSE FINDINGS 19066 NO ADVERSE FINDINGS 19069 NO ADVERSE FINDINGS 19073 NO ADVERSE FINDINGS .................................................................................................................................... DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C28 (PAGE 5): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 19001 NO ADVERSE FINDINGS 19006 NO ADVERSE FINDINGS 19011 NO ADVERSE FINDINGS 19020 DS( 6- 12) BROWN PERIORAL SUBSTANCE DS( 6- 12) RALES DS( 10- 12) URINE-STAINED ABDOMINAL FUR DS( 11 ) COLD TO TOUCH DS( 11- 12) MIOSIS DS( 11- 12) ABDOMINAL DISTENTION DS( 11- 12) EMACIATION DS( 11- 17) LOCALIZED ALOPECIA: HEAD DG( 0- 6) LOCALIZED ALOPECIA: HEAD DG( 7- 20) LOCALIZED ALOPECIA: LIMBS DL( 2- 22) LOCALIZED ALOPECIA: LIMBS a DL( 7- 22) ABDOMINAL DISTENTION a DL( 14- 15) EMACIATION DL( 14- 22) RALES DL( 17 ) DEHYDRATION DL( 18- 22) EMACIATION a 19022 NO ADVERSE FINDINGS 19025 NO ADVERSE FINDINGS 19027 NO ADVERSE FINDINGS 19028 NO ADVERSE FINDINGS 19030 DL( 17- 22) HEAD: SCAB (0.3 CM X 0.3 CM)a DL( 17- 22) LOCALIZED ALOPECIA: HEAD a 19031 NO ADVERSE FINDINGS 19032 NO ADVERSE FINDINGS 19033 NO ADVERSE FINDINGS 19049 NO ADVERSE FINDINGS 19059 NO ADVERSE FINDINGS 19070 --------------------^________ NO ADVERSE FINDINGS .................................................................. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy . DL = DAY OF LACTATION PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C29 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS .................................................................................................................................... DOSAGE GROUP RAT DOSAGE (MG/KG/DAY) NUMBER .................................. I 0 19010 DL 22 P 70 ALL TISSUES APPEARED NORMAL. 19012 DL 22 P 58 ALL TISSUES APPEARED NORMAL. 19019 DL 22 P 59 ALL TISSUES APPEARED NORMAL. 19021 DL 22 P 58 ALL TISSUES APPEARED NORMAL. 19023 DL 22 P 60 ALL TISSUES APPEARED NORMAL. 19041 DL 22 P 59 ALL TISSUES APPEARED NORMAL. 19042 DL 22 P 58 ALL TISSUES APPEARED NORMAL. 19044 DL 22 P 56 ALL TISSUES APPEARED NORMAL. 19050 DL 22 P 60 ALL TISSUES APPEARED NORMAL. 19053 DL 22 P 59 ALL TISSUES APPEARED NORMAL. 19065 DL 22 P 58 ALL TISSUES APPEARED NORMAL. 19068 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19072 DL 22 P 58 ALL TISSUES APPEARED NORMAL. 19074 DL 22 P 59 ALL TISSUES APPEARED NORMAL. 19075 DL 22 P 59 ALL TISSUES APPEARED NORMAL. I1 0.3 19004 DL 22 P 59 ALL TISSUES APPEARED NORMAL. 19009 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19016 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19018 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19026 DL 22 P 60 ALL TISSUES APPEARED NORMAL. 19036 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19037 DL 22 P 58 ALL TISSUES APPEARED NORMAL. 19043 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19047 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19048 DL 22 P 60 ALL TISSUES APPEARED NORMAL. 19052 DL 22 P 56 ALL TISSUES APPEARED NORMAL. 19055 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19061 DL 22 P 56 ALL TISSUES APPEARED NORMAL. 19067 DL 22 P 58 ALL TISSUES APPEARED NORMAL. 19071 DL 22 P 57 ALL TISSUES APPEARED NORMAL. .................................................................................................................................... DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table C27) for external observations confirmed'atnecropsy. P I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I 1 I I I I 1 I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I (dl I I I tnl I 21 II 0 H 1I I 2 ; I I 2 ; I a 1 I I crll I I 01 I I I I I I I I I I I 418-028:PAGE C-52 I I I I I I I I I I I I I I I I 1 1 I I I I I I I I I I I I 1% IS P I P I z P I P I P I P I P I P IzP I P I P I P I P I I I E W I I I I 1 I 1 1 I I I I I I I I I I I I I I I I I I I I I I I I I I PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C29 (PAGE 3): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS IV 3 19002 DL 22 P 69 ALL TISSUES APPEARED NORMAL. 19005 DL 22 P 59 ALL TISSUES APPEARED NORMAL. 19035 DL 22 P 59 ALL TISSUES APPEARED NORMAL. 19039 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19040 DL 22 P 58 ALL TISSUES APPEARED NORMAL. 19045 DL 22 P 57 ALL TISSUES APPEARED NORMAL. 19046 DG 25 NP 39 ALL TISSUES APPEARED NORMAL. 19051 DG 25 NP 52 OVARIES: LEFT, SMALL (0.003 G) . ALL OTHER TISSUES APPEARED NORMAL. 19054 19058 19062 DL 22 P DL 22 P DL 22 P 59 ALL TISSUES APPEARED NORMAL. 57 ALL TISSUES APPEARED NORMAL. 59 ALL TISSUES APPEARED NORMAL. 19063 DL 22 P 58 SPLEEN: OMENTUM ADHERED TO THE LENGTH OF THE OUTER CAPSULE. ALL OTHER TISSUES APPEARED NORMAL. 19066 DL 22 P 59 ALL TISSUES APPEARED NORMAL. 19069 DL 22 P 58 ALL TISSUES APPEARED NORMAL. 19073 DL 22 P 57 ALL TISSUES APPEARED NORMAL. .................................................................................................................................... DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table C27) for external observations confirmed at necropsy. PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T - 7 7 0 6 . 1 ) TABLE C29 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 19020 DL 22 P 59 LARGE AND SMALL INTESTINES: DISTENDED WITH GAS. THYMUS: SMALL. SPLEEN: SMALL. ADRENALS : BILATERAL, LARGE. . STOMACH: FUNDIC MUCOSAL SURFACE CONTAINED APPROXIMATELY 12 BLACK AREAS (PINPOINT TO 0 . 3 CM IN DIAMETER) ALL OTHER TISSUES APPEARED NORMAL. 19022 19025 19027 19028 19030 19031 19032 19033 19049 19059 19070 ...................................... DG = DAY OF PRESUMED GESTATION P = PFLEGNANT NP = NOT PREGNANT a. Refer to the individual clinical DL 22 P 60 ALL TISSUES APPEARED NORMAL. DL 22 P 59 ALL TISSUES APPEARED NORMAL. DL 22 P 59 ALL TISSUES APPEARED NORMAL. DL 22 P 57 ALL TISSUES APPEARED NORMAL. DL 22 P 60 ALL TISSUES APPEARED NORMAL. DL 22 P 56 ALL TISSUES APPEARED NORMAL. DL 22 P 59 ALL TISSUES APPEARED NORMAL. DL 22 P 56 ALL TISSUES APPEARED NORMAL. DL 22 P 57 ALL TISSUES APPEARED NORMAL. DL 22 P 56 ALL TISSUES APPEARED NORMAL. DL 22 P 57 ALL TISSUES APPEARED NORMAL. ..................................................................... DL = DAY OF LACTATION observations table (Table C27) for external observations confirmed at necropsy . 418-028:PAGE C-55 I I I I I I I I I I I I I I I . % I I ..........I I 2~100000000001 I . . . . . . . . . . . . rldrlrlrlrlrlrlrlrl I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I I . . . . . . . . . . . . . . .I 1mm~omr~rlwmwmomm13 ~~~w~wmmw~wwrwr~ lOOOOOOOOOOOOOOOlE I I I I I I I I I I I I I I I I I I I I I I .. I I ki k . ' d o I Ikh d0I I I I I I I I I I . . . . . . . . . .I $g~diirlrlrlrlrlrldl . . . . . . . . . . . . . . .Iwa ? j ~ I o o o o o o o o o o o o o I o oW l ~ w I I I I I I I I I I 1 I I I I I I I I I I I2 I? I8 I gI \ I I0 I I I I I I Iul I I I I I I I I I I I I I I I I I I I I I I I I I H I gI & Id IW Iw IW I$ I0 In 418-028:PAGE C-56 I I I I I I I I . . . . . . . . . . . I Im*m*wmPPmd I0 I - 5I **mm*mmmm* I0 I I Id I w~loooooooooo Ix I I p: o\o I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 418-028:PAGE C-57 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I -rl I rl I 5 I a, I I I I w I I -8 I a, I I I rl I I rl I I k I I JJ I I 4 I 8 I I I I -4 I I 9 I a, I I I I 0 I JJ I 418-028:PAGE C-58 PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T - 7 7 0 6 . 1 ) TABLE C30 (PAGE 5 ) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - FO GENERATION FEMALE RATS .................................................................................................................................... DOSAGE GROUP V 10 MG/KG/DAY .................................................................................................................................... BRAIN LIVER KIDNEY KIDNEY ADRENAL ADRENAL RAT TERMINAL BODY LEFT RIGHT LEFT RIGHT NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW WT. % TBW WT . % TBW WT. % TBW WT. % TBW a WT. % TBW a .................................................................................................................................... 19001 P 278. 2.00 0.72 12.11 4.36 1.36 0.49 1.35 0.48 0.041 14.75 0.031 11.15 19006 P 304. 2.13 0.70 14.39 4.73 1.44 0.47 1.55 0.51 0.057 18.75 0.055 18.09 19011 P 283. 2.05 0.72 14.24 5.03 1.41 0.50 1.36 0.48 0.045 15.90 0.045 15.90 19020 P 210. 2.00 0.95 7.01 3.34 1.14 0.54 1.09 0.52 0.058b 27.62 0.056 26.67 19022 P 294. 2.18 0.74 12.61 4.29 1.45 0.49 1.63 0.55 0.064 21.77 0.041 13.94 19025 P 291. 2.13 0.73 13.99 4.81 1.36 0.47 1.40 0.48 0.061 20.96 0.055 18.90 19027 P 266. 2.27 0.85 12.11 4.55 1.38 0.52 1.35 0.51 0.053 19.92 0.047 17.67 19028 P 339. 2.02 0.60 15.89 4.69 1.38 0.41 1.41 0.42 0.044 12.98 0.041 12.09 19030 P 314. 2.18 0.69 14.52 4.62 1.46 0.46 1.50 0.48 0.062 19.74 0.046 14.65 19031 P 361. 2.24 0.62 19.51 5.40 1.52 0.42 1.47 0.41 0.042 11.63 0.038 10.53 .................................................................................................................................... RAT TERMINAL BODY SPLEEN THYMUS OVARY LEFT OVARY RIGHT UTERUS WITH CERVIX HEART NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % TBW WT. % TBW WT. % TBW a WT. % TBW a WT. % TBW WT. % TBW .................................................................................................................................... 19001 P 278. 0.57 0.20 0.18 0.06 0.048 17.27 0.047 16.91 0.64 0.23 0.92 0.33 19006 P 304. 0.54 0.18 0.15 0.05 0.056 18.42 0.074 24.34 0.46 0.15 1.57 0.52 19011 P 283. 0.50 0.18 0.15 0.05 0.046 16.25 0.036 12.72 0.68 0.24 1.24 0.44 19020 P 210. 0.28b 0.13 O.llb 0.05 0.036 17.14 0.047 22.38 0.50 0.24 0.90 0.43 19022 P 294. 0.53 0.18 0.14 0.05 0.060 20.41 0.035 11.90 0.74 0.25 1.54 0.52 19025 P 291. 0.56 0.19 0.25 0.08 0.053 18.21 0.058 19.93 0.53 0.18 1.38 0.47 19027 P 266. 0.61 0.23 0.23 0.09 0.039 14.66 0.018 6.77 0.44 0.16 1.32 0.50 19028 P 339. 0.54 0.16 0.23 0.07 0.087 25.66 0.053 15.63 0.37 0.11 1.56 0.46 19030 P 314. 0.58 0.18 0.17 0.05 0.045 14.33 0.048 15.29 0.78 0.25 1.37 0 -44 19031 P 361. 0.64 0.18 0.16 0.04 0.076 21.05 0.087 24.10 0.74 0.20 1.50 0.42 19032 P 287. 0.031 10.80 0.043 14.98 0.33 0.11 19033 P 328. 0.069 21.04 0.076 23.17 0.82 0.25 19049 P 330. 0.050 15.15 0.073 22.12 0.50 0.15 19059 P 317. 0.061 19.24 0.063 19.87 0.39 0.12 19070 P 351. 0.051 14.53 0.045 12.82 0.47 0.13 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGANWEIGHT/TERMINAL BODY WEIGHT) x 1 0 0 . P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) a. Value was multiplied by 1000. . b. Dam 1 9 0 2 0 had a large left adrenal, small spleen, and small thymus. See the individual necropsy observations table (Table C28) PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C31 (PAGE 1): ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO BRAIN WEIGHT - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT NUMBER BRAIN WEIGHT - - - - - - - - - -. - - - - - - - - - - - - - - - 19012 P 2.22 19019 P 2.22 19021 P 2.21 19023 P 2.11 19041 P 2.09 19042 P 2.25 19044 P 2.17 19050 P 2.11 19053 P 2.25 19065 P 2.00 LIVER KIDNEY LEFT KIDNEY RIGHT ABS. REL. ABS. REL. ABS. REL. WT . % BRW WT. % BRW WT . % BRW ------------------- .................................... 16.17 728.38 1.53 68.92 1.66 74.77 15.21 685.14 1.43 64.41 1.46 65.76 14.21 642.99 1.50 67.87 1.55 70.14 13.09 620.38 1.23 58.29 1.39 65.88 12.15 581.34 1.24 59.33 1.35 64.59 14.31 636.00 1.60 71.11 1.57 69.78 19.73 909.22 1.51 69.58 1.60 73.73 12.84 608.53 1.32 62.56 1.41 66.82 13.17 585.33 1.26 56.00 1.34 59.56 12.97 648.50 1.30 65.00 1.33 66.50 ADRENAL LEFT ABS. REL. WT. % BRW .------------------ 0.050 0.055 0.045 0.054 0.048 0.056 0.076 0.048 0.049 0.042 2.25 2.48 2.04 2.56 2.30 2.49 3.50 2.27 2.18 2.10 ADRENAL RIGHT SPLEEN ABS. REL. ABS. REL. WT . % BRW WT. % BRW ---_______---_______-------------- 0.047 2.12 0.78 35.14 0.047 2.12 0.64 28.83 0.040 1.81 0.58 26.24 0.028 1.33 0.58 27.49 0.044 2.10 0.54 25.84 0.046 2.04 0.64 28.44 0.053 2.44 0.67 30.88 0.036 1.71 0.74 35.07 0.043 1.91 0.61 27.11 0.044 2.20 0.54 27.00 19012 P 2.22 0.23 10.36 0.092 4.14 0.073 3.29 0.57 25.68 1.32 59.46 19019 P 2.22 0.31 13.96 0.044 1.98 0.064 2.88 0.51 22.97 1.55 69.82 19021 P 2.21 0.11 4.98 0.044 1.99 0.040 1.81 0.53 23.98 1.31 59.28 19023 P 2.11 0.16 7.58 0.039 1.85 0.062 2.94 0.62 29.38 1.62 76.78 19041 P 2.09 0.16 7.66 0.062 2.97 0.037 1.77 0.46 22.01 1.35 64.59 19042 P 2.25 0.33 14.67 0.059 2.62 0.057 2.53 0.68 30.22 1.64 72.89 19044 P 2.17 0.27 12.44 0.070 3.22 0.081 3.73 0.56 25.81 1.45 66.82 19050 P 2.11 0.20 9.48 0.044 2.08 0.048 2.27 0.56 26.54 1.19 56.40 19053 P 2.25 0.17 7.56 0.058 2.58 0.085 3.78 0.47 20.89 1.41 62.67 19065 P 2.00 0.17 8.50 0.068 3.40 0.066 3.30 0.69 34.50 1.13 56.50 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - - - - - -.-------_- -- _- ------ - - - - - _ _ _ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/BRAIN WEIGHT) x 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) 418-028:PAGE C-61 I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I . . . . . . . . . . I I I ImdmPmVdmOm I .3 I dm*mOP*mmm I J&l I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I i sw a &. . I I I I I I I I I I I I I I I I I I I . . . . . . . . . . I 1*wa,wa,Trm*oo Ia,mwa,wmoPma, I lPdwwmwma,mm IwPmmwmmmmm I I I . . . . . . . . . . I im~+mwmomdmm immmdmmdmo~ 1 lddrlddddrldd I I I I I I I I I I I I I I I m d o w m a , m m m w: ~3 a Iwmrwrmwwmm~p I ..........I I 0 0 0 0 0 0 0 0 0 0 I o\o I I I I. II II JW I m I P I I I I I I I I I I w I I I 1 I I 3 I I I I w I I E I I w I 2w I I J I c4 I z I I I z I I I 3 I I I E I W I I IPI II w I I I I I I I I I I 0 I !h I I I I I I s I I I 8 I :GI 3J1 4 :2 I I GI I I I I E3n I I I I I I I I [ I I I I I H I 1 g I I I I E I :!I X I I E I I sw I 14 Iw I I I 2a I I I I I I I I I I I I I I 0 I : k i E I I ;; E I X I I IW E I I I I sw I I I I I I 2 I I I I I 8 I I 1'3. c4 I 0 I IW I I W h I\ I o\o I Y I :" I w Irl I 0 I I H I I I I I I 3 I I I I I I I 1 2 I I I I w I I E I I X I I I I E I I I I I I 2! I I 2 I I I I W I I p: I I 0 I I I I I I I I m h I I I I IH I IH I w IH I W I I P4I I P I II 1 g Y I I I8 rl I m I 1 w u I IW I w I l w 1 4 I 8E 1 0 1 I I I I I I I I I * r r I rl W W rl 0 rl In 0 rl W I I rl cn m 0 cn m m rl I I I I cn r rl In P P 0 m cv I I cv m cv m rl cv cv m cv m I I I I I I I * * I I I cv cv cn m cv 0 rl 0 Ln I I W 03 Tr W WWWm W 1 I 1 I 00 00000000 I I I I I I I I I * * I I I N cn cn cv P CO r P CO I I CO cv 0 m W P CO W cn I I I I N m cv cv cv rl rl cv cv rl I I I I I I I * * * * r I I I 0 rl rl cn W CO cv cn W 0 I I W w m m 0 0 In I 1 000000 00 I I I I 0 000 000000 I I I I I I I I I * * ** * * I I 0 I m rl cv I cn rl W r m 03 I cn 03 m cn I I I I cv m cv cv cv rl cv cv N rl I I I I I I I * * * I I I w r m 0 W cn m cn rl 0 I II 0 m W 0 0 0 m W 0 m0 0 m 0 0 m 0 II I I I 0000000000 I I I I I I I I I I * I cv m P 0 rl * * I m 03 rl 0 rl I cv 0 m cv rl W cv I I * *r I I I W m 0 W r m m r- I I W P W CO m W W P W I I I I I I I * * * * I ld I I I P m W W rl m W cv m m dl m rl 0 9 m 1 I I I I rl rl rl rl rl rl rl rl rl rl I I I I I I I I I I I * I 0 0 cn W 9 rl m rl r- I * I m 0 W cv ul W W 03 r 1 * r r I I I 1 rl m W rl W 0 P rl m P W 03 W W m cv W I I I I I I I I * ** * * * I I rl cv I m W cv I I r9l m m W m cv r m I 1 I I rl rl rl rl rl rl rl rl rl rl I I I I I I I I I I I * I cv cn cv rl CO 0 W m m I * I m 0 03 m W cn ul cn P I I I * * I r m W m Lo 0 W 0 I I * I CO 0 m cn Ln W W W m W r 0 rl cv rW 0W W I I I rl I I I I 1 I rl r W 0 0 cn m m m cv I * * I m 0 P m a\ cn W W I I I * I cv m cv cn cv W cn cv cv I I rl rl rl rl rl rl rl rl rl rl I I I I I I I I I I I I I I I I m W rl m rl m m CO 0 cv I I rl rl 0 0 cv rl cv cn rl 0 I I I I cv cv cv cv cv cv cv rl (\I cv I I I I I I I I PI PI PI PI PI PI PI PI PI PI I I I I I I m P W m m P 9 cn v W I I 0 0 0 rl rl rl cv cv m m I I 0 00 0000 0 00 I I cn cn cn cn cn cn cn cn cn cn I I rl rl rl rl rl rl rl rl rl rl I I I I I I I 418-028:PAGE C-62 1 I I I I I I I I I I I I x I I I I 8k I I I I. u I I0 I I0 2 I Irl I I I I I I I I W CO rl m cv W CO m cv I m rl cv W m cv P r W * I I 0 0 W m W m cv r 1 W W m W W W W rm I I I * * I I 0-l 0 P m rl W 0 03 rl I cv m rl m m m cv I I rl rl rl rl rl rl rl rl rl I I I I I I I bm ( r r cn m m cv (ru co cv m r mcn CO rl I I cv W W CO rl W CO r 0 I cv m cv N N rl rl rl cv I I I * * * * * I I CO cn m CO CO ul cv v I P Ln ul m m I I 0000000 0 0 1 I I I * I I rl W 0 P m 0 m cv I P cn r cn rl a cn cn ul I I m m cv m cv N 0 cv m I I I * * * I I I cn W P W m m 0 CO 03 co 0 co cv m r I 00 00 00000 I I 0 00 0 00 000 1 I I I * * * I I I m ul 9 cn cv m cn W m r m (\1 cn 0 cn ul I I I I I cv cv N m rl N rl cv cv cv I I I I I I I * * * * I I I m 0 03 m m m rl cv rl I I m m m W ul W ul I I 00000 0 00 00 I I I I 000 0000000 I I I I I I I I I * I I I r W W m W ul cv 03 r cn I I m m P 03 0 m 0 m W I m I I W I I W 0 0 W CO CO CO 0 I rl rl rl I I I I I I I * I I I cv r cv w m cn W 03 cv I I rl rl rl N cv rl rl rl rl cv I I I I 0 000 0 0 0000 I I I I I I I I I 1 I I I 1 I I I I m W rl Ln rl m m W 0 cv I I rl d 0 0 cv rl cv cn rl 0 I I I I cv cv cv cv (\I cv cv rl cv cv I I I I I I I I PI PI PI PI PI PI PI PI PI PI I I I * I I I m P CO m ul r v cn W I I 0 0 0 rl rl rl cv cv m m I I 0 00 0 000 0 0 0 I I cn cn cn cn cn cn cn cn cn cn I I rl rl rl rl rl rl rl rl rl rl I I I I I I I 418-028:PAGE C-63 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T - 7 7 0 6 . 1 ) TABLE C31 (PAGE 5 ) : ORGAN WEIGHTS AND RATIOS (2) OF ORGAN WEIGHT TO BRAIN WEIGHT - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS .................................................................................................................................... DOSAGE GROUP V 10 MG/KG/DAY .................................................................................................................................... RAT BRAIN LIVER KIDNEY LEFT KIDNEY RIGHT ADRENAL LEFT ADRENAL RIGHT SPLEEN NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % BRW WT. % BRW WT. % BRW WT. % BRW WT. % BRW WT . % BRW .................................................................................................................................... 19001 P 2.00 12.11 605.50 1.36 68.00 1.35 67.50 0.041 2.05 0.031 1.55 0.57 28.50 19006 P 2.13 14.39 675.59 1.44 67.60 1.55 72.77 0.057 2.68 0.055 2.58 0.54 25.35 19011 P 2.05 14.24 694.63 1.41 68.78 1.36 66.34 0.045 2.20 0.045 2.20 0.50 24.39 19020 P 2.00 7.01 350.50 1.14 57.00 1.09 54.50 0.058a 2.90 0.056 2.80 0.28a 14.00 19022 P 2.18 12.61 578.44 1.45 66.51 1.63 74.77 0.064 2.94 0.041 1.88 0.53 24.31 19025 P 2.13 13.99 656.81 1.36 63.85 1.40 65.73 0.061 2.86 0.055 2.58 0.56 26.29 19027 P 2.27 12.11 533.48 1.38 60.79 1.35 59.47 0.053 2.33 0.047 2.07 0.61 26.87 19028 P 2.02 15.89 786.63 1.38 68.32 1.41 69.80 0.044 2.18 0.041 2.03 0.54 26.73 19030 P 2.18 14.52 666.06 1.46 66.97 1.50 68.81 0.062 2.84 0.046 2.11 0.58 26.60 19031 P 2.24 19.51 870.98 1.52 67.86 1.47 65.62 0.042 1.88 0.038 1.70 0.64 28.57 .................................................................................................................................... THYMUS OVARY OVARY UTERUS HEART RAT BRAIN LEFT RIGHT WITH CERVIX NUMBER WEIGHT ABS. REL. ABS. REL. ABS. REL. ABS. REL. ABS. REL. WT. % BRW WT. % BRW WT. % BRW WT. % BRW WT. % BRW .................................................................................................................................... 19001 P 2.00 0.18 9.00 0.048 2.40 0.047 2.35 0.64 32.00 0.92 46.00 19006 P 2.13 0.15 7.04 0.056 2.63 0.074 3.47 0.46 21.60 1.57 73.71 19011 P 2.05 0.15 7.32 0.046 2.24 0.036 1.76 0.68 33.17 1.24 60.49 19020 P 2.00 O.lla 5.50 0.036 1.80 0.047 2.35 0.50 25.00 0.90 45.00 19022 P 2.18 0.14 6.42 0.060 2.75 0.035 1.60 0.74 33.94 1.54 70.64 19025 P 2.13 0.25 11.74 0.053 2.49 0.058 2.72 0.53 24.88 1.38 64.79 19027 P 2.27 0.23 10.13 0.039 1.72 0.018 0.79 0.44 19.38 1.32 58.15 19028 P 2.02 0.23 11.39 0.087 4.31 0.053 2.62 0.37 18.32 1.56 77.23 P CI 19030 P 2.18 0.17 7.80 0.045 2.06 0.048 2.20 0.78 35.78 1.37 62.84 19031 P 2.24 0.16 7.14 0.076 3.39 0.087 3.88 0.74 33.04 1.50 66.96 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/BRAIN WEIGHT) X 1 0 0 . P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) . a. Dam 1 9 0 2 0 had a large left adrenal, small spleen, and small thymus. See the individual necropsy observations table (Table C28) PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C32 (PAGE 1): PRIMORDIAL FOLLICLE COUNT - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS AVERAGE 19019 AVERAGE 19021 AVERAGE 19023 AVERAGE 19041 37 46 33 34 36.2 4 1 2 0 1 1.6 16 16 21 20 14 17.4 7 14 10 14 11 11.2 7 14 2 5 7 46 34 36 36 34.6 0 5 2 2 7 3.2 4 4 12 17 30 13.4 22 11 13 27 6 15.8 4 3 4 6 9 AVERAGE 19044 AVERAGE 19050 AVERAGE 19053 AVERAGE 19065 0 1 6 3 2.2 3 11 9 13 51 17.4 3 5 2 4 3 3.4 24 11 27 27 25 22.8 4 3 9 3 14 9 9 16 13 13.0 5 8 8 9 17 9.4 1 5 3 6 10 5.0 3 11 16 20 38 17.6 7 15 11 12 16 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C32 (PAGE 2): PRIMORDIAL FOLLICLE COUNT - INDIVIDUAL DATA - FO GENERATION FEMALE RATS AVERAGE 19006 AVERAGE 19011 AVERAGE 19020 AVERAGE 4 9 6 23 9.0 27 11 23 26 21 21.6 6 16 8 16 37 16.6 5 4 15 8 20 10.4 5 6 17 20 10.8 5 6 19 20 20 14.0 1 0 12 24 8 9.0 2 1 2 12 20 7.4 AVERAGE 19027 AVERAGE 19028 AVERAGE 19030 AVERAGE 12 6 6 7 8.4 6 6 17 13 43 17.0 3 2 2 5 7 3.8 9 21 24 22 26 20.4 6 15 25 12 12.4 0 5 8 12 8 6.6 1 7 13 13 10 8.8 3 13 6 7 20 9.8 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C33 (PAGE 1): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS ________________________________________-------------------------------------------------------------------------------------------- _ _ _ _ _ _ _ _ _D_AY_ _ _ _ _1 _ _ _ _2_ _ _ _ _3_ _ _ _ _4_ _ _ _ _5_ _ _ _ _6_ - - - - - -7- - - - - - -8- - - - - - -9- - - - - -1-0- - - - - - -1-1- - - - - -1-2- - - - - -1-3- - - - - -1-4-a- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - _. _ _ _RA_T_ _# _ _ _ _ _ _D_OS_A_GE_ _GR_O_U_P_I_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - -o- -M-G-/-K-G-/-D-A-Y- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 19010 213. 218. 223. 225. 224. 229. 235. 237. 238. 239. 246. 251. 246. 249. 19012 19019 235. 230. 239. 232. 238. 240. 236. 242. 247. 244. 246. 251. 250. 254. 251. 261. 261. 256. 261. 263. 267. 269. 258. 269. 270. 272. 267. 277. 19021 231. 236. 237. 240. 244. 248. 245. 250. 253. 258. 258. 260. 264. 266. 19023 219. 213. 222. 227. 223. 222. 228. 232. 229. 224. 233. 235. 235. 234. 19041 245. 239. 247. 252. 252. 249. 258. 264. 266. 260. 268. 272. 268. 264. 19042 227. 225. 232. 236. 238. 236. 236. 244. 247. 248. 248. 242. 249. 250. 19044 231. 230. 233. 235. 242. 238. 240. 247. 249. 243. 246. 245. 249. 250. 19050 220. 222. 226. 229. 229. 232. 236. 245. 243. 240. 245. 251. 246. 247. 19053 234. 238. 243. 244. 246. 255. 260. 262. 262. 269. 272. 274. 271. 277. 19065 223. 223. 230. 232. 234. 241. 245. 247. 244. 250. 255. 255. 254. 261. 19068 232. 229. 236. 247. 256. 260. 265. 277. 275. 281. 291. 288. 288. 293. 19072 222. 217. 228. 229. 228. 228. 239. 244. 238. 238. 247. 250. 242. 240. 19074 235. 238. 238. 241. 244. 247. 246. 253. 251. 254. 254. 258. 263. 260. 19075 232. 224. 229. 234. 234. 236. 242. 244. 244. 246. 247. 246. 227. 242. ________________________________________-------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I1 0.3 MG/KG/DAY ________________________________________-------------------------------------------------------------------------------------------- 19004 205. 203. 214. 215. 214. 216. 220. 226. 222. 219. 226. 228. 226. 223. 19009 231. 234. 236. 246. 248. 252. 252. 263. 257. 260. 260. 265. 264. 265. 19016 222. 224. 225. 227. 231. 232. 234. 234. 240. 238. 239. 247. 243. 244. 19018 222. 228. 228. 233. 232. 230. 240. 239. 244. 241. 241. 246. 246. 247. 19026 221. 227. 232. 236. 231. 233. 237. 239. 236. 241. 246. 251. 249. 249. 19036 230. 235. 239. 243. 247. 249. 249. 255. 260. 262. 263. 269. 271. 268. 19037 217. 217. 222. 228. 228. 227. 236. 241. 244. 237. 240. 240. 242. 249. 19043 227. 228. 233. 237. 238. 237. 245. 248. 246. 244. 253. 253. 252. 252. 19047 219. 213. 207. 217. 211. 212. 213. 223. 224. 217. 219. 229. 228. 227. 19048 228. 230. 235. 240. 239. 240. 246. 244. 245. 245. 253. 255. 252. 253. 19052 228. 228. 226. 235. 240. 234. 236. 245. 246. 244. 247. 254. 255. 251. 19055 19061 235. 219. 231. 225. 228. 225. 239. 227. 249. 233. 243. 238. 241. 237. 253. 241. 258. 244. 253. 248. 255. 242. 264. 241. 266. 246. 262. 250. 19067 237. 238. 245. 246. 240. 246. 251. 252. 245. 250. 258. 259. 254. 259. 19071 230. 232. 232. 238. 238. 242. 241. 248. 251. 252. 250. 261. 262. 261. ________________________________________------------------------------------------------------------------------------------------ DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . a. Last value recorded before cohabitation. 418-028:PAGE C-68 I I I I 1 1 I I I I I I I I I I I I I I 1 1 I I I I I I I I I I I I I I I I la1 1w1 Irll I I I I I 1 I I I I lml lrll I I I I I 1 I I I I lcvl lrll I I I I I I I 1 I I lrll lrll I I I I I 1 I I I I I01 ldl I I I I I I I I I I lml I I I I 1 1 I I I I I I Ico I 2 I I I I \ I I 2 I I I I \ I I W x ~r I I I I I d I I I I I I I I IW I I I I I I 1 I I I I I I Im I I I I I I I I I I I I I I9 I I I I I I I I I I I I tm I I I I I I H I H I H I I I I I 1 p: I N 1 0 I I I I W I I w I I I I W I I 3 I r l I n I 1 I 0 I I I I I ; zI I I in I I I I I I I I # I I 3 I I I I I I I l I l I l I l I l I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I . . . . . . . . . . . . . . . I I I I I iwmwcvooomwwwmmod I ~ m m w r m w m r m w w m m r mI lmmcvcvcvmmmcvcvcvmcvmm I I I . . . . . . . . . . . . . . . I I I I I imrlwdmmmmowowmoo I Imcomrwmmwmwwmmwm I lmcvcvmmmmmcvcvmmmmm I I I I I .I . . . . . . . . . . . . . . . I I I I I I . . II I am I wco I cvcv I I .. I I I mw I wr I cvm I I .. I I I wr I ww I mm I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I 1 I I I I I I I I I I I I I l I l I l I l I l I I 1 I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I . . . . . . . . . . . . . . . I 1 I I ocowmorrwmmorlm~m I mmmmmmrlmcvmcvwmmm I mcvmmmmcvcvmmmcvmmm I I I I I 1 I I I I I I I I I I I I I I I I I I I PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C33 (PAGE 3): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS ________________________________________-------------------------------------------------------------------------------------------- _ _ _ _ _ _ _ _ _D_AY_ _ _ _ _1_ _ _ _ _2 _ _ _ _3_ _ _ _ _4_ _ _ _ _5_ _ _ _ _6_ - - - - - -7- - - - - - -8- - - - - - -9- - - - - - 1- 0- - - - - - -1-1- - - - - -1-2- - - - - -1-3- - - - - -1- 4- a- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - RAT # DOSAGE GROUP V 10 MG/KG/DAY ________________________________________-------------------------------------------------------------------------------------------- 19001 215. 214. 217. 217. 208. 212. 218. 220. 215. 214. 218. 221. 218. 213. 19006 231. 234. 238. 235. 236. 241. 242. 246. 250. 258. 258. 257. 259. 266. 19011 228. 230. 234. 238. 236. 243. 244. 248. 242. 241. 249. 249. 245. 243. 19020 226. 224. 222. 229. 231. 203. 199. 192. 186. 188. 203. 212. 220. 221. 19022 221. 221. 227. 230. 226. 230. 239. 240. 236. 236. 247. 247. 241. 243. 19025 229. 236. 240. 244. 239. 249. 259. 267. 271. 272. 283. 285. 280. 282. 19027 218. 220. 224. 228. 226. 229. 231. 235. 231. 235. 238. 240. 238. 238. 19028 232. 232. 232. 241. 242. 244. 243. 250. 253. 251. 252. 258. 259. 259. 19030 220. 220. 227. 230. 228. 232. 237. 242. 238. 241. 249. 253. 249. 251. 19031 245. 252. 255. 262. 264. 262. 255. 263. 264. 266. 268. 272. 277. 269. 19032 227. 229. 234. 240. 239. 243. 251. 258. 255. 256. 262. 266. 263. 264. 19033 225. 224. 228. 231. 234. 231. 233. 245. 244. 241. 246. 250. 256. 252. 19049 226. 228. 234. 237. 237. 240. 245. 254. 254. 252. 257. 262. 264. 257. 19059 240. 242. 243. 246. 250. 252. 250. 254. 256. 258. 256. 262. 263. 260. _ _ _ _1_90_7_0_ _ _ _ _ _2_20_._ _ _2_18_._ _ _21_8_._ _ _22_7_._ _ 2_3_1_._ _2_3_3._ - - -2-3-0-.- - - -2-3-9-.- - - -2-3-9-.- - - - 2- 4- 1- -.- - - -2-3-8-.- - - -2-4-6-.- - - -2-4-6-.- - - -2-4-8-.- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. P n 418-028:PAGE C-70 I I I I I 1 I I I I I I I I I I I I I I m I I I I I 3w I I I 3w I I I h I I z Z I 0 I H I I w I I I i3 I I W I I 0 I rh I I I I I n4 I E I 4 I I I I I 2 I E I 2 w n I 3 2 E cwn W 3 E 5 cn I cn E X ::2! * n 0 a i w 3 h rl 2 w W . I Y * m V 3 w i3 I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I . IZ I I I I t I I I I I I I I I I . . . . . I I 1 I ..........I I lmmrloPPPrlPwocvwu)wlml ~mmmmw~mcvom~mocvw~cvt ~mmmmcvmmmmmmmmmcvi I I I I I I I I I I I I I I I I I I I I I ...............I I I Iu)Pwou)mmcvmcv*mPmrll*l I lmcvcvcvw*cvrlo*m*orlwlcvl I ~mmmmcvmmmmmmmmmcv~ I I I I I I I I I I I I I I I I I I I I I I I ...............I I I ~w~w~m~owommw~dcv~m~ I 1cvcvrlrlwmcvoocvcvmorlwlcvI I ~mmmmcvmmmmmmmmmcvi I I I I I I I I I I I I I I I I I I I I I I I ...............I I ~~mrlrlocv~mcvwwmrlu)o~cv~ ~cv~~owm~omd~mmoco~cv~ ~mmmmcvmmmcvmmmcvmcv~ I I I I * I mI rll * I I I I I I I I I I . . . . I I I I I I I ...............I 8 lmcvw*m*wt-omcyrlmwwlrllmmlnlnu, lrlrlooPcvommcvrlmwmwlcvlm**ww Immmmcvmmcvcvmmmcvcvcvi 1***mm I I I .. m * * .. mw o* *m I I ..I *cv I cvw I em I I I I I I I I I I I I I I I 1 f ...............I I I mw*cvrlo*rlcvmoooocvI I rlooo~cvmmcorlocva30~1 I mmmmcvmcvcvcvmmmcvmcvi I I I . . . . . . . . . . . . . . . .I I I I I . . . . . . . . . . . . I I I I I '''1 I oomu)mwmworlwowlnrll I rl00mwrlcnwwrlmcvu)mw1 I mmmcvcvmcvcvcvmcvmcvcvcv~ I I I I f I I I I I I I 1 1 I ...............I I ...............I rcvrlOrlP*mm*mOcvm~*lwlwu)wmlnwPoomrlomPol lrlorl~wrlwwPoorlPm*lrllmwmPcvrlcn~wrlrlrlu)P*l~ ~mmmcvcvmcvcvcvmmmcvcvcvi ~ m m m m m ~ m m m ~ w ~ m m m i c n I I I IW . . . . I i$J I I I w I I. Im Id II m om rlm mcv NO Pcn cvcv I I I I I* Icv Irl II m *. mm ow mcv *. cv* PW cvcv I I I I I. lm I0 II m . . o m ow mcv . . w m ww cvcv I I I I I I I I I 1 I I I I I I I I I : I I I I I I locvmrlmrlcv*Ommwm*ml lrlrlrlcvcv***mmu)wPPPl 10000000000000001 ~mmmmmmmmmmmmmmm lrlrlrlrlrlrlrlrlrlrlrlrlrlrlrll I I I I I I I I 10000000000000001~" ~I m m m m m m m m m m m m m c n m l p l a lrl~rlrlrlrlrlrlrlrlrlrlrlrlrll * I I I1 4 I i pln I I PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C34 (PAGE 2): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS .................................................................................................................................... RAT # DOSAGE GROUP I1 0.3 MG/KG/DAY .................................................................................................................................... PREGNANCY . . . . . . .S.T.A.T.U.S. . . .D.A.Y. . .0. . . . . . . . .1. . . . . . . . .2. . . . . . . . .3. . . . . . . . .4. . . . . . . . . 5. . . . . . . . . 6. . . . . . . . . .7. . . . . . . . .8. . . . . . . . .9. . . . . . . .1.0. . . . . . . .1.1. . . . . . . . 1. 2. . 19004 P 239. 245. 252. 262. 265. 269. 270. 276. 280. 288. 292. 301. 306. 19009 P 19016 P 269. 249. 275. 250. 278. 258. 281. 260. 283. 266. 286. 260. 291. 269. 294. 270. 298. 281. 299. 277. 304. 282. 310. 291. 315. 296. 19018 P 255. 262. 266. 269. 278. 281. 286. 292. 295. 299. 309. 318. 322. 19026 P 19036 P 19037 P 19043 P 19047 P 268. 271. 260. 255. 232. 259. 281. 268. 265. 244. 273. 288. 274. 275. 241. 273. 292. 272. 274. 248. 285. 296. 280. 278. 248. 288. 301. 287. 285. 251. 290. 304. 290. 295. 255. 298. 313. 292. 292. 256. 296. 321. 296. 295. 264. 302. 324. 301. 305. 264. 311. 330. 302. 310. 270. 327. 332. 310. 318. 274. 329. 342. 319. 328. 279. 19048 P 271. 273. 282. 287. 288. 291. 295. 303. 299. 306. 309. 312. 317. 19052 P 259. 269. 262. 274. 272. 285. 288. 288. 293. 299. 305. 309. 318. 19055 P 19061 P 267. 259. 277. 259. 284. 266. 289. 264. 294. 271. 300. 278. 306. 278. 307. 279. 305. 288. 318. 291. 322. 290. 334. 296. 339. 306. 19067 P 273. 270. 268. 283. 288. 289. 289. 294. 299. 300. 308. 312. 321. 19071 P 264. 274. 281. 286. 292. 295. 304. 302. 309. 316. 319. 331. 338. ________________________________________-------------------------------------------------------------------------------------------- . . . . . . . . . . . . . . . .D.A.Y. .1.3. . . . . . . .1.4. . . . . . . . 1. 5. . . . . . . . .1.6. . . . . . . .1.7. . . . . . . .1.8. . . . . . . .1.9. . . . . . . .2. 0. . . . . . . . .2.1. . . . . . . .2.2. . . . . . . .2.3. . . . . . . .2.4. . . . . . . . .2.5 19004 P 310. 320. 325. 334. 349. 361. 383. 396. 19009 P 321. 321. 330. 335. 343. 349. 326. 334. 361. 19016 P- 299. 310. 312. 325. 338. 348. 361. 380. 395. 19018 P 326. 332. 339 * 348. 363. 373. 391. 408. 422. 19026 P 336. 349. 346. 361. 378. 394. 418. 432. 452. 19036 P 343. 348. 357 * 367. 383. 398. 410. 424. 422. 19037 P 321. 325. 328. 342. 351. 364. 377. 393. 399. 5 19043 P 329. 338. 346. 357. 372. 387. 389. 405. 417. 90 19047 P 280. 289. 297. 304. 314. 329. 334 * 364. 380. 19048 P 321. 329. 336. 336. 348. 358. 358. 375. 382. 0 19052 P 322. 330. 338. 350. 366. 377. 391. 409. R) ?? 19055 P 341. 349. 358. 371. 384. 401. 411. 433. 452. 19061 P 308. 315. 325. 338. 345. 351. 367. 382. F 19067 P 324. 325. 340. 344. 351. 370. 379. 395. 414. 8 19071 P 339. 348. 364. 365. 380. 394. 409. 431. 444. .................................................................................................................................... P P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 4 + ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C34 (PAGE 3): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .................................................................................................................................... PREGNANCY . . . . . . .S.T.A.T.U.S. . . .D.A.Y. . .0. . . . . . . . . 1. . . . . . . . . 2. . . . . . . . . .3. . . . . . . . .4. . . . . . . . .5. . . . . . . . .6. . . . . . . . . 7. . . . . . . . . .8. . . . . . . . .9. . . . . . . .1.0. . . . . . . . 1. 1. . . . . . . . .1.2 19003 P 244. 252. 252. 258. 258. 260. 262. 264. 267. 269. 271. 287. 286. 19007 P 293. 297. 302. 304. 312. 310. 315. 318. 324. 332. 340. 345. 350. 19008 P 19013 P 283. 291. 282. 289. 293. 294. 296. 296. 302. 304. 310. 311. 312. 310. 317. 310. 323. 320. 327. 317. 335. 328. 344. 333. 345. 341. 19014 NP 250. 259. 263. 264. 270. 276. 281. 280. 290. 291. 295. 295. 299. 19015 P 250. 259. 262. 264. 266. 265. 259. 264. 275. 275. 287. 291. 298. 19017 P 258. 262. 261. 275. 269. 275. 278. 283. 288. 292. 298. 301. 316. 19024 P 291. 294. 292. 304. 305. 311. 309. 318. 316. 329. 330. 340. 342. 19029 P 266. 267. 278. 275. 277. 283. 284. 284. 280. 285. 290. 294. 306. 19034 P 266. 274. 280. 284. 289. 292. 294. 300. 308. 313. 321. 334. 339. 19038 NP 237. 243. 254. 262. 261. 265. 273. 278. 281. 282. 285. 291. 296. 19056 P 272. 277. 282. 285. 290. 287. 292. 293. 299. 308. 313. 315. 318. 19057 P 269. 271. 269. 281. 279. 282. 289. 290. 296. 301. 307. 310. 318. 19060 P 276. 271. 278. 288. 277. 295. 301. 302. 306. 314. 320. 329. 334. 19064 P 263. 275. 274. 288. 286. 291. 294. 295. 305. 307. 312. 317. 333. .................................................................................................................................... . . . . . . . . . . . . . . . .D.A.Y. .1.3. . . . . . . .1.4. . . . . . . . .1.5. . . . . . . .1.6. . . . . . . .1.7. . . . . . . .1.8. . . . . . . .1.9. . . . . . . .2.0. . . . . . . . 2. .1. . . . . . . .2.2. . . . . . . .2.3. . . . . . . .2.4. . . . . . . . 2. 5. 19003 P 19007 P 19008 P 19013 P 19014 NP 19015 P 19017 P 19024 P 19029 P 19034 P 19038 NP 19056 P 19057 P 19060 P 19064 P 291. 354. 352. 350. 297. 290. 314. 341. 317. 340. 294. 324. 318. 346. 331. 301. 360. 358. 358. 298. 298. 323. 345. 317. 346. 294. 326. 328. 347. 337. 308. 369. 368. 361. 303. 310. 332. 355. 323. 359. 288. 338. 333. 353. 343. 320. 380. 370. 372. 306. 315. 338. 358. 336. 364. 284. 342. 337. 371. 348. 335. 391. 389. 386. 312. 329. 358. 374. 355. 369. 287. 355. 357. 379. 366. 354. 412. 411. 398. 310. 346. 364. 388. 363. 390. 288. 368. 375. 392. 384. 365. 431. 423. 409. 310. 360. 385. 368. 376. 400. 281. 384. 388. 409. 411. 386. 444. 439. 432. 310. 374. 406. 416. 393. 413. 282. 395. 402. 425. 432. 420. 450. 316. 386. 421. 400. 403. 432. 281. 420. 424. 445. 313. 286. 305. 288. 293. 288. 287. 302. *5(7 0 w 00 5 418-028:PAGE C-73 I I I I I I I I : zI I I# I I I I I I I I I I I I I I I I II . . . . . . . I momwP O W I mom03 r l w m I mmcvcv mmcv I ...... I I I Wco o m 0 0 W I m m ( u ( u l n m P cu I (u cvm m m c v m m I I I I I I . . . . . . . I 1 I I mmwrlm * o I cvocnw rlcvw I mmmcv mmcv I I I ...... I * d c v w m o c o m I c n c v r l w c n m rl I cv cvm m m c v m m I I I I I I . . . . . . . . . I I I I I I I W O N w o P m c o u ) co I m I o c v c o w c v o m P * 0 I (u I m m c v cvm m m c v m m I I I I I I I . . . . . . . I I I I OcvPcn 03mw I NcnPWcndP I mcvcvcv cvmcv I ...... I I I o m * o e r l cv I cv w r l O m P * 0 I cv cvm m m c v m m I I I I I I III . . . . . . . I W o m P *mrl I r l c n P W cnrlco I mcvcvcv cvmcv I . . . . . . W m N W W W W P O c n c v w m cn c v m c v m c v m cv I I I I d I cv I I I I I I . . . . . . . I I I I OPN* NWIn I d W W W cncnP I mcvcvcv cvcvcv I .. coo WO cum I I I 0 I0 cn I cv cv I I I I I I . . . . . . . I I I I P w d ww o o I owcow comr I mcvcvcv cvcvcv I . . . . . . w m c n * W c v W W0-l W d W N W c v c v c v m c v m cv I I I I cn I rl I I I I I I . . . . . . . I I I I W I n O O mwco I O W W W WcnW I mcvcvcv NNcv I . . . . . . * P m m o t - * W c n P r l W r l co c v c v c v m c v m cv I I I I W I rl I I I I I I . . . . . . . I I I I m * m w rlmtI O W P I n comw I mcvcvcv cvcvcv I ...... I I I c u m w w c v r l cn I r W W P O W r l P I rl c v c v c u m r u m cv I I I I I I . . . . . . . I I I I cn*cvW rcocn 1 cncoPUl P W W I cvcvcvcv cvcvcv 1 . . . . . . w o * o c o * m m w POLO0 P c v c v c v m c v m cv I I I I W I rl I I I I I I I I I ....... I I I I ...... I I co I W I cu I I I I I I I I I I I I I I W I cv I W I I I I I I I I I I I I I I I m I W I I I I cv I I I I I I I I I I I I I PIPI :nnaaPIn22PIannnnn: ICJmmcnomWrl*wcvmWcnml 100mmw*~InInmWwWwPI 10000000000000001 llncncncncnmcncncncncnrncncncnl lrlrlrlrldrlrlrlrlrlrldrlrldl I I I I I I I I I I I I I I w I P I cv I I I I I I II cwo I cv I I I I I I I 0 I W I N I I I I I I I I I I I I I I * * ** I I cn 0 0 rl I cv 0 0-l I m I I I I I . . I I W cn m c v cv m I 0 W c o w rl cv * * * I m m m I I I I I . . I I * I cn cv P *w m* 0 cv cn 0 * I m m m m m I I I . . I I I I I 0 P wcn co m m c v 0 t- cvn I mmmmmm I I I . . I I I I d W m w t- cv I P m cvcv m W I mmmmmm I I I I I . . I I 0 W t-v m m * I m cv o r l W I mmmmmm I I I I I . . I I cv cn I m rl * I m m P P m m I I I 0 9 m . . . . . . . W W r l P W c o P cvcncnowow mmmwm*m . . . . . . . W m W m r l * o rlcoPcnIncoco mmmmmmm . . . . . . . * I n W c v m m w owmcnmrlw mmmmmwm . . . . . . . r o o m r l w c n om*wcvo* mmmmmvm ....... I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I- II mw 3I w ; g; % Ilk : g I II nw !I4 II Wx !w Im I I I I I I I I 1 I * * I I I dW cn cv I I rl cv I I mm m m I I I I I I I I I I I I I I co W 0 W I I m0 cv m I I mm m m I I I I 1 I I I I I I I I I !2 !2 I P I PI PI PI PI PI PI PI PI PI PI PI PI I I * I c v In m rn 0 In W rl co cv m W 07 m I * * I 0 0 m m w In m m W W W W t- I I 00 000000 0 0 0 0000 I l c n cn 0-l cn ol cn cn cn cn cn cn cn rn cn m I I r l rl rl rl rl d d d rl rl rl rl rl rl rl I I I I I I I I I PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C34 (PAGE 5): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .................................................................................................................................... RAT # DOSAGE GROUP V 10 MG/KG/DAY .................................................................................................................................... PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 11 12 .................................................................................................................................... 19001 P 223. 229. 233. 237. 235. 242. 239. 243. 251. 250. 251. 255. 266. 19006 P 19011 P 268. 254. 280. 261. 281. 260. 282. 263. 292. 264. 293. 264. 296. 275. 298. 278. 306. 275. 311. 287. 318. 297. 323. 289. 333. 292. 19020 P 236. 234. 249. 254. 256. 259. 257. 263. 269. 276. 283. 311. 293. 19022 P 267. 271. 266. 282. 290. 293. 293. 294. 298. 303. 311. 316. 320. 19025 P 293. 303. 303. 307. 314. 313. 316. 324. 329. 333. 342. 351. 352. 19027 P 248. 252. 251. 260. 256. 260. 269. 270. 272. 274. 282. 287. 290. 19028 P 265. 271. 278. 283. 283. 288. 292. 289. 297. 301. 304. 308. 318. 19030 P 273. 271. 272. 286. 278. 284. 284. 288. 286. 292. 299. 307. 315. 19031 P 281. 290. 285. 297. 298. 303. 303. 313. 322. 318. 327. 337. 346. 19032 P 281. 279. 292. 292. 299. 300. 301. 309. 310. 315. 323. 331. 341. 19033 P 249. 260. 258. 263. 262. 267. 272. 278. 279. 285. 288. 292. 304. 19049 P 256. 267. 278. 281. 287. 284. 295. 296. 302. 307. 309. 312. 329. 19059 P 267. 276. 271. 278. 286. 289. 291. 297. 301. 299. 308. 314. 325. 19070 P 252. 259. 259. 270. 266. 275. 272. 276. 286. 287. 290. 296. 301. .................................................................................................................................... . . . . . . . . . . . . . . . .D.A.Y. .1.3. . . . . . . .1.4. . . . . . . . 1. 5. . . . . . . . 1. 6. . . . . . . . .1.7. . . . . . . .1.8. . . . . . . .1.9. . . . . . . .2.0. . . . . . . . 2. 1. . . . . . . . .2.2. . . .. . . . .2.3. . . . . . . .2.4. . . . . . . . 2. 5. 19001 P 271. 278. 283. 289. 303. 309. 320. 334. 343. 19006 P 336. 347. 353. 366. 379. 397. 422. 436. 451. 19011 P 308. 305. 309. 320. 340. 354. 375. 386. 406. 19020 P 293. 300. 303. 306. 321. 332. 349. 362. 19022 P 321. 328. 337. 345. 361. 369. 389. 416. 391. 19025 P 359. 354. 364. 372. 384. 401. 409. 413. 425. 19027 P 296. 299. 303. 308. 326. 341. 353. 373. 386. 19028 P 324. 327. 338. 344. 354. 365. 377. 394. 411. 19030 P 314. 321. 328. 332. 345. 354. 368. 382. 397. 19031 P 348. 357. 366. 371. 389. 398. 414. 436. 19032 P 344. 350. 355. 355. 375. 390. 404. 417. 447. 19033 P 303. 312. 324. 328. 344. 358. 370. 390. 19049 P 332. 338. 338. 346. 369. 373. 386. 410. 425. 19059 P 329. 333. 344. 352. 363. 376. 383. 405. 427. 19070 P 311. 313. 323. 332. 347. 358. 351. 372. 404. .................................................................................................................................... P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 418-028:PAGE C-75 I I I I I I . . . . . 4 I I I II II I I I m I m w o 0 0 cv ID cv c v r l 0 m 0 0 1 I I * I I d I b, P m b, m w m m m W m m w I I I I I I m m m m m m m m m m m m mcv I I I I I I 1 I I I I I I I I I I I I I I . . . . . . 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I I I I I I I I I I I I I I I I I I I I W I r l w o o w ~ ~ o m ~ r l r l ~ c ~ ~ i r l i m m c v ~ r c v u l w rI l ~ n m w ~ m o I I I m b , b , c v r l w b , b , r l w ~ r m b , r l i c v i m ~ ~ o r l w m w * m * w wI ~ n ~ I I I mmmmmmmmmmmmmmmi i m m m m m m m m m ~ m m m m mI I I I I I I I I I I I I I I 1 1 1 I I I I I I I I I I 1 1 I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I I I I I I I lPlrlPmb,wmwob,PNcvb,NmlolPb,**Nmmwwwww*oPl i3.11 l u l b , b , c v r l w b , m o w w w m m o l N l b , I n w m m P w P m w I n w m c O N l 1 8 ; ;mmmmmmmmmmmmmmmi ~mmmmmmmmmmmmmmmi I 1 I l\l I I I I IWI I I I I Igl l\l I I . . . . . . . . . . . . . . . I I I I I I . . . . . . . . . . . . . . . I I I I I 101 I l I l I l I l I I I ~ I I I I I l I l I l I l I I I ~ I I I I I l I l I l I l I l I I I ~ I I I I I l I l I l I l IHI 1 1 ~~ ~~m~rrl~~rlom~wcv~o~rl~~mr~mwwmm~~ ~mmmcvmmmmmmmmmmmi ~ m m m m m m m m m m m ~ m m m ~ l I I I l I I I I I I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ll I I I l I I I I I I I m~rlm~ooc~c~rlomm~mwm~w~oc~~~mmcv~wrlocvrl Ib,NmrlrlmmNOm**mmcOlrllwPmb,NmcOInmwPcOmwrll ~ m m m m m m m m m m m m ~ m Ic v i I~ m m m m m m m m m m m m m m m ~ l I I I I l I I I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ll I I I l I I I I I I ~~~~rw~~ocvowmwmo~~~~mcv~rlcvww~m ~mcvmwom~rlmm~~o~w~rl~ulwwm~~~~~ ImmmNmmmmcvmmmmmNl Immmmmmmmmmmmmmml l I I I l I I I l I I I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . l I 1 I l I I 1 I I 1 I ~~~cvommwmmulw~momm~w~~w~mm~nrlrmowwwwm ~mcv~wm~cvommm~orl~~rl~wwwul~c~~w~w~ I m m m N N m m m c v m m m m m N Il 1I m m m m m m m m m m m m m m m l l I l I I I l I I I IIl . . . . . . . . . . . . . . . II IfI . . . . . . . . . . . . . . . III ~~~omm~~~~om~o~wm~~ul~mrlrlcvr II 8 W I I 1l w4 I I ; Q 1 0 I I I I I I I I I I I I I I I I I I I# I I I I3 I I I I I I I I I I I I ~ m m m c v c v m m m c v m m m ~I m ~~I i m m m m m m m m m m m m ~ m ~ ~ I I I I I I I I . . . . . . . . . . . . . . I l l I I I I I I I w*(U mPmrlmb,cv w r l m w I I I rlcvo r m m o w c v m m d r l w I 1 I m m m cvmmmcvmm mmmcv I I I I I I I .I wl rll m uI I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I ~ m r ml r l N * o m m c O N * U l I I I rldNNb,**LnLnW W P P P I I I 0000000000 0000 I I I m o mmmmmmmmmmmm I I I rlrlrl rlrlrlrlrlrlrl rlrlrlrl I I I I I I I I I I I I I mI PI 01 mI rll I I I I I I I PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C35 (PAGE 2): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .................................................................................................................................... RAT # DOSAGE GROUP I1 0.3 MG/KG/DAY ________________________________________-------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 .................................................................................................................................... 19004 19009 19016 19018 19026 19036 290. 286. 295. 302. 318. 304. 293. 306. 297. 308. 296. 294. 296. 273. 298. 311. 273.a 297. 301. 251.a 306. 311. 320. 296. 303. 301. 310. 318. 314. 296. 317. 312. 320. 320. 311. 309. 310. 324. 323. 325. 326. 319. 316. 318. 321. 327. 331. 303. 330. 325. 323. 336. 333. 318. 331. 331. 318. 347. 347. 334. 333. 329. 324. 339. 341. 335. 334. 342. 328. 332. 338. 347. 330. 343. 329. 342. 355. 352. 19037 267. 272. 277. 269. 288. 292. 296. 301. 318. 321. 251. 230.a 298. 19043 314. 314. 311. 316. 316. 321. 332. 325. 342. 349. 346. 354. 352. 19047 281. 269. 271. 276. 287. 284. 304. 296. 311. 312. 318. 318. 264 .a 19048 306. 300. 305. 302. 307. 315. 320. 327. 336. 343. 336. 351. 345 * 19052 19055 302. 320. 297. 326. 261.a 320. 264.a 332. 285. 332. 315. 332. 316. 348. 317. 332. 331. 353. 330. 363. 339. 354. 350. 364. 340. 360. 19061 309. 290. 297. 293. 309. 307. 315. 323. 277.a 297. 319. 340. 332. 19067 305. 306. 311. 310. 314. 319. 331. 335. 346. 352. 350. 356. 367. 19071 334. 334. 335. 312. 329. 320. 332. 339. 342. 348. 364. 365. 356. .................................................................................................................................... DAY 14 15 16 17 18 19 20 21 22b .................................................................................................................................... 19004 349. 337. 345. 317. 332. 344. 325. 304. 276. 19009 349. 348. 343. 341. 345. 349. 342. 336. 346. 19016 342. 341. 336. 330. 327. 329. 324. 308. 314. 19018 336. 337. 341. 341. 353. 349. 340. 343. 338. 19026 360. 354. 356. 355. 341. 353. 358. 360. 296. 19036 344. 355. 356. 343. 345. 358. 344. 351. 359. 19037 310. 326. 332. 312. 332. 326. 342. 321. 309. 19043 19047 19048 355. 323. 355. 342. 321. 346. 338. 330. 349. 333. 318. 338. 344. 324. 356. 349. 317. 365. 344. 319. 368. 348. 308. 350. 356. 325. 306. 19052 315.c 352. 353 * 349. 345. 351. 346. 346. 320. 19055 331. 348. 363. 366. 372. 367. 368. 350. 336. 19061 338. 335 * 322. 348. 338. 337. 334. 327. 315. 19067 344. 352. 352. 360. 371. 358. 343. 308.a 294. 19071 362. 316.a 369. 365. 371. 367. 372. 348. 374. .................................................................................................................................... DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value appeared incorrectly recorded and was excluded from group averages and statistical analyses. b. Dams were fasted prior to sacrifice. c. Water access was interrupted; value excluded from group averages and statistical analyses. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C35 (PAGE 3): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .................................................................................................................................... 19003 19007 19008 19013 19014 285. 284. 303. 309. 331. 326. 346. 334. NOT PREGNANT 287. 316. 328. 315.a 293. 316. 335. 348. 300. 316. 316.a 349. 308. 337. 341. 354. 312. 339. 350. 350. 317. 348. 355. 357. 316. 347. 351. 359. 330. 354. 361. 372. 274.a 364. 368. 372. 281.a 363. 361. 389. 320. 373. 358. 390. 19015 19017 19024 19029 19034 1903 8 19056 281. 320. 266. 307. 295. 312. 282. 284. 311. 320. NOT PREGNANT 317. 311. 283. 310. 321. 281. 323. 316. 288. 312. 327. 283. 322. 282.a 277. 313. 332. 290. 328. 321. 295. 322. 332. 301. 309.a 319. 302. 324. 342. 305. 354. 331. 296. 339. 344. 309. 345. 311.a 313. 341. 354. 309. 338. 339. 321. 352. 361. 309. 345. 339. 311. 350. 368. 318. 349. 346. 310. 356. 369. 326. 351. 352. 344. 363. 348. 323. 355. 343. 19057 19060 305. 336. 294. 318. 278.a 330. 300. 328. 311. 342. 320. 345. 324. 353, 324. 348. 321. 358. 339. 366. 336. 369. 350. 360. 352. 373. . . . . .1.9.0.6.4. . . . . . . . . .3.1.1... . . . . .3.0.5... . . . . .3.1.9... . . . . .3.1.8... . . . . .3. 0. 0. .. . . . . . 3. 1. 3. .. . . . . . 3. 1. 9. .. . . . . . 3. 2. 1. .. . . . . . .3.4.0... . . . . .3.4.2... . . . . .3.4.0... . . . . .3.4.4... . . . . .3.0.2. ..a DAY 14 15 16 17 18 19 20 21 22b .................................................................................................................................... 19003 344. 336. 335. 333. 330. 335. 315. 338. 284. 19007 369. 331. 359. 354. 377. 369. 352. 355. 303. 19008 361. 370. 358. 340. 360. 356. 350. 350. 292. 19013 19014 391. 379. NOT PREGNANT 387. 369. 380. 386. 381. 374. 389. 19015 339. 323. 322. 304. 312. 323. 316. 312. 274. 19017 336. 343. 366. 343. 363. 373. 358. 360. 350. 19024 364. 326.a 368. 351. 366. 369. 367. 354. 309. 1902 9 323. 326. 332. 299.a 321. 332. 317. 322. 332. 19034 19038 359. 352. NOT PREGNANT 358. 339. 371. 356. 358. 352. 287. 19056 347 * 341. 343. 346. 348. 329. 350. 338. 292. 19057 351. 337. 343. 352. 337. 356. 354. 305. 287. 19060 362. 350. 371. 373. 351. 372. 351. 352. 357. 19064 344 * 330. 349. 354. 363. 359. 366. 352. 289. .................................................................................................................................... DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value appeared incorrectly recorded and was excluded from group averages and statistical analyses. b. Dams were fasted prior to sacrifice. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C35 (PAGE 4) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .................................................................................................................................... RAT # DOSAGE GROUP IV 3 MG/KG/DAY ____--_-_______L_------------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 .................................................................................................................................... 19002 19005 19035 19039 19040 19045 19046 19051 19054 19058 19062 19063 19066 19069 19073 335. 330. 328. 284. 295. 297. 271. 267. 276. 271. 269. 265. 321. 299. 299. 315. 315. NOT PREGNANT 315. NOT PREGNANT; MATING NOT 280. 296. 291. 309. 320. 276.a 304. 308. 318. 331. 332. 341. 290. 280. 288. 332. 329. 332. 318. 315. 310. 337. 307. 285. 270. 314. 318. CONFIRMED 296. 268.a 317. 348. 292. 332. 306, 338. 317. 289. 272. 315. 304.a 297. 300. 326. 354. 297. 330. 303. 340. 322. 286. 254.a 323. 315. 304. 309. 322. 341. 303. 288.a 321. 345. 320. 292. 283. 328. 310. 303. 313. 332. 357. 301. 275.a 328. 356. 325. 292. 278. 339. 318. 312. 319. 334. 351. 314. 320. 330. 338. 328. 309. 292. 350. 352. 312. 329. 347. 370. 313. 333. 340. 348. 328. 309. 288. 350. 346. 357. 328. 305. 293. 348. 348. 368. 340. 316. 300. 356. 368. 367. 336. 332. 306. 356. 376. 313. 340. 351. 368. 315. 288.a 346. 306. 337. 348. 378. 304.a 337. 352. 290.a 351. 349. 385. 322. 353. 362. 333. 353. 368. 381. 351. 365. 290.a _________________-________c_____________-------------------------------------------------------------------------------------------- DAY 14 15 16 17 18 19 20 21 22b .................................................................................................................................... 19002 19005 19035 19039 19040 361. 341. 315. 311. 360. 358. 342. 322. 310. 355. 360. 351. 314. 310. 350. 369. 320. 306. 296. 334. 358. 344. 322. 320. 365. 376. 352. 326. 317. 361. 378. 350. 315. 313. 367. 359. 333. 312. 317. 356. 302. 285. 261. 315. 294. 19045 19046 19051 349. 377. 361. 353. NOT PREGNANT NOT PREGNANT; MATING NOT CONFIRMED 380. 402. 386. 398. 385. 19054 335. 332. 333. 313. 328. 326. 322. 323. 287. 19058 331. 302.c 345. 350. 345. 364. 363. 324. 350. 19062 369. 358. 356. 335. 352. 355. 354. 342. 290. 19063 377. 380. 383. 370. 384. 388. 377. 385. 369. 19066 338. 332. 332. 325. 336. 341. 340. 328. 292. 19069 349. 361. 366. 365. 373. 375. 358. 352. 362. 19073 350. 357. 368. 375. 371. 364. 365. 348. 363. .................................................................................................................................... DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value appeared incorrectly recorded and was excluded from group averages and statistical analyses. b. Dams were fasted prior to sacrifice. c. Water access was interrupted; value excluded from group averages and statistical analyses. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C35 (PAGE 5): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .................................................................................................................................... RAT # DOSAGE GROUP V 10 MG/KG/DAY .................................................................................................................................... . . . . . . . . . . . . . . . .D.A.Y. . .1. . . . . . . . .2. . . . . . . . .3. . . . . . . . .4. . . . . . . . .5. . . . . . . . . 6. . . . . . . . . .7. . . . . . . . .8. . . . . . . . .9. . . . . . . .1.0. . . . . . . .1.1. . . . . . . .1.2. . . . . . . . 1. 3. . 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 19032 19033 19049 19059 19070 256. 308. 291. 260. 278. 310. 281. 313. 307. 316. 317. 295. 310. 312. 290. 257. 308. 286. 268. 300. 318. 283. 302. 301. 324. 330. 292. 304. 313. 297. 254. 313. 290. 279. 310. 326. 278. 307. 304. 289.a 327. 292. 308. 314. 302. 254. 317. 302. 274. 314. 315.a 274. 310. 309. 286.a 319. 299. 305. 312. 302. 234.a 322. 303. 271. 317. 331. 242.a 310. 318. 319. 309. 305. 286.a 313. 305. 262. 324. 308. 243. 322. 349. 276. 311. 320. 329. 307. 308. 302. 317. 312. 261. 328. 302. 269. 335. 358. 294. 315. 327. 340. 326. 320. 318. 322. 313. 265. 330. 321. 266. 343. 347. 283. 315. 331. 340. 335 * 326. 320. 327. 323. 259. 331. 321. 268. 344. 361. 291. 325. 348. 354. 339. 329. 326. 338. 337. 273. 346. 331. 262. 361. 365. 238. 331. 340. 362. 343. 342. 339. 348. 330. 281. 344. 338. 253. 341. 368. 296. 334. 346. 366. 328. 339 * 335. 347. 337. 278. 339. 326. 268. 360. 381. 304. 331. 346. 371. 284.a 335. 331. 352. 357. 286. 348. 345. 259. 355. 379. 322. 338. 348. 371. 354. 344. 348. 350. 355. .----___-------_____------ - - - - - - - - - _ _ - _ _ - - _ _ _ _ _ _ _ _ _ - _ _ _ _ _ _ _ _ _ _ _ _-_-_- -_-_ _ _ _ 19001 287. 288. 286. 278. 304. 302. 296. 292. 278. 19006 367. 366. 366. 356. 373. 373. 378. 373. 304. 19011 332. 294.a 318. 326. 338. 350. 330. 330. 283. 19020 270. 246. 252. 246. 252. 251. 246. 228. 210. 19022 360. 348. 352. 352. 359. 363. 349. 349. 294. 19025 384. 373. 358. 355. 392. 363. 358. 346. 291. 19027 19028 317. 318. 312. 335. 309. 294.c 306. 329. 302. 340. 321. 348. 315. 330. 315. 346. 266. 339. 19030 357. 355. 353. 344. 346. 346. 365. 355. 314. 19031 316.c 362. 365. 364. 375. 373. 381. 371. 361. 19032 351. 336. 359. 343. 356. 319.a 341. 344. 287. 19033 347. 341. 338. 332. 329. 332. 323. 323. 328. 19049 348. 346. 358. 341. 347. 334. 349. 349. 330. 19059 320. 322. 357. 358. 345. 339. 360. 343. 317. P 19070 303.c 337. 358. 365. 351. 358. 363. 352. 351. .................................................................................................................................... M DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value appeared incorrectly recorded and was excluded from group averages and statistical analyses. 4 W b. Dams were fasted prior to sacrifice. c. Water access was interrupted; value excluded from group averages and statistical analyses. PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T - 7 7 0 6 . 1 ) TABLE C36 (PAGE 1 ) : FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C36 (PAGE 2) : FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C36 (PAGE 3): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C37 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C37 (PAGE 2): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENEFLATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C37 (PAGE 3 ) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C38 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C38 (PAGE 2): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 19002 113. 132. 427. 19005 175. 159. 497. 19035 108. 112. 438. 19039 146. 145. 436. 19040 140. 189. 542. 19045 130. 160. 584. 19046 NOT PREGNANT 19051 NOT PREGNANT; MATING NOT CONFIRMED 19054 113. 125. 360. 19058 98. 184. 496. 19062 147. 175. 560. 19063 131. 160. 568. 19066 110. 139. 453. 19069 106. 81. 451. 19073 114. 167. 523. .......................................................... DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C38 (PAGE 3 : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C39 (PAGE 1): MATING AND FERTILITY, ESTROUS CYCLING AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C39 (PAGE 2): MATING AND FERTILITY, ESTROUS CYCLING AND DAYS IN COHABITATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS P n PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C39 (PAGE 3): MATING AND FERTILITY, ESTROUS CYCLING AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C40 (PAGE 1) : FUNCTIONAL OBSERVATIONAL BATTERY - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS HOME CAGE BEHAVIOR 3 2 3 3 3 3 3 3 3 1 ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 1 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 1 1 REARS IN OPEN FIELD 10 12 12 19 7 13 15 7 14 11 DEFECATION IN OPEN FIELD 1 1 1 2 1 1 2 1 1 1 URINATION IN OPEN FIELD 1 1 2 2 1 2 2 1 1 1 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 1 1 1 1 1 1 1 1 1 1 GAIT ABNORMALITY, SEVERITY 1 1 1 1 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE 1 1 1 1 1 1 1 1 1 1 LACRIMATION 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION 0 0 0 0 0 0 0 0 0 0 ABNORMAL RESPIRATION 0 0 0 0 0 0 0 0 0 0 APPEARANCE 1 1 la 1 lb 1 1 1 1 lc VISUAL REACTION 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION 2 2 2 2 2 2 2 2 2 2 AUDITORY REACTION 3 3 3 3 3 3 3 3 3 3 TAIL-PINCH REACTION 2 2 2 3 2 3 2 2 2 2 AIR RIGHTING RESPONSE 1 1 1 1 1 1 1 1 1 1 PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 41 325 240 550 3 70 655 330 2 75 505 430 FORELIMB GRIP TEST #2 (G) 465 650 345 500 3 55 845 3 75 350 545 600 HINDLIMB GRIP TEST #1 (G) 245 365 450 335 675 565 52 0 2 95 405 3 15 HINDLIMB GRIP TEST #2 (G) 335 445 420 300 670 4 75 350 350 530 415 LANDING FOOT SPLAY #1 (CM) 12.1 8.8 8.0 4.4 6.4 6.5 9.5 4 .O 9.3 7.0 LANDING FOOT SPLAY #2 (CM) 9.6 8.0 8.3 6.0 5.7 5.7 8.0 6.4 9.4 7.7 .-B-O--D-Y--W-E-I-G--H-T--(-G--)-------------------3-7-2--------3-7-4---------3-1-4--------3-2-7--------3--8-2--------3-6-6---------3-4-2--------3-4-2--------3-5--1--------3-6-4----------------------------------- VALUES ARE THE QUANTITY, CATEGORY "MBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. a. Localized alopecia: head (1.0 cm x 0.4 cm) . b. Localized alopecia: left forepaw and forelimb (1.5 cm x 0.6 cm) and right forelimb (0.6 cm x 0.7 cm). c. Ears torn: both pinnae. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C40 (PAGE 2): FUNCTIONAL OBSERVATIONAL BATTERY - INDIVIDUAL DATA - FO GENERATION FEMALE RATS HOME CAGE BEHAVIOR 3 1 3 2 1 3 2 1 3 1 ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 1 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 1 1 REARS IN OPEN FIELD 10 12 12 9 11 14 14 15 18 15 DEFECATION IN OPEN FIELD 1 1 1 1 1 1 1 1 1 1 URINATION IN OPEN FIELD 1 2 1 1 1 1 1 2 1 2 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 1 1 1 1 1 1 1 1 1 1 GAIT ABNORMALITY, SEVERITY 1 1 1 1 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE 1 1 1 1 1 1 1 1 1 1 LACRIMATION 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION 0 0 0 0 0 0 0 0 0 0 ABNORMAL RESPIRATION 0 0 0 0 0 0 0 0 0 0 APPEARANCE 1 1 la 1 1 1 1 1 1 1 VISUAL REACTION 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION 2 2 2 2 2 2 2 2 2 2 AUDITORY REACTION 3 3 3 3 3 3 3 3 3 3 TAIL-PINCH REACTION 2 2 2 2 2 2 2 2 3 2 AIR RIGHTING RESPONSE 1 1 1 1 1 1 1 1 1 1 PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 165 510 2 85 275 325 155 120 400 220 305 FORELIMB GRIP TEST #2 (G) 390 333 140 450 530 145 4 75 535 325 310 HINDLIMB GRIP TEST #1 (G) 335 365 375 385 365 270 420 255 320 410 HINDLIMB GRIP TEST #2 (G) 315 380 525 3 55 285 365 665 3 75 240 325 LANDING FOOT SPLAY #1 (CM) 6.0 8.7 7.9 5.2 6.3 6.2 6.5 5.2 6.2 5.8 LANDING FOOT SPLAY #2 (CM) 7.9 9.0 6.9 6.8 7.4 6.7 7.0 5.4 5.7 6.5 ._B_OD-Y_-W_EI_G_H_T _(-G_)______---------3-1-7--------3-5-6---------3-5-2--------3-5-0---------3-3-4--------3-6-2--------3--3-5--------3-5-6--------3--4-5--------3-6-5------------------------------------ VALUES ARE THE QUANTITY, CATEGORY NUMBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. a. Localized alopecia: head (0.4 cm x 0.5 cm) . PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C40 (PAGE 3): FUNCTIONAL OBSERVATIONAL BATTERY - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS HOME CAGE BEHAVIOR 3 3 1 3 ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 1 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 .1 1 REARS IN OPEN FIELD 14 9 10 8 6 13 14 10 16 16 DEFECATION IN OPEN FIELD 1 1 1 1 1 1 1 3 1 2 URINATION IN OPEN FIELD 1 1 1 1 2 1 1 1 2 2 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 1 1 1 1 1 1 1 1 1 1 GAIT ABNORMALITY, SEVERITY 1 1 1 1 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE 1 1 1 1 1 1 1 1 1 1 LACRIMATION 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION 0 0 0 0 0 0 0 0 0 0 ABNORMAL RESPIRATION 0 0 0 0 0 0 0 0 0 0 APPEARANCE 1 1 la 1 lb 2 1 lc 1 1 VISUAL REACTION 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION 2 2 2 2 2 2 2 2 2 2 AUDITORY REACTION 3 3 3 3 3 3 3 3 3 3 TAIL-PINCH REACTION 2 2 2 2 2 3 2 2 2 3 AIR RIGHTING RESPONSE 1 1 1 1 1 1 1 1 1 1 PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 325 975 85 200 420 330 695 320 610 255 FORELIMB GRIP TEST #2 (G) 245 535 75 365 455 720 665 3 95 505 300 HINDLIMB GRIP TEST #1 (G) 295 280 370 4 95 445 450 465 435 455 290 HINDLIMB GRIP TEST #2 (G) 445 3 15 320 470 375 530 440 520 605 365 LANDING FOOT SPLAY #1 (CM) 5.4 7.0 6.2 10.8 6.8 7.7 8.8 6.2 7.4 8.3 LANDING FOOT SPLAY #2 (CM) 6.3 8.5 5.8 10.6 7.2 6.4 10.4 8.0 7.5 8.4 ._B-O_DY__WE_I_G_HT__(. G-.I--_____---------3-2--3--------3-6-5--------3-6--0--------3-8-6--------3-2-5---------3-6-9--------3-5-9---------3-1-6--------3-4-4---------3-5-9------------------------------------ ~ ~~ ~ VALUES ARE THE QUANTITY, CATEGORY NUMBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. a. Chromorhinorrhea. b. Localized alopecia: head (0.3 cm in diameter). c. Localized alopecia: both forepaws and forelimbs (2.8 cm x 0.5 cm) and head (0.3 cm x 0.5 cm). PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C40 (PAGE 4): FUNCTIONAL OBSERVATIONAL BATTERY - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS HOME CAGE BEHAVIOR 3 1 2 3 1 2 3 2 1 1 ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 1 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 1 1 REARS IN OPEN FIELD 11 7 15 12 13 10 10 11 8 10 DEFECATION IN OPEN FIELD 3 1 1 1 2 1 1 1 1 1 URINATION IN OPEN FIELD 1 1 1 1 1 1 1 1 1 1 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 1 1 1 1 1 1 1 1 1 1 GAIT ABNORMALITY, SEVERITY 1 1 1 1 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE 1 1 1 1 1 1 1 1 1 1 LACRIMATION 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION 0 0 0 0 0 0 0 0 0 0 ABNORMAL RESPIRATION 0 0 0 0 0 0 0 0 0 0 APPEARANCE 1 1 1 1 la 3b 1 lc 1 1 VISUAL REACTION 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION 2 2 2 2 2 2 2 2 2 2 AUDITORY REACTION 3 3 3 2 3 3 3 3 3 3 TAIL-PINCH REACTION 2 2 2 2 2 2 2 2 2 2 AIR RIGHTING RESPONSE 1 1 1 1 1 1 1 1 1 1 PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 415 355 445 4 90 560 525 260 310 545 4 95 FORELIMB GRIP TEST #2 (G) 685 360 3 95 465 555 605 435 225 530 390 HINDLIMB GRIP TEST #1 (G) 465 255 190 355 340 350 220 400 465 270 HINDLIMB GRIP TEST #2 (G) 470 295 3 15 430 435 325 365 460 485 365 LANDING FOOT SPLAY #1 (CM) 8.0 5.5 5.4 6.5 8.0 7.5 7.4 7.5 7.0 4 .O LANDING FOOT SPLAY #2 (CM) 8.3 5.8 6.3 7.2 9.4 9.6 9.0 7.0 7.5 5.0 ._B-O-D--Y__WE-_IG_H_T__(_G)_-__----------------3--3-0----------3-2-2----------3-1-7----------3-5-3----------3-8-0----------3~ -3-9-~ --. -------3-5--5---------3-5--1---------3--6-8---------3--3-7---------------------- VALUES ARE THE QUANTITY, CATEGORY NUMBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. a. Localized alopecia: left axilla (4.0 cm x 3.0 cm). b. Localized alopecia: right forepaw and forelimb (1.7 cm x 0.6 cm). c. Tip of tail missing. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C40 (PAGE 5): FUNCTIONAL OBSERVATIONAL BATTERY - INDIVIDUAL DATA - Fo GENERATION FEMALE FLATS HOME CAGE BEHAVIOR 2 2 2 3 2 3 1 3 3 3 ALTERATIONS (HOME CAGE) 1 1 1 1 1 1 1 1 1 1 REACTION TO REMOVAL 1 1 1 1 1 1 1 1 1 1 REACTION TO HANDLING 1 1 1 1 1 1 1 1 1 1 REARS IN OPEN FIELD 12 9 9 9 9 10 14 8 15 14 DEFECATION IN OPEN FIELD 1 1 1 1 1 1 1 2 1 2 URINATION IN OPEN FIELD 1 1 1 1 1 1 2 2 1 1 LEVEL OF AROUSAL 3 3 3 3 3 3 3 3 3 3 ALTERATIONS (OPEN FIELD) 1 1 1 1 1 1 1 1 1 1 GAIT PATTERN 4a 1 1 4a 1 1 1 1 1 1 GAIT ABNORMALITY, SEVERITY 2 1 1 2 1 1 1 1 1 1 PALPEBRAL CLOSURE 1 1 1 1 1 1 1 1 1 1 PROMINENCE OF THE EYE LACRIMATION 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 SALIVATION 1 1 1 1 1 1 1 1 1 1 PILOERECTION ABNORMAL RESPIRATION APPEARANCE VISUAL REACTION 0 0 0 0 0 0 0 0 0 0 0 0 0 lc 0 0 0 0 0 0 1 lb 1 Id 1 1 1 1 le 1 2 2 2 2 2 2 2 2 2 2 TACTILE REACTION AUDITORY REACTION TAIL-PINCH REACTION 2 2 2 2 2 2 2 2 2 2 2 3 3 3 3 3 3 3 3 3 2 2 2 2 2 2 2 2 2 3 AIR RIGHTING RESPONSE 1 1 1 1 1 1 1 1 1 1 PUPIL RESPONSE TO LIGHT 1 1 1 1 1 1 1 1 1 1 FORELIMB GRIP TEST #1 (G) 170 785 525 205 560 535 3 15 270 595 270 FORELIMB GRIP TEST #2 (G) 290 735 465 240 645 540 600 525 4 95 440 HINDLIMB GRIP TEST #1 (G) 385 4 10 330 435 385 520 265 3 05 4 05 410 HINDLIMB GRIP TEST #2 (G) 380 530 490 370 630 430 4 10 480 53 0 445 LANDING FOOT SPLAY #1 (CM) 4.8 6.3 9.0 8.8 10.5 10.0 6.3 8.7 6.2 10.4 LANDING FOOT SPLAY #2 (CM) 5.8 7.0 9.4 8.7 8.5 9.0 6.5 9.2 7.0 9.2 .-B""O"D"Y""W"E""I"G"H"T"""("G")""-------------------2-5--0---------3-6-5---------3-2-0---------2--5-4---------3-6-6----------3-6-2---------3-1-8---------3--4-3---------3-3--2---------3-7-3---- VALUES ARE THE QUANTITY, CATEGORY NUMBER, QUANTAL RESPONSE OR SCORE RECORDED FOR EACH ITEM IN THE BATTERY. a. Tip-toe walk. b. Ear torn: right pinna. c. Rales. d. Localized alopecia: both forepaws and forelimbs (1.0 cm x 0.4 cm); Dehydration; Abdominal distention. e. Scab: head (0.3 cm in diameter); Localized alopecia: head (0.5 cm in diameter). BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 84 77 76 63 74 73 73 69 60 71 86 17 67 0 66 52 61 59 65 65 83 30 65 0 72 64 18 64 63 45 70 7 25 2 23 3 33 9 108 6 65 3 43 2 49 4 54 9 33 13 76 3 11 0 57 2 34 10 6 40 39 41 20 0 4 1 3 4 4 0 32 53 23 0 0 11 24 1 10 39 12 37 7 0 13 55 54 29 1 6 20 41 4 20 1 5 71 8 1 25 17 4 10 1 2 0 30 0 23 5 71 5 10 2 1 1 19 9 59 4 57 4 21 1 1 1 2 2 27 5 40 1 9 0 37 2 8 4 8 51 67 2 2 1 75 6 3 3 7 56 83 2 3 0 47 5 1 39 22 30 81 4 5 37 79 4 0 70 39 3 59 12 10 0 36 4 0 59 74 5 1042 343 411 129 637 2 93 488 448 610 469 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C41 (PAGE 2): MOTOR ACTIVITY - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DOSAGE GROUP I 0 MG/KG/DAY FLAT "MBER 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 BLOCK 1 191 138 161 108 167 180 157 185 125 136 BLOCK 2 157 34 147 0 116 84 100 147 85 83 BLOCK 3 119 41 75 0 96 98 29 81 71 47 BLOCK 4 113 7 25 3 17 2 27 10 101 1 BLOCK 5 83 4 48 0 43 1 105 8 31 13 BLOCK 6 111 1 6 0 75 1 37 12 1 31 BLOCK 7 37 50 23 0 1 1 2 1 2 0 BLOCK 8 35 93 28 0 0 8 31 0 12 47 BLOCK 9 5 60 6 0 14 67 62 41 0 0 BLOCK 10 26 50 5 36 0 4 95 7 0 29 BLOCK 11 22 6 10 2 4 0 29 0 43 2 BLOCK 12 74 3 10 3 0 1 18 12 103 5 BLOCK 13 68 2 21 0 2 0 0 0 42 5 BLOCK 14 42 1 9 0 36 1 5 1 1 52 BLOCK 15 77 1 0 0 119 5 1 4 5 89 BLOCK 16 71 1 0 0 74 4 0 79 17 24 BLOCK 17 100 2 3 38 92 3 0 113 56 0 BLOCK 18 114 21 7 0 40 6 0 106 84 4 P c-r TOTAL 1445 515 584 190 8 96 466 698 807 779 568 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C41 (PAGE 3): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP I1 0.3 MG/KG/DAY RAT NUMBER 19004 19009 19016 19018 19026 19036 19037 19043 19047 19048 BLOCK 1 65 68 53 60 75 73 73 65 77 62 BLOCK 2 36 75 61 55 55 64 68 27 16 79 BLOCK 3 31 104 3 6 57 58 50 3 22 78 BLOCK 4 49 41 7 1 54 20 27 16 10 82 BLOCK 5 29 46 0 9 30 12 8 5 0 73 BLOCK 6 22 45 1 7 3 0 30 28 0 58 BLOCK 7 3 18 4 22 3 29 9 12 0 78 BLOCK 8 13 59 2 2 0 2 29 8 0 17 BLOCK 9 0 79 0 1 1 3 10 5 2 4 BLOCK 10 1 22 4 12 5 2 14 2 0 3 BLOCK 11 0 1 6 1 54 0 1 4 0 3 BLOCK 12 7 1 5 11 10 0 7 2 0 16 BLOCK 13 4 6 1 7 38 12 4 6 2 70 BLOCK 14 40 2 14 8 41 0 30 0 4 51 BLOCK 15 30 3 0 6 17 5 50 2 10 7 BLOCK 16 42 32 1 6 3 4 42 3 9 8 BLOCK 17 4 63 13 13 0 4 6 2 10 2 BLOCK 18 6 41 3 1 0 27 25 0 4 1 90 TOTAL 3 82 706 178 228 446 3 15 483 190 166 6 92 0 h) 00 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C41 (PAGE 4): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP I1 0.3 MG/KG/DAY RAT NUMBER 19004 19009 19016 19018 19026 19036 19037 19043 19047 19048 BLOCK 1 BLOCK .2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 131 213 85 76 152 181 120 120 133 2 04 41 168 93 49 96 136 14 0 41 19 157 47 135 3 7 82 92 91 1 35 108 67 56 4 0 116 12 60 18 8 139 32 55 0 9 51 15 19 1 0 93 20 76 0 7 0 0 60 25 0 122 1 11 3 31 3 46 8 9 0 102 12 84 2 1 0 2 36 4 0 14 0 119 0 0 1 1 12 3 0 2 0 29 1 11 4 1 15 0 0 1 0 0 6 3 70 0 2 0 0 2 6 0 4 7 9 0 6 1 0 9 2 3 0 4 42 19 3 10 2 62 33 0 17 13 72 0 36 0 1 69 43 1 0 3 22 1 71 1 8 5 62 36 0 4 1 2 55 0 9 7 2 91 12 11 0 3 11 0 6 1 4 40 0 0 0 28 22 0 1 1 503 1117 230 236 721 539 767 2 34 222 1098 0 0 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C41 (PAGE 5 ) : MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP I11 1 MG/KG/DAY RAT NUMBER 19003 19007 19008 19013 19015 19017 19024 19029 19034 19056 BLOCK 1 76 74 55 71 69 70 74 69 58 67 BLOCK 2 61 40 63 70 68 59 85 81 68 71 BLOCK 3 65 65 70 19 30 37 59 48 48 61 BLOCK 4 40 60 48 3 61 19 61 56 6 85 BLOCK 5 18 27 89 5 64 25 60 70 43 35 BLOCK 6 37 49 78 1 84 36 58 65 9 9 BLOCK 7 46 50 70 1 74 40 55 44 22 43 BLOCK 8 49 57 64 2 40 38 53 38 34 5 BLOCK 9 9 53 57 2 8 51 24 65 10 2 BLOCK 10 5 85 74 0 2 27 24 38 27 12 BLOCK 11 0 57 64 1 1 29 58 53 1 5 BLOCK 1 2 3 1 71 10 3 16 59 41 2 5 BLOCK 1 3 45 9 78 0 3 27 26 50 1 4 BLOCK 1 4 58 1 77 0 2 32 6 64 1 3 BLOCK 1 5 52 7 59 1 1 61 10 39 4 2 BLOCK 1 6 43 1 85 19 6 39 1 46 0 5 BLOCK 1 7 51 7 60 0 8 43 1 70 1 6 BLOCK 1 8 47 4 42 1 5 10 28 39 1 2 TOTAL 705 647 1204 206 52 9 659 742 976 336 422 izI PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C41 (PAGE 6): MOTOR ACTIVITY - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DOSAGE GROUP I11 1 MG/KG/DAY RAT NUMBER 19003 19007 19008 19013 19015 19017 19024 19029 19034 19056 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 180 153 118 150 153 201 174 97 186 132 96 69 13 9 109 138 12 9 123 85 119 137 128 109 143 19 21 66 176 30 136 57 64 99 85 2 78 22 98 66 9 179 23 27 136 11 123 49 84 88 53 40 59 75 102 0 119 53 81 59 2 11 65 55 140 0 154 63 70 61 24 58 69 90 123 0 78 65 56 49 69 8 13 68 109 0 7 87 14 71 10 0 4 122 112 0 1 36 27 47 26 12 0 77 100 1 0 41 93 84 0 1 3 0 93 10 1 22 136 73 4 2 71 7 158 0 2 29 24 90 0 3 60 0 125 0 1 47 4 106 0 2 80 9 109 0 0 97 11 56 2 0 69 0 116 24 9 57 0 83 0 3 91 9 138 0 12 47 0 77 0 4 56 1 52 0 0 7 22 42 1 0 1131 970 2098 326 897 1118 1193 1264 641 649 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C41 (PAGE 7): MOTOR ACTIVITY - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DOSAGE GROUP IV 3 MG/KG/DAY RAT NUMBER 19005 19035 19039 19040 19045 19054 19058 19062 19063 19066 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 59 82 75 62 55 68 67 49 71 69 63 77 58 18 66 75 48 69 37 60 52 51 63 11 67 75 27 41 32 74 51 33 64 2 79 62 13 40 32 14 54 37 73 1 50 52 5 33 6 28 64 7 15 15 56 56 2 54 12 69 78 5 4 57 66 48 2 57 11 55 66 34 28 29 44 75 22 49 36 62 57 27 9 1 52 43 39 55 33 24 55 0 9 0 59 43 18 49 22 4 50 6 5 0 59 35 3 47 26 4 54 3 3 32 53 6 16 35 14 2 68 1 0 3 55 37 3 21 7 3 60 0 9 11 53 52 3 53 3 0 46 5 3 1 69 25 1 44 1 3 58 4 39 2 50 4 1 31 81 2 54 19 17 4 57 2 0 10 52 4 50 3 2 0 76 0 14 24 17 4 1039 3 94 4 76 249 1066 758 2 84 761 4 93 481 TOTAL = SUM OF BLOCKS; EACH BLOCK CONSISTS OF A 5 MINUTE PERIOD. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C41 (PAGE 8): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP IV 3 MG/KG/DAY RAT NUMBER 19005 19035 19039 19040 19045 19054 19058 19062 19063 19066 BLOCK 1 241 163 158 215 236 181 154 143 150 169 BLOCK 2 167 129 110 23 188 184 67 158 63 106 BLOCK 3 106 63 100 26 196 155 45 69 41 143 BLOCK 4 130 42 89 1 192 133 23 52 43 25 BLOCK 5 108 29 87 0 14 0 58 16 60 4 18 BLOCK 6 120 5 29 22 131 64 5 82 20 89 BLOCK 7 117 2 2 79 150 73 1 149 12 68 BLOCK 8 108 42 45 37 76 73 60 98 56 106 BLOCK 9 112 31 14 0 194 65 56 109 68 17 BLOCK 10 83 0 10 0 154 39 29 79 21 1 BLOCK 11 83 2 3 0 137 31 5 77 31 3 BLOCK 12 63 1 2 38 66 2 24 47 22 1 BLOCK 13 135 0 0 0 112 33 11 36 5 2 BLOCK 14 100 0 4 6 101 40 6 73 1 0 BLOCK 15 58 4 0 0 150 9 1 107 0 1 BLOCK 16 83 2 54 1 81 1 2 62 98 2 BLOCK 17 105 23 13 1 119 0 0 10 77 1 BLOCK 18 73 1 1 0 152 0 24 27 14 1 * P TOTAL 1992 539 72 1 449 2575 1141 52 9 1438 726 753 0 P PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C41 (PAGE 9): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP V 10 MG/KG/DAY RAT NUMBER 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 BLOCK 1 72 76 61 75 63 83 52 78 65 59 BLOCK 2 30 4 59 28 59 45 27 45 64 64 BLOCK 3 20 8 63 3 63 9 41 7 58 39 BLOCK 4 7 2 52 0 56 5 42 1 5 35 BLOCK 5 5 1 64 0 46 27 31 2 4 6 BLOCK 6 6 18 59 0 39 29 17 6 10 13 BLOCK 7 6 1 57 1 23 7 23 16 3 3 BLOCK 8 61 1 61 48 63 49 19 3 3 18 BLOCK 9 57 4 61 3 44 54 0 9 14 33 BLOCK 10 43 2 30 35 28 4 4 7 2 0 BLOCK 11 43 4 65 1 41 1 7 24 5 6 BLOCK 12 52 7 64 48 38 1 34 30 9 14 BLOCK 13 46 4 62 5 46 1 1 51 2 3 BLOCK 14 16 4 38 1 47 2 0 40 3 17 BLOCK 15 7 3 44 0 40 3 0 48 0 6 BLOCK 16 5 1 52 39 36 32 2 17 6 2 BLOCK 17 1 2 67 50 46 10 0 2 2 3 BLOCK 18 6 24 52 44 39 47 1 5 4 4 8 TOTAL 483 166 1011 381 817 409 301 3 91 259 325 I ? PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C41 (PAGE 10): MOTOR ACTIVITY - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP V 10 MG/KG/DAY RAT NUMBER 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 BLOCK 1 BLOCK 2 BLOCK 3 BLOCK 4 BLOCK 5 BLOCK 6 BLOCK 7 BLOCK 8 BLOCK 9 BLOCK 10 BLOCK 11 BLOCK 12 BLOCK 13 BLOCK 14 BLOCK 15 BLOCK 16 BLOCK 17 BLOCK 18 TOTAL 162 12 8 132 104 152 151 148 147 134 2 04 17 1 120 16 157 119 121 104 123 188 13 14 122 0 125 6 58 4 107 74 6 1 60 0 126 1 66 0 2 84 3 3 87 0 86 29 55 0 2 11 6 25 100 0 49 56 35 6 14 19 1 1 94 0 21 2 25 20 1 0 52 0 84 54 106 76 23 1 2 19 78 5 94 2 81 72 0 3 16 50 34 0 39 19 35 1 2 11 0 0 54 2 98 0 53 0 5 17 4 13 90 7 81 65 68 0 39 20 4 13 49 4 121 4 86 0 0 55 1 7 28 2 51 0 89 0 0 47 1 38 3 2 58 0 54 0 0 51 0 5 1 0 65 27 34 45 0 9 3 1 0 1 86 61 61 4 0 2 1 1 1 34 78 35 50 69 0 3 3 3 598 230 1570 387 1433 631 577 500 418 730 0 m PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C42 (PAGE 1): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA FO GENERATION FEMALE RATS/F~GENERATION LITTERS PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C42 (PAGE 2): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS/Fl GENERATION LITTERS .................................................................................................................................... DURATION OF LITTER DELIVERED NUMBER OF LIVE PUPS AT TOTAL RAT/ GESTATION LIVE STILL- TOTAL COMPLETION OF DAY POSTPARTUM IMPLAN- LITTER (DAYS) BORN BORN BORN 1 8 15 22 TATIONS NUMBER N N N N MF MF MF MF N .................................................................................................................................... MATERNAL DOSAGE GROUP I11 1 MG/KG/DAY .................................................................................................................................... 19003 22 16 0 16 88 88 88 88 17 19007 22 18 0 18 7 11 7 10 7 10 7 10 18 19008 22 15 0 15 78 77 77 77 16 19013 23 15 0 15 96 96 96 96 15 19014 NOT PREGNANT 19015 23 15 0 15 96 96 96 96 15 19017 23 14 0 14 68 68 68 68 14 19024 23 15 0 15 96 96 96 96 16 19029 23 16 0 16 97 87 87 87 17 19034 23 13 1 14 67 67 67 67 15 19038 NOT PREGNANT 19056 23 14 0 14 77 77 77 77 14 19057 23 17 0 17 98 98 98 98 17 19060 22 14 0 14 86 86 86 86 15 19064 23 18 0 18 99 99 98 98 17 .................................................................................................................................... MATERNAL DOSAGE GROUP IV 3 MG/KG/DAY .................................................................................................................................... 19002 23 lO(1) 1 11 36 36 36 36 14 19005 23 16 0 16 6 10 6 10 6 10 6 10 16 19035 23 14 0 14 68 68 68 68 16 19039 22 15 0 15 69 59 59 59 15 19040 22 17 0 17 7 10 7 10 7 10 7 10 17 19045 23 15 0 16 [ 1 1 69 69 69 69 16 19046 NOT PREGNANT 19051 NOT PREGNANT; MATING NOT CONFIRMED 19054 23 5 0 5 32 32 32 32 6 19058 23 13 0 13 85 85 85 85 14 19062 22 15 0 15 96 96 96 96 15 19063 23 13 1 14 2 11 2 11 2 11 2 11 16 19066 23 13(1) 1 14 57 57 57 57 15 19069 23 17 0 17 6 11 6 11 6 10 6 10 17 19073 23 14 0 14 86 76 76 76 15 .................................................................................................................................... M = W E F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. [ ] = NUMBER OF PUPS IN WHICH CANNIBALIZATION AND/OR AUTOLYSIS PRECLUDED THE DETERMINATION OF VIABILITY. I I I I I I * I I E I U I x H I I 0 E I I I I I I I I I I I I I I I I I I I I I I w H X E I x w ul W 0 P P I I I I I 1 I 1 I I I I I I I t I 1 I I I I I I I I I I I I I I I I I I I I I I I 1 I I I 1 I I I I I I I I I I I 1 I I I I 418-028:PAGE C-109 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 19-11 I .. I ;~;PNwmWt'oPFPPm*mmI I rl rl rl I I l\l I I I ~ ~ ~ j * w t ' w m rml * m w w t ' rwlrlr l oI w rI l 1 I01 I I lrll I I I I I I I I I I I 1 I I I I I I 1Hl I 1 I I t I 0 E I h m W N 0 I W *rl 131 19-1 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C43 (PAGE 1): PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION LITTERS 19010 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 19068 19072 19074 19075 6.8 6.6 6.6 16.4 15.9 16.1 28.4 27.7 27.9 41.9 40.0 40.6 6.6 6.4 6.5 13.8 13.6 13.7 26.5 25.5 25.8 42.7 40.6 41.2 6.6 6.2 6.4 15.4 14.4 14.9 28.6 26.5 27.6 40.0 37.2 38.7 6.4 5.9 6.2 12.7 11.9 12.4 23.4 22.1 22.9 35.8 33.8 35.1 6.3 6.2 6.2 16.8 16.5 16.7 32.8 32.1 32.5 46.1 44.4 45.4 6.2 6.0 6.1 13.0 13.0 13.0 24.1 23.8 23.9 32.6 32.5 32.5 6.5 6.4 6.4 14.1 13.6 13.8 26.4 26.3 26.4 36.4 35.6 35.9 5.7 6.0 5.8 13.8 13.0 13.4 26.8 25.2 26.0 42.6 40.5 41.5 6.6 6.2 6.3 14.6 13.7 14.0 27.1 25.7 26.2 36.4 35.1 35.6 6.4 6.4 6.4 15.6 15.4 15.5 29.3 28.2 28.8 35.9 35.5 35.7 6.0 5.7 5.8 13.2 12.2 12.7 24.8 23.4 24.0 32.4 30.8 31.6 6.8 6.4 6.6 17.2 15.8 16.3 29.7 27.4 28.2 47.7 45.1 46.0 5.9 5.5 5.7 12.3 11.9 12.1 22.7 21.6 22.2 31.7 30.0 30.8 7.1 6.7 6.8 17.7 16.8 17.0 27.4 27.4 27.4 35.2 34.0 34.4 6.2 6.3 6.2 12.9 13.0 12.9 22.3 23.1 22.6 36.1 36.1 36.1 19004 19009 19016 19018 19026 19036 19037 19043 19047 19048 19052 19055 19061 19067 19071 5.7 5.4 5.5 13.5 11.9 12.7 24.5 22.3 23.3 31.1 28.8 29.9 7.2 7.0 7.0 15.4 15.5 15.5 34.4 33.5 33.8 56.4 53.2 54.1 6.4 6.1 6.4 14.6 12.9 14.2 27.7 23.1 26.8 46.3 38.1 44.6 7.1 6.5 6.8 15.3 14.4 14.9 28.3 25.6 27.2 45.6 41.6 43.8 7.1 6.5 6.8 13.7 12.1 13.0 25.0 22.7 24.0 31.8 29.4 30.7 6.2 5.6 5.8 11.6 9.6 10.4 26.9 24.1 25.4 41.1 36.8 38.8 5.6 5.4 5.5 11.5 11.2 11.4 18.3 18.0 18.2 31.2 29.2 30.3 6.6 6.2 6.4 14.0 13.4 13.7 29.4 28.3 28.9 46.4 43.7 45.2 7.2 6.8 7.0 15.3 14.4 14.9 28.2 27.0 27.6 38.2 37.3 37.8 6.4 6.2 6.3 16.1 16.2 16.1 33.1 32.4 32.8 49.0 46.4 47.8 6.2 5.9 6.0 11.0 10.8 10.8 22.6 22.8 22.8 35.8 35.9 35.9 6.7 6.4 6.6 14.3 13.1 13.9 26.4 25.1 26.0 39.8 37.2 38.9 5.8 5.6 5.7 13.8 12.8 13.4 24.8 24.6 24.8 32.2 33.8 32.8 7.0 6.5 6.7 13.7 13.3 13.5 29.4 28.1 28.7 40.2 38.8 39.4 6.5 6.7 6.6 11.4 11.7 11.5 24.8 24.5 24.7 40.0 39.7 39.9 M = MALE F = FEMALE T = TOTAL (SUM OF PUP WEIGHTS/NUMBER OF LIVE PUPS) DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C43 (PAGE 2): PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION LITTERS .................................................................................................................................... RAT/ LITTER DAY 1 DAY 8 DAY 15 DAY 22 NUMBER M F T M F T M F T M F T .................................................................................................................................... MATERNAL DOSAGE GROUP I11 1 MG/KG/DAY .................................................................................................................................... 19003 19007 19008 19013 19014 19015 19017 19024 6.6 6.0 6.3 6.3 5.8 6.0 6.2 5.7 6.0 7.0 6.4 6.8 NOT PREGNANT 6.7 6.2 6.5 7.4 6.9 7.1 6.2 5.8 6.1 13.5 13.7 14.7 15.2 12.1 12.3 14.1 13.9 12.8 12.9 14.4 14.7 14.0 14.8 14.5 13.1 14.4 13.4 13.7 14.5 14.1 23.2 24.5 29.2 27.9 21.3 21.7 28.1 25.1 22.2 22.9 28.6 26.8 26.3 27.6 27.4 24.9 25.8 25.4 25.8 26.6 26.6 30.4 33.6 41.5 47.0 28.1 30.5 39.0 42.3 29.2 31.8 40.2 45.1 38.4 45.8 40.0 35.9 44.2 37.5 37.4 44.9 39.0 19029 6.3 5.8 6.1 14.1 12.2 13.2 26.3 22.2 24.4 43.7 34.6 39.5 19034 6.8 6.4 6.6 15.3 14.3 14.8 30.4 28.6 29.4 41.2 39.0 40.0 19038 NOT PREGNANT 19056 6.8 6.6 6.7 15.1 14.8 14.9 29.6 28.8 29.2 41.4 39.8 40.6 19057 6.6 6.4 6.5 13.1 12.9 13.0 23.6 23.2 23.4 32.7 32.0 32.4 19060 6.9 6.4 6.7 14.8 13.4 14.2 29.2 26.4 28.0 46.6 42.8 45.0 19064 6.7 6.2 6.5 12.1 11.5 11.8 22.8 22.2 22.5 27.8 30.2 28.9 .................................................................................................................................... MATERNAL DOSAGE GROUP IV 3 MG/KG/DAY ________________________________________-------------------------------------------------------------------------------------------- 19002 7.6 7.1 7.3 19.1 17.9 18.3 35.5 33.1 33.9 48.0 45.4 46.3 19005 6.5 5.8 6.0 13.2 13.0 13.1 24.2 23.7 23.9 33.2 31.7 32.2 19035 7.2 6.7 6.9 12.9 12.5 12.6 23.3 22.5 22.8 33.8 32.3 32.9 1903 9 5.9 5.4 5.6 11.1 10.2 10.6 22.9 21.2 21.8 35.1 32.4 33.4 19040 5.4 5.1 5.2 11.6 10.9 11.2 25.2 23.4 24.1 31.7 29.3 30.3 19045 6.5 6.6 6.6 11.7 12.0 11.9 24.0 23.6 23.8 37.6 36.2 36.7 19046 , NOT PREGNANT 19051 NOT PREGNANT; MATING NOT CONFIRMED 19054 7.4 7.5 7.4 20.1 21.2 20.6 41.2 40.4 40.9 57.1 56.6 56.9 19058 6.6 6.5 6.6 14.9 14.6 14.8 27.8 27.2 27.6 46.4 45.1 45.9 19062 6.9 6.6 6.8 16.7 15.8 16.3 30.7 29.4 30.2 42.4 40.1 41.4 19063 7.0 6.9 6.9 14.6 15.2 15.1 29.8 30.0 29.9 47.6 46.2 46.4 19066 6.5 6.3 6.4 15.8 15.9 15.9 30.9 30.8 30.8 43.8 43.0 43.3 19069 5.9 5.7 5.8 10.3 9.7 9.9 19.3 20.4 20.0 30.1 33.4 32.2 19073 7.2 6.8 7.0 13.9 12.8 13.4 27.7 25.6 26.8 44.3 40.9 42.8 .................................................................................................................................... M = MALE F = FEMALE T = TOTAL (SUM OF PUP WEIGHTS/NUMBER OF LIVE PUPS) DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C43 (PAGE 3): PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION LITTERS .................................................................................................................................... RAT/ LITTER DAY 1 DAY 8 DAY 15 DAY 22 . . N. U. M. B. E. R. . . . . . . . .M. . . . . .F. . . . . .T. . . . . . .M. . . . . .F. . . . . .T. . . . . . .M. . . . . .F. . . . . .T. . . . . . . M. . . . . . .F. . . . . T. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . MATERNAL DOSAGE GROUP V 10 MG/KG/DAY .................................................................................................................................... 19001 6.8 6.6 6.6 14.0 14.3 14.2 26.6 26.6 26.6 44.1 44.0 44.1 19006 6.5 6.1 6.2 13.8 13.1 13.3 25.4 23.7 24.2 35.2 32.7 33.4 19011 6.9 6.6 6.8 13.9 13.6 13.8 26.2 25.5 25.9 36.4 35.5 36.0 19020 5.8 5.5 5.7 9.2 9.1 9.2 15.7 15.4 15.6 15.4 15.5 15.5 19022 6.0 5.7 5.9 15.3 14.9 15.2 29.0 27.0 28.3 36.3 34.5 35.7 19025 5.4 5.0 5.3 12.9 11.0 12.3 24.2 20.4 22.9 30.8 26.8 29.4 19027 7.3 6.5 6.8 12.6 12.1 12.2 23.6 21.9 22.4 33.0 31.1 31.6 19028 6.7 6.3 6.4 13.1 13.2 13.2 25.9 25.9 25.9 39.8 40.1 40.0 19030 6.4 6.1 6.2 15.0 14.3 14.6 29.5 28.1 28.7 40.1 38.2 39.1 19031 5.9 5.7 5.8 12.3 11.9 12.1 27.6 25.7 26.6 44.8 41.1 43.0 19032 7.3 6.9 7:l 14.6 13.2 13.9 25.9 24.4 25.2 35.9 34.2 35.1 19033 6.6 6.0 6.2 13.8 12.6 13.1 27.4 25.2 26.2 44.1 41.0 42.4 19049 6.7 6.2 6.6 12.6 11.0 12.2 25.4 22.8 24.7 36.8 32.9 35.7 19059 7.0 6.5 6.9 12.3 11.0 11.9 24.9 21.3 23.7 34.6 30.4 33.2 _ _ _1_9_07_0_ _ _ _ _6_.5_ _ _6_._3 _ _6_._4_ _ _15_._4 _ _14_._8 _ _15_._2_ _ _29_._8 _ -2-8-.-0- - -2-9-.-2- - - -4-9-.-6- - -4- 6- -.-8- - 4- 8- -.-7- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - . M = MALE F = FEMALE T = TOTAL (SUM OF PUP WEIGHTS/NUMBER OF LIVE PUPS) DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T - 7 7 0 6 . 1 ) TABLE C44 (PAGE 1): PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS 19010 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 19068 19072 19074 19075 6.8 7.0 7.4 6.0 6.9 6.8 7.3 6.9 5 . 1 6.5 6.8 7.0 6.3 6.0 6.5 6.3 6.5 6.9 7.0 6.4 6 . 1 6.5 6.8 6.2 6.6 6.2 6.7 6.6 6 . 1 6.4 6.8 6.2 6.5 6.6 6.4 6.9 6.7 6.8 6.3 6.4 6.3 6.0 6.2 6.3 6.0 6.7 6.4 6.5 6.7 6.7 6 . 1 6.4 6.4 6.2 5.6 5.5 6 . 1 6.2 6.0 6.9 6.2 6.2 5.8 6 . 1 6.2 6.5 6.0 6.1 6.2 6 . 1 6.2 6.3 6.3 5.8 6 . 1 6.3 6.1 6.6 6.5 5.9 6.0 6.6 6.4 6.3 5.9 5.8 6.0 6.0 5.4 5.6 6.2 7.2 6.9 6.3 6.5 6 . 1 6.3 6.4 6.4 7.0 6.5 6 . 1 6.3 6.5 6 . 1 5.8 5.0 5.7 5.8 5.9 6 . 1 6 . 1 5.3 5.4 6.3 6 . 1 6.7 6.0 6.6 6.3 5.4 5 . 1 6.9 6.6 6.9 6.5 6.2 5.9 6.3 5.7 6.2 6.5 6.0 6 . 1 6 . 1 6.4 6.5 6.5 6.6 6.5 6 . 1 6.2 6.9 6.5 6.8 6.0 6.2 6.5 6.4 6.3 6.6 6.3 6.3 5.9 5.8 5.6 5.9 6.0 5.9 6 . 1 6.0 6.7 5.2 5.7 6.0 6.2 5.8 5.4 5.4 5.9 5.7 5.6 5 .7 7.0 6.6 6.6 7.0 6.8 7.0 6.2 6.8 6.7 6.3 6.5 6.0 6.0 6.6 6.7 5.9 5.7 6 . 1 6.0 5.5 6.0 6 . 1 6.0 5.5 5.7 5.5 5.4 5.6 5.6 5.2 5.5 6.9 7 . 2 7.3 7.0 MD 1 7 . 0 6 . 8 7 . 1 6.6 7 . 1 7.0 6.0 6 . 6 6.3 6 . 7 6.4 6 . 1 5.5 6.4 5.8 6.4 6.4 6.3 6.5 6.5 6.2 6.4 6.3 6.3 6.0 6.3 RAT/ LITTER # MATERNAL DOSAGE GROUP I1 0.3 MG/KG/DAY POSTPARTUM DAY 1 19004 5.6 5.9 5.9 5.8 5.9 5.2 5.5 5.8 5.0 5.5 5.7 5.3 6 . 1 5.2 5.3 4.9 5.8 5.4 19009 7.5 7.0 6.7 6.9 7 . 1 6.8 7.4 19016 6.6 6.3 6.5 6 . 1 6.9 6.7 6.4 6.7 6.3 6.0 5.9 6.8 5.7 6.2 6.5 19018 7.2 6.7 7.2 6.9 7.2 7.2 7.7 6.9 6.1 6.6 6.4 6.4 6.9 6.5 19026 6.7 7.1 7.3 7.3 6.9 6.7 7.4 7 . 1 7.4 7.0 6.7 6.9 6.6 6.4 6 . 1 6.7 5.7 6.6 6.5 19036 6 . 1 6.4 6 . 6 6 . 0 6 . 2 5 . 6 6 . 6 5 . 7 4 . 9 5 . 8 4 . 6 5.5 5.6 5 . 9 5 . 7 FD 1 P w 19037 5.7 5.9 5.6 5.6 5.9 5.6 5.6 4.9 5 . 1 5.8 5.9 5.3 5.0 5.4 5.3 5.4 00 19043 6.8 6.7 6 . 1 6.9 6.8 6.7 6.6 6.3 5.8 6.2 6.4 6.2 6.0 19047 7 . 1 7.7 7.9 7.6 6.6 6.5 7 . 1 6.8 6.5 6.8 6.7 6.7 7.5 19048 6.7 6.6 6.4 6.3 5.9 6.2 6.2 6.4 5.9 19052 6.0 6.1 6.3 6.3 6.3 6.1 6.1 5.6 5.8 6.5 5.4 6.3 6.1 5.6 6.0 5.7 5.7 19055 6 . 8 6 . 5 7 . 2 6 . 6 7.0 6 . 8 6.4 6 . 6 6 . 7 6 . 4 MD 1 6 . 8 6 . 6 6 . 2 6.3 6 . 0 19061 5.9 5.9 5.8 5.6 5.7 6.3 5.7 5.6 5.8 5.6 5.7 5.6 5.3 19067 7.2 6.8 7.3 6.9 7 . 1 7.0 6.5 7.0 6.2 6.2 6.6 6.8 6.5 6.2 19071 6.6 6.3 5.4 7.4 6.6 6.9 6.7 6.4 7.0 6.5 6.5 6.5 6.9 6.9 ................................................................................................................. ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C44 (PAGE 2): PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS 19002 7.7 7.6 7.5 MS 7.2 7.1 6.9 7.2 7.3 7.1 FD 1 19005 6.9 6.2 7.0 5.9 6.4 6.4 6.2 5.9 5.6 5.4 5.6 6.0 5.7 6.0 6.0 5.7 19035 7.3 7.3 7.4 7.2 6.9 6.9 6.6 6.6 6.6 7.0 6.7 6.5 6.7 6.8 19039 6.4 5.8 5.4 5.9 5.7 6.4 5.4 5.9 5.5 5.5 5.5 4.8 5.4 6.0 5.1 19040 19045 5.5 5.1 5.8 4.9 5.7 5.3 5.3 5.2 5.1 5.2 5.3 5.4 5.4 5.4 4.8 4.8 6.5 7.1 6.5 7.1 5.4 6.5 6.2 6.7 6.5 6.8 6.8 6.3 6.3 6.4 7.2 uu 19046 NOT PREGNANT 19051 NOT PREGNANT; MATING NOT CONFIRMED 19054 7.0 7.5 7.7 7.6 7.4 19058 6.2 7.0 6.9 6.3 6.6 6.6 6.3 7.1 6.7 6.4 6.1 6.8 6.6 19062 7.2 6.9 6.9 6.9 6.9 6.9 7.0 7.3 6.4 6.1 6.6 6.6 6.6 6.7 6.7 19063 6.7 7.3 7.6 6.8 6.5 6.5 6.8 6.6 6.5 7.3 6.7 7.1 7.4 FS 19066 6.4 7.2 6.6 6.1 6.2 6.3 MD 1 6.2 5.9 6.0 6.7 6.3 6.5 FS 19069 6.1 5.8 6.1 5.1 6.1 6.0 6.3 5.7 5.2 5.8 5.7 6.2 4.5 5.8 5.8 5.8 19073 7.2 7.0 6.8 7.8 6.9 7.2 7.7 7.4 7.0 6.7 6.4 6.6 7.1 6.7 ...................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN, U-UNCERTAIN,D-DIED,M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C44 (PAGE 3): PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS .................................................................................................................................... PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 .................................................................................................................................... RAT/ LITTER # MATERNAL DOSAGE GROUP V 10 MG/KG/DAY POSTPARTUM DAY 1 .................................................................................................................................... 19001 6.9 6.4 6.8 7.0 6.7 6.1 6.5 6.6 6.8 6.5 6.7 19006 7.0 6.3 6.6 7.0 6.1 5.8 5.9 6.1 6.4 6.5 5.6 6.5 6.2 6.0 6.7 5.9 6.0 5.9 FD 1 FS 19011 7.4 7.3 6.3 7.0 6.9 6.7 6.8 7.1 6.7 6.5 6.2 6.6 6.3 19020 6.3 6.0 6.1 6.7 4.5 5.4 5.7 5.6 4.9 5.2 5.5 5.4 6.3 19022 6.2 6.0 5.8 6.1 5.7 6.5 5.9 6.0 6.1 MD 1 5.4 5.5 5.4 5.9 5.9 6.0 19025 5.1 5.0 5.2 5.4 5.4 5.4 5.6 6.1 5.1 5.6 5.7 5.5 4.8 5.2 4.8 5.2 5.2 4.9 5.0 5.0 19027 7.3 6.9 8.1 6.9 7.2 6.0 6.4 6.8 6.8 6.6 5.9 6.3 6.9 6.5 19028 6.8 7.1 6.1 6.8 6.6 6.2 5.9 6.6 6.5 6.5 6.5 5.9 FS 19030 6.6 5.9 6.8 6.9 6.0 6.4 5.9 6.3 4.9 6.5 6.3 6.1 6.7 19031 6.2 6.3 6.2 5.0 5.9 5.8 5.9 5.9 6.1 5.7 5.5 5.5 5.5 5.9 5.9 19032 7.6 7.4 7.2 7.4 7.2 7.1 7.4 6.9 6.9 7.2 7.0 7.1 6.7 6.7 19033 6.4 6.7 6.5 6.9 6.4 6.6 6.0 5.9 5.9 6.1 6.0 5.8 6.4 5.9 19049 7.0 6.4 7.0 6.4 7.3 6.6 6.8 6.4 6.7 6.3 7.2 6.8 6.0 6.1 5.9 19059 7.2 7.0 6.6 6.8 7.2 7.0 7.4 7.0 7.3 6.8 6.5 6.5 6.4 6.5 6.8 19070 6.8 6.6 6.0 6.5 6.5 6.4 6.3 6.7 6.2 5.7 6.6 6.7 6.4 FD 1 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE! PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED,M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-028:PAGE C-116 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I 031 I W *rl . . * w m* rlrl I In I n I I h I I I m I I m I rl I I I I I I I I I I I I I I ..... I w w W P 9 mI .I rl m * m rl m I rl rl rlrl rl rll I I wml-wmwrlm+w~ ........*.I mmommowrlmm1 rlrlrlrlrlrlrlrlrlrll . . . . . . . . . . . . . , P I l w * * r l l - r l m m m w * m m P o l lrlrlrlrlrlrlrlrlrlrlrlrlrlrlrll I I I I I l-Pmommml-wl-wwovml I ...............I I wmmml-mmrlmmmmml-41 I rlrlrlrlrlrlrlrlrlrlrlrlrlrlrll I I I I I Ommmdrlm*ommwrlmml I ...............I I wm*mw*mmmInmmrlm*l I rlrlrlrlrlrlrlrldrlrlrlrlrlrll I 2 : n t . . . . . . . . . \ I WI 1 mmw*wl-mrlrlmo*wwm1 .....I wmmrlmmmmm~owrll-ml rlrlrlrlrlrlrlrlrl"rlrlrlrlrll \I I " : . . . . . . . . . . . . . . . W I r l * o w m r l m m w o o l - r l w m II I 0 1 m m m r l w w ~ O m w m m m w mI I rlrlrlrlrlrlrlrlrirlrlrlrlrlrl I I I . . . . . . . . . . . . . . . I I I mrll-mol-Pmwwmmwmw I I I 1 wm*mwmmmrlmmwmwo I I rlrlrlrlrlrlrlrlrlrlrlrlrlrlrl I I I I I I I I I I I I I I I . . . . . . . . . . . . . I I lommol-mmrll-mw*mrlol I .( ~l-mwmmmvmm*rl~mnm~ lddrldrldrlrlrlrlrlrld~rll HI I I I rl 0 rl . . . . . ~ w r m m wmrlrlm rlrlrlrl . . . . . . . o m r l w * m m *mmPrlm* rlrlrl rlrlrl . . . . . . . o w o w t - m o mmmwomw rlrlrl rlrlrl . . . . . . . w w m * w w w Trmmwrlmm rlrlrl rlrlrl . . . . . . . m r l * m w m m *mmmom* rlrlrlrlrlrlrl . . . . . . . . . . . . . . U I .I ~X~inmmml-mmm~mml-mwmi mlrlrlrlrlrlrlrlrlrlrlrlrlrlrlrll I dl . . . . . . . . . . . . . . . I ~w~mmwwmmm~~rrlwm~ 1 ...............I 2tm*mmwom*mmql-owwi lrlrlrlrlrlrlrlrlrldrlrlrlrlrll I I I I I #I I #I m R . . 0 0 mm rl . . w m dm rlrl I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I. I VL YL .... I omwrlm I h *I0 rl*m*m I rlrlrlrl lrn rlrlmmc) rlrlrlrlrl w I I3 I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I' I I I I I PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C44 (PAGE 5): PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS _________________________________^______-------------------------------------------------------------------------------------------- _ _ _ _P_UP_ _# _ _ _ _1_ _ _ _2_ _ _ _ 3_ _ _ _ _4_ _ _ _5_ _ _ _6_ _ _ _7_ _ _ _ -8- - - - - -9- - - - -1-0- - - - -1-1- - - - -1- 2- - - - - -1-3- - - - -1-4- - - - -1-5- - - - -1-6- - - - -1-7- - - - - -1-8- - - - -1-9- - - - -2-0- - - - - - - - - - - - - - - - - RAT/ _L_I_TT_E_R_#_ _ _ _M_A_TE_R_NA_L_ D_O_S_AG_E_G_R_OU_P_ _I1_1_ _ _ _ _ _ _ _ _ _ _ _ 1_ -M-G-/-K-G-/-D-A- Y- - - - - - - - - - - - - - P- -O-S-T-P-A-R-T-U-M- -D-A-Y- - -8- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 19003 13.9 13.6 14.3 11.1 13.5 14.1 13.5 13.7 11.7 12.9 12.3 11.1 10.9 11.5 14.2 12.5 19007 19008 14.1 13.7 13.3 13.7 12.5 14.8 13.9 11.7 13.5 11.2 12.7 11.9 12.2 12.9 11.7 12.8 12.5 FD 2 13.7 14.8 15.4 15.6 15.1 14.7 13.7 14.8 14.0 13.6 14.6 15.2 14.0 12.5 FM 2 19013 15.8 14.9 14.0 15.1 15.1 15.3 15.9 15.4 15.6 14.2 12.6 15.3 12.7 13.4 15.0 19014 19015 NOT PREGNANT 13.9 14.5 14.3 14.2 15.2 13.5 14.1 12.9 13.9 11.1 13.2 13.8 13.8 13.3 13.2 19017 14.1 15.7 15.7 14.4 15.0 13.6 13.6 15.3 12.3 15.3 15.1 15.2 14.2 13.8 19024 19029 19034 14.6 14.3 15.1 14.2 14.1 15.0 14.7 13.5 15.5 13.8 14.3 15.4 14.3 14.9 14.4 13.7 15.7 15.0 14.0 13.8 14.1 15.1 14.9 14.9 MM 2 12.7 14.6 13.6 12.7 14.8 12.7 10.9 15.2 13.3 12.6 14.0 15.5 11.7 14.8 13.5 12.1 13.0 11.8 FS 12.7 19038 NOT PREGNANT 19056 15.0 14.9 16.0 14.6 15.4 14.3 15.4 14.1 15.4 14.5 15.5 15.2 14.5 14.3 19057 13.2 13.1 12.6 13.5 12.8 13.0 13.2 12.4 13.8 14.0 14.2 12.1 11.9 12.1 12.0 13.9 12.8 19060 16.1 14.9 16.2 14.9 13.2 14.1 14.0 14.6 12.9 12.2 14.5 13.0 15.8 12.0 19064 11.1 11.6 14.3 10.1 10.8 10.9 13.8 11.7 14.9 9.7 11.5 10.7 11.3 13.8 9.3 13.6 10.3 13.4 ________________________________________-------------------------------------------------------------------------------------------- RAT/ _.L_I_T_TE_R_#_ _ _ _M_A_TE_R_NA_L_ D_O_S_AG_E_G_R_OU_P_ _IV_ _ _ _ _ _ _ _ _ _ _ _ _3_ -M-G-/-K-G-/-D-A-Y- - - - - - - - - - - - - -P-O-S- T- P- A- R- T- U- -M- -D-A-Y- - -8- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 19002 19.8 19.2 18.4 MS 18.3 17.9 17.3 17.9 17.9 18.2 FD 1 19005 13.6 11.9 14.8 12.2 12.5 14.0 13.4 12.2 12.8 13.9 13.3 13.9 13.1 12.9 12.3 12.7 19035 19039 13.0 13.2 12.9 13.2 12.3 12.6 12.2 11.0 13.5 12.7 12.1 13.6 12.9 11.8 10.4 11.2 12.1 11.6 10.1 MD 2 9.0 8.9 11.5 9.8 9.9 9.9 10.8 11.2 11.3 19040 12.5 10.5 11.8 13.0 9.7 12.0 11.8 9.8 11.3 8.5 11.2 12.8 9.9 12.7 12.0 11.1 10.1 f: 19045 19046 12.0 12.8 11.5 11.7 9.8 12.4 12.3 12.7 11.9 11.1 11.3 13.1 12.9 11.8 11.0 UU NOT PREGNANT 19051 NOT PREGNANT; MATING NOT CONFIRMED 19054 19058 20.7 20.4 19.2 21.5 21.0 14.4 13.9 14.8 14.6 16.6 14.8 14.6 15.2 15.9 14.2 14.2 13.7 15.1 19062 19063 19066 16.2 15.7 16.6 17.3 13.4 15.7 17.3 15.3 15.5 17.1 14.0 15.9 16.7 16.3 15.5 17.4 15.9 16.5 16.4 16.7 MD 1 15.0 13.4 15.7 16.7 14.8 17.1 15.7 15.3 16.3 16.9 15.1 15.3 16.2 16.1 15.0 15.5 13.8 15.4 15.6 FS FS 15.2 19069 10.6 10.2 11.4 10.1 8.3 11.0 9.8 10.1 10.4 8.4 10.8 10.3 10.7 6.2 9.4 10.0 10.4 19073 15.4 13.4 13.6 13.5 13.5 14.5 13.6 MM 3 12.9 13.3 12.4 13.3 12.7 11.9 ________________________________________-------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED,M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C44 (PAGE 6): PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS ________________________________________-------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 .................................................................................................................................... RAT/ _L_I_TT_E_R_#_ _ _ _M_A_TE_R_NA_L_ D_O_S_AG_E_G_R_OU_P_ _V _ _ _ _ _ _ _ _ _ _ _ _ _ 1_0- -M-G-/-K-G-/- D- A- -Y- - - - - - - - - - - - P- O- -S-T-P-A-R-T-U-M- -D-A-Y- - -8- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 19001 14.3 13.6 13.2 14.8 14.4 14.3 14.0 15.3 14.1 14.3 13.7 19006 12.9 13.5 13.8 14.8 14.2 MD 2 13.4 14.0 13.0 12.7 12.8 12.1 13.7 11.5 12.8 15.4 12.8 13.4 FD 1 FS 19011 13.9 14.2 13.8 14.3 13,3 14.1 13.7 13.0 13.4 13.2 14.9 13.2 14.1 19020 8.5 9.3 9.1 8.8 8.6 8.4 10.8 9.8 7.7 8.3 9.4 9.5 10.8 19022 15.8 15.3 16.2 14.8 15.3 15.2 14.8 15.6 14.5 MD 1 14.5 15.7 14.1 14.3 16.0 FM 2 19025 12.5 10.6 14.5 12.7 12.3 13.7 12.6 15.0 9.9 12.1 12.4 15.3 14.4 11.5 11.9 12.3 10.5 10.5 10.3 10.2 19027 12.0 13.2 13.1 12.2 10.8 11.8 11.8 11.6 12.2 13.5 12.6 12.8 12.6 10.9 19028 13.1 13.7 13.3 11.7 13.8 13.2 12.9 13.7 12.6 14.5 13.4 12.4 FS 19030 15.6 14.5 14.9 15.7 13.7 15.3 15.0 14.4 14.5 11.9 13.3 15.5 15.4 19031 12.7 10.7 12.4 11.3 13.6 12.9 12.6 MM 2 13.1 11.8 11.0 13.8 12.4 10.3 10.8 19032 13.6 15.2 15.0 14.6 15.2 13.4 14.9 12.5 14.0 14.1 13.0 12.5 13.4 13.1 19033 13.5 13.9 13.5 14.7 14.0 13.3 12.7 12.3 13.5 12.2 12.6 12.1 13.0 12.4 19049 11.5 13.7 13.1 11.0 14.1 12.8 12.5 12.1 14.8 12.7 10.4 12.5 12.7 9.3 9.4 19059 10.3 11.5 12.8 13.1 12.5 13.0 12.3 13.0 12.1 12.4 9.9 11.5 11.4 11.9 10.5 . 19070 ...... . . . . . . 15.8 ..... . 13.2 ..... . 14.3 ..... . 15.2 ..... . 16.1 ..... . 16.4 ..... . .1.5...8. . . 1.6...0. . . 1. 4. .. 7. . . 1. 6. .. 9. . . 1. 4. .. .8. . .14. .. 9. . . 1. 2. ...9. . F. D. . .1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN,U-UNCERTAIN, D-DIED,M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/D.EVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T - 7 7 0 6 . 1 ) TABLE C44 (PAGE 7 ) : PUP BODY WEIGHTS FROM BIRTH TO DAY 2 2 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS ________________________________________------------------------------------------------------------------------------------------- _ _ _ _P_UP_ _#_ _ _ _1_ _ _ _2_ _ _ _ 3_ _ _ _ _4 _ _ _ _5_ _ _ _6_ _ _ _ 7_ _ _ _ -8- - - - - -9- - - - -1-0- - - - -1-1- - - - -1- 2- - - - - -1-3- - - - -1-4- - - - -1-5- - - - -1-6- - - - - 1- -7- - - - -1-8- - - - -1-9- - - - -2-0- - - - - - - - - - - - - - - - - RAT/ _L_IT_T_E_R _#_ _ _ _M_A_TE_R_NA_L_ D_O_S_AG_E_G_R_OU_P_ _I _ _ _ _ _ _ _ _ _ _ _ _o_M_G-/-K-G-/-D-A-Y- - - - - - - - - - - - - - P- O- S- T- -P-A-R-T-U-M- -D-A-Y- -1-5- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 19010 28.0 2 8 . 9 2 6 . 7 2 9 . 8 28.3 3 0 . 1 2 7 . 6 2 7 . 1 27.0 2 8 . 6 28.2 2 8 . 9 23.5 FD 3 19012 26.2 25.8 26.8 27.4 26.2 26.1 26.2 22.3 26.7 25.2 23.7 25.9 28.9 23.9 23.3 28.0 19019 28.9 29.3 25.2 30.6 28.4 .29.1 27.7 29.7 27.2 25.4 22.9 28.6 25.5 28.8 27.0 19021 23.3 23.1 24.8 22.8 21.4 23.5 23.2 24.0 24.1 21.3 22.8 21.8 22.6 22.2 19023 3 1 . 0 3 1 . 6 3 4 . 7 3 3 . 2 3 2 . 3 3 4 . 8 3 1 . 8 3 1 . 0 32.8 3 2 . 8 3 2 . 6 3 1 . 1 FD 4 19041 26.0 21.6 22.9 26.1 23.7 24.3 24.3 27.1 25.4 20.5 25.6 25.0 25.8 23.6 23.4 23.2 19.6 22.4 19042 26.9 26.5 28.3 26.1 23.4 27.3 26.3 29.4 28.3 26.2 27.0 22.5 26.2 24.8 26.1 19044 2 3 . 6 3 0 . 7 2 2 . 2 2 6 . 1 2 6 . 2 2 9 . 8 2 9 . 8 2 5 . 9 2 5 . 6 2 7 . 2 2 1 . 6 2 5 . 3 2 6 . 1 2 5 . 8 2 6 . 9 2 3 . 1 FM 2 19050 26.3 26.8 28.3 28.3 25.8 26.9 25.0 24.8 23.1 27.5 25.5 26.4 26.0 27.7 23.9 19053 2 9 . 0 3 1 . 0 2 8 . 4 2 8 . 6 2 7 . 1 3 1 . 1 2 9 . 6 2 9 . 5 MD 5 2 7 . 1 2 6 . 9 2 7 . 7 2 8 . 7 2 8 . 8 2 8 . 1 3 0 . 4 19065 2 9 . 2 2 5 . 1 2 3 . 3 2 5 . 1 2 2 . 5 2 4 . 2 1 9 . 6 2 7 . 8 26.8 2 7 . 2 2 5 . 0 2 4 . 6 2 2 . 9 2 5 . 8 1 9 . 9 2 0 . 8 2 5 . 5 2 2 . 2 1 9 . 6 FD 5 19068 29.4 2 9 . 5 28.3 3 0 . 0 31.3 MD 2 2 9 . 2 2 6 . 5 28.2 2 7 . 6 28.4 25.6 26.7 2 7 . 3 27.5 19072 20.6 23.4 21.2 23.3 24.0 25.6 20.0 23.8 23.1 20.4 21.3 22.3 20.7 21.4 21.9 21.5 19074 2 9 . 1 2 8 . 8 2 7 . 6 2 4 . 0 MD 1 28.2 26.2 2 8 . 2 27.6 26.3 26.8 27.2 25.8 2 8 . 2 29.0 . 1 . 9 . 0 . 75 ... . . . . . . 23. ... 7 . . . 23. ... 8 . . . 21. ... 6 . . . .2.0.. .5 . . .2.3. ..0. . . 1. 9. ..4. . . 2. 3. .. 1 . . . 2. 4. ..0. . . 2. 1. ..9. . . 2. 2. ..2. . . 2. 2. ..6. . . 2. 4. ..5. . . 2. 2. .. 7 . . . 2. .2 .. 2 . . . .2 .1.. 0 . . . .2 .4.. 9 . . . . . . . . . . . . . . . . . . . . . . . . . RAT/ _L_I_TT_E_R _#_ _ _ _M_A_TE_R_NA_L_D_O_S_AG_E_G_R_OU_P_ _I1_ _ _ _ _ _ _ _ _ _ _ _0_.3_ -M-G-/-K-G-/-D-A- Y- - _ - - - - - - - - -P-O-S-T-P-A-R-T-U-M- -D-A-Y- -1-5- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 19004 2 2 . 4 2 5 . 9 2 5 . 7 2 4 . 5 2 0 . 9 2 6 . 3 2 3 . 6 2 6 . 5 1 8 . 7 2 1 . 1 1 9 . 0 2 1 . 5 2 6 . 8 2 3 . 0 2 2 . 6 2 3 . 2 2 4 . 6 FM 6 19009 34.4 34.4 32.6 32.8 33.2 35.0 33.9 19016 27.5 29.1 27.8 23.7 28.5 29.6 30.0 27.1 28.2 26.3 26.1 29.0 25.4 1 7 . 1 26.7 19018 29.6 27.0 2 8 . 1 30.5 26.6 2 7 . 1 28.8 28.8 26.3 23.1 28.4 26.4 24.9 24.6 19026 29.2 24.0 2 3 . 0 27.3 2 6 . 1 24.3 2 2 . 3 2 4 . 0 23.9 2 5 . 8 2 4 . 9 22.2 23.6 2 2 . 9 21.3 2 1 . 8 2 3 . 2 2 1 . 7 FD 8 19036 2 5 . 4 2 7 . 0 2 8 . 0 2 7 . 2 2 7 . 1 2 6 . 8 2 5 . 0 1 8 . 3 2 5 . 6 2 9 . 0 2 0 . 2 2 7 . 3 2 3 . 3 FMll FD 2 FD 1 19037 1 8 . 8 1 8 . 6 1 6 . 8 1 9 . 2 1 5 . 6 1 9 . 5 2 0 . 2 1 7 . 9 1 9 . 3 1 9 . 7 1 5 . 9 1 7 . 8 1 9 . 6 1 7 . 5 1 5 . 9 FD15 19043 29.4 30.6 28.6 28.3 29.7 29.1 30.2 27.8 27.9 27.5 28.3 28.9 29.3 19047 29.5 28.7 29.2 29.5 26.9 28.0 25.8 26.5 23.3 29.0 28.8 29.3 24.8 19048 31.8 33.9 31.9 33.8 34.0 30.9 31.9 33.9 33.1 19052 23.4 21.5 22.0 24.4 25.7 18.7 23.3 24.8 24.3 2 1 . 1 24.9 22.7 23.3 23.3 19.3 21.5 FD 5 19055 26.2 27.8 27.4 26.5 2 4 . 7 2 6 . 1 2 8 . 0 26.5 23.7 27.5 MD 1 25.0 24.4 2 3 . 6 2 6 . 8 25.6 19061 2 4 . 5 2 4 . 8 2 5 . 5 2 5 . 1 2 4 . 5 2 5 . 0 2 4 . 6 MD 3 2 4 . 9 2 4 . 3 2 4 . 8 2 4 . 1 2 5 . 1 19067 2 9 . 5 2 9 . 9 2 9 . 9 2 9 . 4 2 9 . 0 2 8 . 9 2 9 . 4 2 7 . 9 26.8 2 7 . 7 2 8 . 6 2 7 . 5 2 8 . 6 FD 8 19071 26.1 24.7 24.9 27.3 26.0 22.2 22.8 24.7 25.0 18.1 27.0 26.7 22.7 27.4 ............................................................................................................................. . ALL WEIGHTS WERE RECORDED IN GRAMS (G) MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVEl S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER'' INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C44 (PAGE 8): PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS ________________________________________-------------------------------------------------------------------------------------------- . . . . . P. U. .P. .#. . . . . .1. . . . . .2. . . . . .3. . . . . .4. . . . . .5. . . . . .6. . . . . .7. . . . . . 8. . . . . . 9. . . . . 1. 0. . . . . 1. 1. . . . . .1.2. . . . .1.3. . . . .1.4. . . . .1.5. . . . .1.6. . . . .1.7. . . . .1.8. . . . .1.9. . . . . 2. 0. RAT/ . L. I. T. T. E. .R. .#. . . . . .M.A.T.E.R.N.A.L. .D.O.S.A.G.E. .G.R.O.U.P. .I.1.1. . . . . . . . . . . . . . . . 1. .M.G./.K. G. /. D. A. Y. . . . . . . . . . . . . . P. O. S. T. .P.A.R.T.U.M. .D.A.Y. .1.5. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19003 19007 19008 19013 19014 19015 19017 19024 19029 19034 19038 19056 19057 19060 19064 24.4 21.2 23.5 24.8 25.5 25.5 31.1 28.7 29.9 29.0 27.3 27.4 NOT PREGNANT 27.5 25.0 29.2 27.1 28.9 28.4 26.0 27.7 28.8 26.9 27.4 23.1 30.5 30.5 30.8 NOT PREGNANT 28.7 30.6 30.8 21.9 24.4 22.8 27.7 31.8 29.9 21.6 25.0 21.1 25.1 25.1 26.9 28.4 23.2 27.9 29.8 26.1 29.8 29.5 24.6 30.3 23.9 23.5 25.3 28.3 28.8 25.9 27.8 26.4 27.0 30.9 27.8 22.2 27.2 26.5 24.8 24.1 29.2 29.4 24.7 25.7 26.3 25.8 29.8 31.3 24.6 29.0 21.9 24.5 21.5 30.5 28.3 27.7 28.2 26.7 26.7 27.6 28.2 24.1 28.5 22.6 18.8 23.4 29.2 27.9 26.7 28.1 25.7 27.4 28.2 29.7 24.3 29.3 20.8 21.2 21.4 27.8 25.0 27.2 26.4 28.9 MM 2 27.8 27.6 23.3 28.7 22.0 19.1 23.6 26.8 24.3 28.2 26.4 27.9 20.1 29.8 27.7 22.8 29.5 21.9 23.3 23.9 29.8 24.1 25.0 28.4 25.2 20.6 29.9 30.1 23.5 25.1 26.4 19.6 21.3 26.9 26.6 20.5 26.4 24.7 20.9 26.9 30.1 24.2 23.5 16.8 21.9 19.6 29.9 28.0 24.9 18.1 26.1 22.4 29.8 27.9 24.3 27.1 17.4 20.3 20.2 26.2 25.8 26.5 24.8 24.9 24.5 FS 28.3 22.9 24.6 21.6 21.3 21.0 FM 2 22.0 24.2 23.8 23.3 21.9 24.3 23.4 21.1 23.8 22.9 26.7 21.4 22.8 22.1 FD 2 FMll __________________________________^_____-------------------------------------------------------------------------------------------- RAT/ _L_I_TT_E_R_#_ _ _ _M_A_TE_R_NA_L_ _DO_S_AG_E_G_R_O_UP_ _IV_ _ _ _ _ _ _ _ _ _ _ _3_M_G-/-K-G-/-D-A-Y- - - - - - - - - - - - - -P-O-S-T-P- A- R- T- U- M- - -D-A-Y- -1-5- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 19002 34.5 36.4 35.7 MS 33.1 33.6 32.9 32.4 33.7 33.0 FD 1 19005 23.1 26.5 23.4 24.8 22.1 25.2 24.8 23.7 24.2 24.0 24.0 25.1 21.1 24.7 22.5 23.0 19035 19039 23.6 23.5 23.2 23.0 22.4 24.1 23.3 22.5 21.3 22.7 23.1 23.4 21.6 21.9 24.7 19.3 24.0 23.5 23.0 MD 2 18.8 20.6 19.4 23.6 17.0 21.8 22.6 22.1 24.8 19040 19045 19046 19051 26.1 26.1 26.3 27.4 22.4 24.2 23.4 24.9 21.7 25.4 24.5 24.0 NOT PREGNANT NOT PREGNANT; MATING NOT CONFIRMED 23.9 23.0 22.4 22.7 27.3 26.3 24.6 25.0 22.6 21.9 21.4 24.6 27.5 23.7 25.6 22.0 24.4 23.2 20.1 UU 18.1 19054 39.0 41.2 43.5 40.1 40.8 19058 30.7 26.7 29.1 27.6 24.9 27.1 28.4 28.0 26.5 26.3 26.2 28.0 29.0 19062 31.4 31.3 30.4 31.2 31.2 30.0 31.3 28.9 30.3 30.3 30.0 29.9 29.0 28.7 28.7 19063 28.3 31.3 31.6 29.9 29.2 32.0 30.0 27.6 29.2 31.9 30.7 26.8 30.8 FS 19066 31.4 31.2 30.0 32.0 30.1 30.0 MD 1 30.8 30.2 31.9 30.9 29.5 32.1 FS 19069 16.8 21.7 19.4 19.6 18.2 19.9 19.5 21.8 17.0 21.4 22.3 21.5 20.9 20.0 19.8 19.9 FDlO 19073 26.7 27.5 29.3 27.1 28.0 28.1 27.3 MM 3 25.3 26.1 26.3 25.6 25.2 25.3 ________________________________________-------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN, U-UNCERTAIN,D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T - 7 7 0 6 . 1 ) TABLE C44 (PAGE 9 ) : PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS ________________________________________-------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 1 2 13 14 15 16 17 18 19 20 ________________________________________-------------------------------------------------------------------------------------------- RAT/ LITTER # MATERNAL DOSAGE GROUP V 10 MG/KG/DAY POSTPARTUM DAY 1 5 ________________________________________-------------------------------------------------------------------------------------------- 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 19032 19033 19049 19059 19070 27.5 23.2 27.0 17.0 30.9 25.6 23.5 28.5 29.1 25.3 25.9 27.4 27.2 26.4 30.1 26.3 26.7 25.9 13.8 27.6 24.5 24.2 22.7 29.3 24.6 26.9 28.2 26.1 24.3 29.4 26.2 26.2 27.1 15.3 30.0 24.4 25.0 27.8 29.7 30.7 26.4 27.1 25.8 28.1 31.0 26.2 25.7 26.2 14.8 29.7 23.7 21.9 25.5 30.9 28.8 23.9 27.0 26.1 25.6 30.4 26.6 25.4 25.8 16.6 28.7 26.2 21.7 25.1 29.3 27.8 26.7 27.1 24.5 23.8 29.7 26.2 MD 2 24.3 15.1 25.7 23.4 23.2 26.3 28.5 26.9 26.2 27.9 26.6 25.5 28.5 26.1 22.5 27.5 17.0 29.3 18.7 22.1 26.6 29.4 29.2 25.3 25.8 20.8 23.3 29.5 28.0 25.2 25.8 15.9 27.5 24.0 21.6 25.8 28.8 MM 2 25.5 25.3 25.2 22.7 30.1 26.4 27.5 24.2 14.9 31.4 23.8 22.2 27.4 24.0 26.4 24.5 25.4 24.0 24.1 30.5 25.8 23.6 25.7 15.8 MD 1 21.4 20.0 25.1 29.7 23.6 26.5 24.8 27.6 25.2 25.1 27.3 24.9 25.1 15.2 25.6 25.3 22.7 24.6 29.0 26.5 24.6 26.6 25.1 22.1 27.3 26.3 24.6 16.2 27.0 26.8 21.4 25.5 28.0 26.2 23.4 24.3 26.1 17.4 29.3 22.1 27.5 15.1 29.2 27.4 24.0 FS 28.0 27.0 23.7 24.7 20.3 22.2 FD13 22.0 25.1 25.1 23.6 20.5 28.3 20.8 25.1 22.9 24.5 24.5 20.4 FD 1 25.1 20.1 24.3 23.2 FM 2 20.0 21.3 22.4 21.2 17.7 FD 1 FS 19.2 19.1 ^_______________________________________-------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN,U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C44 (PAGE 10): PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS .................................................................................................................................... PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 .................................................................................................................................... RAT/ LITTER # MATERNAL DOSAGE GROUP I o MG/KG/DAY POSTPARTUM DAY 22 .................................................................................................................................... 19010 43.0 44.2 41.1 39.4 39.0 39.5 40.7 33.4 42.7 40.0 41.6 41.6 41.4 FD 3 19012 43.0 43.7 40.3 44.2 42.2 37.9 41.6 41.1 39.0 36.9 41.8 36.7 42.6 41.0 42.1 45.4 19019 39.6 41.8 36.2 39.0 39.9 42.5 41.2 39.7 37.6 35.4 38.6 39.6 36.3 37.6 35.4 19021 34.5 34.4 39.4 35.8 35.0 35.6 38.3 36.0 33.2 34.1 34.9 32.1 34.5 33.4 19023 44.0 45.6 47.2 44.0 48.9 46.4 46.9 44.6 44.2 45.4 44.6 43.1 FD 4 19041 37.0 31.1 34.9 29.4 32.6 30.4 31.4 33.3 33.9 33.4 35.4 37.9 29.6 35.7 26.8 35.8 25.3 31.8 19042 38.7 37.4 37.5 36.4 35.0 33.1 34.6 37.6 35.3 35.7 34.4 37.2 35.7 32.8 36.7 19044 48.1 48.4 25.8 43.9 45.5 43.1 36.4 49.3 43.6 41.6 41.1 37.8 36.6 40.0 43.1 40.2 FM 2 19050 37.9 37.4 37.1 35.0 34.7 34.5 38.4 34.3 32.4 37.1 38.5 34.3 34.5 32.6 34.6 19053 34.8 39.9 37.9 33.8 39.2 32.3 35.3 34.1 MD 5 34.7 35.2 34.3 38.3 37.1 31.7 37.1 19065 41.2 25.4 30.1 36.9 33.3 30.9 31.5 33.6 29.0 32.8 30.9 31.5 32.4 35.7 31.7 28'.9 23.9 33.0 26.8 FD 5 19068 47.9 46.2 48.8 50.0 45.5 MD 2 45.4 45.9 45.9 46.8 42.5 44.2 47.1 45.0 43.1 19072 34.0 33.7 27.0 29.9 33.7 33.5 32.9 28.6 28.0 28.9 33.0 33.7 30.3 28.6 28.6 29.1 19074 35.4 37.2 38.8 29.2 MD 1 32.9 35.1 32.5 33.6 33.5 34.7 35.2 34.7 33.0 35.1 19075 34.4 38.5 35.5 37.1 30.8 35.5 38.5 35.4 40.7 36.0 34.5 33.8 37.4 36.0 38.6 34.5 .................................................................................................................................... RAT/ LITTER # MATERNAL DOSAGE GROUP I1 0.3 MG/KG/DAY POSTPARTUM DAY 22 ________________________________________-------------------------------------------------------------------------------------------- 19004 33.4 34.4 26.5 33.6 29.0 30.4 33.2 28.5 33.3 26.3 32.4 24.0 28.9 29.9 26.4 29.4 29.1 FM 6 19009 57.1 55.6 51.3 51.4 57.7 53.8 51.8 19016 46.5 47.4 48.5 44.5 44.2 44.4 45.8 47.6 49.3 50.3 41.3 45.4 45.1 25.8 43.4 19018 48.4 47.0 45.9 46.3 44.8 43.4 44.6 44.0 45.6 45.0 39.2 38.1 41.0 40.6 19026 32.3 32.5 39.1 30.7 30.1 34.9 29.3 29.6 28.7 30.7 29.1 31.2 29.3 27.6 34.6 28.6 27.1 27.5 FD 8 8 19036 39.7 40.9 43.1 42.9 42.1 38.1 29.1 35.7 42.7 43.2 39.2 30.5 37.0 FMll FD 2 FD 1 P w 19037 33.1 25.3 30.5 31.8 31.6 34.6 31.0 31.9 30.8 29.1 32.9 27.9 31.9 27.0 24.9 FD15 00 19043 46.0 46.4 47.2 46.2 48.4 44.9 45.8 43.2 44.6 43.7 45.2 42.9 42.6 1 19047 38.9 39.1 34.4 37.1 39.3 39.5 39.5 41.3 39.7 36.9 40.5 33.0 32.2 19048 46.5 51.0 49.7 47.1 50.5 47.0 48.0 45.8 45.0 19052 38.6 38.6 31.7 33.6 37.3 35.2 40.1 35.4 31.1 34.0 37.8 40.5 36.9 37.5 29.9 36.2 FD 5 19055 40.7 40.1 41.3 40.2 39.6 38.1 42.9 38.5 36.3 40.3 MD 1 38.9 39.4 34.5 37.8 35.6 19061 33.8 30.7 32.2 31.1 34.4 30.2 32.7 MD 3 35.9 31.5 34.2 32.7 34.6 19067 41.4 39.9 40.2 40.8 39.6 39.3 40.5 39.4 37.5 39.1 37.0 40.2 37.7 FD 8 19071 40.7 42.3 42.1 40.5 44.4 34.9 40.0 35.3 36.9 41.3 43.3 42.6 43.1 31.1 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN,U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T - 7 7 0 6 . 1 ) TABLE C44 (PAGE 1 1 ) : PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS .................................................................................................................................... PUP # 1 2 3 4 5 6 7 8 9 10 11 1 2 13 14 15 16 17 18 19 20 .................................................................................................................................... RAT/ LITTER # MATERNAL DOSAGE GROUP I11 1 MG/KG/DAY POSTPARTUM DAY 22 .................................................................................................................................... 19003 35.4 31.5 30.7 26.0 32.3 31.9 24.2 31.3 28.4 30.2 25.0 28.6 26.6 31.6 28.2 26.1 19007 34.7 35.5 35.5 30.4 27.9 36.0 35.0 31.2 29.3 28.0 31.0 29.0 31.8 3 0 . 1 33.8 27.4 33.8 FD 2 19008 4 3 . 6 4 0 . 5 4 2 . 9 4 1 . 5 4 0 . 8 3 8 . 6 4 2 . 5 3 7 . 2 4 1 . 3 3 8 . 4 3 8 . 4 4 2 . 5 3 9 . 9 3 5 . 5 FM 2 19013 45.7 45.8 49.6 42.0 49.1 46.6 50.2 45.5 48.4 41.8 39.5 42.8 46.1 37.9 45.5 19014 NOT PREGNANT 19015 39.1 39.8 41.8 40.0 37.1 33.5 38.6 37.3 38.4 38.8 36.2 38.1 35.8 30.1 36.4 19017 49.7 42.2 45.8 47.5 48.8 40.5 47.3 48.8 44.4 45.0 47.1 41.9 30.8 48.3 19024 39.0 39.8 39.7 38.5 38.2 43.4 37.3 42.8 41.4 37.1 41.3 35.6 35.6 38.5 36.8 19029 4 6 . 5 4 1 . 8 4 4 . 0 4 1 . 8 4 6 . 1 4 7 . 6 4 4 . 3 3 7 . 7 MM 2 3 8 . 6 3 2 . 0 3 1 . 7 3 8 . 0 3 7 . 6 3 4 . 2 3 0 . 3 19034 4 1 . 4 4 0 . 9 4 1 . 2 4 1 . 4 4 0 . 6 4 1 . 9 3 8 . 9 4 0 . 2 4 0 . 4 4 0 . 4 3 8 . 7 3 7 . 5 3 7 . 0 FS 19038 NOT PREGNANT 19056 39.3 41.4 41.4 39.5 42.6 41.6 44.1 38.5 41.5 41.2 39.6 41.6 38.0 38.1 19057 34.2 33.7 32.2 33.7 30.9 32.3 32.1 32.5 33.0 31.5 34.6 31.1 33.9 30.7 33.9 29.7 30.3 19060 47.4 47.0 49.2 45.8 48.8 46.4 45.5 42.8 46.7 42.7 48.8 40.6 39.5 38.5 19064 3 . 0 3 4 . 7 2 8 . 0 3 6 . 4 2 9 . 4 3 3 . 0 2 9 . 4 2 8 . 2 2 8 . 4 3 6 . 9 2 8 . 4 3 0 . 3 3 4 . 2 3 5 . 6 2 7 . 5 2 4 . 0 2 4 . 6 FMll .................................................................................................................................... RAT/ LITTER # MATERNAL DOSAGE GROUP IV 3 MG/KG/DAY POSTPARTUM DAY 22 ________________________________________-------------------------------------------------------------------------------------------- 19002 47.2 47.4 49.4 MS 44.4 46.3 4 5 . 3 46.0 45.4 45.0 FD 1 19005 31.3 36.4 34.3 31.3 32.7 33.1 28.9 32.0 32.7 31.5 29.6 33.0 33.2 33.4 28.1 34.5 19035 32.8 35.2 33.6 33.2 34.6 33.3 32.2 30.6 33.9 30.6 34.7 29.9 34.4 3 2 . 1 19039 3 6 . 0 39.4 3 6 . 2 3 4 . 0 3 0 . 1 MD 2 3 2 . 8 3 5 . 6 3 1 . 7 36.8 3 1 . 7 2 8 . 5 32.7 3 3 . 7 27.8 19040 32.8 30.6 32.9 32.0 33.6 31.6 28.3 26.9 27.1 28.3 31.7 30.0 32.9 35.5 32.1 24.7 23.7 19045 3 7 . 1 4 1 . 2 3 7 . 1 3 3 . 5 3 9 . 4 3 7 . 1 3 8 . 3 3 5 . 8 3 6 . 0 3 7 . 8 3 6 . 4 3 0 . 6 3 6 . 0 3 9 . 9 3 4 . 8 UU 19046 NOT PREGNANT 19051 NOT PREGNANT; MATING NOT CONFIRMED 19054 57.5 59.1 54.8 55.5 57.6 19058 51.3 44.7 41.2 47.9 45.6 47.6 47.5 45.7 4 4 . 1 47.5 45.8 42.7 45.3 19062 43.0 43.6 41.0 42.9 41.0 40.1 44.0 42.9 42.7 41.3 40.4 41.2 38.2 39.9 39.5 19063 4 8 . 7 4 6 . 5 4 6 . 1 4 8 . 6 4 5 . 9 4 9 . 4 4 8 . 4 4 4 . 0 4 2 . 8 5 0 . 1 4 7 . 2 3 9 . 7 4 6 . 2 FS 19066 4 3 . 5 4 4 . 3 4 3 . 8 4 4 . 3 4 2 . 9 4 5 . 7 MD 1 4 3 . 5 4 3 . 5 4 0 . 9 4 2 . 5 4 1 . 4 4 3 . 2 FS 19069 3 1 . 7 2 8 . 5 2 8 . 1 3 0 . 5 3 2 . 5 2 9 . 3 3 3 . 2 33.4 3 5 . 7 2 7 . 5 3 3 . 0 3 7 . 2 3 6 . 8 3 2 . 3 3 3 . 8 3 1 . 4 FDlO 19073 44.5 44.2 44.0 43.8 45.6 46.2 42.0 MM 3 39.5 38.6 42.7 41.3 42.2 41.2 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C44 (PAGE 12): PUP BODY WEIGHTS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS .................................................................................................................................... PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 .................................................................................................................................... RAT/ LITTER # MATERNAL DOSAGE GROUP V 10 MG/KG/DAY POSTPARTUM DAY 22 .................................................................................................................................... 19001 19006 43.5 45.6 43.0 44.3 45.4 43.3 44.0 43.0 42.9 44.2 45.6 33.2 36.3 35.4 34.5 36.6 MD 2 31.6 32.2 31.7 32.0 30.9 36.9 34.9 34.8 34.4 28.9 29.9 34.2 FD 1 FS 19011 37.9 38.1 33.7 35.8 38.9 34.9 35.5 35.6 37.9 35.7 35.0 33.7 34.9 19020 19022 19025 19027 19028 15.1 32.3 31.2 34.8 39.9 15.8 33.5 30.6 33.3 43.1 16.0 37.9 29.9 32.3 38.4 16.2 39.1 30.5 31.8 44.4 14.4 39.3 27.0 29.3 33.2 16.0 35.4 35.9 32.4 39.6 15.3 35.3 25.0 32.5 40.4 14.8 37.0 31.7 29.0 40.6 14.5 37.2 30.2 31.1 39.5 18.1 MD 1 34.5 30.6 35.6 14.9 34.6 30.6 31.1 42.5 15.9 36.3 31.8 31.7 42.4 14.2 35.6 31.5 30.1 FS 32.5 30.5 33.0 33.3 24.4 FM 2 27.6 26.8 31.0 23.8 23.8 19030 19031 19032 39.1 43.4 37.3 38.9 45.6 34.6 40.8 45.0 36.4 43.3 50.8 33.5 40.0 40.4 36.7 38.4 40.9 37.4 39.8 47.8 35.6 42.0 MM 2 32.4 39.6 44.3 34.5 32.2 39.5 33.8 38.7 41.2 35.2 37.6 39.2 36.1 37.9 44.0 32.9 38.4 34.9 40.9 19033 45.7 40.7 43.6 45.8 45.1 43.8 41.2 44.2 42.6 39.1 41.0 41.0 40.2 39.0 19049 37.8 34.6 35.3 37.1 36.1 39.9 39.6 33.9 39.1 31.2 39.9 36.2 38.6 32.4 24.4 19059 35.2 33.3 33.8 37.4 37.4 33.6 35.5 33.7 35.0 31.0 33.8 27.5 25.6 32.2 32.9 19070 48.6 51.7 54.1 47.6 49.5 52.1 46.9 46.5 47.5 46.8 43.9 48.8 FD13 FD 1 .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN,U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 4 1 8 - 0 2 8 : ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T - 7 7 0 6 . 1 ) TABLE C 4 5 (PAGE 1 ) : PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 2 2 POSTPARTUM - INDIVIDUAL DATA - F 1 GENERATION PUPS - - - - - - - _ _. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19010 M A M A M A M A F A F A F A F A F A F A F A F A F A F D 3 19012 M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A F A 19019 M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A 19021 M A M A M A M A M A M A M A M A M A F A F A F A F A F A 19023 M A M A M A M A M A M A M A F A F A F A F A F A F D 4 19041 M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A F A F A 19042 M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A 19044 M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A F M 2 19050 M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A 19053 M A M A M A M A M A M A M A M A M D 5 F A F A F A F A F A F A F A 19065 M A M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A F D 5 19068 M A M A M A M A M A M D 2 F A F A F A F A F A F A F A F A F A 19072 M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A 19074 M A M A M A M A M D 1 F A F A F A F A F A F A F A F A F A F A 19075 M A M A M A M A M A M A M A M A M A M A M A F A F A F A F A F A RAT/ LITTER # MATERNAL DOSAGE GROUP I1 0 . 3 MG/KG/DAY .................................................................................................................................... 19004 M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F M 6 19009 19016 19018 19026 MA MA F A F A F A F A F A MA MA MA MA MA MA MA MA MA MA MA MA F A F A F A MA MA MA MA MA MA MA MA F A F A F A F A F A F A MA MA MA MA MA MA MA MA MA MA F A F A F A F A F A F A F A F A F D 8 19036 M A M A M A M A M A M A F A F A F A F A F A F A F A F M l l F D 2 F D l 19037 M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A F D 1 5 19043 M A M A M A M A M A M A M A F A F A F A F A F A F A 19047 M A M A M A M A M A M A M A F A F A F A F A F A F A 19048 M A M A M A M A M A F A F A F A F A 19052 M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A F D 5 19055 19061 MA MA MA MA MA MA MA MA MA MAMD1 F A F A F A F A F A MA MA MA MA MA MA MAMD3 F A F A F A F A F A 19067 M A M A M A M A M A M A F A F A F A F A F A F A F A F D 8 19071 M A M A M A M A M A M A M A M A F A F A F A F A F A F A .................................................................................................................................... F I R S T LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C45 (PAGE 2): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS .................................................................................................................................... P U P # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 .................................................................................................................................... RAT/ LITTER # MATERNAL DOSAGE GROUP I11 1 MG/KG/DAY .................................................................................................................................... 19003 M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A 19007 M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A F D 2 19008 M A M A M A M A M A M A M A F A F A F A F A F A F A F A F M 2 19013 M A M A M A M A M A M A M A M A M A F A F A F A F A F A F A 19014 NOT PREGNANT 19015 M A M A M A M A M A M A M A M A M A F A F A F A F A F A F A 19017 M A M A M A M A M A M A F A F A F A F A F A F A F A F A 19024 M A M A M A M A M A M A M A M A M A F A F A F A F A F A F A 19029 M A M A M A M A M A M A M A M A M M 2 F A F A F A F A F A F A F A 19034 M A M A M A M A M A M A F A F A F A F A F A F A F A F S 19038 NOT PREGNANT 19056 M A M A M A M A M A M A M A F A F A F A F A F A F A F A 19057 M A M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A 19060 M A M A M A M A M A M A M A M A F A F A F A F A F A F A 19064 M A M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A F M l l .................................................................................................................................... RAT/ LITTER # MATERNAL DOSAGE GROUP IV 3 MG/KG/DAY .................................................................................................................................... 19002 M A M A M A M S F A F A F A F A F A F A F D 1 19005 M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A 19035 M A M A M A M A M A M A F A F A F A F A F A F A F A F A 19039 M A M A M A M A M A M D 2 F A F A F A F A F A F A F A F A F A 19040 M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A 19045 M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A U U 19046 NOT PREGNANT 19051 NOT PREGNANT; MATING NOT CONFIRMED 19054 M A M A M A F A F A 19058 M A M A M A M A M A M A M A M A F A F A F A F A F A 19062 M A M A M A M A M A M A M A M A M A F A F A F A F A F A F A 19063 M A M A F A F A F A F A F A F A F A F A F A F A F A F S 19066 M A M A M A M A M A F A M D 1 F A F A F A F A F A F A F S 19069 M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A F D l O 19073 M A M A M A M A M A M A M A M M 3 F A F A F A F A F A F A .................................................................................................................................... FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN,U-UNCERTAIN,D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C45 (PAGE 3): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS .................................................................................................................................... P U P # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 .................................................................................................................................... RAT/ LITTER # MATERNAL DOSAGE GROUP V 10 MG/KG/DAY ________________________________________-------------------------------------------------------------------------------------------- 19001 M A M A M A M A F A F A F A F A F A F A F A 19006 M A M A M A M A M A M D 2 F A F A F A F A F A F A F A F A F A F A F A F A F D 1 F S 19011 M A M A M A M A M A M A M A F A F A F A F A F A F A 19020 M A M A M A M A M A M A M A M A F A F A F A F A F A 19022 M A M A M A M A M A M A M A M A M A M D 1 F A F A F A F A F A F M 2 19025 M A M A M A M A M A M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A 19027 M A M A M A M A F A F A F A F A F A F A F A F A F A F A 19028 M A M A M A M A M A F A F A F A F A F A F A F A F S 19030 M A M A M A M A M A M A F A F A F A F A F A F A F A 19031 M A M A M A M A M A M A M A M M 2 F A F A F A F A F A F A F A 19032 M A M A M A M A M A M A M A F A F A F A F A F A F A F A 19033 M A M A M A M A M A M A F A F A F A F A F A F A F A F A 19049 M A M A M A M A M A M A M A M A M A M A M A F A F A F A F A 19059 M A M A M A M A M A M A M A M A M A M A F A F A F A F A F A 19070 M A M A M A M A M A M A M A M A F A F A F A F A F D 1 3 F D 1 .................................................................................................................................... FIRST LETTER - - M-MALE, F-FEMALE, U-SEX UNDETERMINABLE SECOND LETTER - - A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C46 (PAGE 1): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 22 POSTPARTUM - INDIVIDUAL DATA - F1 GENERATION PUPS I 0 19010 1- 2 1/14 PUPS: PALE IN APPEARANCE. 19044 1- 2 1/17 PUPS: BACK, BRUISE (2.5 CM X 2.0 CM) . 19050 1 1/15 PUPS: PALE IN APPEARANCE. 19072 18-22 1/16 PUPS: RIGHT EYE, CORNEAL 0PACITY.b I1 0.3 19004 8 1/17 PUPS: COLD TO TOUCH; NOT NESTING OR NURSING. 19036 1- 4 1/14 PUPS: BACK, BRUISE (2.0 CM IN DIAMETER). 19037 18 4/15 PUPS: DEHYDR74TION. 19071 11 1/14 PUPS: COLD TO TOUCH. I11 1 19003 17 1/16 PUPS: I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I 1 I 1 I I I I I I I I I I I I 1 I I I I 1 I I I I I I I I I I I I I I I I I I I I I I I I I rdI I I 1 UlI I 21 . .. . .. .. I 01 I H I m m m ul I 2 : 3! I PI 2 2 2 I PI I 2 ; I a 1 I UII I I 01 I I I I I I I 1 I I I I I I I I I I I I I I I I I & A 0 H 3 0 4 0 V m H n x n w 0 E E 2 m PI P m rl rl \ \ rl cv I I I -1 ull *I 01 1 UI I rll *I 01 I n~ nH uI -1 I U2 I' UII I .. I . 418-028:PAGE C-129 * CD 0 cn 0 cv 0 0 0 cn m cn rl rl rl I I0 IU PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C47 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - F1 GENERATION PUPS 19012 22 16 PUPS: APPEARED NORMAL. 19019 22 15 PUPS: APPEARED NORMAL. 19021 22 14 PUPS: APPEARED NORMAL. 19023 4 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 22 12 PUPS: APPEARED NORMAL. 19041 22 18 PUPS: APPEARED NORMAL. 19042 22 15 PUPS: APPEARED NORMRL. 19044 22 16 PUPS: APPEARED NORMAL. 19050 22 15 PUPS: APPEARED NORMAL. 19053 5 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 22 15 PUPS: APPEARED NORMAL. 19065 5 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 22 19 PUPS: APPEARED NORMAL. 19068 2 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 22 14 PUPS: APPEARED NORMAL. 19072 22 16 PUPS: APPEARED NORMAL. 19074 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 22 14 PUPS: APPEARED NORMAL. 19075 22 16 PUPS: APPEARED NORMAL. - - - - -. _ _ _ _ - - - - _ - - - - - - - - _ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table C 4 4 ) for external clinical observations confirmed at necropsy. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C47 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - F1 GENERATION PUPS 19026 8 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 22 18 PUPS: APPEARED NORMAL. 19036 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 22 13 PUPS: APPEARED NORMAL. 19037 15 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 22 1 PUP: KIDNEYS: RIGHT, PELVIS, MODERATE DILATION. ALL OTHER TISSUES APPEARED NORMAL. 14 PUPS: APPEARED NORMAL. 19043 22 13 PUPS: APPEARED NORMAL. 19047 22 13 PUPS: APPEARED NORMAL. 19048 22 9 PUPS: APPEARED NORMAL. 19052 5 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 22 16 PUPS: APPEARED NORMAL. .................................................................................................................................... a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table C44) for external clinical observations confirmed at necropsy. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 22 15 PUPS: APPEARED NORMAL. 19061 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 22 12 PUPS: APPEARED NORMAL. 19067 8 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 22 13 PUPS: APPEARED NORMAL. 19071 22 14 PUPS: APPEARED NORMAL. b w 00 PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C47 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - F1 GENERATION PUPS 19007 2 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 22 17 PUPS: APPEARED NORMAL. 19008 19013 19015 19017 19024 19029 22 14 PUPS: APPEARED NORMAL. 22 15 PUPS: APPEARED NORMAL. 22 15 PUPS: APPEARED NORMAL. 22 14 PUPS: APPEARED NORMAL. 22 15 PUPS: APPEARED NORMAL. 22 15 PUPS: APPEARED NORMAL. 19034 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 22 13 PUPS: APPEARED NORMAL. 19056 22 14 PUPS: APPEARED NORMAL. 19057 19060 22 17 PUPS: APPEARED NORMAL. 22 14 PUPS: APPEARED NORMAL. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _1_9_06_4_ _ _ _ _ _ _ _ _ _ _ _2_2- - - - - - - - - - - - - - - -1-7- -P-U- P- S- :- - -A-P-P-E-A- R- E- D- - N- O- R- -M-A-L-.- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table C44) for external clinical observations confirmed at necropsy. PROTOCOL 418-028: ORAL (GAVAGE)COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) MATERNAL DOSAGE GROUP LITTER MATERNAL DOSAGE (MG/KG/DAY) NUMBER ........................................... -- IV 3 19002 DAY POSTPARTUM 1 22 OBSERVATIONS a 1 PUP: 1 PUP: 9 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. STILLBORN. AUTOLYSIS PRECLUDED FURTHER EVALUATION. APPEARED NORMAL. 19005 19035 22 16 PUPS: APPEARED NORMAL. 22 14 PUPS: APPEARED NORMAL. 19039 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 22 14 PUPS: APPEARED NORMAL. 19040 19045 19054 19058 19062 22 17 PUPS: APPEARED NORMAL. 22 15 PUPS: APPEARED NORMAL. 22 5 PUPS: APPEARED NORMAL. 22 13 PUPS: APPEARED NORMAL. 22 15 PUPS: APPEARED NORMAL. 19063 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 22 13 PUPS: APPEARED NORMAL. 19066 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 22 12 PUPS: APPEARED NORMAL. + P 19069 10 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 22 16 PUPS: APPEARED NORMAL. 19073 22 13 PUPS: APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table C44) for external clinical observations confirmed at necropsy. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'S STUDY NUMBER: T-7706.1) TABLE C47 (PAGE 6): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - F1 GENERATION PUPS 19006 19011 19020 19022 19025 19027 19028 19030 19031 19032 19033 19049 19059 19070 ------ 11 PUPS : APPEARED NORMAL. 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 22 17 PUPS: APPEARED NORMAL. 22 13 PUPS: APPEARED NORMAL. 22 13 PUPS: APPEARED NORMAL. 1 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 22 14 PUPS: APPEARED NORMAL. 22 20 PUPS: APPEARED NORMAL. 22 14 PUPS: APPEARED NORMAL. 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 22 12 PUPS: APPEARED NORMAL. 22 13 PUPS: APPEARED NORMAL. 22 14 PUPS: APPEARED NORMAL. 22 14 PUPS: APPEARED NORMAL. 22 14 PUPS: APPEARED NORMAL. 22 15 PUPS: APPEARED NORMAL. P w 22 15 PUPS: APPEARED NORMAL. 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 13 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 22 12 PUPS: APPEARED NORMAL. PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST (SPONSOR'SSTUDY NUMBER: T-7706.1) TABLE C48 (PAGE 1): PUP LIVER WEIGHTS - INDIVIDUAL DATA - F1 GENERATION PUPS 19009 19016 19018 19026 19036 19037 19043 19047 2.350 1.874 1.768 1.062 1.596 1.170 2.098 1.510 2.356 1.633 1.752 1.214 1.596 1.182 2.074 1.462 2.354 1.784 1.760 1.138 1.596 1.176 2.086 1.486 19007 19008 19013 19015 19017 19024 19029 19034 19056 1.324 1.516 1.976 1.416 2.102 1.504 2.076 1.550 1.500 1.278 1.732 1.736 1.510 2.188 1.550 1.456 1.670 1.646 1.301 1.624 1.856 1.463 2.145 1.524 1.766 1.610 1.573 ALL WEIGHTS WERE RECORDED IN GRAMS (G). 56.35 45.32 45.50 32.30 40.74 30.66 46.34 38.78 53.20 38.10 41.70 30.04 34.30 29.16 43.80 36.62 54.10 42.61 43.60 31.17 37.52 29.91 45.07 37.70 35.34 41.82 46.36 38.06 44.96 40.04 43.22 41.24 40.88 29.70 40.10 42.36 35.84 43.88 37.62 33.24 38.76 39.48 32.52 40.96 44.36 36.95 44.42 38.83 38.23 40.00 40.18 418-028:PAGE C-137 : I I 0 m co co c*v 0 cn * * 03 rl m Lo m m 0 m r m W In W * *rl W * . . . . . . . cv 0 w Inmmwmcncn m mrlmmmmm * I W 03 co m 03 W 0 *cvWOWOW * cv rl m 0 co rl co * m * r * r l r l . . . . . . . rl 0 r W m cn W * m m m m w m cv m m * w r l o c n m mrlmmmdcm I * co W m 0 0 0 0c o o o o m o * * . . . . . . . I rl W rl a, In W cv m m w o o w c n m rl r r r m r * * * m m m Ln In W m W o m c n c n m mrlmmmmm I * 0 r0rW W W 0 r 03 m 03 0 * Tr W m m m rl rl rl rl cv m rl cv * . . . . . . . m c v r r l c n * w m m Wcnrlmco*rl m*mrlmww rl r l o r l r l r l r l r l * . . . . . . . I I 0m0m 0 m m co a3 m cn rl 0 m cv * O W O W c v c v rl m * a ) w c o * m mm r rl r rl In * m c v r l c v r W rl rl rl cv cv rl cv rl r l o r l r l r l r l r l I I * * 0 W r 00 03 o * \ o c o m o * * * cv cv m W r In c n m m c n c o m c n * m m W rl m 0 m cv*mocvmm . . . . . . . I I rl rl 4 cv cv rl cv rl r l o r l r l r l r l r l APPENDIX D PROTOCOL AND AMENDMENTS 418-028:PAGE D-1 905 Ymhy Drive, 6&. A Hotshorn, PA I904 4 Telephone: (2 IS) 44347I 0 T&@: (215) 4438587 -xw ARGUS RESEARCH Chudes River Laborotcines Discovery und Development Services PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 STUDY TITLE: Oral (Gavage) Combined Repeated Dose Toxicity Study of T-7706 with the ReproductiodDevelopmentalToxicityScreening Test PURPOSE: The purpose of this study is to provide information on the possible health hazards that may result fiom repeated exposure of Crl:CD@(SD)IGSBR VAFPlusO male and female rats to a test substance beginning before cohabitation,throughmating and continuingfor at least 42days (male rats) or through parturition until day 21 of lactation (femalerats). This repeated dose study incorporates a reproductioddevelopmentaltoxicity screeningtest that can be used to provide initial infomation on possible effects on male and female reproductive performance (e.g., gonadal function, mating behavior, conception,developmentof the conceptus and parturition). The study also places emphasis on neurological effects as a specific endpoint and should identify the neurotoxic potential of a test substance,which may warrant further in-depth investigation. TESTING FACILITY: Because of the selectivity of the endpoints and the short duration of the study, the screening test will not provide evidence for definitive claims of no reproductioddeveloprnentaleffects. In particular, it offers only limited means of detecting postnatal manifestationsof prenatal exposure or effects that may be induced during postnatal exposure. Argus Research 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 Telephone: (2 15) 443-87 10 Telefax: (215)443-8587 STUDY DIRECTOR: Raymond G. York, Ph.D., DABT Associate Director of Research Email: raymond.york@criver.corn Address as cited above for Testing Facility SPONSOR. 3M Corporate Toxicology 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 418-028:PAGE D-2 STUDY MONITOR: John Butenhoff,Ph.D., DABT, C M 3M Corporate Toxicology 3M Medical Department Telephone: (651) 733-1962 Telefax: (651) 733-1773 Email: j lbutenhoff'@mmm.com Protocol 4 18-02 8 Page 2 REGULATORY CITATIONS: Organisationfor Economic Co-operation and Development (1996). OECD Guidelinefor Testing of Chemicals. Section 4, No. 422: Combined Repeated Dose Toxicity Study with the ReproductiodDevelopmental Toxicity ScreeningTest, adopted 22 March 1996. Organisation for Economic Co-operation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Final]. U.S.Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFRPart 58. Japanese Ministry of Health and Welfare (1997). GoodLaboratoly Practice Standardfor Safety Studies on Drugs,MHW OrdinanceNumber 21, March 26,1997. This studywill be conducted in compliancewith the Good LaboratoryPractice (GLP) regulations cited above. All changes or revisions of this protocol shall be documented, signed by the Study Director and the Sponsor, dated and maintained with the protocol. The Testing Facility`s Quality Assurance Unit (QAU) will audit the protocol, the raw data and the report, and will inspect critical phases of those portions of the study conducted at the Testing Facility in accordancewith the Standard OperatingProcedures of the Testing Facility. The final report will include a compliancestatement signedby the StudyDirector that the report accuratelyreflects the raw data obtained during the performance of the study and that all applicableGLP regulations were followedin the conduct of the study. Should significant deviations fkom GLP regulations occur, each will be described in detail, together with how the deviationmight affect the quality or integrity of the study. Should any portion of the study be conducted by a subcontractor or by the Sponsor, the Study Director will ensure that a qualified Principal Investigatoris identifiedby the facility conducting that portion of the study. The QAU for this facility will conduct critical phase inspections and audit respectiveresults and reports for that study portion accordingto the SOPSofthat facility. Such critical phase inspection reports and report audits will be submitted by the facility to the Principal Investigator and the Study Director. The dates of the inspections and report submissions will be incorporated into a QAU Statement generated by that facility and provided 418-028:PAGE D-3 Protocol 418-028 Page 3 to the TestingFacility for inclusion in the final report. In addition, this facility will provide a statement of GLP compliance, as described above, signed by the Principal Investigator for inclusion in the final report. SCHEMATICOF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT 1 to the protocol. TEST SUBSTANCEA N D VEHICLE: Identification: Test Substance: T-7706 [PerfluorohexaneSulfonatePotassium Salt (PFHS)J Lot identificationwill be documented in the raw data. The Sponsor will provide to the Testing Facility documentationor certificationof the identity, composition, method of synthesis, strength and activity/purity of the test substance. This documentationwill be included in the finalreport. Vehicle: Aqueous 0.5% carboxymethlycellulose(CMC) (mediumviscosity) prepared using reverse osmosis membrane processed deionized water (R.O. deionized water). Lot identification and Supplier will be documented in the raw data. Neither the Sponsornor the StudyDirector is aware of anypotential contaminantslikelyto be present in the vehicle that would interfere with the results of this study. Therefore, no analyses other than those mentioned in this protocol will be conducted. Safety Precautions: Gloves, dust-mist/HEPA-filteredmask, appropriate eye protection and Unifodlab coat to be worn during formulationpreparation and dosage. The Material SafetyData Sheet (MSDS)is attached to the protocol (ATTACHMENT2). Storape: Bulk Test Substance: Bulk Vehicle Components: Prepared Test Substance and Vehicle Formulations: Room temperature. Room temperature. Refiigerated (2OC to 8OC). All test substance shipments should be addressed to the attention of Julian Gulbinski, Manager of Formulation Laboratory, at the previously cited Testing Facility address and telephone number. 418-028:PAGE D-4 Protocol 4 18-028 Page 4 Shipmentsshould include informationconcerningstorage conditions and shippingcartons should be labeled appropriately. The recipient should be notified in advance of shipment. FORMULATION: Frequency of Preparation: Formulations (suspensions)will be prepared weekly at the TestingFacility. Detailed preparation procedures will be attached to this protocol (ATTACHMENT 3). Adiustment for Puritv: The test substancewill be considered 100%pure for the purpose of dosage calculations. Testiw Facilitv Reserve Samples: The Testing Facility will reserve a sample of each lot of bulk test substance (approximately 1 g) and bulk vehicle components(approximately 1g or 5 d)used duringthe course of the study. Sampleswill be stored under the previously cited conditions. ANALYSES: Results of required analyses will be provided to the Testing Facility for inclusion in the study report. Samples additional to those described below may be taken if deemed necessary during the course of the study. Additional analyses, if required, will be documentedby protocol amendment. Bulk Test Substance Sampling: A sample of approximately 1 g of the test substance will be taken on the last day of treatment and sent (ambient conditions) to: Principal Investigator: Lisa Clemen 3M EnvironmentalTechnology and Safety Services Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone: (651) 778-5568 Telefax: (651) 778-6176 Email: laclernen@mmm.com The recipient will be notified in advance of sample shipment. 418-028:PAGE D-5 Analvses of Prepared Formulations: Protocol 4 18-028 Page 5 Concentrationand Homogeneity: Concentrationand homogeneity of the prepared formulationswill be verified during the course of thisstudy. Quadruplicatesamples (2 mL each) will be taken &omthe top, middle and bottom of each concentration on the first day of preparation. Two samples from each quadruplicate set will be shipped for analysis; the remaining samples will be retained at the Testing Facility as backup samples. Quadruplicate samples will be taken from each concentration on the last day of preparation. Two samples fi-omeach quadruplicateset will be shipped for analysis;the remaining samples will be retained as backup samples. Backup samples will be stored under the previously cited conditions and discarded at the Testing Facility upon the request of the Sponsor. Stability: Stabilityof the prepared fonnulationswill be documentedduring this study. Two sets of duplicate samples (2 mL each) fiom each concentrationwill be taken on the first day of preparation. One sample of each duplicate set will be shipped on the day of preparation. These samples will be analyzed at the following time points: as soon after preparation as possible and ten days after the first analysis. The remainingsampleswill be retained at the Testing Facility as backup samples. Backup samples will be stored under the previously cited conditions and . discarded at the Testing Facility upon the request of the Sponsor. Shippinp Instructlons: Samplesto be analyzed will be shipped (reliigerated)to: Principal Investigator: Lisa Clemen 3M EnvironmentalTechnology and Safety Services Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone: (651)778-5568 Telefax: (651)778-6176 Email: laclemen@mmm.com The recipient will be notified in advance of sample shipment. DISPOSITION: Prepared formulations will be discarded at the Testing Facility. All remaining bulk test substance will be returned to: Dan Hakes 3M EHSR - Auto & Chm Grp 3M Center, Building 236-1B-10 St. Paul, Minnesota 55144-1000 Telephone: (651)733-2392 418-028:PAGE D-6 TEST SYSTEM: Protocol 4 18-028 Page 6 Species/Strain and Reason for Selection: The Crl:CD@(SD)IGSBR VAF/Plus@rat was selected as the Test System because: 1) it is one mammalian species accepted for use in toxicity studies and it has been widely used throughout industry; 2) this strain of rat has been demonstrated to be sensitive to reproductive and developmental toxins; and 3) historical data and experience exist at the Testing Number: Initial population acclimated: 100male and 100virgin female rats. Population selected for main study: 75 male and 75 virgin female rats (15 per sex per dosage group). Population selectedfor toxicokinetic study: 15 male and 15female rats (threeper sex per dosage group). Bodv Weight and Age: Male rats will be ordered to weigh fiom 275 g to 300 g each at receipt, at which time they will be expected to be at least 60 days of age. Female rats will be ordered to weigh fiom 200 g to 225 g each at receipt, at which time they will be expected to be at least 56 days of age. Actual body weights will be recorded the day after receipt and will be documented in the raw data. The weight ranges will be included in the final report. At study initiation, the weight variation of the rats will not exceed *20% of the mean weight of each sex. - Sex: Both male and female rats will be evaluated. - Source: CharlesRiver Laboratories,Inc. The rats will be shipped in filtered cartonsby air freight andor truck fkom Charles River Laboratories, Inc., to the Testing Facility. Identification: Rats are permanently identified using Monela self-piercing ear tags (Gey Band and Tag Co., Inc., No. MSPT 20101). Male and female rats are assigned temporary numbers at receipt and given unique permanent identificationnumbers when assigned to the studybefore administration of the first dosage. Pups will not be individually identifiedduring lactation; all parameterswill be evaluated in terms of the litter. 418-028:PAGE D-7 Protocol 4 18-028 Page 7 ANIMAL HUSBANDRY: All cage sizes and housing conditions are in compliancewith the Guidefor the Care and Use of Laboratory animal^'^'. Argus Research is an AAALAC-accredited facility. Fo generationrats will be individually housed in stainless steel wire-bottomedcages except during the cohabitation and postpartum periods. During cohabitation, each pair of rats will be housed in the male rat's cage. Beginning no later than day 20 of presumed gestation, Fo generation female rats will be individually housed in nesting boxes. Each dam and delivered litter will be housed in a common nesting box during the postparhim period. Nesting Material: Nesting material (bed-o'cobs@)will be provided. Bedding will be changed as often as necessary to keep the animals dry and clean. Analyses for possible contamination are conducted semi-annually and documented in the raw data. Room Air, Temperature and Humidity: The animal room is independentlysupplied with at least ten changesper hour of 100% fkesh air that has been passed through 99.97% HEPA filters. Room temperaturewill be maintained at 64OF to 79OF (18OC to 26OC) and monitored constantly. Room humidity will also be monitored constantly and maintained at 30% to 70%. Light: An automaticallycontrolled 12-hour light:12-hourdark fluorescent light cycle will be maintained. Each dark period will begin at 1900hours EST. The light cycle may be adjusted by the StudyDirector or designee if deemed necessary to accommodatescheduled laboratory activities. Any such adjustment will be documented in the raw data. - Diet: Rats will be given Certified Rodent Diet@#SO02 (PMI Nutrition International) available ad libitum fiom individual feeders. Feed will be removed the evening prior to the scheduled sacrifice. - Water: Water will be available ad libitum from individual bottles attached to the cages or from an automaticwatering access system. All water will be fiom a local source and passed through a reverseosmosis membrane before use. Chlorinewill be added to the processed water as a bacteriostat; processed water is expected to contain no more than 1.2 ppm chlorine at the time of analysis. Water is analyzed monthly for possible bacterial contaminationand twice annually for possible chemical contamination. 418-028;PAGE D-8 Protocol 418-026 Page 8 Contaminants: Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the certified diet, in the drinking water or in the nesting materials at levels that would interferewith the results of this study. Therefore, no analyses other than those routinely performed by the feed supplier or those mentioned in this protocol will be conducted. DAY NUMBERING SYSTEM: Gestation day 0 is defined as the day spermatozoaare observedin a smear of the vaginal contents andor a copulatory plug observed in situ. - The day of birth is designatedlactation day 0 (postpartumday 0) in the Health Effects Test Guidelines - Reproduction and Fertility Effects (Office of Prevention, Pesticides and Toxic Substances870.3800,August, 1998)and in the OECD Guideline for the Testing of Chemicals Combined Repeated Dose ToxicityStudywith the ReproductiodDevelopmentalToxicity ScreeningTest (Section4, No.422,22 March 1966). This same day is designated day 1 postpartum (day 1 of lactation) in the Standard Operating Procedures of the Testing Facility. Throughout this protocol, the day of birth will be designated day 1 postpartum (day 1 of lactation) and all subsequent ages of the F1 generation rats and days of the lactation period will be determined and cited accordingly. RANDOMIZATION AND COHABITATION: Upon anival, rats will be assigned to individual housing on the basis of computer-generated random units. During an acclimation period of at least five days, male and female rats will be selected for study on the basis of physical appearance and body weights recorded during acclimation. The rats will be assigned to dosage groups based on computer-generated (weightordered) randomization procedures. Withineach dosage group, consecutiveorder will be used to assign rats to cohabitation, one male rat per female rat. The cohabitationperiod will consist of a maximum of 14 days. Female rats with spermatozoaobserved in a smear of the vaginal contentsandor a copulatoryplug observed in situ will be considered to be at day 0 of presumed gestation and assigned to individual housing. Female rats not mated within the first seven days of cohabitation will be assigned alternatemale rats that have mated (same dosage group) and will remain in cohabitation for a maximum of seven additional days. Day 1 of lactation (postpartum) is defined as the day of birth and is also the first day on which all pups in a litter are individually weighed (pup body weights will be recorded after all pups in a litter are delivered and groomed by the dam). Litters will not be culled during the lactation period, because random selection of pups for culling could result in potential biases in pup viabilities and body weight gains over this period. 418-028:PAGE D-9 Protocol 4 18-028 Page 9 Within each dosage group, consecutive order will be used to assign the first 10 male and the first 10 femalerats to a hctional observationalbattery (FOB)and motor activity assessment., blood sample collection for clinical chemistry and hematology (CC&H), and histological evaluations. On day 22 postpartum, a table of random units will be used to select five male and five female pups per litter for blood sample and liver collection;these pups will only be selected from the ten dams selectedfor FOB,motor activity, CC&H and histological evaluation. ADMINISTRATION: Route and Reason for Choice: The oral (gavage) route was selected for use because: 1) in comparison with the dietary route, the exact dosage can be accurately administered; and 2) it is one of the possible routes for environmental exposure. Method and Frequency: Dosageswill be adjusted daily for body weight changes and given at approximatelythe same time each day. The first day of dosage is designated as day 1 of study. Male rats will be given the test substance once daily beginning 14 days before a cohabitation period that consists of a maximum 14 days. Dosage will continue through the day before sacrifice, after completionof the cohabitationperiod, after a minimum of 42 days of administration. Femalerats will be given the test substanceonce dailybeginning 14 days before a cohabitation period that consists of a maximum of 14 days. Dosage will continue through the day before scheduled sacrifice (day 2 1 of lactation). Rationale for Dosage Selection: Dosages were selected by the Sponsor based on previous studies conducted with the test substance,taking into account possible differencesin sensitivitybetween pregnant and nonpregnant rats. The highest dosage will be expected to cause toxic effects but not mortality or obvious suffering. The descending sequence of the lower dosage levels will be selected for the purpose of demonstrating any dosage-related response, with no adverse effects expected at the lowest level. Dosape Levels, Concentrationsand Volumes: 418-028:PAGE D-10 Protocol 418-028 Page 10 a. The test substancewll be considered 100%pure for the purpose of dosage calculations. b. Three additional ratsper sex per dosagegroup will be assig&d to toxic&etic sarnple collection. TESTS, ANALYSES AND MEASUREMENTS - FOGENERATION: Viabilitv - Male and Female Rats: All Periods: At least twice daily. - Clinical Observationsand/or General Amearance Male and Female Rats: Acclimation Period: Weekly. Dosage Period: Daily before dosage. On the first day ofdosage, postdosageobservationswill be recorded at approximately hourly intervals for the first four hours and at the end ofthe normal working day. Subsequentpostdosageobsenrations will be recorded at intervals deemed appropriateby the StudyDirector or designee after determination ofpeak toxicologic effects. Maternal Behavior: Days 1,5,8,15 and 22 postpartum. Observed abnormal behavior recorded daily. Clinical observationsmay be recorded more fiequentlythan cited above, if deemed appropriate by the StudyDirector a d o r StudyMonitor. Data collected for rats assigned to toxicokinetic sample collectionwill not be summarized or analyzed statistically. - Detailed Clinical Observations Male and Female Rats: Once before the b t dosage and at least once weekly thereafter, detailed clinical observations will be conducted for all male and female rats. These observationswill be made outsidethe cage in a standard arena at the same time each day of conduct. Effort will be made to ensure that variations in the test conditionsare minimal and that observationsare conductedby observers unaware of treatment groups. Signs noted should include,but not be limited to: changesin skin, fur, eyes, mucous membranes, occurrence of secretions and excretionsand autonomic activity (e.g., lacrimation, piloerection, pupil size, unusual respiratorypattern). Changes in gait, posture 418-028:PAGE D-11 Proiocol4 18-028 Page 11 and response to handling as well as the presence of clonic or tonic movements, sterotypic behavior (e.g., excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behavior (e.g., self-mutilation,walking backwards) should also be recorded. Bodv Weiphts - Male and Female Rats: Acclimation Period: Weekly. Dosage Period: Daily. Sacrifice: Terminal weight. Feed Consumotion Values - Male Rats (recorded and tabulated): Dosage Period: Weekly. Feed Consumotion Values - Female Rats (recorded and tabulated): Dosage Period: Weekly to cohabitation. Gestation Period: Days 0,7,10, 12, 15, 18,20 and 25 (if necessary). Postpartum Period Days 1,5,8 and 15. Feed consumption not tabulated after day 15 postpartum, when it is expected that pups will begin to consume maternal feed. Feed Consumption Values - Male and Female Rats: Feed consumptionvalues may be recorded more fiequentlythan cited above if it is necessary to replenish the feed. During cohabitation,when two rats occupy the same cage with one feedjar, replenishmentof the feedjars will be documented. Individual values will not be recorded or tabulated. Toxicokinetic Sample Collection: On day 14 and 42 of study, blood samples (approximately1mL each) will be collected fkom each male rat assignedto the toxicokineticsample collectionportion ofthe study (3 pel group). On day 14 of study and day 21 of presumed gestation, blood samples (approximately 1 mL each) will be collected from each female rat assigned to the toxicokinetic sample collectionportion of the study (3 per group). Samples will be collected prior to dosage on day 14 of study. The time of eachblood collection will be recorded in the raw data. 418-028:PAGE D-12 Protocol 4 18-028 Page 12 Blood will be collected fiom the orbital sinus. If necessary, blood may be collected fiom an alternate site; if so, the alternate site will be documented in the raw data.) The samples will be transferred into EDTA-coated (purple top) tubes and spun in a centrifige. The resulting serum will be transferred into polypropylenetubes labeled with the protocol number, Sponsor study number, animal number, sex, group number, dosage level, day of study, collection interval, date of collection, species, generation and storage conditions. All samples will be immediately fiozen on dry ice and maintained fkozen (<-7O"C) until shipment for analysis. After the last blood sample collection, rats will be sacrificed and samples of the liver will be collected for analysis. Shippiup Instructions: Samplesto be analyzed will be shipped on dry ice to: Principal Investigator: Lisa Clemen 3M Environmental Technology and Safety Services Building 2-3E-09 St. Paul, Minnesota 55 133-3331 Telephone: (651) 778-5568 Telefax: (65 1) 778-6176 Email: laclemen@mrnm.com The recipient will be notified in advance of sample shipment. Estrous Cvclinp and Mating: Estrous cycling will be evaluated by examination of vaginal cytologybeginning with the day after the first administrationand then until spermatozoaare observed in a smear of the vaginal contents andlor a copulatoryplug is observed in situ during the cohabitationperiod. - Caesarean-Sectioninp Toxicokinetic Studv: On day 21 of presumed gestation, blood and liver sampleswill be collected fkom all female rats designated for toxicokinetic sample collection. Blood samples will be collected from the rats as previously described. After sacrifice, the liver of each rat will be excised and the liver weight will be recorded. The median liver lobe will be frozen and stored (5-20C) until shipment for possible analysis. The fetuses will be removed firom the uterus and blood samples will be collected from each fetus via decapitation. Blood will be placed into tubes, pooled per litter, allowed to clot and spun in a centrifuge; the resulting s e m will be transferred into labeledpolypropylenetubes. All samples will be immediatelyfkozen on dry ice and maintained fkozen (5-70C) until shipment for analysis. 418-028:PAGE D-13 Protocol 4 18-028 Page 13 The liver ftom each fetus will be collected, pooled per litter and placed into labeled tubes. The samples will be fkozen and stored (<-2OoC) until shipment for analysis. Samples will be shipped on dry ice to Lisa Clemen at the previously cited address. Natural Deiiverv: Female rats will be evaluated for: Adverse Clinical Signs Observed During Parturition. Duration of Gestation (day 0 of presumed gestation to the time the first pup is observed). Litter Size (defined as all pups delivered). Pup Viability at Birth. Functional Observational Battery: On one occasion during the course of the study, a functional observational battery will be conducted on 10male and 10 femalerats per group. For male rats, this assessmentwill be conducted shortly before scheduled sacrifice, but prior to blood sample collection for hematology and clinical chemistry evaluations. Female rats should be tested during the lactation period, shortly before scheduled sacrifice. The FOB,to be conductedby an observerunaware of the group assignmentof the rat, will assess the followingparameters: 1. Lacrimation, salivation, palpebral closure, prominence of the eye, pupillary reaction to light, piloerection, respiration, and urination and defecation (autonomic functions). 2. Sensorimotorresponses to visual, auditory,tactile and painful stimuli (reactivity and sensitivity). 3. Reactions to handling and behavior in the open field (excitability). 4. Gait pattern in the open field, severity of gait abnormalities, air righting reaction, and landing foot splay (gait and sensorimotorcoordination). 5. Forelimb and hindlimb grip strength. 6. Abnormal clinical signs including but not limited to convulsions, tremors and other unusual behavior, hypotonia or hypertania, emaciation, dehydration, unkempt appearance and deposits around the eyes, nose or mouth. 418-028:PAGE D-14 Protocol 418-028 Page 14 Evidence of the ability of this battery to detect the effects of positive control substances will be provided (Testing Facility Positive Control Data). Data will also be provided to document interobserverreliability if more than one observer is involved in the testing. Motor Activitv Test: Motor activity will be evaluated on 10 male and 10 female rats per group once during the course of the study. This assessment will be conducted shortly before scheduled sacrifice, but prior to blood sample collection. The movements of each rat will be monitored by a passive infrared sensor mounted outside a stainless-steelwire-bottomed cage (40.6 x 25.4 x 17.8 cm). Each test session will be 1.5 hours in duration with the number of movements and time spent in movement tabulated at each fiveminute interval. The apparatus will monitor a rack of up to 32 cages and sensors during each session, with each rat tested in the same location on the rack across test sessions. Groups will be counterbalancedacross testing sessions and cages. Data will be provided to demonstrate that the test system is capable of detecting increases in activity produced by positive control substances (Testing Facility Positive Control Data). HEMATOLOGY AND CLINICAL CHEMISTRY: At scheduled sacrifice, the rats (fasted) assigned to hematology and clinical chemistry (H&CC) sample collection will be exsanguinatedfiorn the inferior vena cava following sacrificeby carbon dioxide asphyxiation. Approximately 5 niL of blood will be collected and processed as describedbelow. Determinationsadditional to those describedbelow may be conducted if the known properties of the test substancemay, or are suspectedto, affect related metabolicprofiles (e.g., calcium, phosphate, fasting triglycerides and fasting glucose, specific hormones, and cholinesterase). Hematolow: Approximately 1 mL of blood will be collected into EDTA-coated tubes and maintained on wet ice or refiigerated until shipment for analysis of the followinghematologicparameters: Erythrocyte Count (RBC) Hematocrit (HCT) Hemoglobin (HGB) Mean Corpuscular Hemoglobin (MCH) Mean Corpuscular Hemoglobin Concentration (MCHC) Mean Corpuscular Volume (MCV) Leukocyte Count, Total (WBC) Leukocyte Differential Platelet Count (PLAT) Mean Platelet Volume (MPV) Cell Morphology Two blood smear slides will be prepared at the Testing Facility for each sample for measurementsof differentialleukocyte count. All samples (on wet ice) and slides (ambient conditions) will be shipped to Redfield Laboratories at the following address. Approximately 1.8 mL of blood will be added to a tube containing 0.2 mL of sodium citrate (0.129 M). The contents will be mixed and maintained on wet ice until the tubes are centrifbged 418-028:PAGE D-15 Protocol 4 18-028 Page 15 (within30 minutes of the collectiontime). The resultingplasma will be transferred 2.0 mL polypropylenetubes labeled with study number, Sponsor's study number, rat number, dosage level, day of study, collection interval, date of collection, species, generation and storage. All sampleswill be frozen on dry ice and maintained frozen (570C) until shipment on dry ice by overnight courier for measurement ofprothrombin time (PT) and activated partial thromboplastintime (APTT). Approximately2 mL of blood will be collected into serum separator tubes and centrifuged. The resulting sera samples will be immediately fiozen on dry ice and maintained fiozen (570C) until shipment for analysis of the followingparameters: Total Protein (TP) Triglycerides (TRI) Albumin (A) Globulin (G) Albumin/Globulin Ratio (NG) Glucose (GLU) Cholesterol (CHOL) Total Bilirubin (TBILI) Urea Nitrogen (BUN) Creatinine (CREAT) CreatinineKinase (CK) Alanine h i n o t r a n s f e r a s e (ALT) Aspartate Aminotransferase (AST) Alkaline Phosphatase (ALK) Calcium (CA) Phosphorus (PHOS) Sodium (NA) Potassium (K) Chloride (CL) Sampleswill be shipped (on dry ice) to Redfield Laboratories at the following address. ShiDDinP Instructions: Sampleswill be shippedto arrive on Monday through Friday according to the conditions described above to: Principal Investigator: Ms.Phyllis Powell Redfield Laboratories A Division of CRL-DDS 100East Boone Street P.O.Box 308 Redfield, Arkansas 72132 Telephone: (501) 397-2540 Telefax: (501) 397-2002 The recipient will be notified in advance of sample shipment. Urinalysiswill not be conductedunless indicated based on expected or observed toxicity of the test substance. 418-028:PAGE D-16 METHOD OF SACRIFICE: Protocol 4 18-028 Page 16 Fo generation rats will be sacrificed by carbon dioxide asphyxiation. GROSS NECROPSY AND - HISTOPATHOLOGY FOGENERATIONRATS: - ScheduledSacrifice Toxicokinetic Studv: Scheduledsacrificeof male rats will be conducted on day 42 of study. Scheduledsacrificeof female rats will be conducted on day 21 of presumed gestation. Blood samples will be collected fi-omthe rats as previously described. After sacrifice, the liver of each rat will be excised and the liver weight will be recorded. The median liver lobe will be fiozen and stored (5-20"C) until shipment for analysis. Fetal samples will be collected as previously described. Carcasses will be discarded without fiuther evaluation. Sampleswill be shipped on dry ice to Lisa Clemen at the previously cited address. Scheduled Sacrifice - Main Studv: Scheduledsacrificeofmale rats will be conducted on the day followingthe last dosage administration, after a minimum of42 days of dosage. Scheduled sacrificeof female rats will be conductedon day 22 oflactation. Gross necropsy of all male and female rats will include an initial physical examination of external surfaces and all orifices, as well as the cranial, thoracic and abdominal cavities and their contents. Special attention will be paid to the organs of the reproductive system. The number of implantation sites and corpora lutea will be recorded. Male and female rats will be examined for gross lesions. Gross lesions will be retained in neutral buffered 10% formalin and examined histologically. Tissue trimming and histopathology will be performed under the supervisionof or by a Board-Certified VeterinaryPathologist. The ovaries and the uterus with cervix of each female rat will be weighed, and ovaries, uterus, vagina and a mammary gland will be retained in neutral buffered 10% fomalin. Uteri of apparently nonpregnant rats will be examined after being pressed between glass plates to confirm the absence of implantation sites, and retained in neutral buffered 10% formalin. 418-028:PAGE D-17 Protocol 4 18-028 Page 17 Sperm Evaluations of Male Rats: To assess the potential toxicity of the test article on the male reproductive system, the endpoints listed below will be evaluated fiom the first 10male rats in each dosage group. Organ Weights: The following organs will be individually weighed: right testis, left testis, left epididymis (whole and cauda), right epididymis, seminal vesicles (with and without fluid) and prostate. Sperm Evaluations: Sperm concentration and motility will be evaluated using computerassisted sperm analysis (CASA). Motility will be evaluated by the Hamilton Thome IVOS by collection of a sample fkom the left vas deferens. A homogenate will be prepared fiom the left cauda epididymisfor evaluationby the Hamilton Thorne NOS to determine sperm concentration (sperm per gram of tissue weight). The remaining portion of the left cauda epididymis will be used to manually evaluate sperm morphology. Sperm morphology evaluations will include the following: 1) determination of the percentage of normal sperm in a sample of at least 200; and 2) qualitative evaluation of abnormal sperm, including such categories as abnormal head, abnormal tail, and abnormal head and tail. See ATTACHMENT4 for additional tissues to be weighed and retained fiom the ten rats per sex per group assigned to histological sample collection and evaluation. All other tissues will be discarded. Scheduled Sacrifice of Female Rats that Do Not Deliver Litters: Rats that do not deliver a litter will be sacrificed on day 25 of presumed gestation. Gross necropsy, examination and tissue retention will be conducted as described previously for rats at scheduled sacrifice. Dams with No Survivinp Pups: Damswith no survivingpups will be sacrificed after the last pup is found dead, missingor presumed cannibalized. Gross necropsy, examination and tissue retention will be conducted as described above for rats at scheduled sacrifice. 418-028:PAGE D-18 Rats Found Dead or Moribund: Protocol 4 18-028 Page 18 Rats that die or are sacrificedbecause of moribund condition, abortion or premature deliverywill be examined for the cause of death or moribund condition on the day the observation is made. The rats will be examined for gross lesions. Testes and epididymides of male rats will be excised and paired organ weights will be recorded. The epididymides will be retained in neutral buffered 10% formalin. The testes will be fixed in Bouin's solution for 48 to 96 hours and then retained in neutral buffered 10%formalin. Pregnancy status and uterine contents of female rats will be recorded. Aborted fetuses and/or delivered pups will be examined to the extent possible. Uteri of apparently nonpregnant rats will be examined aRer being pressed between glass plates to confirm the absence of implantationsites. Ovaries and uteri will be retained in neutral buffered 10% formalin. TESTS. ANALYSESA N D MEASUREMENTS - F1 GENERATION: Viabilitv: Preweaning Period: Litters will be observed for dead pups at least twice daily. The pups in each litter will be counted once daily. Clinical Observations and/or General Appearance: Preweaning Period: Once daily. Pups will be observed if they are warm and clean, for evidence of a nest and if pups are grouped together and nursing or have milk in stomach. Each pup will be examined for general shape of the head, trunk,limbs, tail and presence of anus. Clinical observations may be recorded more fiequently than cited above, if deemed appropriate by the StudyDirector and/or the StudyMonitor. Body Webhts: Preweaning Period: Days 1(birth), 8,15 and 22 postpartum. Sacrifice: Terminal weight. Feed Consumption Values (recorded and tabulated): Preweaning Period: Not recorded. METHOD OF SACRIFICE- F1 GENERATIONPUPS: F1 generationpups will be sacrificedby carbon dioxide asphyxiation. 418-028:PAGE D-19 Protocol 4 18-028 Page 19 NECROPSY - F1 GENERATION: Gross lesions will be retained in neutral buffered 10% formalin for possible fbture evaluation. Unless specificallycited below, all other tissues will be discarded. Pups Found Dead on Dav 1 Postnartum: Pups that die before examinationof the litter for pup viability will be evaluated for vital status at birth. The lungs will be removed and immersed in water. Pups with lungs that sink will be identified as stillborn; pups with lungs that float will be identified as liveborn, and to have died shortly after birth. Pups with gross lesions will be preserved in Bouin's solution for possible future evaluation. Pups Found Dead or Moribund on Days 2 to 4 Postpartum: Pups found dead or sacrificedbecause of moribunditywill be examined for gross lesions and for the cause of death or the moribund condition. Pups with gross lesions will be preserved in Bouin's solution for possible fbture evaluation. Scheduled Sacrifice: On dayr 22 postparh~np, ups will be will be sacrificedand examined for gross lesions; gross lesions will be preserved in neutral buffered 10% formalin. Necropsy will include a single crosssectionof the head at the level ofthe frontal-parietalsuture and examination of the crosssectioned brain for apparent hydrocephaly. Blood samples will be collected from each selected pup (5 per sex per litter from the 10females per group selected for FOB and motor activity assessment,blood sample collection for H&CC, and histological evaluations)from the vena cava. The blood will be placed into tubes, pooled per litter, allowed to clot and spun in a centrifuge; the resulting serum will be transferred into labeled polypropylene tubes. All samples will be immediatelyfiozen on dry ice and maintained frozen (5-70"C)until shipment for analysis. The liver fkom each selectedpup will be collected excised and the organ weight recorded. The median lobe will be ftozen and stored (520C) until shipment for possible analysis. Frozen samples will be shipped to Lisa Clemen at the previously cited address. The remaining portion of each liver will be retained in neutral buffered 10% formalin for possible histological evaluation. The livers will be processed and evaluated histologically as described for the Fo generation rats in Histological Evaluation in ATTACHMENT 4. 4 18-028:PAGE D-20 PROPOSED STATISTICAL The following schematicrepresents statistical analyses of the data. Protocol 4 18-028 Page 20 I. Parametric A. Bartletrs Test TvDe of Testa 11. N o n D a r a m e t r i c b A. Kruskai-Wallis Test g 75%liesat anyconcentration) I I Significant at p~0.001 Not Significant I Nonparametric I Analysis of Variance A Significant at ~ 5 0 . 0 5 Not Significant I Dunnett's Test Significant at p50.05 I Dunn's Test Not Significant B. Fisher's Exact Test on Proportionof Ties (+75% Yea al any concenbrtlpn) B. + Analysis of Variance with RepeatedMeasures . Significant at ~ 5 0 . 0 5 Not Significant A (Dosage) (Dosage x Block interaction) Dunnett's Test One-way ANOVA for each block I Significant at ~ 0 . 0 5 I Dunnett's Test I Not Significant 111. Test for ProDortion Data Variance Test for Homogeneity of the Binomial Distribution ---------------------------------- a. Statisticallysignificant probabilitiesare reported as eitherp 5 0.05 o r p 5 0.01. b. Proportion data are not included in this category. c. Test for homogeneity of variance. 418-028:PAGE D-21 Protocol 418-028 Page 21 Test items in the FOB using interval scales, such as the grip-strengthtests and the landingfoot splay test, as well as body weight data and feed consumptionvalues will be analyzed as described under the Parametric heading of the schematic. Bartlett's Test of Homogeneity of Variances(g)will be used to estimate the probabilitythat the groups had different variances. A nonsignificant result (p>O.OOl) will indicatethat an assumptionof homogeneityof variance is not inappropriate, and the data will be compared using the Analysis of Variance Test("). If that test is significant(p<0.05), the groups exposedto the test article/substancewill be comparedwith the control group using Dunnett's Test("). If Bartlett's Test is significant (pSO.OOl),the Analysis of Variance Test is not appropriate, and the data will be analyzed as described under the Nonparametricheading of the schematic. When 75% or fewer of the scores in all the groups are tied, the Kruskal-WallisTest(I2)will be used to analyze the data, and in the event of a significant result @10.05), Dunn's Test(13)will be used to compare the groups exposed to the test articlehbstance with the control group. When more than 75% of the scores in any group are tied, Fisher's Exact Test(14)will be used to compare the proportion of ties in the groups. Data fiom the motor activity test, with repeated measurementswithin a session, will be analyzed using an Analysis of Variance with Repeated Measures('*),as described under that heading in the schematic. A significant effect (p10.05) in that test can appear as effect of Concentration (a difference between groups in the total across all measurements in a session) or as an interaction between Concentrationand Block (a differencebetween groups at specific measurement periods). If the Concentrationeffect is significant,the totals for the control group and the groups given the test article/substancewill be compared using Dunnett's Test. If the Concentrationx Block interactionis significant, an Analysis of Variance Test will be used to evaluatethe data at each measurement period, and a significantresult (p3.05) will be followedby a comparison of the groups using Dunnett's Test. Test items in the FOB having graded or count scores will be analyzed using the procedures described under the Nonparametricheading of the schematic. Clinical observation incidencedata, as well as the descriptiveand quanta1data fiom the FOB, will be analyzed as contingencytables using the Variance Test for Homogeneityof the Binomial Distribution(16). Alternate or additionalstatistical evaluations may be performed if deemed necessaryor appropriate. 418-028:PAGE D-22 Protocol 4 18-028 Page 22 DATA ACOUISITION, VERIF'ICATIONAND STORAGE: Data generated during the course of this studywill be recorded either by hand or using the Argus AutomatedData Collectionand Management System,the Vivarium Temperatureand Relative Humidity Monitoring System,the CoulbournInstruments PassiveInpared Motor Activity System, the Codbourn Instruments Auditory Startle System, the CoulbournInstruments Spatial Delayed Alternation System, and/or the passive avoidance software. All data will be tabulated, summarized and/or statistically analyzed using the Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, MicrosoJtExcel [part ofMicrosoft Office 97 (version SR-2)] and/or me SASSystem (version 6.12). Records will be reviewed by the StudyDirector and/or appropriatemanagementpersonnel within 21 days after generation. All original records will be stored in the archives of the Testing Facility. All originaldata will be bound and indexed. A copy of all raw data will be suppliedto the Sponsor upon request. Preserved tissues will be stored at the Testing Facility at no charge for one year after mailing of the draft finalreport, after which time the Sponsor will be contacted to determinethe dispositionof these materials. KEY PERSONNEL: Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Director of Research: Alan M. Hoberman, Ph.D., DABT Associate Director of Research and Study Director: Raymond G. York, Ph.D., DABT Director of Operations and Compliance: Barbara J. Patterson, B.A. Director of Laboratory Operations: John F. Bamett, B.S. Director of StudyManagement: ValerieA. Sharper, M.S. Manager of Animal Operations: Dena C. Lebo, V.M.D. Chairperson, InstitutionalAnimal Care and Use Committee: Douglas B. Learn, Ph.D. Consultant, VeterinaryPathology: W. Ray Brown, D.V.M., Ph.D., ACVP 418-028:PAGE D-23 RECORDS TO BE MAINTAINED: Protocol418-028 Page 23 Protocol and Amendments. Test Substance, Vehicle and/or Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (if prescribed by Staff Veterinarian). General Comments. Clinical Observationsand/or General Appearance. Body Weights. Feed Consumption Values. Natural Delivery Observations. FOB and Motor Activity Observations. Blood Sample Collection, Processing and Shipment. Gross Necropsy Observations. Organ Weights. Photographs (if required). StudyMaintenance(room and environmentalrecords). Feed and Water Analyses. Packing andlor Shipment Lists. FINAL REPORT: The Study Director will provide periodic updates of study progress to the Sponsor. Draft summary tables of unaudited computer-recordeddata may accompany these updates. Statistical analyses will not be performed on these interim data. A comprehensive draft final report will be prepared on completion of the study and will be finalized followingconsultationwith the Sponsor. The report will include the following: S u m m q and Conclusion. ExperimentalDesign and Method. Evaluation of Test Results. Appendices: Figures, Summary and Individual Tables Summarizingthe Above Data, Protocol and Associated Amendments and Deviations, StudyDirector's GLP Compliance Statement, Reports of SupportingData (if appropriate) and QAU Statement. The Sponsorwill receive one copy of the draft report and two copies of the final report. Data will be hand- andor computer-recorded. Records will be reviewed by the Study Director and/or appropriatemanagementpersonnel within 21 days after generation. All originalrecords will be stored in the archives of the Testing Facility. All original data will be bound and indexed. A copy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the Testing Facility at no charge for one year after mailing of the draft final report, after which time the Sponsor will be contacted to determine the disposition of these materials. 418-028:PAGE D-24 Protocol 418-028 Page 24 INSTITUTIONALANIMAL CARE AND USE COMMITTEESTATEMENT: The procedures described in this protocol have been reviewed by the Testing Facility's Institutional Animal Care and Use Committee. All procedures described in this protocol that involve study animals will be conducted in a manner to avoid or minimize discomfort, distress or pain to the animals. The Sponsor's signature below documentsthe fact that informationconcerning the necessity for conducting this study and the fact that this is not an unnecessarily duplicative study may be obtained from the Sponsor. No alternative(in vitro)procedureswere available for meeting the stated purposes of the study. REFERENCES: 1. Christian, M.S.and Voytek, P.E. (1982). In VivoReproductive and Mutagenicity Tests. Environmental ProtectionAgency, Washington,D.C. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 2. Christian, M.S.(1984). Reproductivetoxicity and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November 7,1983), J. Clin. Psychiat. 45(9):7-10. 3. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) ControlData in the CharlesRiver Crl:CD@BRRat. Charles River Laboratories,Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 4. Institute of Laboratory Animal Resources (1996). Guidefor the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Haggerty, G.C. (1989). Development of Tier I neurobehavioraltesting capabilities for incorporation into pivotal rodent safety assessment studies. J. h e r . Col. Toxicol. 8 5 3 70. 6. Irwin, S. (1968). Comprehensiveobservational assessment: Ia. A systemic quantitative procedure for assessing the behavioral and physiologic state of the mouse. Psychophannacologia(Berlin) 13:222-257. 7. Moser, V.C. (1989). Screeningapproachesto neurotoxicity: A functional observational battery. J. h e r . Col. Toxicol. 8:85-94. 8. O'Donoghue, J.L. (1989). Screening for neurotoxicityusing a neurologically based examination and neuropathology. J. h e r . Col. Toxicol. 8:97-116. 9. Sokal, R.R. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances. Biometry, W.H. Freeman and Co., San Francisco, pp. 370-371. 418-028:PAGE D-25 Protocol 418-028 Page 25 10. Snedecor,G.W. and Cochran,W.G. (1967). Analysis of Variance. StatisticalMethods, 6th Edition, Iowa State University Press, Ames, pp. 258-275. 1 1. Dunnett, C.W. (1955). A multiple comparisonprocedure for comparingseveral treatmentswith a control. J. h e r . Stat. Assoc. 50:1096-1121. 12. Sokal,R.R. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometry, W.H.Freeman and Co., San Francisco, pp. 388-389. 13. Dunn, O.J.(1964). Multiplecomparisonsusingrank s m s . Teclmometrics6(3):241-252. 14. Siegel, S. (1956). Nonpararnetric Statisticsfor the Behavioral Sciences,McGraw-Hill, New York, pp. 96-104. 15. SAS Institute, Inc. (1988). Repeated measures analysis of variance. S A S / S T A F User's Guide,Release 6.03 Edition, Cary, NC, pp. 602-609. 16. Snedecor, G.W. and Cochran, W.G.(1967). Variance test for homogeneityof the binomial distribution. Statistical Methods, 6th Edition,Iowa State UniversityPress, Ames, pp. 240-241. PROTOCOL APPROVAL: FOR THE TESTING FACILITY Alan M.Hobeman, Ph.D., DABT Director of Research A s s w t e Director of kaeadch Study Director Thegsa H. Woodard, D.&. Member,Institutional Animal Care and Use Committee FOR THE SPONSOR 4 18-028:PAGE D-26 Protocol 418-028 Page 26 418-028:PAGE D-27 ATTACHMENT 1 SCHEMATICOF STUDY DESIGN A N D STUDY SCHEDULE 418-028:PAGE D-28 ATTACHMENT 1 STUDY SCHEMATIC Protocol 4 18-028 page 1 of3 COMBINED REPEAT DOSE AND REPRODUCTIVE/DEVELOPMENTALTOXICITY SCREENa First Day of Test Substance Dosage I Premating I Last Day of Test Substance Dosage Cohabitation I Last Day of Test Substance Dosage' Female Rats Motor Activity/F0Bb 1 2 Weeks 2 Weeks Natural Delivery Presumed Gestation 3 Weeks Postpartum Period I I. I I I Day21 Day22' Motor Activity/FOBd =Dosage Period a. For additional details see "Tests, Analyses and Measurements"section ofthe protocol. b. FOB and motor activityevaluationsconductedon ten males per group. c. Male rats sacrificed after completion of at least 42 days of dosage; necropsy and retention of male reproductiveorgans. Hematology, clinical biochemistry and histological samples collected fiom ten male rats per group. d. Ten female rats per group assigned to FOB evaluation and motor activity evaluation. e. Ten female rats per group and their litters sacrificedon day 22 postpartum; necropsy and retention of female reproductive organs Hematology, clinical biochemistry and histologicai samples collected. Remaining female rats sacrificed on day 22 postpartum and discarded. ATTACHMENT 1 4 18-028:PAGE D-29 Protocol 4 18-028 Page 2 of 3 SCHEDULE' 26 MAR 02 01 APR02 01 APR 02 - 12 JUN 02 - 14 APR 02P M 21 APR 02AM 21 APR02 PM - 28 APR 02 AM 14 APR 02 15 APR 02 28 APR 02 06 MAY 02 19 MAY 02 06 MAY 02 - 07 MAY 02 Animal Receipt - Acclimation Begins. Start of Dosage Period - Male Rats (14 days before cohabitation and through a 14-daycohabitation period until the day before sacrifice after at least 42 days ofdosage). Dosage Period - Female Rats (14 days before cohabitation through Day 22 of lactation). CohabitationPeriod (Maximumof 14 days). Male 1 (7 days) Male 2 (7days) Day 14 Toxicokinetic Sample Collection First Possible Day 0 of Presumed Gestation. Last Possible Day 0 of Presumed Gestation. First Possible Day 21 of Presumed Gestavion Toxicokinetic Sample Collection and Sacrifice Toxicokinetic Study Female Rats Last Possible Day 2 1 of Presumed Gestation Toxicokinetic Sample Collection and Sacrifice Toxicokinetic Study Female Rats FOB and Motor Activity Evaluation- 10Male Rats per Group a. The start date of the study is the day the Study Director signs the protocol. b. Throughout this schedule, the day of birth is designated day 1 postpartum (day 1 of lactation) and all subsequent ages of the F1 generation rats and days of the lactation period will be determined and cited accordingly, 3s described above the protocol section, "Day Numbering System." 418-028:PAGE D-30 ATTACHMENT 1 06 MAY 02 23 MAY 02 10MAY 02 23 MAY 02 12MAY 02 13 MAY 02 23 MAY 02 - 09 JUN 02 27 MAY 02- 13 JUN 02 24 SEP 02 Protocol 4 18-028 Page 3 of 3 First Possible Delivery (Day 21 of presumed gestation). Last Possible Delivery (Day 25 of presumed gestation). First Possible Day 25 of Presumed Gestation Female Sacrifice. Last Possible Day 25 of Presumed Gestation Female Sacrifice. Day 42 Toxicokinetic Sample Collection and Scheduled Sacrifice - Toxicokinetic Study Male Rats Scheduled Sacrifice - Main Study Male Rats (Earliestpossible date). Hematology, Clinical Biochemistry and Histological Sample Collection of Selected Male Rats. FOB and Motor Activity Evaluation - 10 Female Rats per Group. Day 22 Postpartum - Sacrifice Female Rats and Pups. Hematology, Clinical Chemistry and Histological Sample Collection of Selected Female Rats and Pups. DrafFinal Report 418-028:PAGED m 3 1 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 418-028:PAGE D-32 MATERIAL SAFETV DATA S H E T (Experimental) 3M 3~ center S t . ~ a u l ,Minnesota 55144- 1000 1-800-364-3577 o r (651) 737-6501 (24 hours) Copyright, 1999, Minnesota Mining and Manufacturing Company. - A U rights reserved. Copying and/or downloadrng of t h i s information f o r the purpose o f properly u t i l i z i n g 3M products is allowed provided that: 1) the information is copied i n full w i t h no changes unless p r i o r agreement is obtained from 3M, and 2) neither the copy nor the original i s resold o r otherwise distributed w i t h the intention of earning a p r o f i t thereon. OIVISION: 3M SPECIALTY MATERIALS MATERIAL : L-9051 DEVELOPMENTAL PRODWT ISSUED: December 0 7 , WQ9 SUPERSEDES: flay 17, 1999 DOCUMENT: 04-5470-2 . . POTASSIUM PERFLWROHEXANE SULFONATE. ..... RESIDUAL ORGANIC FtUOROCHEMICALS... 3871 -99-6 Mixture 100.0 Unknown T h i s material is n o t l i s t e d on the TSCA inventory and should be used f o r research and development purposes only under the direct supervision of a technically qualified individual. ............. BOILING PQfNT:.... ............ VAPOR PRESSURE:.... VAPOR DEMSIfV:................. N/A Negligible NjA ............. EVAPORATION RATE:. Negligible ........... ............. SOLUBILITY IN WTER: SPECIFIC GRAVITY:. sLight ca.-l.O Waterrl PERCENT VOLATlLEs. ............. (Bulk) NegligiUe vprHs:.co.s..m.:............................................NN//DA MELTING P o r w : . . . . . . . . . . . . ..... N/D APPEARANCE AND ODOR: Off -white crystalline solid, sharp odor. - MSDS: L 9051 OEVELOPMENTAL PRODUCT December 07, 1999 418-028:PAGE D-33 PAGE 2 ..... FLASH PuPNI:.............. ........ - FLAMMABLE LIMITS LEL: - .. .... FLAMMAsLE LIMITS UEL: *. .. AUTOIGNITION TEMPERATURE: * . 212 li setaflash Setaflash Closed Cup N/A N/A N/D EXTINGUISHING MEDIA: Water, Carbon dioxide, Dry chemical, Foam SPECIAL FIRE FIGHTING PROCEDURES: Wear f u l l p r o t e c t i v e clothing, i n c l u d i n g helmet, self-contained, p o s i t i v e pressure o r pressure demand breathing apparatus, bunker coat and pants, bands around arms, klaist and legs, face mask, and p r o t e c t i v e c o v e r i n g f o r exposed areas of the head. UNUSUAL FIRE AND EXPLOSION HAZARDS: See Hazardous Decomposition section f o r products o f combustion. STABILITY: Stable INCOMPATIBILInC MATERIALS/CONDITIONS TO AVOID: None known. HAZARDOUS POLYMERIZATION: Hazardous polymerization w i l l not occur. HAZARDOUS DECOMPOSITION PRODUCTS: May produce fluorocarbon gases if exposed t o very high temperatures (over 300 C). SPILL RESPONSE: Observe p r e c a u t i o n s from other sections. C o l l e c t s p i l l e d material. Use wet sweeping compound o r water t o avoid dusting. Clean up residue. Place i n a closed container. RECOMMENDED DISPOSAL: Incinerate i n an industrial o r commercial f a c i l i t y in the presence o f a combustible material. Combustion produots w i l l include HF. Disposal a l t e r n a t i v e : Oispose of waste product in a f a c i l i t y permitted t o accept chemical waste. -. I MSDS: -L 9051 DEVELOPMENTAL PRODUCT December 0 7 , 1999 418-028:PAGE D-34 PAGE 3 tNVPRZlj#MENI Ai OAT A : Not determined. REGULATORY INFORMATION: V o l a t i l e Organic Compounds: N/D. VOC Less H20 & Exempt Solvents: N/D. Since regulations vary, consult applicable regulations or authorities before disposal. U.S. EPA Hazardous Haste Number = None (Not U.S. EPA Hazardous) OTHER ENVIRONMENTAL INFORMATION: This product may contain one o r more organic fluorochemicals that have the p o t e n t i a l t o r e s i s t degradation and p e r s i s t i n the environment. EPCRA HAZARD CLASS: FIRE HAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: No EYE CONTACT: Immediately flush eyes with large amounts of water. Get immediate medical attention. SKIN CONTACT: Flush s k i n with l a r g e amounts of water. I f i r r i t a t i o n persists, get medical attention. INHALATION: Ifsigns/symptoms occur, reflove person t o fresh a i r . If signs/symptoms continue, c a l l a physician. I F SWALLOWED: If swallowed, c a l l a physician imediately. Only induce vomiting a t the instruction o f a physician. #ever give anything by mouth t o afi unconscious person. EYE PROTECTTON: Avolid eye contact. Avoid eye contact w i t h vapor, spray, o r m i s t . The following should be worn alone o r i n combination, as appropriate, t o prevent eye contact: Wear vented goggles. --- Ab ----- brev - i - a - t ----- ions - : - -* N --- /D' - - - - ---- Not ---------*--- Determined - ---- N/A - - - - ---- Not ----------.-- Applicable - - --. CA * - - - - A p p r o x i m a t e l y 418-028:PAGE D-35 MSDS: L - 9 0 5 1 DEVELOPMENTAL PRODUCT December 0 7 , 1993 PAGE 4 ----.--------------------------------------------------- 7. PRECAUTIONARY INFORMTION (continued) SkIN PRBTECTZOPC; Avoid s k i n contact. Wear appnpriate gloves when handling t h i s m a t e r i a l . A p a i r o f gloves made from the following naterial(s) are recommended: butyl rubber, pQlyethylene/polyvinylidene c h l o r i d e (Saranex). use one o r more of the following personal p r o t e c t i o n items as necessary to prevent s k i n contact: head covering, coveralls. P r o t e c t i v e garments father than gloves) should be made of e c ithe hlor r id o e f - the follow (Saranex) i n g materials: polyethylene/polyvinylidene RECOMMENDED VENTILATION: Use w i t h appropriate local exhaust ventilation. Provide appropriate l o c a l exhaust v e n t i l a t i o n a t transfer p o i n t s . Use ]in a wellv e n t i l a t e d area. Provide s u f f i c i e n t v e n t i l a t i o n t o r a i n t a i n emissions below recommended exposure l i m i t s . If exhaust v e n t i l a t i o n is not adequate, use appropriate respiratory protection. Provide . v e n t i l a t i o n adequate t o control vapor concentrations below recommended exposure limits andlor control spray o r M i s t . Local exhaust v e n t i l a t i o n is recornmended where the mes airborne. m a t e r i a l beco RESPZRATORY PROTECTION: Avoid breathing o f airborne material. Select one o f t h e following NIOSH approved r e s p i r a t o r s based on airborne concentration of contaminants and i n accordance w i t h OSHA regulations: h a l f -mask supplied a i r respirator, f ull-face supplied a i r respirator. PREVENTION OF ACCIDENTAL INGESTION: Do n o t eat, drink o r smoke when using t h i s product. Wash exposed areas thoroughly w i t h soap and water. Wash hands a f t e r handling and before eating. RECOMMENDED STORAGE : Store a t room temperature. closed &ten not i n use. Keep container dry. Keep container F I R E AplO EXPLOSION AVOEOMCE: Not determined. EXPOSURE LIMITS MSOS: L-9051 DEVELOPMENTAL PRODUCT Oecember 0 7 , 1999 EXPOSURE LIMITS (continued) 418-028:PAGE D-36 PAGE 5 -SOURCE 3M: OF EXPOSURE LIMIT DATA: 3M Recommended Exposure Guidelines YE CONTACT: No information was found regarding effects from eye contact. Single exposure May cause: Mild Eye I r r i t a t i o n : signs/symptoms can include redness, swelling , pain and tearing. SKIN CONTACT: No information was found regarding effects from skin contact. May be absorbed through the skin and persist i n the body f o r an extended time. Single exposure may cause: , Moderate Skin I r r i t a t i o n : signs/symptoms can include redness swelling, itching, and dryness. INHALATION : No information was found regarding effects from inhalation exposure. May be absorbed by inhalation and persist i n the body f o r an extended the. Single overexposure above recommended guidelines, may cause: I r r i t a t i o n (upper r e s p i r a t o r y ) : signs/symptoas can include soreness o f t h e nose and throat, coughing and sneezing. IF SWALLOWED: Ingestion is not a l i k e l y route of exposure t o t h i s product. NO i n f o r m a t i o n was found regarding effects from swallowing, Animal studies conducted on Organic flMorOch@mlCalSwhich may be present i n t h i s product indicate effects including l i v e r disturbances, weight 1 0 ~ 5 ,loss o f appetite, lethargy, and neurological, pancreatic, adrenal and hematologic effects. Thew are no known human health I . MSDS: L-9051 DEVELOPMENTAL PRODUCT December 07, 1399 418-028:PAGE D-37 PAGE 6 effects from a n t i c i p a t e d exposure t o these organic fluorechsaicaPs when used as intended and instructed. OTHER HEALTH W R D INFORMATION: This product may c o n t a i n one o r more organic fluorochemicals t h a t have t h e p o t e n t i a l t o be absorbed and remain i n t h e body f o r long periods o f time, e i t h e r as the parent molecule or as metabolites, and may accumulate w i t h repeated exposures. There are no known human health e f f e c t s from a n t i c i p a t e d exposure t o these organic fluorochemicals when used as intended and instructed. The presence o f organic fluorochemicals in the blood of the general population and subpopfflations, such a s HOrkerS, has been published dating back t o the 1970's. 3M's epidemiological study of i t s own workers i n d i c a t e s no adverse e f f e c t s . SECTION CHANGE DATES PRECAUTIONARY INFO. SECTION CHANGED SINCE flay 17, 1999 ISSUE The i n f o r m a t i o n i n t h i s Material Safety Data Street (MSDS) is believed t o be correct as o f the date issued. 3M MAKES NO M M T I E S , EXPRESSEO OR IMPLIED, INCLWDING, BUT NOT LIMITED TO, ANY CIPLIED GIARffA$JNOF wmxuwrmr~rwOR FITNESS FOR A PARTICULAR PURPOSE OR COURSE OF PERFORMANCE OR USAGE OF TRADE. User is responsible for determining whether t h e 3M product is f i t f o r a p a r t i c u l a r purpose and suitable f o r user's method o f use o r application. Given the v a r i e t y of factors t h a t can a f f e c t t h e use and a p p l i c a t i o n of a 3M product, some o f which a r e uniquely w i t h i n the user's knowledge and control, i t is essential that t h e user evaluate the 3M product t o determine whether it is fit f o r a p a r t i c u l a r purpose and s u i t a b l e f o r user's Method of use or application. 3M provides information in electronic form as a service t o its custoeers. Due t o t h e remote p o s s i b i l i t y t h a t e l e c t r o n i c t r a n s f e r may have resulted i n e r r o r s , omissions o r alterations i n t h i s information, 3M makes no representations as t o i t s completeness or accuracy. f n addition, information obtarned from a database may not be as Current as the information in t h e MSDS available d i r e c t l y from 3M. 418-028:PAGE D-38 ATTACHMENT 3 TEST SUBSTANCE PREPARATION PROCEDURE 418-028:PAGE D-39 ATTACHMENT 3 Protocol 418-028 Version: 418-028(19 MAR 02) Page I of 2 TEST SUBSTANCE PREPARATlON PROCEDURE Test Substance: T-7706 Vehicle: Aqueous 0.506 CMC (medium viscosity) A. Purpose: The purpose of this procedure is to provide a method for the preparation of dosage suspensions of T-7706 for oral (gavage) administration to rats on Argus Research Study number 418628. B. General Information: 1. Afl suspension containers will be labeled and cofor-coded. Each label wiil specify the protocol number, test subsfanee identification, Argus batch number, concentration, dosage level, preparation date, expiration date and storage conditions. - - 2. Suspensions wifl be prepared: - Daily X Weekly - Approximately every ten days - For- days of use By Sponsor 3. Suspensions will be administered at a final dosage volume of I 0 mUkg. 4. Safety: -X Gloves, uniformhab coat, goggles or safety glasses with side shields - X Dust-mistlHEPA-filteredMask - Half-Face Respirator if not used in a chemical fume hood - Full-Face Respirator/Positive Pressure Hood - Tyvek Suit or fyvek apron and sleeves - - 5. Dosage suspensions adjusted for % Activity/Purity or Correction Factor: - - Yes X No (Calculations based on 100%) - % Activity %Purity Correction Factor 6 Sampling requirements: Cited in protocol 7. Storage: Cited in protocol 4 18-028:PAGE D-40 ATTACHMENT 3 Protocol418-028 Version: 418-028(19 MAR 02) Page 2 of 2 TEST SUBSTANCE PREPARATION PROCEDURE NOTE: Prior to test substance preparation accurately measure the required amount of the appropriate vehicle (R.O. deionized water should be used for calibration purposes) in a graduated cylinder, pour the required amount of vehicle into a beaker. Carefully mark each beaker at the meniscus. This mark will be used during the preparation to bring the test substance slurry up to volume. C. Dosage Suspension Preparation: I. Weigh the required amount of test substance on a piece of weigh paper or into an.appropriately sized mortar (see PREPARATION CALCULATIONS). 2. If weigh paper is used, transfer the test substance to an appropriately sized mortar. If necessary, grind the test substance into a fine powder. Slowly add a small amount of vehicle and grind. Continue to add vehicle slowly and grind the vehicle and the test substance together to form a fine slurry. Transfer the vehicle/test substance slurry to a marked beaker. 3. Rinse the mortar and pestle with additional vehicle to remove any remaining test substance. Transfer rinse to beaker. 4. Add additional vehicle to the beaker to bring volume up to the mark. Place on magnetic stir plate and agitate prior to and during sampling, aliquotting and/or administration. 5. Repeat steps (It)hrough (4) for each concentration. Approved b CIarification: Lo "- Yes Date: WP$+%i?& [see attached clarification form] 418-028:PAGE D-41 ATTACHME" 4 TISSUES TO BE WEIGHED, RETAINED AND EXAMINED HlSTOLQGICALLY 418-028:PAGE D-42 ATTACHMENT 4 Protocol 4 18-028 Page 1 of 2 TISSUES TO WEIGHED AND RETAINED FOR POSSIBLEEXAMINATION FROM TEN RATS PER SEX PER GROUP The ten rats per sex per group assigned to firnctional observationalbattery and motor activity tests will be assignedto hematology,clinical biochemistryand histologicalevaluations. Tissues to be Weiphed: The followingorganswill be excised,trimmed and individuallyweighed as soon as possible after excision to avoid drying. liver spleen kidneys brain adrenals heart thymus ovaries testes uterus (with cervix) right epididymis prostate left epididymis (whole and cauda) seminal vesicles (with and without fluid) Tissues to be Retained: The followingtissues or representativesampleswill be retained in neutral buffered 10% fonnalin. brain (representative regions including cerebrum, cerebellum, pons) small and large intestines(includingPeyer's patches) lungs (perfused with neutral buffered 10% fonnalin) lymph nodes (submandiblar and mediastinal) peripheral nerve (sciatic or tibial) gross lesions spinal cord (cervical, thoracic and lumbar) stomach liver kidneys adrenals spleen heart thymus thyroidparathyroid trachea uterus urinary bladder bone marrow (sternum) testes* ovaries prostate uterus seminal vesicles (with coagulating gland) vagina mammary gland (female rats only) * Testes will be fixed in Bouin's solution for 48 to 96 hours before being retained in neutral buffered 10% fonnalin. Additionally, the remaining portion of the left epididymis (corpus and caput), as well as the right epididymiswill be fixed in neutral buffered 10%formalin. 418-028:PAGE D-43 ATTACHMENT 4 Protocol 4 18-028 Page 2 of 2 Histological Examination: Histological examination of retained tissues, includingreproductiveorgans, will be coriducted for the assigned ten rats per sex &om the control and high dosage groups and from the F1 generation pups (livers) &om the control and high dosage groups. If lesions attributed to the test substanceare observedin the rats exposed to the high test substanceconcentration,the same tissues will be examined &om the assigned ten rats per sex exposed to the lower test substance concentrations. Should results warrant examination of the lower dosage groups and conduct of quantitativeevaluation, scheduled report date and prices will be adjusted accordingly. The postlactional ovary should containprimordial and growing follicles as well as the large corpora lutea of lactation. Histopathologicalexaminationmay detect qualitativedepletion of the primordial follicle population. A quantitative evaluation of primordial follicles will be conducted for Fo generation female rats; the number of rats, ovarian section selection and section sample size will be statisticallyappropriate for the evaluationprocedureused. Examinationwill include enumerationof the number of primordialfollicles,which can be combined with small growing follicles, for comparison of ovaries of rats assigned to treated and control groups. Shipping Instructions: Tissues to be examinedhistologicallywill be shipped (ambient conditions)to: Principal Investigator: W. Ray Brown, D.V.M., Veterinary Pathologist Research Pathology Services, Inc. 438 E. Butler Avenue New Britain, Pennsylvania 18901 Telephone: (215) 345-7070 Telefax: (215) 345-4326 Email: WRBRP S@concentric.net Ph.D., ACVP The recipient will be notified in advance of sample shipment. 418-028:PAGE D-44 PROTOCOL 418-028 ORAL,(GAVAGE) COMBINEDREPEATED DOSE TOXICITYSTUDY OF T-7706WITH THE REPRODUCTIONDEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER. T-7706.1 Amendment 1 - 17 April 2002 1. Detailed Clinical Observations - Male and Female Rats (page 10 of the protocol): [EffectiveDate: I April 20023 Detailed clinical observationswill not be recorded for male and female rats assigned to toxicokinetic sampling. Reason for Change: This change was made because sufficient data for evaluation of detailed clinical observationswill be available fiom the rats assigned to the main study. 2. Feed Consumption Values - Male Rats and Female Rats (page 11ofthe protocol): [EffectiveDate: 1 April 20021 Feed consumptionvalues will not be recorded or tabulated for male and female rats assigned to toxicokinetic sampling. Reason for Change: This change was made because sufficient data for evaluation of feed consumption will be available from the rats assigned to the main study. revisions made to this finaIized amendment must be made by subsequent amendment, 418-028:PAGE D-45 3. Estrous Cvclinn and Mating (page 12 of the protocol): Protocol 418-028 Amendment 1 Page 2 [Effective Date: 1 April 20021 For the rats assigned to toxicokinetic sampling, estrous cyclingwill be evaluated during the cohabitationperiod until spermatozoa are observed in a smear of the vaginal contents andor a copulatoryplug is observed in situ, but not during the dosage period, prior to cohabitation. Reason for Change: This change was made because sufficient data for evaluation of estrous cycling will be available fkom the rats assigned to the main study. 4. Scheduled Sacrifice - Toxicokinetic Studv (page 16 of the protocol): [EffectiveDate: 5 April 20021 The number of implantationsites and corpora lutea will be recorded. Carcasses will be discarded without fiuther evaluation. Reason of Channe: This change was made in order to provide more idormation about possible toxicity of the test substance in pregnant rats. 5. Scheduled Sacrifice - Main Studv (page 16 ofthe protocol): [EffectiveDate: 27 March 20021 The number of implantationsites will be recorded, rather than the number of implantation sites and corpora lutea will be recorded. Reason of Change: The number of corpora lutea will not be recorded because corpora lutea regress at a rapid rate and are not counted on studies at weaning. 6. Scheduled Sacrifice (page 19 of the protocol): [EffectiveDate: 4 April 20021 The liver from each selectedpup will be excised and the organ weight recorded, rather than the liver fkom each selected pup will be collected excised and the organ weight recorded. Any revisions made to this finalized amendment must be made by subsequent amendment. 4 18-028:PAGE D-46 Protocol 418-028 Amendment 1 Page 3 Reason for Chanpe: This change clarifiesthe protocol by removing an extraneous word. Theresa H. Woodard, D.V.M. Member, Institutional Animal Care and Use Committee Date JohnButenhoff, PbD., DABT, C M Date Study Monitor and Sponsor's Representative Any revisions made to this finalized amendment must be made by subsequent amendment. ---418-028:PAGED-47 ARGUS RESEARCH Churls River Laboratories Dizovety and DeveCopment Semikes PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 Amendment 2 - 15 July 2002 1. Safetv Precautions (page 3 of the protocol and Attachment 3 page 1 of the protocol): [EffectiveDate: 29 April 20021 A half face respirator will be worn in addition to gloves, appropriateeye protection and a unifodlab coat during formulation preparation of the bulk test substance. Reason for Change: This change was made to match the bulk test substance text with the text located within the Material Safety Data Sheet. 2. Study Schedule (Attachment 1 page 2 of the protocol): - [EffectiveDate: 29 April 20021 The dates for FOB and motor activity evaluations have been extended to four days (06 MAY 02 - 09 MAY 02) rather than two days (06MAY 02 07MAY 02). Any revisions to this finalized amendment must be made by subsequent amendment. 418-028:PAGE D-48 Protocol 4 18-028 Amendment 2 Page 2 Reason for Change: This change was made because more time is needed to evaluate animals assigned to FOB and motor activity evaluations. 0 Study Director Any revisions to this finalized amendment must be made by subsequent amendment. 418-028:PAGE D-49 -/* ARGUS RESEARCH Charfes River Laboratories Discovery und Development Services PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706WITH THEREPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 Amendment 3 - 17 July 2002 - 1. Bulk Test Substance Sampling, Shitmina Instructions, ShirpingInstructions, Caesarean-Sectioning - Toxicokinetic Study, Scheduled Sacrifice Toxicokinetic Studv, and Scheduled Sacrifice, (pages 4,5,12,13, 16 and 19, respectively of the protocol) [Effective Date: 29 May 2002) Samples for perfluorohexanesulfonate(PFHS) analysis shipped to Lisa Clemen at 3M Environmental Technology and Safety Serviceswill be reshipped and remaining samples, that were to be shipped to Lisa Clemen and have not yet been shipped, will be shipped to: John Flaherty, Ph.D. (PrincipalInvestigator) Exygen Research 3058 Research Drive State College, Pennsylvania 16801 Telephone: (814)272-1039,ext. 122 Telefax: (814) 231-1580 Email: john.flaherty@exygen.com Any revisions to this finalized amendment must be made by subsequent amendment. 418-028:PAGE D-50 Protocol 4 18-028 Amendment 3 Page 2 These samples include: Bulk test substance sampling (page 4 of the protocol) Concentration and homogeneity(page 5 of the protocol) Stability(page 5 of the protocol) Plasma toxicokinetic samples (page 11to 12 and page 16of the protocol) Plasma samples, rather than serum samples, were retained. Liver toxicokinetic samples (pages 12 and 16 of the protocol) Pooled fetal serum toxicokineticsamples (pages 12 and 16of the protocol) Pooled fetal liver toxicokinetic samples (pages 13 and 16 of the protocol) Pooled pup serum toxicokineticsamples (page 19 of the protocol) Pup liver toxicokinetic samples (page 19 of the protocol) The aqalyses will be subcontractedto Exygen by the Sponsor and the Quality AssuranceUnit for Exygen Research will conduct critical phase inspectionsand audit the respective results and reports accordingto the Standard Operating procedures of that facility. Such critical phase inspection reports and audit reports will be submittedby that facility to the StudyDirector, Raymond G. York. The date of the inspections and report submissions will be incorporatedinto a QAU statement generated by Exygen Research for inclusion in the final report for Protocol 418-028. Reason for Change: This change was made at the request of the Sponsor because 3M is not able to complete the formulation analysis with their current staffing. /'.Director of Research Thered H. Woodard, B.V.M. Member, Institutional Animal Care and Use Committee Date John Butenhoff, Ph.D., DABT, C M Date Study Monitor and Sponsor's Representative Any revisions to this finalized amendment must be made by subsequent amendment, 418-028:PAGE D-51 PROTOCOL 4 18-028 ORAL (GAVAGE) COMBINEDREPEATED DOSE TOXICITY STUDY OF T-7706WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 Amendment 4 - 20 March 2003 1. Bulk Test Substance Sampling,ShippingInstructions, Shipping Instructions, Caesarean-Sectioning - Toxicokinetic Studv, Scheduled Sacrifice - Toxicokinetic Studv, and Scheduled Sacrifice, (pages4,5, 12, 13, 16 and 19,respectively of the protocol) and Amendment 3. Item 1: Bulk Test SubstanceSampling;,ShipDing Instructions, ShippingInstructions,Caesarean-Sectioning- ToxicokineticStudv, Scheduled Sacrifice - Toxicokinetic Studv, and Scheduled Sacrifice (page 1 of Amendment 3) [Effective Date: 27 February 20031 The analyses performed by Exygen Research will be done according to Exygen Method ExM-023-071 Revision 1, entitled "Method of Analysis for the Determination of Perfluorohexanesulfonate(PFHS), Perfluorooctanesulfonate(PFOS) and PentadecafluorooctanoicAcid (PFOA) in Rat Liver, Serum and Urine Revision 1". , Any revisions to this finalized amendment must be made by subsequent amendment. 418-028:PAGE D-52 Protocol 418-028 Amendment 4 Page 2 Reason for Change: This change was made at the request of the Exygen Research in order to clarify the method of analysis. Chair, Institutional Animal Care and Use Committee Study Monitor and Sponsor's Representative Any revisions to this finalized amendment must be made by subsequent amendment. APPENDIX E DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 4 18-028: PAGE E- 1 DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 1. The following rats were not fasted overnight prior to necropsy. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter and there were no indications of toxicity in clinical chemistry parameters for the female rats. 2. Postdosage observations were performed out of the range of 60 -I- 10 minutes on the following dates. Dosage Group I IV Dosage (mgkglday) 0 3 Date 09 APR 02 03 JUN 02 03 JUN 02 Number of Rats Male Rats Female Rats 4 1 1 Range of Time Deviated (minutes) +1 to +2 +8 +7 These deviations did not adversely affect the outcome or interpretation of the study because the extent of the deviation was less than 10 minutes. 3. On day 39 of study (DS 39) (9 May 2002), a postdosage clinical observation was not recorded for one male rat in the 0.3 mg/kg/day dosage group (19153). This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 4. On DS 44 (14 May 2002), dosage volume was not recorded for one male rat in the 3 mg/kg/day dosage group (19156). This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 4 18-028: PAGE E-2 5. On DS 46 (16 May 2002), postdosage clinical observations and dosage volumes were not recorded for two male rats in the 0 mg/kg/day dosage group (19161 and 19167). These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 6. Detailed clinical observations were not recorded weekly for the following rats. Dosage Dosage Group (mg/kg/day) Date Sex Rat Numbers I 0 09 MAY 02 Female 19023, 19041, 19050 I1 0.3 09 MAY 02 Female 19004 I11 1 09 MAY 02 Female 19008 I IV I 3 I 09MAY02 I Female I 19054.19062 I v l 10 I 09MAY02 I Female 19020,19030 These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 7. On DS 13 (13 April 2002), feed left values were not recorded for all rats before they were placed into cohabitation. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 8. On days 1 and 2 of lactation (DLs 1 and 2) (10 May 2002 and 11 May 2002, respectively), one pup in the 0 mg/g/day dosage group (litter 19050) was recorded as having an adverse clinical observation without the sex being recorded. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 9. On DL 3 (12 May 2002), one litter in the 10 mg/kg/day dosage group (19022) was not recorded as having been observed. All pups appeared normal on DL 4. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 10. On DL 5 (12 May 2002), maternal observation was not recorded for one rat in the 0.3 mg/kg/day dosage group (19018). Maternal behavior on DL 8 was normal for this rat. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 11. On 19 May 2002, the frozen liver sample for one female rat with a non-confirmed date of mating in the 0 mg/kg/day dosage group (19077) was lost before sample shipment. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 4 18-028: PAGE E-3 12. On DL 22 (31 May 2002) the liver weight of one pup in the 1mg/kg/day dosage group (pup 9 in litter 19024) was not recorded. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. All deviations are documented in the raw data. Associate Director o&Res&kch Study Director APPENDIX F CERTIFICATE OF ANALYSIS 418-028:PAGE F-1 Overall Quantitative Compositional Results by LCMS, "F-NMR, and 'H-NMR Analyses Component Identities * Relative Wt.% Concentrations (single trial measurements, excluding any water) CF3CF2CF2CF2CF2CF2SO3"' IS'+' 88.93% (NMR) (CF3)2-CF-(CF2)3-S0tmK' (+' 6.83% (NMR) CF3CF2-CF(CF3)-CF2CF2-S03(-) K(+' CF3CF2CF2CFz-CF(CF3)-S0t-IK"' CF$F2CF2CF( CF3)-CF2-S0$-) K'+) 2.75%(NMR) 0.83% (NMR) 0.56% (NMR) (CF3)3-C-CF2-CF2S03'-) K"' 0.078% (NMR) C812HS03") K"' 0.009%(LCMS) C7F1sSO$-' K'+' 0.006% (LCMS) C4FgSO3(-)K(+' 0.004% (LCMS) Probable CnH2,,+2hydrocarbons 0.0034% (NMR) Other components Sum of CsF&O3(-) K(+)Isomers Sum of Known ImDurities * Trace amounts of other unassigned components were also detected. <0.001% (LCMS) 99.98% 0.021 % Page 1 of 1 File Reference: Co4-PFHS-127498-80.doc 418-028:PAGE F-2 3~ SPECIALTY MATERLALS MANUFACTURING DMSION ANALYTICAL LABORATORY - To: From: Request No. GID:32537 Dan Hakes - (3-2392) - EHSR - Auto & Chem Group - 236-1B-10 Tom Kestner - (3-5633) - SMMD Analytical Lab - 236-2B-11 Subject: - Date: Chemical Characterization of PFHS (127498-80) by 'H-NMR & "F-NMR Spectroscopy April 15,2002 ~~ SAMPLE DESCRIPTION: 127498-80: PFHS made by George Moore and to be used for toxicologicaltesting. Nominal product =C6F13-S03(')K(+)(white powder). OBJECTIVE: This sample was subjected to 'H-NMRand 19F-NMRspectral analyses to determine the purity of the nominal product and to characterizeas many impurity componentsas possible. Joel Miller also performed an LCMS analysis and his results were reported to you previously. EXPERIMENTAL: A portion ofthe sample(-400 mg) was totally dissolved in deuterated dimethylsulfoxide(DMSO-d6) and then the solution was spiked with a small amount of 1,4-bis(trifluoromethyl)benzene (p-HFX) for NMR analysis. A 400 MHz 'H-NMR spectrum (# h32537.GID.401) and a 376 MHz "F-NMR spectrum (# f32537.GID.401) were acquired at room temperatureusing a Varian UNITYplus 400 FT-NMRspectrometer. The p-HFX was used as a 'H/'%NMR cross integration standard to permit the cross correlationof the relative 'H and '4:signal intensities for evaluation of the overall sample composition. RESULTS: The combined N M R spectral data indicated the sample of 127498-80consisted of a high purity form of the nominal isomeric product mixture, C,,F2n+l-S03c)M", where 'n' was mainly 6 and where the metal cation ['M'+'']was assumed to be K(+).Trace-levels ofprobable aliphatic hydrocarbon impurity components were also observed and quantified. The qualitative and quantitativecompositional results that were derived fiom the single trial 'W'9F-NMR cross integration analysis are summarizedin TABLE4 on the followingpage. Any water that may have been present in the samplewas ignored for calculationpurposes. The relative weight percent concentrations shown in TABLE-1 should be very close to their respective absolute weight percent values. Trace amounts of other unidentified impurity componentswere also detected in the NMR spectra, but additional work would be needed in an effort to assign or quanti@ the unassigned impurities. Copies of the N M R spectra are attached with the paper copy of this report for your reference. If you have any questions about these results, or if any furtherwork is needed, please let me know. Tom Kestner c: JoelMiller George Moore Rick Payfer File Reference:dh32537.GID.docf93 Page 1 of 2 April 15,2002 418-028:PAGE F-3 3M SMMD Analytical Lab Request ## GID:32537 Identified Components * CF3CF2CF2CF2CF2CF~SO~I(-P' ' "F/'H-NMR Relative Wt.% Concentrations (single tial measurement) 88.94% (CF3)2-CF-( CF2)3-S0$-) K") 6.83% CF3CF2-CF(CF3)-CFzCFz-S03(-) K'" 2.75% CF3CF$F2CF2-CF( CF3)-SOJ-' K'+' 0.83% CF3CF;!CF2CF(CF3)-CF2-SO$' IS'+' 0.56% (CF3)3-C-CF2-CF2SOt-'K"' 0.078% Probable C,,Hzn+2hydrocarbons 0.0034% * Trace amounts of other unassigned components were also detected in the NMR spectra. Study NO.FACT-TCROOS Page 2 of2 APPENDIX G ANALYTICAL AND BIOANALYTICAL REPORT 418-028:PAGE G-1 ANALYTICAL PHASE STUDY TITLE Oral (Gavage) Combined Repeated Dose Toxicity Study of T-7706 with the ReproductiodDevelopmental Toxicity Screening Test DATA REOUIREMENTS OECD Principles on Good Laboratory Practice, ENV/MC/CHEM(98)17, U.S. Food and Drug Administration 21 CFR Part 58, MHW Good Laboratory Practice Standard for Safety Studies on Drugs Ordinance Number 21, March 26,1997 STUDY DIRECTOR Raymond G.York ANALYTICAL REPORT COMPLETION DATE July 21,2003 PERFORMXNG LABORATORY Exygen Research 3058 Research Drive State College, PA 16801 Phone: 814-272-1039 TESTING FACILITY Argus Research 905 Sheehy Drive, Building A Horsham, PA 19044-1297 Phone: 215-443-8710 STUDY SPONSOR 3M Corporate Toxicology Building 220-2E-02 St. Paul, MN 55144-1000 PROJECT Protocol Number: 418-028 Sponsor's Study Number: T-7706.1 Exygen Study Number: 023-072 Total Pages: 153 418-028:PAGE G-2 Exygen Study No.: 023-072 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Exgyen Study Number 023-072, entitled "Oral (Gavage) Combined Repeated Dose Toxicity Study of T-7706 with the ReproductiodDevelopmental Toxicity Screening Test," conducted for 3M Corporate Toxicology, was performed in compliance with U.S. Food and Drug Administration Good Laboratory Practice Regulations 21 CFR Part 58, OECD Principles on Good Laboratory Practice. ENV/MC/CHEM(98)17 and MHW Good Laboratory Practice Standards for Safety Studies on Drugs Ordinance Number 21 by Exygen Research. /John Flaherty / Principal Investigator Exygen Research Argus Research ~ John Butenhoff Study Monitor * 3M CorporateToxicology Exygen Research Date Date Date Page 2 of 153 418-028:PAGE G-3 Exygen StudyNo.:023-072 OUALITY ASSURANCE STATEMENT Exygen Research's Quality Assurance Unit reviewed Exygen Study Number 023-072, entitled, "Oral (Gavage) Combined Repeated Dose Toxicity Study of T-7706with the ReproductiodDevelopmental Toxicity Screening Test". All reviewed phases were inspected for conduct according to Exygen Research's Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Exygen Principal Investigator and Management and to the Study Director. Phase Date Inspected Date Reported to Date Reported to Principal Exygen Date Reported to Investigator Management Study Director 1. Protocol Review 09/24/02 11/20/02 11/22/02 01/20/03 2. Extraction, Fortification 09/24/02 11/21/02 12/18/02 0 1/20/03 3. Raw Data Review 02/03 -07/03 and Draft Analytical and 02/10- 11/03 Report Review 02/24/03 02/28/03 03/07/03 4. Final Analytical Report Review 07/21/03 07/2 1/03 07/21/03 07/21/03 Exygen Research Page 3 of 153 4 18-028:PAGE G-4 Exygen Study No.:023-072 CERTIFICATION OF AUTHENTICITY This report, for Exygen Study Number 023-072, is a true and complete representation of the raw data for the study. Submitted by: Exygen Research 3058 Research Drive State College, PA 16801 (814) 272-1039 Principal Investigator, Exygen: Vice President Exygen Research Exygen Research Facility Management: Exygen Research /lGe Study Di ctor, Argus Research: fl 3-JUL - 03 Date Argus Research Study Monitor, 3M: John Butenhoff Date 3M Corporate Toxicology Exygen Research Page 4 of 153 418-028:PAGE G-5 Exygen Study No.:023-072 STUDY IDENTIFICATION Oral (Gavage) Combined Repeated Dose Toxicity Study of T-7706 with the ReproductiodDevelopmenta1Toxicity Screening Test PROTOCOL NUMBER: 418-028 SPONSOR'SSTUDY NUMBER: T-7706.1 EXYGEN STUDY NUMBER: 023-072 TYPE OF STUDY: Residue SAMPLE MATRIX: Rat Liver, Serum, and Plasma TEST SUBSTANCE: Perfluorohexanesulfonate (PFHS) SPONSOR: 3M Corporate Toxicology Building 220-2E-02 St. Paul, MN 55144-1000 STUDY DIRECTOR: Raymond G. York Argus Research 905 Sheehy Drive, Building A Horsham, PA 19044-1297 TESTING FACILITY: Argus Research 905 Sheehy Drive, Building A Horsham, PA 19044-1297 STUDY MONITOR: John Butenhoff 3M Corporate Toxicology Building 220-2E-02 St. Paul, MN 55144-1000 PERFORMING LABORATORY: Exygen Research 3058 Research Drive State College, PA 16801 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: 03/26/02 Analytical Start Date: 09/16/02 Analytical Termination Date: 1I /13/02 Analytical Report Completion Date: 07/21/03 Exygen Research Page 5 of 153 418-028:PAGE G-6 Exygen Study No.:023-072 PROJECT PERSONNEL The Study Director for this project was Raymond G. York at Argus Research. The following personnel fkom Exygen Research were associated with various phases of the study: Name John Flaherty Karen Risha Paul Connolly Xiaoming Zhu Lawrence Ord Rickey Kelley Arny Sheehan Emily Decker Mark Neeley Title Vice President Scientist Technical Lead-LC/MS Technician Sample Custodian Sample Custodian Technician Scientist Scientist Exygen Research Page 6 of 153 . 418-028:PAGE G-7 Exygen Study No. 023-072 TABLE OF CONTENTS Page TITLE PAGE ....................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCESTATEMENT.............................. 2 QUALITY ASSURANCE STATEMENT .......................................................................... 3 CERTIFICATIONOF AUTHENTICITY........................................................................... 4 STUDY IDENTIFICATION...............................................................................................5 PROJECT PERSONNEL ....................................................................................................6 TABLE OF CONTENTS..................................................................................................... 7 LIST OF TABLES............................................................................................................... 8 LIST OF FIGURES ............................................................................................................. 9 LIST OF APPENDICES....................................................................................................10 1.0 SUMMARY................................................................................................................11 2.0 OBJECTIVE............................................................................................................... 11 3.0 INTRODUCTION ......................................................................................................11 4.0 ANALYTICAL TEST SAMPLES............................................................................. 12 5.0 REFERENCE MATERIAL........................................................................................ 13 6.0 DESCRIPTIONOF ANALYTICAL METHOD.......................................................1.3 6.1. Extraction Procedure ...............................................................................................13 6.2 Preparationof Standardsand Fortification Solutions............................................... 14 6.3 6.4 CInhsrtroummaetongt rSaepnhsy.it.l...w....t.y............................................................................................................................................................................................... 14 14 6.5 Description of LC/MS/MS Instrument and Operating Conditions........................... 15 6.6 Quanitation and Example Calculation..................................................................... 15 7.0 EXPERIMENTAL DESIGN ...................................................................................... 17 8.0 RESULTS ...................................................................................................................18 9.0 CONCLUSIONS......................................................................................................... 18 10.0 RETENTION OF DATA AND SAMPLES ............................................................. 19 Exygen Research Page 7 of 153 . 418-028:PAGE G-8 Exygen StudyN o. 023-072 LIST OF TABLES Page Table I. Summary of PFHS in Control Rat Plasma Samples....................................... 21 Table I1. Summary of PFHS in Control Rat Serum Samples........................................ 21 Table I11. Summary of PFHS in Control Rat Liver Samples.......................................... 21 Table IV. Summary of PFHS Fortification Recoveries in Rat Plasma...........................22 Table V. Summary of PFHS Fortification Recoveries in Rat Serum............................ 22 Table VI. Summary of PFHS FortificationRecoveries in Rat Liver..............................23 Table VII. Summary of PFHS Residues in Rat Plasma Samples..................................... 24 Table VIII. Summary of PFHS Residues in Rat Serum Samples...................................... 26 Table IX. Summary of PFHS Residues in Rat Liver Samples........................................ 28 Table X. Summary of PFHS Residues in Dosing Solutions..........................................40 Table XI. S u m m v of PFHS Residues in Stability Samples.......................................4..1 Table XI1. Summary of PFHS Residues in Homogeneity Samples................................. 42 Exygen Research Page 8 of 153 418-028:PAGE G-9 Exygen StudyNo.:023-072 Figure 1. Figure 2. LIST OF FIGURES Pane Typical Calibration Curve for PFHS .............................................................. 44 Chromatogram Representing a 0.1 ng/mL standard for PFHS ....................... 45 Figure 3. Chromatogram Representing a Control Rat Plasma Sample for PFHS (Exygen ID 0202913 Control, Data Set: 091602B)........................................46 Figure 4. Chromatogram Representing a Control Rat Serum Sample for PFHS (Exygen ID: 0202987 Control, Data Set: 091902B) ......................................47 Figure 5. Chromatogram Representing a Control Rat Liver Sample for PFHS, (Exygen ID: 0202877 Control ,Data Set: 100202A)............................,,,.,,...48 Figure 6. Chromatogram Representing Control Rat Plasma Sample Fortified with 10ppb of PFHS (Exygen ID: 0202913 Spk A, Data Set: 091602B)......49 Figure 7. Chromatogram Representing Control Rat Serum Sample Fortified with 10 ppb of PFHS (Exygen ID: 0202987 Spk A, Data Set: 091902B)......50 Figure 8. Chromatogam Representing Control Rat Liver Sample Fortified with 10ppb of PFHS (Exygen ID: 0202877 Spk A, Data Set: 100202A).....51 Figure 9. ChromatogramRepresenting Rat Plasma SampleAnalyzed for PFHS (Exygen ID: 0201816, Sponsor ID: 19176 GROUP I, Data Set: 091602B) ..52 Figure 10. Chromatogram Representing Rat Serum Sample Analyzed for PFHS, DF=lO (Exygen ID: 0201863, Sponsor ID: 19079 GROUP 11, Data Set: 091902BR)......................................................................................53 Figure 11. Chromatogram Representing Rat Liver Sample Analyzed for PFHS (Exygen ID: 0202050, Sponsor ID: 19018 GROUP I1 Male 1, Data Set: 100202A).................................................................................................54 Exygen Research Page 9 of 153 4 18-028:PAGE G- 10 Exygen Study No.:023-072 LIST OF APPENDICES Page Appendix A Study Protocol 418-028 (Exygen Study No. 023-072) and Amendments, Deviation andNote to File..................................................55 Appendix B Analytical Method: Method of Analysis for the Determination of Perfluorohexanesulfonate (PFHS),Perfluorooctanesulfonate (PF0S) and Pentadecafluorooctanoic Acid (PFOA) in Rat Liver, Serum and Urine Revision 1 (Exygen Method No. ExM-023-071Revision 1) .................110 Exygen Research Page 10 of 153 418-028:PAGE G-11 Exygen StudyNo.:023-072 1.0 SUMMARY Exygen Resarch extracted and analyzed rat liver, serum, and plasma samples for the determination of perfluorohexanesulfonate (PFHS) according to Exygen Method ExM- 023-071 Revision 1 (Appendix B). The limit of quantitation for PFHS in rat liver was 10 ng/g and 10 ng/mL in rat serum and plasma. The LOQ for each matrix was determined in a method validation study performed at Exygen (Exygen Study No: 023-073). PFHS in the rat plasma samples ranged fi.om non-detected levels to 237,000 ng/mL. PFHS in the rat serum samples ranged fiom non-detected levels to 189,000 ng/mL. PFHS in the rat liver samples ranged .fromnon-detected levels to 675,000 ng/g. The average percent recoveries k standard deviations for PFHS in rat plasma, serum, and liver samples were 84% k 7%, 95% It 12%, and 86% f 13%, respectively. PFHS residues in the dosing solutions, stability samples and homogeneity samples were all within 70% to 125% of their known concentrations. 2.0 OBJECTIVE The objective of the analytical part of this study was to determine levels of perfluorohexanesulfonate (PFHS) in specimens of rat liver, serum and plasma samples, bulk test substance, concentration and homogeneity formulations, and stability solutions according to Protocol 418-028 (Appendix A). 3.0 INTRODUCTION This report details the results of the analysis for the determination of PFHS in rat liver, serum and plasma samples, bulk test substance, concentration and homogeneity formulations, and stability solutions using the analytical method entitled, "Method of Analysis for the Determination of Perfluorohexanesulfonate (PFHS), Perfluorooctanesulfonate (PFOS) and Pentadecafluorooctanoic Acid (PFOA) in Rat Liver, Serum and Urine Revision 1." The study was initiated on March 26, 2002, when the study director signed protocol number 418-028. The analytical start date was September 16, 2002, and the analytical termination date was November 13,2002. Exygen Research Page 11 of 153 418-028:PAGE G-12 Exygen StudyNo.:023-072 4.0 ANALYTICAL TEST SAMPLES The control rat liver (Exygen ID 0202877) used for the matrix blanks and matrix fortifications was received fi-ozen on dry ice on August 7, 2002 fiom Pel-Freez Biologicals, Rogers, Arkansas. The control rat plasma (Exygen ID 0202912 and 0202913) used for the matrix blanks and matrix fortifications was received fiozen on dry ice on August 8, 2002 from Pel-Freez Biologicals, Rogers, Arkansas. The control rat serum (Exygen ID 0202987) used for the matrix blanks and matrix fortifications was received fiozen on dry ice on August 15, 2002 fiom Pel-Freez Biologicals, Rogers, Arkansas. Dosage solutiodsuspension samples (Exygen ID 0201753-0201755) were received refrigerated at Exygen from Argus Research on May 29, 2002. Dosage solutiodsuspension samples (Exygen ID 0201757-0201758)were received refkigerated at Exygen fiom Argus Research on June 11, 2002. These samples were logged in by Exygen personnel and placed in refkigerated storage. Bulk T N S sample (Exygen ID 0201756) was received ambient at Exygen Erom Argus Research on June 11,2002. This sample was logged in by Exygen personnel and placed in refiigerated storage. Median liver lobe (Exygen ID 0201759-0201787), pooled fetal liver (Exygen ID 0201788-0201800), plasma (Exygen ID 0201801-0201860),pooled fetal serum (Exygen ID 0201861-0201874), pooled pup serum (Exygen ID 0201875-0201924), and pup livers (Exygen ID 0201925-0202408) were received fiozen on dry ice at Exygen Erom Argus Research on June 18, 2002. These samples were logged in by Exygen personnel and placed in fi-ozenstorage. Median liver lobe (Exygen ID 0203650) and pooled fetal liver (Exygen ID 0203651) were received fi-ozen on dry ice at Exygen from Argus Research on September 11,2002 and logged in by Exygen personnel and placed in fi-ozenstorage. Prepared formulations (Exygen ID 0203923-0203962) were received fkozen fkom 3M on September 19,2002 and logged in by Exygen personnel and placed in fiozen storage. Sample log-in and chain of custody information can be found in the raw data package associated with this study. Storage records will be kept at Exygen Research. Exygen Research Page 12 of 153 418-028:PAGE G-13 Exygen StudyNo.:023-072 5.0 REFERENCE MATERIAL The analytical standard PFHS was received at Exygen on January 26, 2000 firom 3M Environmental Technology and Services. The available infonnation for the reference material is listed below. The reference material was stored frozen. Compound Exygen Inventory No.. PFHS SP244 TCR No. SE036 Purity (%I Expiration Date 99.99 01/01/10 The molecular structures of test substance is given below: Name: PFHS Chemical Name: Perfluorohexanesulfonate Molecular Weight: 399 F F F F F F ,,/,/\F- 503 - F F F 6.0 DESCRIPTION OF ANALYTICAL RlETHOD The analytical method ExM-023-071Revision 1 was used forthis study. 6.1. Extraction Procedure A 1 mL aliquot of the serum and plasma and 1 g of liver were used for the extraction procedure for the laboratory controls and fortifications. Due to insufficient sample size, a 100 pL aliquot of the serum and plasma and 0.1 g of the liver were used for the extraction procedure for the study samples. After fortification of appropriate samples, the serum samples were brought up to 20 mL with Type 1Water and the liver sampleswere brought - up to 10 mL with Type I Water. The serum samples were vortexed for 1 minute and - the liver samples were homogenized with a tissuemizer for 1 minute. An aliquot of one - milliliter was transferred fiom each sample and 5 mL of acetonitrile was added and the samples were shaken for 20 minutes. The samples were centrifbged and the supernatant was decanted onto a conditioned SPE column. Then the samples were eluted with 2 mL of methanol. Each sample was analyzed by LC/MS/MS electrospray. An extraction procedure was not necessary for the bulk test substance, concentration and homogeneity formulations, and the stability solutions. 10 mg of the bulk test substance was weighed and brought to volume with methanol in a 100-mL volumetric flask. The Exygen Research Page 13 of 153 4 18-028:PAGE G- 14 Exygen Study No.:023-072 sample was then diluted 100000 times with methanol to fit in the range of the analytical curve and analyzed by LCIMSIMS electrospray. The concentration and homogeneity formulations and the stability solutions were all diluted appropriately with methanol and analyzed by LC/MS/MS electrospray. 6.2 Preparation of Standards and Fortification Solutions A stock standard solution of PFHS was prepared on August 20, 2002 as specified in Exygen method ExM-023-071 Revision 1. The stock standard solution was prepared at a concentration of 100 pg/mL by dissolving 10 mg of each standard (corrected for purity only) in methanol. From this solution, a 1.0 pg/mL fortification standard solution was prepared by taking 1 mL of the stock and bringing the volume up to 100 mL with methanol. A 0.1 pg/mL fortification standard was prepared by taking 10 mL of the 1.0 pg/mL fortification standard and bringing the volume up to 100mL with methanol. A set of standards containing PFHS was prepared via dilution of the 0.1 pg/mL and various calibration solutions in the following manner: Initial Conc. (pg/mL)' 0.1 0.1 0.1 0.005 0.002 0.001 ofPFHS Volume (mL) 5 2 1 10 10 10 Diluted to (mL) 100 100 100 100 100 100 Final Conc. ( p g / m v 0.005 0.002 0.00 1 0.0005 0.0002 0.0001 The stock standard solution and all fortification and calibration standard solutions were stored in a reiiigerator (4" & 2C) when not in use. Documentation of standard preparation can be found in the raw data associated with this report. 6.3 Chromatography Quantification time of PFHS - of PFHS was accomplished by LC/MS/MS electrospray. was 8.3 min. Peaks above the LOQ were not detected The retention in any of the control samples corresponding to the analyte retention time. 6.4 Instrument Sensitivity The smallest standard amount injected during the c,iromatographic run hac a concentration of 0.0001 pg/mL of PFHS. Exygen Research Page 14 of 153 418-028:PAGE G-15 Exygen Study No.: 023-072 6.5 Description of LC/MS/MS Instrument and Operating Conditions Instrument: Micromass Quattro Ultima (Micromass) Interface: Electrospray (Micromass) Computer: COMPAQ Professional Workstation AP200 Software: Windows NT, Masslynx 3.3 HPLC: Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven HPLC Co1umn:Genesis C, (Jones Chromatography), 2.1 mm x 50 m,41.1 Column Temp.: 35" C Injection Vol.: 15 pL Mobile Phase (A): 2 mM Ammonium Acetate in type I water Mobile Phase (B): Methanol Time %A 0.0 90 2.0 90 5.0 10 9.0 10 9.5 0 14.0 0 14.5 90 20.0 90 %B Flow Rate (mL/min) 10 0.3 10 0.3 90 0.3 90 0.3 100 0.3 100 0.3 10 0.3 10 0.3 Ions monitored: Analyte PFHS Mode Negative Transition Monitored 399 -+ 80 Approximate Retention Time (min) -8.3 min. 6.6 Quantitation and Example Calculation Fifteen microliters of sample or calibration standard were injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using l/xfit weighted linear regression) by Masslynx software using six concentrations of standards. The concentration was determined fiom the equations below. Equation 1 calculated the amount of analyte found (in ng/mL, based on peak area) using the standard curve (linear regression parmeters) generated by the Masslynx software Exygen Research Page 15 of 153 418-028:PAGEG-16 Exygen Study No.: 023-072 program. Then Equation 2 calculated the amount of analyte found in ng/g for liver and ng/mL for serum and plasma. Equation 1: Analyte found (ng/mL) = (Peak area - intercept) x DF x AF slope Where: AF = Aliquot Factor DF = Dilution Factor Equation 2: Analyte found (ppb, ng/g for liver and ng/mL for serum and plasma) = analyte found (ng/mL) x FV (mL) sample volume (mL) or sample weight (8) Where: FV = Final Volume For samples fortified with known amounts of PFHS prior to extraction, Equation 3 calculated the percent recovery. Equation 3: Recovery (%) = ((analyte found (ppb) - analvte in control (ppb)) x 100% amount added (ppb) To find the total analyte found corrected for the salt content Equation 4 was used. Equation 4: Total Analyte Found Corrected (ppb) = analyte found (ppb) x salt correction factor Where the salt correction factor = 0.91. An example of a calculation using an actual sample follows: Rat liver sample Exygen ID 0202877 Spk A (Set: 100202A), fortified at 10 ng/g where: peak area intercept slope dilution factor - 2125 - - - 41.0365 4728.12 -- 1 ppb added (fort level) = 10 avg. amt in controls = final volume - - aliquot factor - - sample weight - - 0 (Not Detected) 2 mL 10 1.0 g Exygen Research Page 16 of 153 4 18-028:PAGE G- 17 Exygen Study No.:023-072 From equation 1: Analyte found (ng/mL) From equation 2: Analyte found (ppb) From equation 3: YORecovery 12125 - 41.03651x 1 x 10 4728.12 4.41 ng/mL (4.41 ng/mL x 2 mL) (1.0 9 8.82 ppb From equation 4: Analyte Found Corrected (ppb) 88% 8.82 ppb x 0.91 8.03 ppb 7.0 EXPERIMENTAL DESIGN - Each set of samples (liver, serum or plasma) consisted of one matrix blank, two matrix blanks fortified at known concentrations, and 20 samples. Each sample was extracted using the method and then analyzed in duplicate. Exygen Research Page 17 of 153 418-028:PAGE G-18 Exygen StudyNo.:023-072 8.0 RESULTS The PFHS found in the control rat plasma, serum, and liver samples are listed in Tables 1-111. Peaks were not detected in any of the control plasma or serum samples corresponding to the analyte retention time. Some peaks were detected in the control liver samples corresponding to the analyte retention time; however, the peaks detected were all below the LOQ of 10 ppb. Fortification recoveries for PFHS in the rat plasma, serum, and liver samples are detailed in Tables IV-VI. The average percent recoveries k standard deviations for PFHS in rat plasma, serum, and liver samples were 84% +, 7%, 95% k 12%, and 86% +, 13%, respectively. PFHS in the rat plasma samples ranged from non-detected levels to 237,000 ng/mL. Individual results are listed in Table VII. PFHS in the rat st=t.um samples ranged from non-detected levels to 189,000 ng/mL. Individual results are listed in Table VIII. PFHS in the rat liver samples ranged from non-detected levels to 675,000 ng/g. Individual results are listed in Table IX. PFHS residues in the dosing solutions ranged fi-om 88% to 125% of their known concentrations. The results are listed in Table X. PFHS residues found in the stability samples ranged from 91% to 122% of their known concentrations. The results are listed in'Table XI. PFHS residues found in the homogeneity samples ranged fiom 70% to 116% of their known concentrations. The results are listed in Table XII. 9.0 CONCLUSIONS The rat liver, serum, and plasma samples were successfblly extracted and analyzed for PFHS according to analytical method ExM-023-071 Revision 1. The bulk test substance, concentration and homogeneity formulations, and stability solutions were also ' successllly analyzed for PFHS according to analytical method ExM-023-071 Revision 1. Exygen Research Page 18 of 153 418-028:PAGE G-19 Exygen StudyNo.:023-072 10.0 RETENTION OF DATA AND SAMPLES When the ha1 analytical report is complete, all original paper data generated by Exygen Research will be shipped to the sponsor. This does not include facility-specific raw data such as instrument or temperature logs. Exact copies of all raw data, as well as a signed copy of the final analytical report and all original facility-specific raw data, will be retained in the Exygen Research archives for the period of time specified in 21 CFR Part 58, OECD ENV/MC/CHEM(98)17, and MHW OrdinanceNumber 21. Retained samples of reference substances are archived by the sponsor. Exygen Research Page 19 of 153 418-028:PAGE G-20 Exygen StudyNo.: 023-072 TABLES Exygen Research Page 20 of 153 Table I. 418-028:PAGE G-21 Exygen Study No.: 023-072 Summary of PFHS in Control Rat PIasma Samples Sponsor LD LOT 17824 LOT 17824 LOT 05024 LOT 17824 Exygen ID 0202913 Control 0202913 Control 0202912 Control 02029 13 Control Set Number 091602AR 091602B 091802A 091902A PFHS Found (ppb) ND ND ND ND Table 11. Summary of PFHS in Control Rat Serum Samples Sponsor m 361 19-3 361 19-3 361 19-3 361 19-3 361 19-3 Exygen ID 0202987 Control 0202987 Control 0202987 Control 0202987 Control 0202987 Control Set Number 0919028 092002A 092002B 092302A 101102A PFHS Found (ppb) ND ND ND ND ND Table 111. Summary of PFHS in Control Rat Liver Samples Sponsor ID LOT 21824 LOT 21824 LOT 21824 LOT 2 1824 LOT 21824 LOT 21824 LOT 21824 LOT 21824 LOT 21824 LOT 2 1824 LOT 21824 LOT 2 1824 LOT 21824 LOT 21824 LOT 21824 LOT 21824 Exygen ID 0202877 Control 0202877 Control A 0202877 Control A 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control Set Number 092802A 092602A 092602B 092702A 092702B 100202A 100102A 100302A 100402A 100402BR 100902AR 100902BR 101002A 101002B 101102AR 101402A PFHS Found (ppb) ND ND 2-32 CNQ) 3-84 (NQ) 3-85CNQ) ND ND ND ND 2-09 (NQ) 3.95 WQ) 2-01 (NQ) 1-85 (NQ) 3-95 VQ) 4-73 CNQ) 3.37 (NQ) ND = Not Detected (Area less than the lowest concentration of the calibration standards (0.1 ng/mL)) NQ = Not Quantifiable (Area is greater than 0.1 ng/mL but less than LOQ (10 ng/mL)) Exygen Research Page21 of 153 418-028:PAGE G-22 Exygen StudyNo.:023-072 Table IV. Summary of PFHS Fortification Recoveries in Rat Plasma Sponsor ID LOT 17824 LOT 17824 19176GROW 1 LOT 17824 LOT 17824 19176 GROUP I LOT 05024 LOT 05024 19076GROUP I LOT 17824 LOT 17824 19076 GROUP I Exygen ID 0202913 SpkA 0202913 Spk B 0201801 Spk C 0202913 SpkA 0202913 SpkB 0201816 Spk C 0202912 Spk A 0202912 Spk B 0201846 Spk C 0202913 Spk A 0202913 Spk B 0201831 Spk C Set Number Amount Added (ppb) 09 1602A 10 09 1602A 50 091602A 09 1602B 09 1602B 09 1602B 09 1802A 091802A 09 1802A 09 1902A 09 1902A 09 1902A 5000 10 50 5000 10 50 5000 10 50 5000 Average: Standard Deviation: Relative Standard Deviation: YO Recovery 91 84 77 86 84 78 88 86 89 71 96 82 84 7 8 Table V. Summary of PFHS Fortification Recoveries in Rat Serum Sponsor ID 361 19-3 361 19-3 19076 GROUP I 361 19-3 361 19-3 19012 GROUP I 361 19-3 361 19-3 19036 GROUP I1 361 19-3 361 19-3 19005 GROUP IV 361 19-3 361 19-3 Exygen ID 0202987 Spk A 0202987 Spk B 0201861 Spk C 0202987 Spk A 0202987 Spk B 0201875 Spk C 0202987 Spk A 0202987 Spk B 0201890 Spk C 0202987 Spk A 0202987 Spk B 0201905 Spk C 0202987 Spk A 0202987 Spk B Set Number Amount Added (ppb) 091902B 10 09 1902B 50 09 1902B 092002A 092002A 092002A 092002B 5000 10 50 5000 10 092002B 50 092002BR 5000 092302A 10 092302A 50 101102A 200000 101102A 10 101 102A 50 Aver age: Standard Deviation: Relative Standard Deviation: YO Recovery 92 96 94 1 06 105 90 97 97 120 90 101 88 78 72 95 12 12 Exygen Research Page 22 of 153 418-028:PAGE G-23 Exygen Study No.: 023-072 Table VI. Summary of PFHS Fortification Recoveries in Rat Liver Sponsor ID LOT 21824 LOT 2 1824 19176 GROUP I LOT 2 1824 LOT 21824 19076 GROUP I LOT 2 1824 LOT 21824 19076 GROUP I LOT 2 1824 LOT 21824 19012 GROUP I, Male 1 LOT 21824 LOT 2 1824 19021 GROUP I, Male 1 LOT 2 1824 LOT 21824 19018 GROUP 11,Male 1 LOT 2 1824 LOT 21824 19041 GROUP I, Male 1 LOT 2 1824 LOT 21824 19036 GROUP 11, Male 1 LOT 2 1824 LOT 21824 19003 GROUP 111,Male 1 LOT 21824 LOT 2 1824 19008 GROUP 111, Male 1 LOT 21824 LOT 2 1824 19015 GROUP 111,Male 1 LOT 21824 LOT 2 1824 19035 GROUP IV,Male 1 LOT 21824 LOT 2 1824 19040GROUP IV,Male 2 LOT 21824 LOT 21824 19006GROUP V,Male 1 LOT 21824 LOT 2 1824 19020 GROUP V, Male 2 LOT 21824 LOT 2 1824 19025GROUP V,Male 1 Exygen ID 0202877 Spk A 0202877 Spk B 0201759 Spk C 0202877 Spk A 0202877 SpkB 0201774 Spk C 0202877 Spk A 0202877 SpkB 0201788 Spk C 0202877 Spk A 0202877 Spk B 0201925 Spk C 0202877 SpkA 0202877 Spk B 0201945 Spk C 0202877 Spk A 0202877 Spk B 0202050 Spk C 0202877 Spk A 0202877 Spk B 0201965 Spk C 0202877 Spk A 0202877 Spk B 0202070 Spk C 0202877 Spk A 0202877 Spk B 0202119 Spk C 0202877 Spk A 0202877 Spk B 0202139 Spk C 0202877 Spk A 0202877 SpkB 0202159 Spk C 0202877 Spk A 0202877 Spk B 0202229 Spk C 0202877 Spk A 0202877 Spk B 0202249 Spk C 0202877 Spk A 0202877 Spk B 0202320 Spk C 0202877 Spk A 0202877 SpkB 0202340 Spk C 0202877 Spk A 0202877 Spk B 0202360 Spk C Exygen Research Set Number Amount Added (ppb) 092802A 10 092802A 50 092802A 5000 092602A 10 092602A 50 092602A 5000 092602B 10 092602B 50 092602B 5000 092702A 10 092702A 50 092702A 5000 092702B 10 092702B 50 092702B 5000 100202A 10 100202A 50 100202A 5000 100102A 10 100102A 50 100102A 5000 100302A 10 100302A 50 100302A 5000 100402A 10 100402A 50 100402A 5000 100402BR 10 100402BR 50 100402BR 5000 100902AR 10 100902AR 50 100902AR 5000 100902BR 10 100902BR 50 100902BR 5000 101002A 10 10 1002A 50 101002AR 5000 101002B 10 101002B 50 101002BR 5000 101102AR 10 101102AR 50 101102AR 25000 101402A 10 101402A 50 10 1402ARR 5000 Average: Standard Deviation: Relative Standard Deviation: YO Recovery 83 87 96 125 110 100 104 87 111 75 93 94 83 100 109 88 71 76 74 77 76 98 81 77 81 79 62 70 78 86 72 81 79 72 70 72 96 93 84 97 72 90 88 75 83 95 77 110 86 13 16 Page 23 of 153 418-028:PAGE G-24 Exygen Study No.: 023-072 Table VII. Summary of PFHS Residues in Rat Plasma Samples Sponsor ID 19176GROUP I 19176GROUP I* 19177 GROUP I 19177 GROUP I* I9178 GROUP I 19178GROUP I* 19179 GROUP II 19179GROUP II* 19180 GROUP II I9180 GROUP It* 19181 GROUPII 19181 GROUP LI* 19182 GROUP III 19182 GROUP LU* 19183 GROUP III 19183 GROUP III* 19184 GROUP IlI 19184 GROUP LU* 19185GROUP IV 19185 GROUq IV* 19186 GROUP IV 19186GROUP IV* 19187 GROUP N 19187 GROUP IV* 19188 GROUP V 19188 GROUP V* 19189 GROUP V 19189 GROUP V* 19190 GROUP V 19190 GROUP'V 19176 GROUP I 19176 GROUP I* 19177 GROUP I 19177 GROUP I* 19178 GROUP I 19178 GROUP I* I9179 GROUP 11 19179 GROUP II* 19180 GROUP I1 19180GROUP II* 19181 GROUP Il 19181 GROUP 11' 19182 GROUP 111 19182 GROUP III* 19183 GROUP III 19183 GROUP III* 19184 GROUP IU 19184 GROUP III* 19I85 GROUP IV 19185GROUP IV* 19186 GROUP IV 19186 GROUP IV* 19187 GROUP IV 19187GROUP IV* 19188 GROUP V 19188 GROUP V* 19189 GROUP V 19189 GROUP V* 19190 GROUP V 19190 GROUP V* Erygen ID 0201801 0201801 Dup Inj. 0201802 0201802 Dup Inj. 0201803 0201803 Dup hj. 0201804 0201804 Dup Inj. 0201805 0201805 Dup Inj. 0201806 0201806 Dup Inj. 0201807 0201807 Dup Inj. 0201808 0201808 Dup Inj. 0201809 0201809 Dup Inj. 0201810 0201810 Dup Inj. 0201 81 1 0201811 Dup Inj. 0201812 0201812 Dup Inj. 0201813 0201813 Dup Inj. 0201814 0201814 Dup Inj. 0201815 M018l5 Dup Inj 0201816 0201816 Dup Inj 0201817 0201817 Dup Inj 0201818 0201818 Dup Inj 0201819 0201819 Dup Inj 0201820 0201820 Dup Inj 0201821 0201821 Dup Jnj 0201822 0201822 Dup Inj 0201823 0201823 Dup Inj 0201824 0201824 Dup Inj 0201 825 0201825 Dup Inj 0201826 0201826 Dup Inj 0201827 0201'827 D up Inj 0201828 0201828 Dup Inj 020 1829 0201829 Dup Inj 0201830 0201830 Dup Inj Matrix PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA Collection Date 4/14/02 4/14/02 4/14/02 4114/02 4/14/02 4/14/02 4/14/02 414/02 4/14/02 4/1 4/02 4114/02 414/02 4114/02 411 4/02 4/14/02 4/14/02 4114/02 4/14/02 4/14/02 411 4/02 4/14/02 4/14/02 4114/02 4/14/02 4/14/02 411 4/02 4114/02 4114/02 4114/02 4/1 4/02 5/12/02 5llUo2 5/12/02 5/12/02 5/12/02 5112/02 5/1uo2 5112/02 5/12/02 5112/02 5/12/02 5/12/02 511 2/02 5112/02 5112/02 smmz 5112/02 5112/02 5/12/02 5112/02 5112/02 5/12/02 5112/02 511uO2 5/12/02 5112/02 5112/02 5/12/02 511 2/02 5/12/02 Set Number 09 1602A 09160% 09360% 09 1602A 09160% 09 1602A 09160091602AR 09 1602AR 09 1 602AR 091602AR 09 1602AR 09 1602AR 091602AR 091602AR 091602AR 09 1602AR 09 1602AR 091602AR 091602AR 091602AR 091602AR 091602AR 091602AR 09 160091602AR 091602AR 09 1602AR 09 I602AR 091602AR 091602B 09 1602B 091 602B 091602B 091602B 091 602B 09 1602BR 09 1602BR 09 1602BR 091602BR 09 1602BR 09 1602BR 091602BR 09 1602BR 09 1602BR 09 1602BR 09 1602BR 09 1602BR 09 1602BR 091602BR 091602BR 09 1602BR 09 1602BR 09 1602BR 09 1602BR 09 1602BR 09 1602BR 09 1602BR 09 1602BR 09 1 602BR PFES Found (ng/mL) 182 172 103 108 46.P 43.3M 18300 1 9400 15700 15300 20000 20400 58000 58000 68300 69500 1 17000 1 15000 70400 7 1900 126OOO 125000 237000 228000 197000 182000 187000 183000 177000 170000 245 256 382 376 74.7M 65.4M 57400 60200 35700 36400 37 100 38500 91000 85900 90300 87500 90700 89300 119m 119000 137000 142000 128000 127000 188OOO 188000 224000 225000 191000 193000 /\"Although the method used allows for alternate sample volumes or weights and all the peak area responses are within the calibration curve limits, the precision for the 0.1 mL or 0.1 g samplesbelow 100 ppb has not been validated. * Duplicate Injection Exygen Research Page 24 of 153 418-028:PAGE G-25 Exygen Study No.:023-072 Table VII (cont'd). Summary of PFHS Residues in Rat Plasma Samples Sponsor ID 19076 GROUP I 19076 GROUP I* 19077 GROUP I 19077 GROUP I* 19078 GROUP I 19078 GROUP I* 19079 GROUP I1 19079 GROUP 11' 19080 GROUP I1 19080 GROUP 11* 19081GROUP II 19081 GROUP 11* 19082 GROUP III 19082 GROUP III* 19083 GROUP 111 19083 GROUP III* 19084 GROUP 111 19084 GROUP 111' 19085 GROUP IV 19085 GROUP IV* 19086 GROUP IV 19086GROUP IV* 19087 GROUP IV 19087 GROUP IV* 19088 GROUP V 19088 GROUPV* 19089 GROUP V 19089 GROUP V* 19090 GROUP V 19090 GROUP V* 19076 GROUP I 19076 GROOP I* 19077 GROUP I 19077 GROUP I* 19078 GROUP I 19078 GROUP I* 19079 GROUP 11 19079 GROUP 11* 19080 GROUP I1 19080 GROUP 1' 19081GROUP I1 19081GROUP 1. 19082 GROUP I11 19082 GROUP Ill* 19083 GROUP IIl 19083 GROUP Ill* 19084 GROUP 111 19084 GROUP III* 19085 GROUP IV 19085 GROUP IV* 19086 GROUP IV 19086 GROUP IV* 19087 GROUP N 19087 GROUP IV* 19088 GROUP V 19088 GROUP V* 19089 GROUP V 19089 GROUPV* 19090 GROUP V 19090 GROUP V* * Duplicate Injection Exygen ID 020 1846 0201846Dup Inj 0201847 0201847 Dup Inj 0201848 0201848 Dup Inj 020 1849 0201849Dup Inj 0201850 0201850Dup Inj 0201851 0201851Dup Iqj 0201852 0201852 Dup Inj 0201853 0201853 Dup Iqj 0201854 0201854Dup Inj 0201855 0201855 Dup Inj 0201856 0201856 Dup hj 0201857 0201857 Dup Inj 0201858 0201858 Dup Inj 0201859 0201859 h p Inj 0201860 0201860Dup Inj 020183 1 0201831 Dup Inj 020 1832 0201832 Dup Inj 0201833 0201833 Dup Inj 020 1834 0201834Dup Inj 0201835 0201835 Dup hj 0201836 0201836 Dup Inj 0201837 0201837 Dup Inj 0201838 0201838 Dup Iaj 020 1839 0201839 Dup Inj 020 1840 020 1840 Dup Inj 020 184 1 0201841Dup Inj 020 1842 0201842 Dup Inj 0201843 0201843 Dup Inj 020 1844 020I844 Dup Inj 020 1845 0201845 Dup Inj Matrix GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA OD 21 PLASMA GD 21 PLASMA GD 21PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA Collection Date 5/9/02 5/9/02 5/19/02 5/19/02 5/7/02 5/7/02 5/9/02 5/9/02 5/9/02 5/9/02 5110102 5110102 5/9/02 5/9/02 5/7/02 5/7/02 5/9/02 5/9/02 5/6/02 5/6/02 5/6/02 5/6/02 5/6/02 5/6/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 4/14/02 4114/02 4114/02 4/14/02 4/14/02 4114/02 4/14/02 4/14/02 41 14/02 4114/02 4/14/02 4/14/02 4114/02 4/14/02 4/14/02 4114/02 4114/02 4114/02 4/14/02 4/14/02 4/14/02 4114/02 4114/02 4114/02 41 14/02 4/14/02 4114/02 41 14/02 4114/02 4/14/02 set Number 09 1802A 091802A 091802A 09 1802A 091802A 091802A 09 1802AR 091802AR 091802AR 09 1802AR 091802AR 09 1802AR 09 1802AR 09 1802AR 091802AR 091802AR 09 l802AR 091802AR 091802AR 09 1802AR 091802AR 09 1802AR 09 1802AR 09 1802AR 091802AR 09 1802AR 09 1802AR 091802AR 09 1802AR 09 1802AR 091902A 091902A 091902A 091902A 091902A 09 1902A 091902AR 09 1902AR 09 1902AR 091902AR 09 1902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 09 1902AR 09 1902AR 091902AR 091902AR 091902AR 09 1902AR 091902AR 09 1902AR 091902AR 091902AR 09 1902AR 09 1902AR 091902AR PFWS Found (ng/mL) ND ND ND ND ND ND 4150 4000 3240 3230 2640 2670 7150 7280 18000 18100 6490 6900 24800 25400 40700 40800 32500 32300 72600 73600 53200 54400 52200 52800 ND ND 386 398 ND ND 3800 3530 2600 2630 2040 2090 10400 10100 13100 14000 5540 5980 16900 17900 24800 25200 19200 20000 43900 43000 47100 48200 34000 35900 { ND = Not Detected (Area less than lowest calibrationstandard of 0.1 ng/mL) Exygen Research Page 25 of 153 418-028:PAGE G-26 Exygen Study No.:023-072 Table VIII. Summary of PFHS Residues in Rat Serum Samples Sponsor ID 19076 GROUP I 19076 GROUP I* 19078 GROUP I 19078 GROUP I* 19079 GROUP A 19079 GROUP ll* 19080 GROUP 11 19080 GROUP II* 19081 GROUP II 19081 GROUP 1I* 19082GROUP III 19082 GROUP ID* 19083 GROUP III 19083 GROUP ID* 19084 GROUP III 19084 GROUP ID* 19085 GROUP IV 19085 GROUP XV* 19086 GROUP IV 19086 GROUP IV* 19087 GROUP 1V 19087 GROUP IV* 19088GROUP V 19088GROUP V* 19089 GROUP V 19089GROUP V* 19090GROUP V 19090GROUP V* 19012 GROUP I 19012 GROUP I* 19019 GROUP I 19019 GROUP I* 19021 GROUPI 19021 GROUP I* 19023 GROUP r 19023 GROUP I* 19041 GROUPI 19041 GROUP I* 19042 GROUP I 19042 GROUP I* 19044 GROUP 1 19044 GROUP I* 19050 GROUP I 19050 GROUP I* 19053 GROUP I 19053 GROUP I* 19065 GROUP I 19065 GROUP I" 19004GROUP I1 19004 GROUP IP 19009GROUP II 19009 GROUP II* 19016 GROUP II 19016 GROUP II* 19018 GROUP Il 19018 GROUP A* 19026GROUP I1 19026 GROUP II* 19036 GROUP II 19036 GROUP II* Exygen ID 0201861 0201861 Dup Inj 0201862 0201862 Dup Inj 0201863 0201863 Dup Inj 02018 64 0201864 Dup Jnj 0201865 0201865 Dup Inj 0201866 0201866 Dup Iuj 0201867 0201867 Dup Inj 0201868 0201868 Dup Inj 0201 8 69 0201869 Dup Jnj 0201870 0201870 Dup Jnj 02018 71 0201871 D q I n j 0201872 0201872 Dup Inj 0201873 0201873 Dup Jnj 0201874 0201874 Dup Inj 0201 8 75 0201875 Dup Jnj 0201 8 76 0201876 Dup Inj 0201 8 77 0201877 Dup Inj 0201878 0201878 Dup Inj 0201 8 79 0201879 Dup Inj 0201880 0201880 Dup Inj 0201881 0201881 Dup Inj 0201882 0201882 Dup Inj 0201883 0201883 Dup Inj 0201884 0201884 Dup Inj 0201 8 85 0201885 Dup Inj 0201886 0201886 Dup Inj 0201 8 87 0201887 Dup Inj 0201888 0201888 Dup Xnj 0201889 0201889 Dup Inj 0201890 0201890 Dup Inj Matrix Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal S R U ~ Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Fetal Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup Serum Pooled Pup senmr Pooled Pup Serum Pooled Pup Serum Pooled Pup serum Pooled Pup Serum Collection Date 5/9/02 5/9/02 5/7/0 2 5/7/02 5/9/02 5/9/0 2 5/9/02 5/9/02 5/10102 5/10/02 5/9/02 5/9/02 5/7/02 5/7/02 5/9/02 5/9/02 5/6/0 2 5/6/02 5/6/0 2 51610 2 51610 2 51610 2 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/29/02 5/29/02 5/30/02 5/30/02 5/29/02 5/29/02 5B 1/02 5/3 1/02 5/31/02 5/3 1/02 5/30/02 5/30/02 5/28/02 5/28/02 5/3 1102 5J31/02 5/30/02 5/30102 5130m2 5/30/02 5/31/02 5/31/02 5/28/02 5/28/02 5/28/02 5/28/02 5/29/02 5/29/02 5/31102 5B 1/02 5/28/02 5/28/02 Set Number 091902B 09 1902B 0919028 09 1902B 091902BR 091902BR 09 1902BR 091902BR 091902BR 091902BR 091902BR 091 902BR 09 1902BR 091902BR 09 1902BR 09 1902BR 091902BR 091902BR 091 902BR 09 1902BR 091 902BR 09 1902BR 091902BR 091902BR 09 1902BR 091902BR 091902BR 091902BR 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092002AR 092002AR 092002A R 092002A R 092002A R 092002AR 092002AR 092002AR 092002A R 092002A R 092002BR 092002BR PFHS Found (@rnL) ND ND ND ND 6460 6800 3940 4040 5280 5370 12600 13500 15700 15700 11700 1 1600 38600 40200 33000 34700 36900 39200 35700 38400 44900 47900 47900 51 200 ND ND 65.3M 68.9M 81.1m 86.0M 45.6M 42. 7M ND ND ND ND ND ND ND ND ND ND ND ND 9650 9560 7940 7940 8490 8200 8630 8720 5490 5420 6300 6370 MAlthough the method used allows for alternate sample volumes or weights and all the peak area responses are within the calibrationcurve limits, the precision for the 0.1 mL or 0.1 g samples below 100 ppb has not been validated. * Duplicate Injection ND =Not Detected (Area less than lowest calibrationstandard of 0.1 ng/mL) Exygen Research Page 26 of 153 418-028:PAGE G-27 Exygen Study No.: 023-072 Table VIII (cont'd). Summary of PFHS Residues in Rat Serum Samples Sponsor ID 19037 GROUP I1 19037GROUP II* 19043 GROUP I1 19043 GROUP II* I9047 GROUP I1 19047 GROUP II* 19048 GROUP II 19048 GROUP II* 19003 GROUP 1x1 19003 GROUP III* 19007 GROUP 111 19007 GROUP III* 19008 GROUP III 19008 GROUP HI* 19013GROUP Ill 19013 GROUP UI* 19015 GROUP I11 19015 GROUP IIX* 19017GROUP Ill 19017 GROUP III* 19024 GROUP III 19024 GROUP UI* 19029 GROUP Ill 19029 GROUP III* 19034 GROUP III 19034 GROUP III* 19056G R O W III 19056 GROUP III* 19005GROUP IV 19005GROUP IV* 19035 GROUP IV 19035 GROUP lV* 19039 GROUP IV 19039GROUP N* 19040 GROUP IV 19040 GROUP IV* 19045 GROUP IV 19045GROW lV* 19054 GROUP IV 19054GROUP N* 19058 GROUP IV 19058 GROUP lV* 19062 GROUP IV 19062GROUP IV* 19063GROUP Tv 19063 GROUP IV* 19066 GROUP lV 19066GROUP Nf 19001 GROUPV 19001 GROUP V* 19006 GROUP V 19006 GROUP V* 19011 GROUP V 19011 GROUP V* 19020 GROUP V 19020GROUP V* 19022 GROUP V 19022 GROUP VL 19025 GROUP V 19025GROUP V* 19027 GROUP V 19027 GROUP V* 19028 GROUP V 19028 GROUP V* 19030 GROUP V 19030GROUP V* 19031 GROUP V 19031 GROUP V* * Duplicate Injection Exygen ID 0201891 0201891 Dup Inj 0201892 0201892 Dup Inj 020 1893 0201893Dup Inj 0201894 0201894 Dup Inj 0201895 0201895 Dup Iaj 0201896 0201896 Dup Inj 0201897 0201897Dup Inj 0201898 0201898 Dup Inj 0201899 0201899 Dup Xnj 0201900 0201900 Dup Inj 0201 90 I 0201901 Dup Inj 020 1902 0201902 Dup Inj 0201903 0201903 Dup Iaj 0201904 0201904 Dup Inj 0201905 0201905 Dup Inj 0201906 0201906 Dup Inj 0201907 0201907 Dup In. 0201908 0201908 Dup Inj 0201909 0201909 Dup Inj 0201910 0201910 Dup Inj 0201911 020191 1 Dup Inj 02019 12 0201912 Dup Inj 0201913 0201913 Dup Iaj 0201914 0201914 Dup Inj 0201915 0201915 Dup Inj 0201916 0201916 Dup Inj 0201 9 1 7 0201917 Dup Inj 0201918 0201918 Dup Inj 0201919 0201919 Dup Inj 0201920 0201920 Dup Inj 0201921 0201921 Dup Inj 0201922 0201922 Dup Inj 0201923 0201923 h r p Inj 020 1 924 0201924 Dup Inj Matrix Pooled Pup saum Pooled Pup serum Pooled Pup serum Pooled Pup serum Pooled Pllp serum Pooled Pup serum Pooled Pup serum Pooled Pup saum Pooled Pup serum Pooled Pup saum Pooled Pup saum PooIed Pup saum Pooled Pup saum Pooled Pup serum Pooled Pup saum Pooled Pup serum Pooled Pup serum Pooled Pup saum Pooled Pup serum Pooled Pup SenUn Pooled Pup serum Pooled Pup saum Pooled Pup saum Pooled Pup saum Pooled Pup saum Pooled Pup saum Pooled Pup saum Pooled Pup saum PooledPup saum Pooled Pup serum Pooled Pup senun Pooled Pup serum Pooled Pup serum Pooled Pup serum Pooled Pup serum Pooled Pup serum Pooled Pup saum Pooled Pup serum Pooled Pup senun Pooled Pup saum Pooled Pup saum Pooled Pup serum Pooled Pup serum Pooled Pup saum Pooled Pup serum Pooled Pup saum Pooled Pup saum Pooled Pup saum Pooled Pup serum Pooled Pup serum Pooled Pup S m Pooled Pup saum Pooled Pup serum Pooled Pup saum Pooled Pup saum Pooled Pup saum Pooled Pup saum Pooled Pup saum Pooled Pup senun Pooled Pup saum Pooled Pup saum Pooled Pup saum Pooled Pup senun Pooled Pup serum Pooled Pup saum Pooled Pup saum Pooled Pup serum Pooled Pup Saum Collection Date 5/29/02 5/29/02 5/28/02 5/28/02 5/28/02 5/28/02 513 1/02 513 1/02 5/30/02 5/30/02 5/30/02 5/30/02 513 1/02 513 1/02 5/29/02 5/29/02 513 1/02 5/31/02 5/28/02 5/28/02 5 01/02 513 1/02 5/29/02 5/29/02 5/30/02 5/30/02 513 1/02 5/31/02 5/31/02 513 1/02 513 1/02 513 1/02 5/29/02 5/29/02 5/30/02 5/30/02 5/28/02 5/28/02 513 1/02 513 1/02 5/28/02 5/28/02 513 1/02 513 1/02 5/29/02 5/29/02 5/31/02 513 1/02 5/13/02 5/13/02 5/13/02 5/13/02 5/13/02 5/13/02 51 14/02 5/14/02 513 1/02 5/31/02 5130/02 5/30/02 513 1/02 513 1/02 5/28/02 5/28/02 513 1/02 513 1/02 5/28/02 5/28/02 Set Number 092002BR 092002BR 092002 BR 092002BR 092002BR 092002BR 092002BR 092002BR 092002 BR 092002BR 092002BR 092002 BR 092002BR 092002BR 092002BR 092002BR 092002BR 092002BR 092002BR 092002BR 092002BR 092002BR 092002 BR 092002BR 092002BR 092002BR 092002 BR 092002BR 092302AR 092302AR 0923OZAR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR PFHS Found (ng/mL) 12800 13000 5750 5970 8990 8810 11700 1 I700 37300 37400 47300 46300 44000 45400 24300 24600 36900 37000 20800 21200 42900 45000 24900 22600 45000 43700 19700 20500 30900 30800 34600 33500 22600 22800 40700 38400 27800 29300 42900 43800 25800 25800 41200 41100 19800 19100 36800 39300 29000 32700 47900 52400 162000 164000 72800 76200 84100 89400 50200 53900 57200 53700 162000 163000 184000 189000 71200 76200 Exygen Research Page 27 of 153 418-028:PAGE G-28 Exygen Study No.: 023-072 Table IX. Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19176GROUP I 19176 GROUP I* 19177 GROUP I 19177GROUP I* 19178 GROUP I 19178 GROUP I* 19179 GROUP II 19179GROUP II* 19180 GROUP I1 19180GROUP II* 19181 GROUP II 19181 GROUP 11* 19182GROUP I11 19182GROUP III* 19183 GROUP I11 19183 GROUP III* 19184 GROUP III 19184 GROUP III* 19185 GROUP lV 19185 GROUP IV* 19186 GROUP IV 19186 GROUP IV* 19187GROUP IV 19187 GROUP IV* 19188 GROUP V 19188 GROUP V* 19189 GROUP V 19189 GROUP V* 19190GROUP V 19190 GROUP V* 19076 GROUP I 19076 GROUP I* 19077 GROUP I 19077 GROUP I* 19078 GROUP I 19078GROUP I* 19081 GROUP II Female 1 19081 GROUP IIFemale 1* 19079 GROUP I1 19079 GROUP 11* 19080 GROUP I1 19080 GROUP 11* 19082 GROUP III 19082GROW ILI* 19083 GROUP I11 19083 GROUP III* 19084 GROUP 111 19084 GROUP III* 19085 GROUP IV 19085 GROUP IV* 19086 GROUP IV 19086 GROUP IV* Exygen ID 0201759 0201759D u p Inj 020 1760 0201760 Dup Inj 0201 761 0201761 Dup Inj 020 1 762 0201762Dup Inj 020 1 763 0201763 Dup Inj 020 1764 0201764Dup Inj 020 1 765 0201765 Dup Inj 020 1 766 0201766Dup Inj 0201767 0201767 Dup Inj 0201768 0201768 Dup Inj 020 1769 0201769 Dup Inj 0201 770 0201770 Dup Inj 0201771 0201771 Dup Inj 020 1 772 0201772Dup Inj 020 1 773 0201773 Dup Inj 0201 774 0201774Dup In. 020 1775 0201775Dup Inj 020 1 776 0201776 Dup Inj 0203650 0203650Dup Inj 020 1777 0201777Dup lnj 0201778 0201778 Dup Inj 020 1779 0201779 Dup Inj 0201 780 0201780Dup Inj 0201781 0201781 Dup Inj 0201 782 0201782Dup Inj 020 1783 0201783 Dup Inj Matrix Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Collection Date 5/12/02 5/12/02 5/12/02 51 1 2/02 5/12/02 5/12/02 51 12/02 511 2/02 5/12/02 511 2/02 5/12/02 511 2/02 51 12/02 5/12/02 5/12/02 5/12/02 5/12/02 511 2/02 5/12/02 511 2/02 5/12/02 5/12/02 511 2/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 511 2/02 5/12/02 5/9/02 5/9/02 511 9/02 511 9/02 5/7/02 5/7/02 511 0102 5/10/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/7/02 5/7/02 5/9/02 5/9/02 5/6/02 5/6/02 5/6/02 5/6/02 Set Number 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092602A 092602A 092602A 092602A 092602A 092602A 092602A 092602A 092602AR 092602AR 092602AR 092602AR 092602AR 092602AU 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR PFHS Found (ng/mL) 606 576 138 141 326 335 40100 42300 36800 37900 52900 52800 149000 154000 124000 121000 172000 177000 194000 187000 408000 407000 4 1 6000 420000 50 1 000 51 1000 659000 675000 6 12000 603000 ND ND 822M 82.4M ND ND 628 633 1040 974 762 725 1960 2080 3070 3090 2700 2750 6360 6640 9740 9370 MAlthough the method used allows for alternate sample volumes or weights and all the peak area responses are within the calibration curve limits, the precision for the 0.1 mL or 0.1 g samples below 100 ppb has not been validated. * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 28 of 153 418-028:PAGE G-29 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19087 GROUP IV 19087GROUP IV* 19088 GROUP V 19088GROUP V* 19089 GROUP V 19089GROUP V* 19090 GROUP V 19090GROUP V* 19076 GROUP I 19076GROUP I* 19078 GROUP I 19078 GROUP I* 19081GROUP 11Female 2 19081 GROUP 11Female 2* 19079 GROUP 11 19079 GROUP 11* 19080GROUP I1 19080 GROUP II* 19082GROUP III 19082 GROUP III* 19083GROUP ILI 19083 GROUP LIP 19084 GROUP III 19084GROUP III* 19085GROUP IV 19085 GROUP IV* 19086 GROUP IV 19086 GROUP IV* 19087GROUP IV 19087 GROUP IV* 19088 GROUP V 19088 GROUP V* 19089 GROUP V 19089GROUP V* 19090 GROUP V 19090GROUP V* 19012 GROUP I, Male 1 19012 GROUP I, Male 1* 19012 GROUP I, Male 2 19012GROUP I, Male 2* 19012GROUP I, Male 3 19012 GROUP I, Male 3* 19012 GROUP I, Male 4 19012 GROUP I, Male 4* 19012GROUP I, Male 5 19012 GROUP I, Male 5* 19012 GROUP I, Female 7 19012 GROUP I, Female 7* 19012 GROUP I, Female 8 19012 GROUP I, Female 8* 19012 GROUP I, Female 14 19012 GROUP I, Female 14* Exygen ID 0201784 0201784 Dup Inj 0201785 0201785 Dup In, 0201786 0201786Dup Inj 0201787 0201787 Dup Inj 0201788 0201788Dup Inj 020 1789 020 1789 Dup Inj 0203651 0203651 Dup Inj 020 1790 0201790Dup Inj 0201791 0201791 Dup Inj 020 1792 0201792 Dup Inj 020 1793 0201793 Dup Inj 0201794 0201794 Dup Inj 020 1795 0201795 Dup Inj 020 1796 0201796Dup In. 0201797 0201797Dup Inj 020 1798 0201798Dup Inj 0201799 0201799Dup Inj 020 1800 0201800Dup Inj 0201925 0201925 Dup Inj . 0201926 0201926 Dup Inj 0201927 0201927Dup Inj 0201928 0201928 Dup Inj 020 1929 0201929Dup h j 020 1930 0201930Dup Inj 0201931 0201931 Dup Inj 020 1932 0201932 Dup Inj Matrix Median Liver Lobe MedianLiver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Median Liver Lobe Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livas Pup Livers Puu Livers Collection Date 5/6/02 5/6/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/7/02 5/7/02 511 0102 51 10102 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/7/02 5/7/02 5/9/02 5/9/02 5/6/02 5/6/02 5/6/02 5/6/02 5/6/02 5/6/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 Set Number 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 0926028 092602B 092602B 092602B 092602B 092602B 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A PFHS Found (ng/mc) 7500 7210 19000 20000 15100 15300 14700 15100 27.7M 25.7M ND ND 789 82 1 1460 1480 1810 1880 2180 2290 4560 4550 3020 3150 7320 7170 8510 8590 5630 5920 22200 22800 20 100 19800 14700 13600 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND MAlthough the method used allows for alternate sample volumes or weights and all the peak area responses are within the calibrationcurve limits, the precision for the 0.1 mL or 0.1 g samplesbelow 100 ppb has not been validated. * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 29 of 153 418-028:PAGE G-30 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19012GROUP I, Female 15 19012 GROUP I, Female 15* 19012GROUP I, Female 16 19012 GROUP I, Female 16* 19019 GROUP I, Male 1 19019 GROUP 1, Male 1* 19019 GROUP I, Male 2 19019 GROUP I, Male2* 19019GROUP I, Male 3 19019 GROUP I, Male 3* 19019 GROUP I, Male 7 19019 GROUP I, Male 7* 19019 GROUP I, Male 8 19019 GROUP I, Male 8* 19019GROUP I, Female 11 19019 GROUP I, Female 11* 19019GROUP I, Female 12 19019 GROUP I, Female 12* 19019 GROUP I, Female 13 19019 GROUP I, Female 13* 19019GROUP I, Female 14 19019 GROUP I, Female 14* 19019GROUP I, Female 15 19019 GROUP I, Female 15* 19021 GROUP I, Male 1 19021 GROUP I, Male 1* 19021 GROUP I, Male 2 19021 GROUP I, Male 2' 19021 GROUP I, Male 6 19021 GROUP I, Male 6* 19021 GROUP I, Male 8 19021 GROUP I, Male 8* 19021 GROUP I, Male 9 19021GROUP I, Male 9* 19021 GROUP I, Female 11 19021 GROUP I, Female 11* 19021 GROUP I, Female 12 19021 GROUP I, Female 12* 19021GROUP I, Female 13 19021 GROUP I, Female 13* 19021 GROUP I, Female 14 19021 GROUP I, Female 14" 19023 GROUP I, Male 1 19023GROUP I, Male 1* 19023 GROUP I, Male 2 19023 GROUP I, Male 2* 19023GROUP I, Male 3 19023 GROUP I, Male 3* 19023 GROUP I, Male 6 19023 GROUP I, Male 6* 19023 GROUP I, Male 7 19023 GROUP I, Male 7* Ex.yg-en ID 0201933 0201933Dup Inj 0201934 0201934 Dup Inj 0201935 0201935Dup Inj 0201936 0201936 Dup Inj 0201937 0201937 Dup Inj 020 1938 0201938 Dup In. 0201939 0201939 Dup Inj 0201940 0201940Dup lnj 0201941 0201941 Dup Inj 020 1942 0201942Dup Inj 020 1943 0201943 Dup lnj 020 1944 0201944Dup Inj 020 1945 0201945 Dup Inj 0201946 0201946Dup Inj 020 1947 0201947 Dup Inj 020 1948 0201948 Dup InJ 0201949 0201949Dup Inj 0201951 0201951 Dup Inj 0201952 0201952 Dup Inj 0201953 0201953Dup l j 0201954 020 1954 Dup Inj 0201955 0201955Dup Inj 0201956 0201956 Dup Inj 020 1957 0201957 Dup Inj 020 1958 0201958 Dup Inj 0201959 0201959 Dup Inj Matrix Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection Date 5129102 5129102 5129102 5129102 513OlO2 5130102 5130102 5130102 513 0102 5130102 5130102 5130102 5130102 5130102 5130102 513 0102 5130102 5130102 5130102 5130102 5130102 5130102 5130102 5130102 5129102 5129102 5129102 5/29/02 5129102 5/29/02 5/29/02 5129102 5129102 5129102 5/29/02 5/29/02 5129102 5129102 5/29/02 5129102 5/29/02 5/29/02 5131102 513 1102 5131102 5131/02 5131I02 5131102 5131102 5131I02 5131I02 513 1102 Set Number 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 09270249 092702A 092702A 092702B 092702B 0927028 0927028 092702B 092702B 092702B 0927028 092702B 0927028 092702B 0927028 0927028 0927028 092702B 092702B 0927028 0927028 092702B 0927028 092702B 092702B 0927028 092702B 0927028 092702B 092702B 092702B PFHS Found (ng/mL) ND ND ND ND ND ND ND ND 21.7M 18.3M 20SM 18.gM ND ND ND ND ND ND 34.0M 30.6M ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 50.6M 53.F 34.0M 37.0M 19.6M 24.1M ND ND 30.aM 34.0M MAlthough the method used allows for alternate sample volumes or weights and all the peak area responses are within the calibrationcurve limits, the precision for the 0.1 mL or 0.1 g samplesbelow 100 ppb has not been validated. * Duplicate Injection ND =Not Detected (Area less than lowest calibrationstandard of 0.1 ng/mL) Exygen Research Page 30 of 153 418-028:PAGE G-31 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 9 19023 GROUP I, Female 8* 19023 GROUP I, Female 9 19023 GROUP I, Female 9* 19023 GROUP I, Female 10 19023 GROUP 1, Female 10* 19023 GROUP I, Female 11 19023 GROUP I, Female 11* 19023 GROUP I, Female 12 19023 GROUP I, F a a l e 12* 19021 GROUP I, Female 10 19021 GROUP I, Female 10* 19041GROUP I, Male 1 19041GROUP I, Male 1* 19041 GROUP I, Male 2 19041GROUP I, Male 2* 19041 GROUP I, Male 3 19041 GROUP I, Male 3* 19041GROUP I, Male 4 19041 GROUP I, Male 4* 19041 GROUP I, Male 5 19041GROUP I, Male 5* 19041 GROUP I, Female 8 19041GROUP I, Female 8' 19041 GROUP I, Female 9 19041 GROUP I, Female 9* 19041 GROUP I, Female 15 19041 GROUP I, Female IS* 19041 GROUP I, Female 16 19041 GROUP I, Female 16* 19041 GROUP I, Female 17 19041 GROUP I, Female 17* 19004GROUP II,Male 2 19004GROUP 11,Male 2* 19004 GROUP 11,Male 3 19004GROUP 11,Male 3* 19004 GROUP 11, Male 4 19004GROUP 11,Male 4* 19004 GROUP II, Male 5 19004 GROUP 11, Male 5* 19004 GROUP 11, Male 6 19004 GROUP 11, Male 6" 19004 GROUP 11, Female 9 19004GROUP 11, Female 9* 19004 GROUP 11, Female 11 19004GROUP XI, Female 11* 19004 GROUP 11, Female 15 19004GROUP II,Female 15* 19004GROUP 11, Female 16 19004 GROUP 11, Female 16* 19004 GROUP 11, Female 17 19004 GROUP II, Female 17* Exygen ID 0201960 0201960 Dup Inj 0201961 0201961Dup In. 020 1962 0201962Dup I j 020 1963 0201963 Dup Inj 0201964 0201964 Dup In. 020 1950 0201950 Dup Inj 0201965 0201965 Dup Inj 020 1966 0201966 Dup hj 0201967 0201967 Dup Inj 0201968 0201968 Dup Inj 020 1969 0201969Dup Inj 020 1970 0201970 Dup Inj 0201971 0201971 Dup Inj 0201972 0201972 Dup In. 0201973 0201973 Dup Inj 020 1974 020 1974 Dup Inj 0202025 0202025 Dup Inj 0202026 0202026 Dup Inj 0202027 0202027 Dup In. 0202028 0202028 Dup In. 0202029 0202029 Dup Inj 0202030 0202030 Dup Inj 020203 1 020203 1Dup In, 0202032 0202032 Dup Inj 0202033 0202033 Dup Inj 0202034 0202034 Dup Inj Matrix Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Liven Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers hy, Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection Date 5131/02 513 1/02 5131/02 5/31/02 513 1102 5/31/02 5131/02 513 1/02 5131/02 5131102 5/29/02 5/29/02 5/31/02 5131/02 513 1/02 5/31/02 513 1/02 5131/02 5131/02 513 1/02 5/31/02 513 1/02 5131/02 5/31/02 513 1/02 5131/02 5131102 5131102 5/31/02 513 1/02 5/31/02 513 1/02 5/31/02 5/31/02 5131/02 513 1/02 513 1/02 513 1/02 5131/02 5131/02 513 1/02 513 1/02 5131/02 5131/02 5/31/02 513 1102 513 1/02 5/31/02 5131/02 513 1/02 513 1/02 5131/02 set Number 0927mB 092702B 092702B 092702B 0927028 0927028 092702B 092702B 092702B 092702B 092702BR 092702BR 100102A 100102A 100 102A 100102A 100102A 100 102A 100102A 100102A 100102A 100102A 100 102A 100102A 100 102A 100102A 100102A 100102A 100102A 100102A 100102A 100102A 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR PFHS Found (ng/mL) ND ND ND ND ND ND ND ND ND ND ND ND ND ND 24.4M 26.6M 26.0M 24.7M ND ND ND ND ND ND 30.6M 30.8M ND ND ND ND ND ND 1220 1160 1160 I120 2380 2480 1670 1670 2070 2000 1870 1880 1740 1710 1070 1060 1450 1530 946 983 "Although the method used allows for alternate sample volumes or weights and all the peak area responses are within the calibration curve limits, the precision for the 0.1 mL or 0.1 g samples below 100 ppb has not been validated. * Duplicate Injection ND =Not Detected (Arealess than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 31 of 153 418-028:PAGE G-32 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19018 GROUP 11, Male 1 19018 GROUP II,Male 1* 19018 GROUP 11,Male 4 19018GROUP II,Male 4* 19018 GROUP 11, Male 5 19018 GROUP 11, Male 5* 19018 GROUP 11, Male 6 19018 GROUP 11, Male 6* 19018 GROUP II, Male 7 190I 8 GROUP II, Male 7* 19018 GROUP II, Female 9 19018 GROUP II, Female 9* 19018 GROUP 11, Female 10 19018 GROUP 11, Female 10* 19018 GROUP II, Female 11 19018 GROUP II, Female 11* 19018 GROUP IX, Female 12 19018 GROUP II, Female 12* 19018 GROUP 11, Female 14 19018 GROUP XI, Female 14* 19026 GROUP II, Male 5 19026 GROUP II, Male 5* 19026 GROUP II, Male 1 19026 GROUP 11, Male 1* 19026 GROUP I &Male 3 19026 GROUP 11, Male 3* 19026 GROUP 11, Male 4 19026 GROUP II, Male 4* 19026 GROUP II, Male 7 19026 GROUP II, Male 7" 19026 GROUP 11, Female 12 19026GROUP 11,Female 12* 19026 GROUP 11, Female 13 19026GROUP XI, Female 13" 19026 GROUP 11, Female 14 19026 GROUP 11, Female 14* 19026 GROUP 11, Female 15 19026 GROUP 11, Female 15* 19026 GROUP 11, Female 18 19026 GROUP II, Female 18* 19036 GROUP 11, Male 1 19036 GROUP 11, Male I* 19036 GROUP 11, Male 2 I9036 GROUP II, Male 2* 19036 GROUP 11, Male 4 19036 GROUP II, Male 4* 19036 GROUP 11, Male 5 19036 GROUP II, Male 5* 19036 GROUP II, Male 6 19036 GROUP II, Male 6* 19036 GROUP 11, Female 7 19036 GROUP II, Female 7* Exygen ID 0202050 0202050 Dup Inj 020205 1 0202051Dup Inj 0202052 0202052 Dup Inj 0202053 0202053 Dup lnj 0202054 0202054 Dup Inj 0202055 0202055 Dup hj 0202056 0202056 Dup Inj 0202057 0202057 Dup Inj 0202058 0202058 Dup Inj 0202059 0202059 Dup Inj 0202063 0202063 Dup In, 0202060 0202060 Dup hj 0202061 0202061 Dup Inj 0202062 0202062 Dup Inj 0202064 0202064 Dup Inj 0202065 0202065 Dup Inj 0202066 0202066 Dup Inj 0202067 0202067 Dup Inj 0202068 0202068 Dup Inj 0202069 0202069 Dup Inj 0202070 0202070 Dup Inj 0202071 0202071 Dup Inj 0202072 0202072 Dup Inj 0202073 0202073 Dup Inj 0202074 0202074 Dup Inj 0202075 0202075 Dup Inj Matrix Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Liven Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection Date 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 513 1102 513 1/02 5131I02 513 1I02 513 1/02 513 1102 513 1/02 513 1/02 513 1/02 513 1102 513 1I02 513 1102 513 1102 513 1/02 513 1102 513 1/02 513 1/02 513 1/02 513 1/02 513 1/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 Set Number 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A PFHS Found (ng/mL) 839 816 788 818 886 872 868 888 523 514 556 596 804 785 649 664 743 75 1 788 815 937 928 1130 1130 1170 1150 1140 1130 1050 1010 946 956 928 919 1040 1140 1780 1830 1270 1310 746 748 169 174 707 693 965 946 667 649 622 620 * Duplicate Injection N D =Not Detected (Area less than lowest calibrationstandard of 0.1 ng/mL) Exygen Research Page 32 of 153 418-028:PAGE G-33 Exygen Study No.: 023-072 Table IX, (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19036 GROUP 11, Female 8 19036GROUP 11, Female 8* 19036GROUP 11, Female 1 1 19036GROUP II, Female 11* 19036 GROUP 11, Female 12 19036GROUP 11, Female 12* 19036 GROUP 11, Female 13 19036 GROUP II, Female 13" 19037GROUP II,Male 6 19037GROUP II, Male 6* 19037 GROUP 11, Female 15 19037GROUP II, Female 15* 19037GROUP II, Male 2 19037GROUP II, Male 2* 19037GROUP 11, Male 3 19037GROUP 11, Male 3* 19037GROUP 11, Male 7 19037GROUP 11, Male 7* 19037GROUP II, Male 8 19037GROUP II, Male 8* 19037 GROUP 11, Female 10 19037GROUP I& Female 10* 19037GROUP11, Female 11 19037GROUP 11, Female I 1* 19037 GROUP 11, Female 13 19037 GROUP 11, Female 13* 19037 GROUP 11, Female 14 19037GROUP II, Female 14* 19003 GROUP III, Male 1 19003 GROUP IU,Male 1* 19003 GROUP III, Male 3 19003 GROUP 111, Male 3* 19003 GROUP 111, Male 5 19003 GROUP 111, Male 5* 19003 GROUP 111, Male 7 19003 GROUP 111, Male 7* 19003 GROUP 111, Male 8 19003 GROUP 111, Male 8* 19003 GROUP 111, Female 9 19003 GROUP I , Female 9* 19003 GROUP 111, Female 10 19003G R O W 111, Female 10* 19003 GROUP III, Female 1 1 19003 GROUP 111, Female 11* 19003 GROUP 111, Female 15 19003 GROUP 111, Female 15' 19003GROUP UI,Female 16 19003 GROUP UU, Female 16* 19007GROUP III, Male 1 19007GROUP ID, Male 1* 19007 GROUP 111, Male 2 19007 GROUP In, Male 2* EKygen ID 0202076 0202076Dup Inj 0202077 0202077Dup Inj 0202078 0202078Dup In. 0202079 0202079 Dup Inj 0202082 0202082Dup Inj 0202089 0202089Dup Inj 0202080 0202080Dup Inj 020208 1 0202081 Dup Inj 20831983 0202083 Dup Inj 0202084 0202084Jhp I j 0202085 0202085Dup Inj 0202086 0202086Dup Inj 0202087 0202087Dup Inj 0202088 0202088Dup Inj 0202119 0202119 Dup Inj 0202 120 0202120Dup Inj 0202121 0202121 Dup Inj 0202122 0202122Dup Inj 0202 123 0202123Dup Inj 0202124 0202124Dup Inj 0202125 0202125 Dup Inj 0202126 0202126Dup Inj 0202127 0202127Dup Inj 0202 128 0202128Dup Inj 0202129 0202129 Dup Inj 0202 130 0202130Dup Inj Matrix Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection Date 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 513 0102 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 Set Number 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100302AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR PFHS Found (ng/mL) 668 650 554 543 676 642 494 490 906 900 956 965 1670 1670 1430 1380 1280 1270 1760 1770 1150 1160 1300 1320 1680 1670 1330 1350 4640 4730 6250 6320 4490 4650 4540 4440 5820 5 840 4100 4170 4400 4340 3270 3240 4810 4920 5430 5630 3030 3090 43 10 4400 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 33 of 153 418-028:PAGE G-34 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor 19007 GROUP III, Male 3 19007 GROUP 111, Male 3* 19007 GROUP III, Male 6 19007GROUP III, Male 6* 19007GROUP III, Male 7 19007 GROUP 111, Male 7* 19007 GROUP 111, Female 9 19007GROUP III,Female 9* 19007GROUPm,Female 11 19007GROUP III, Female 11* 19007 GROUP III, Female 12 19007 GROUP III, Female 12* 19007GROUP In, Female 13 19007GROUP 111,Female 13* 19007GROUP III, Female 16 19007 GROUP III, Female 16* 19008 GROUP 111, Male 1 19008GROUP In, Male 1* 19008GROUP 111,Male 3 19008 GROUP In, Male 3* 19008GROUP 111,Male 4 19008 GROUP 111, Male 4* 19008 GROUP 111, Male 6 19008GROUP 111,Male 6* 19008 GROUP 111, Male 7 19008 GROUP 111, Male 7* 19008 GROUP 111, Female 9 19008 GROUP In, Female 9* 19008 GROUP 111, Female 10 19008GROUP III, Female 10* 19008 GROUP 111, Female 11 19008GROUP 111,Female 11* 19008 GROUP In, Female 12 19008 GROUP 111, Female 12* 19008 GROUP 111,Female 13 19008 GROUP 111, Female 13* 19013 GROUP 111, Male 1 19013 GROUP Il& Male 1* 19013 GROUP 111,Male 4 19013GROUP 111,Male 4* 19013GROUP III, Male 5 19013 GROUP IU, Male 5* 19013GROUP 111,Male 6 19013GROUP In, Male'6 19013 GROUP 111, Male 9 19013GROUP III, Male 9* 19013GROUP 111,Female 1 1 19013 GROUP 111, Female 11* 19013 GROUP 111, Female 12 19013 GROUP III, Female 12* 19013 GROUP 111, Female 13 19013 GROUP III, Female 13* Exygen ID 0202131 0202131 DupInj 0202 132 0202 132 Dup Inj 0202133 0202133Dup Inj 0202 134 0202 134 Dup Inj 0202 135 0202135Dup Inj 0202136 0202136Dup Inj 0202137 0202137 Dup Inj 0202138 0202138 Dup I j 0202139 0202139 Dup Inj 0202 140 0202 140 Dup Inj 0202141 0202 141 Dup Inj 0202 142 0202142Dup Inj 0202 143 0202143 Dup Inj 0202 144 0202144Dup Inj 0202 145 0202145Dup Inj 0202146 0202146Dup Inj 0202 147 0202147Dup Inj 0202148 0202148 Dup Inj 0202149 0202149 Dup Inj 0202150 0202150 Dup Inj 020215 1 0202151 Dup Inj 0202 152 0202152Dup Inj 0202153 0202153 Dup Inj 0202 154 0202154 Dup Inj 0202155 0202155 Dup Inj 0202156 0202156 Dup Inj Matrix Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5131102 5131102 5131102 5131102 513 1/02 5131102 5131102 5131I02 5131102 513 1102 5131102 5131102 513 1/02 513 1/02 5131I02 513 1/02 5131102 513 1/02 513 1 I02 513 1/02 5/29/02 . 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 Set 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR PFHS 4450 4370 4430 4600 5630 5720 6340 6330 3850 3820 5230 5130 6630 6870 5720 5690 3920 3810 5080 5070 4090 4070 4810 4750 4690 4380 5190 5050 3070 2770 4120 3760 2790 2480 2440 2340 1590 1790 1950 2050 2030 2090 2420 2480 2180 2280 2130 2150 2000 2010 1680 1780 * Duplicate Injection ND =Not Detected (Area less than lowest calibration standard of0.1ng/mL) Exygen Research Page 34 of 153 418-028:PAGE G-35 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19013GROUP III, Female 14 19013GROUP 111,Female 14* 19013GROUP III, Female 15 19013 GROUP 111, Female 15* 19015 GROUP LII, Male 1 19015 GROUP III, Male 1* 19015 GROUP III, Male 3 19015 GROUP III, Male 3* 19015GROUP 111, Male 6 19015 GROUP III, Male 6* 19015GROUP III, Male 7 19015GROUP III, Male 7* 19015GROUP 111, Male 8 19015GROUP III,Male 8* 19015GROUP III,Female 10 19015 GROUP 111, Female 10* 19015GROUP ZU,Female 12 19015GROUP 111, Female 12* 19015GROUP 111,Female 13 19015GROUP III, Female 13* 19015GROUP 111,Female 14 19015GROUP 111, Female 14* 19015GROUP III, Female 15 19015 GROUP III, Female 15* 19005GROUP IV, Male 1 19005GROUP IV, Male 1* 19005GROUP IV,Male 2 19005GROUP N,Male 2* 19005GROUP IV, Male 3 19005GROUP N,Male 3* 19005GROUP IV,Male 5 19005GROUP IV, Male 5* 19005GROUP IV,Male 6 19005GROUP IV, Male 6* 19005GROUP IV, Female 8 19005GROUP IV,Female 8* 19005 GROUP N,Female 10 19005GROUP IV, Female lo* 19005GROUP IV, Female 11 19005GROUP W ,Female 11* 19005GROUP IV, Female 12 19005GROUP IV,Female 12* 19005GROUP IV,Female 15 19005GROUP IV,Female 15" 19035GROUP IV, Male 1 19035GROUP IV, Male 1* 19035GROUP IV, Male 2 19035GROUP IV, Male 2* 19035GROUP IV, Male 4 19035 GROUP IV, Mate 4+ 19035 GROUP IV, Male 5 19035GROUP IV,Male 5* Exygen ID 0202157 0202157 Dup Inj 0202 158 0202158 Dup Inj 0202 159 0202159Dup Inj 0202 160 0202 160 Dup Inj 0202161 0202161 Dup Inj 0202 162 0202162 Dup Inj 0202 163 0202163 Dup Inj 0202 164 0202164Dup Inj 0202 165 0202165Dup Inj 0202 166 0202166Dup Inj 0202 167 0202167Dup Inj 0202168 0202168Dup Inj 0202219 0202219Dup Inj 0202220 0202220 Dup Inj 0202221 0202221 Dup Inj 0202222 0202222 Dup Inj 0202223 0202223 Dup Inj 0202224 0202224 Dup Inj 0202225 0202225 Dup Inj 0202226 0202226 Dup Inj 0202227 0202227 Dup In. 0202228 0202228 Dup Inj 0202229 0202229 Dup Inj 0202230 0202230 Dup Inj 020223 1 0202231 Dup Inj 0202232 0202232 Dup Inj Matrix Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection Date 5/29/02 5/29/02 5/29/02 5/29/02 5131102 513 1/02 513 1/02 5131102 513 1/02 5131102 513 1/02 513 1/02 5131102 513 1/02 5/31/02 513 1/02 513 1/02 5131I02 5131102 513 1/02 5131/02 5131102 513 1/02 5131102 5131102 513 1/02 5131102 5131102 513 1/02 5131102 513 1/02 513 1/02 5131102 5131102 5131/02 513I 102 513 1/02 513 1/02 5131102 5131/02 5131102 5131/02 5131102 513 1/02 513 1102 5131102 513 1/02 5131102 513 1/02 513 1/02 513 1/02 513 1/02 Set Number 100402BR 100402BR 100402BR 100402BR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR PFHS Found (ng/mL) 2290 2300 2000 2070 2950 2840 2780 2840 2860 2940 3150 3220 4680 4710 3800 3 800 4280 4180 4000 3950 3800 3890 5330 5420 8860 8800 9280 9370 10500 11100 8340 9100 11200 10900 11400 11500 12300 12800 12600 12800 9010 9100 11800 12000 11500 12000 14500 14500 13700 13600 11300 10800 * Duplicate Injection ND =Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 35 of 153 418-028:PAGE G-36 Exygen StudyNo.:023-072 Table IX,(cont'd), Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19035GROUP W ,Male 6 19035GROUP IV,Male 6* 19035GROUP IVYFemale 8 19035GROUP IVYFemale 8* 19035GROUP IV,Female 10 19035GROUP IVYFemale lo* 19035GROUP IVYFemale 11 19035GROUP IV,Female 11* 19035GROUP IVYFemale 12 19035GROUP IVYFemale 12* 19035GROUP IV, Female 13 19035GROUP IVYFemale 13* 19039GROUP W ,Male 1 19039GROUP N,Male 1* 19039GROUP IV, Male 2 19039GROUP IV, Male 2* 19039 GROUP IV, Male 3 19039GROUP IVYMale 3* 19039GROUP IV,Male 4 19039GROUP IV,Male 4* 19039GROUP N,Male 5 19039GROUP IVYMale 5* 19039 GROUP N, Female 8 19039GROUP IVYFemale 8* 19039GROUP IVYFemale 11 19039GROUP IVYFemale 11* 19039GROUP IVYFemale 12 19039GROUP IV,Female 12* 19039GROUP IVYFemale 14 19039GROUP IV, Female 14* 19039GROUP IVYFemale 15 19039GROUP IVYFemale 15* 19040GROUP IVYMale 2 19040GROUP IVYMale 2* 19040GROUP IV,Male 3 19040GROUP IV, Male 3* 19040 GROUP IV, Male 4 19040GROUP N,Male 4* 19040GROUP W ,Male 5 19040 GROUP N ,Male S* 19040 GROUP IV, Male 7 19040GROUP IV, Male 7* 19040GROUP IVYFemale 11 19040GROUP IVYFemale 11* 19040GROUP IV, Female 12 19040GROUP IVYFemale 12' 19040GROUP IVYFemale 13 19040GROUP IVYFemale 13* 19040GROUP IV,Female 15 19040 GROUP IV, Female 15* 19040GROUP IV, Female 16 19040 GROUP IVYFemale 16* Exygen ID 0202233 0202233 Dup Jnj 0202234 0202234 Dup In. 0202235 0202235 Dup In. 0202236 0202236 Dup Inj 0202237 0202237 Dup Knj 0202238 0202238 Dup Inj 0202239 0202239 Dup Inj 0202240 0202240 Dup Inj 0202241 0202241 Dup Inj 0202242 0202242 Dup Inj 0202243 0202243 Dup Inj 0202244 0202244 Dup Inj 0202245 0202245 Dup Inj 0202246 0202246 Dup Inj 0202247 0202247 Dup Inj 0202248 0202248 Dup Inj 0 2 0 2 2 4 9 0202249 Dup Inj 0202250 0202250 Dup Inj 020225 1 0202251 Dup Inj 0202252 0202252 Dup Inj 0202253 0202253 Dup Inj 0202254 0202254 Dup Inj 0202255 0202255 Dup Inj 0202256 0202256 Dup Inj 0202257 0202257 Dup Xnj 0202258 0202258 Dup Inj Matrix Pup Livers Pup Livers Pup Livers Pup Liven Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection Date 5131102 513 1/02 513 1/02 513 1 102 513 1102 513 1/02 5l3 1102 5131102 513 1/02 513 1102 513 1/02 5131102 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/30/02 5/30/02 5/30/02 513 0102 5/30/02 5/30/02 5/30/02 5J30102 5/30/02 513 Of02 5/30/02 5/30/02 513OlO2 5/30/02 5130102 5/30/02 5/30/02 5/30/02 s/30/02 5/30/02 Set Number 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 101002AR 10 1002AR 10 1002AR 10 1002AR 10 1002AR 101002AR 101002AR 10 1002- 10 1002AFt 101002AR 101002AR 101002AR 10 1002AR 10 1002AR 10 1002AR 10 1002AR 10 1002AR 101002AR 10 1002AR 101002AR PFHS Found (ng/mL,) 10600 10500 12000 13300 10000 10400 18500 18400 12600 12000 17300 17300 5320 5200 6160 6310 5300 5150 5870 6130 6500 6430 4910 5010 3510 3710 4550 4620 4310 4180 51 10 4920 6480 7690 11500 11200 7540 7750 13000 12200 1 1 100 11100 12100 1 1200 10600 10700 14100 14000 12400 12300 14900 15700 * Duplicate Injection ND =Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 36 of 153 418-028:PAGE G-37 Exygen Study No.: 023-072 Table IX.(cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19045 GROUP IV, Male 1 19045GROUPIV, Male 1* 19045 GROUP IV, Male 2 19045GROUP N,Male 2* 19045 GROUP IV, Male 4 19045GROUP IV,Male 4* 19045 GROUP IV, Male 5 19045GROUPW , Male 5* 19045GROUP W , Male 6 19045GROUP N,Male 6* 19045GROUP N,Female 8 19045GROUP N,Female 8* 19045GROUP IV,Female 9 19045GROUP N,Female 9* 19045GROUP N ,Female 10 19045GROUP IV,Female 10* 19045GROUP ~,Female 11 19045GROUP IV,Female 11* 19045GROUP N ,Female 14 19045GROUP N,Female 14* 19006GROUP V, Male 1 19006GROUP V,Male 1* 19006GROUP V,Male 2 19006GROUP V,Male 2* 19006 GROUP V, Male 3 19006 GROUP V, Male 3* 19006 GROUP V, Male 4 19006 GROUP V, Male 4* 19006 GROUP V, Male 5 19006 GROUP V, Male 5* 19006 GROUP V,Female 8 19006 GROUP V, Female 8* 19006 GROUP V, Female 10 19006 GROUP V, Female 10* 19006 GROUP V, Female 11 19006GROUP V, Female 11* 19006 GROUP V, Female 13 19006 GROUP V, Female 13* 19006GROUP V,Female 18 19006 GROUP V, Female 18* 19011 GROUP V, Male 2 19011 GROUP V, Male 2* 19011 GROUP V,Male3 19011GROUP V, Male 3* 19011 GROUP V, Male 4 19011 GROUP V, Male 4* 19011 GROUPV,Male5 19011 GROUP V, Male 5' 19011 GROUP V,Male 6 19011 GROUP V, Male 6* 19011 GROUP V, Female 9 19011 GROUP V, Female9* Exygen m 0202259 0202259 Dup Inj 0202260 0202260 Dup Inj 0202261 0202261 Dup Inj 0202262 0202262 Dup Inj 0202263 0202263 Dup Inj 0202264 0202264 Dup Inj 0202265 0202265 Dup Inj 0202266 0202266 Dup Inj 0202267 0202267 Dup Inj 0202268 0202268 Dup Inj 0202320 0202320 Dup Inj 0202321 0202321Dup Inj 0202322 0202322 Dup In. 0202323 0202323 Dup Inj 0 2 0 2 3 2 4 0202324Dup Inj 0202325 0202325 Dup Inj 0202326 0202326 Dup Inj 0202327 0202327 Dup Inj 0202328 0202328 Dup Inj 0202329 0202329 Dup Inj 0202330 0202330 Dup Inj 0202331 0202331 Dup Inj 0202332 0202332 Dup Inj 0202333 0202333 Dup Inj 0202334 0202334 Dup Inj 0202335 0202335 Dup Inj Matrix Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection Date 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5128102 5/28/02 512 8/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 512 8102 5/28/02 5/28/02 5f30102 5130f02 5130102 5130f02 5f30102 5f30102 5/30/02 5f30102 5130f02 5130102 5130/02 513Of02 5130/02 5130102 5f30102 5130102 5f30102 5130102 5130102 5130102 5131I02 513 1/02 513 1102 513 1102 513 1/02 513 1102 513 1/02 513 1/02 513 1102 513 1102 5131I02 513 1102 Set Namber 101002AR 101002AR 101002AR 101002AR 101002AR 101002AR 101002AR 101002AR 10 1002AR 101002AR 101002AR 101002AR 101002AR 101002AR 101002AR 101002AR 10 1002AR 10 1002AR 101002AR 101002AR 101002BR 101002BR 10 1002BR 101002BR 10 1002BR 101002BR 101002BR 101002BR 10 1002BR 10 1002BR 10 1002BR 101002BR 10 1002BR 10 1002BR 101002BR 10 1002BR 10 1002BR 101002BR 10 1002BR 101002BR 10 1002BR 10 1002BR 10 1002BR 101002BR 10 1002BR 10 1002BR 101002BR 10 1002BR 101002BR 10 1002BR lOlOO2BR 101002BR PFHS Foand (ng/mL) 6200 6210 4830 4900 7820 7510 5700 5750 4840 4970 7350 7350 6510 6740 8350 8010 6520 6380 6030 5870 13500 13700 16500 16700 25800 26100 30100 29400 30200 29800 23300 22500 17700 16400 25000 25000 16700 16500 20700 20200 19200 20000 I8700 19900 19300 19000 18600 19300 14000 14100 20800 21 100 * Duplicate Injection N D = Not Detected (Area less than lowest calibrationstandard of 0.1 nglmL) Exygen Research Page 37 of 153 418-028:PAGE G-38 Exygen Study No.:023-072 Table IX.(cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19011GROUP V, Female 10 19011 GROUP V, Female 10* 19011 GROUP V, Female 11 19011 GROUPV,Femalell+ 19011GROUP V, Female 12 19011 GROUP V, Female 12' 19011 GROUP V, Female 13 19011 GROUP V, Female 13' 19020 GROUP V, Male 2 19020 GROUP V, Male 2* 19020 GROUP V, Male 4 19020GROUP V, Male 4* 19020 G R O W V, Male 5 19020GROUP V,Male 5" 19020GROUP V, Male 7 19020GROUP V, Male 7* 19020 GROUP V, Male 8 19020 GROUP V, Male 8* 19020 GROUP V, Female 9 19020GROUP V, Female 9* 19020 GROUP V, Female 10 19020 GROUP V, Female 10* 19020GROUP V, Female 11 19020GROUP V, Female 11* 19020GROUP V, Female 12 19020 GROUP V, Female 12' 19020 GROUP V, Female 13 19020 GROUP V, Female 13* 19022 GROUP V, Male 1 19022GROUP V, Male I* 19022 GROUP V, Male 4 19022GROUP V,Male4* 19022 GROUP V, Male 5 19022 GROUP V, Male 5* 19022 GROUP V, Male 7 19022GROUP V, Male 7' 19022 GROUP V, Male 8 19022GROUP V, Male 8* 19022 GROUP V, Female 11 19022GROUP V, Female 11* 19022 GROUP V, Female 12 19022 GROUP V, Female 12* 19022 GROUP V, Female 13 19022 GROW V, Female 13* 19022 GROUP V, Female 14 19022GROUP V,Female 14* 19022 GROUP V, Female 15 19022 GROUP V, Female 15* 19025 GROUP V, Male 2 19025GROUP V, Male 2* 19025 GROUP V, Male 7 19025 GROUP V. Male 7* Exygen ID 0202336 0202336 Dup Inj 0202337 0202337 Dup Inj 0202338 0202338 Dup Inj 0202339 0202339 Dup Inj 0202340 0202340 Dup Inj 020234 1 0202341 Dup In. 0202342 0202342 Jhp Inj 0202343 0202343 Dup Inj 0202344 0202344 Dup Inj 0202 1345 0202345 Dup Inj 0202346 0202346 Dup In. 0202347 0202347Dup Inj 0202348 0202348 Dup Inj 0202349 0202349 Dup Inj 0202350 0202350 Dup h j 0202351 0202351 Dup Inj 0202352 0202352 Dup Inj 0202353 0202353 Dup Inj 0202354 0202354 Dup Inj 0202355 0202355 Dup Inj 0202356 0202356 Dup Inj 0202357 0202357 Dup Inj 0202358 0202358 Dup Inj 0202359 0202359 Dup hj 0202361 0202361 Dup Inj 0202362 0202362 Dup In. Matrix Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection Date 513 1102 513 1102 5131102 513 1102 513 1102 513 1/02 5131102 5131102 513 1102 5131102 513 1102 513 1102 513 1/02 513 1102 513 1102 5131102 5131102 513 1102 5131102 5131102 5131102 5131102 513 1102 5131102 513 1102 5131102 5131102 5131102 513 1 102 513 1102 5131102 5131102 5131102 5131102 513 1102 5131102 513 1102 5131102 5131102 5131102 513 1102 513 1102 5 01102 5131102 513 1102 5131102 513 1102 5131102 5130/02 5130102 513 0102 513 0102 Set Number 101002BRR 101002BRR 101002BRR 101002BRR 101002BRR 101002BRR 101002BRR 10 1002BRR 101 102AR 101102AR 101102AR 101102AFt 101102AR 101102101 102101102AR 101102AR 101102AR 101 102AR 101102101102AR 101102AR 101102AR 101 102AR 101 102101 102AR 101102AR 101102AR 101 102AR 101102AR 101102AR 101102AR 101 102AR 101102AR 101102AR 101 102AR 101102AR 101 102AR 101 102AR 101102AR 101 102AR 101102AR 101 102AR 101102AR 101102AR 101102AR 101102AR 101 102AR 101402AR 10 1402AR 10 1402AR 101402AR PFHS Found (ng/mL) 18700 20000 23500 22400 2 1900 22500 21000 2 1400 11800 11300 9460 9190 8130 8510 9280 9050 8750 8750 14800 14300 12200 12600 14400 14300 15600 15300 20900 21300 11100 11000 12700 12400 12500 12800 1 1800 11600 14400 14300 13700 14100 17400 17500 12200 11600 15700 15300 14100 14200 2 1700 21900 18600 18600 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 38 of 153 418-028:PAGE G-39 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID T O 2 5 GROUP V, Male 8 19025 GROUP V, Male 8* 19025 GROUP V, Male 13 19025 GROUP V, Male 13" 19025 GROUP V, Female 14 19025 GROUP V, Female 14* 19025 GROUP V, Female 15 19025 GROUP V, Female 15* 19025 GROUP V, Female 18 19025 GROUP V, Female 18* 19025 GROUP V, Female 19 19025 GROUP V, Female 19* 19025 GROUP V, Female 20 19025 GROUP V, Female 20* 19025 GROUP V, Male 1 19025GROUP V. Male I* Exygen ID 0202363 0202363 Dup Inj 0202364 0202364 Dup In. 02021365 0202365 Dup Inj 0202366 0202366 Dup Inj 0202367 0202367 Dup Inj 0202368 0202368 Dup Xnj 0202369 0202369 Dup Inj 0202360 0202360 Dup Inj Matrix Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Collection Date 5130102 5130102 5130102 5130102 5130102 5130102 513OlO2 5130102 5/30/02 5130102 5/30/02 5130102 5130102 5130102 5/30/02 5130102 Set Number 101402AR 101402AR 101402AR 101402AR 101402AR 101402AR 101402AR 101402AR 101402AR 101402AR 101402AR 101402AR 101402AR 101402AR 101402ARR 101402ARR PFHS Found (ng/mL,) 21400 2 1800 18300 19400 14700 15300 26400 26500 19800 19500 18900 18800 16800 17600 13300 14400 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 39 of 153 418-028:PAGE G-40 Exygen Study No.: 023-072 Table X. Summary of PFHS Residues in Dosing Solutions Sample Description B-418-028-A (07.Jun.02) 0 mg/mL 1 of 4 B-4 18-028-A (07.Jun.02) 0 mg/mL 1 Of 4 B-418-028-B (24.May.02) 0.03 mg/mL 1 of 4 B-418-028-B (24.May.02) 0.03 mghL 1 of 4 B-418-028-C (24.May.02) 0.1 mg/mL 1 of4 B-418-028-C (24.May.02) 0.1 mg/mL 1 of4 B-418-028-D (07.Jun.02) 0.3 mg/mL 1 of4 B-418-028-D (07.Jun.02) 0.3 mg/mL 1 of 4 B-418-028-E (24.May.02) 1 mg/mL 1 of 4 B-418-028-E (24.May.02) 1 mg/mL 1 of4 Bulk TPJS Sample (09.Jun.02) Bulk TA/S Sample (09.Jun.02) Set Number 1 10702C 110702C 110702C 110702C 110702C 110702C 1 10702C 110702C 110702CR 110702CR 110702C 1 10702C PFHS Known Concentration(ng/-mL) F 0 0 30000 30000 100000 100000 300000 300000 1000000 1000000 100000 100000 PFHS ND ND 26500 27300 88900 9 1000 268000 278000 1010000 1000000 125000 122000 ND = Not Detected (Area less than lowest calibration standard of 0.1 ng/niL,) PFHS 88 91 89 91 89 93 101 100 125 122 Exygen Research Page 40 of 153 418-028:PAGE G-41 Exygen Study No.: 023-072 Table XI. Summary of PFHS Residues in Stability Samples 3M ID E02-0.5 14-3861 1 EO2-0514-38611* E02-05 14-38612 E02-05 14-38612, E02-05 14-38613 EO2-05 14-38613* E02-05 14-38614 E02-05 14-38614* E02-05 14-38615 EO2-05 14-386155 E02-05 14-38616 EO2-05 14-38616* E02-0514-38617 EO2-05 14-38617* EO2-05 14-38618 EO2-05 1438618* E02-0514-38619 E02-05 14-38619' E02-05 14-38620 EO2-05 14-38620* Exygen ID 0203923 0203923 Dup hj 0203924 0203924Dup hj 0203925 0203925 Dup In. 0203926 0203926 Dup Inj 0203927 0203927Dup In. 0203928 0203928Dup Inj 0203929 0203929hrp Inj 0203930 0203930 Dup Iaj 0203931 0203931 Dup Inj 0203932 0203932 Dup Inj Sample Description B418-028 A (29 Mar 02) 0 mdmL, 1of 4 B-418-028A (29 Mar 02) 0 mg/mL 1of 4 B418-028 A (29 Mar 02) 0 mg/mL 2 of 4 8-418-028A (29 Mar 02) 0 mgmL 2 of 4 B418-028B (29 Mar 02) 0.03 mghL 1 of 4 B-418-028B (29 Mar 02) 0.03 m@mL1of 4 B-418-028 B (29 Mar 02) 0.03 m&mL 2 of4 B418-028B (29 Mar02) 0.03 m g / d2 of 4 B418-028 C (29 Mar 02) 0.1 mg/mL 1of 4 B418-028 C (29 Mar 02) 0.1 mg/mL 1of 4 B-418-028C (29 Mar 02) 0.1 mgmL 2 of 4 B-418-028C (29 Mar 02) 0.1 m@mL2 of 4 -18-028 D (29 Mar 02) 0.3 m@mL1of 4 B-418-028D (29 Mar 02) 0.3 mglmL 1of4 8-418-028D (29 Mar 02) 0.3 +UIL 2 Of4 8-418-028D (29 Mar 02) 0.3 WmL 2 Of4 B-418-028E (29 Mar 02) 1m g / d 1of 4 B 418-028E (29 Mar02) 1mumL 1 of4 B418-028 E (29 Mar02) 1mg/mL 2 of 4 B-418-028E (29 Mar02) 1m&mL2 of 4 Set Number 110502A 110502A 110502A 110502A 110502A 110502A 110502A 110502A 110502A 110502A 1 10502A 110502A 110502A 110502A 110502A 110502A 110702AR 110702AR 110702AR 110702AR PrnS Known Cone. (add) 0 0 0 0 30000 30000 30000 30000 100000 100000 100000 100000 300000 300000 3m 300000 1000000 1000000 1000000 1000000 PFHS Found (ndmL) ND ND ND ND 29700 29600 29700 29500 122000 1 17000 100000 100000 308000 316000 326000 324000 910000 9 19000 1030000 1010000 PFHS Recovery (%) 99 99 99 98 122 117 100 100 103 105 109 108 91 92 103 101 * Duplicate Injection ND =Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 41 of 153 418-028:PAGE G-42 Exygen Study No.:023-072 Table XII. Summary of PFHS Residues in Homogeneity Samples 3M ID E02-0514-38621 EO2-0514-38621* EO2-0514-38622 E02-0514-38622* EO2-0514-38623 EO2-0514-38623* E02-0514-38624 EO2-0514-38624* EO2-0514-38625 E02-0514-38625* EO2-0514-38626 EO2-0514-38626* EO2-0514-38627 E02-0514-38627* EO2-0514-38628 EO2-0514-38628* EO2-0514-38629 E02-051438629* E02-0514-38630 E02-0514-38630* E02-0514-38631 EO2-0514-38631* E02-0514-38632 EO2-0514-38632* EO2-0514-38633 EO2-0514-38633* EO2-0514-38634 EO2-0514-38634* EO2-0514-38635 EO2-0514-38635* EO2-0514-38636 E02-0514-38636* EO2-051438637 E02-0514-38637* E02-0514-38638 E02-0514-38638* Eo2-0514-38639 EO2-0514-38639* E02-0514-38640 EO2-0514-38640* E02-0514-38641 E02-0514-38641* EO2-0514-38642 EO2-0514-38642* E02-0514-38643 E02-0514-38643* E02-0514-38644 EO2-0514-38644* EO2-0514-38645 EO2-0514-38645* EO2-0514-38646 E02-051438646' EO2-0514-38647 EO2-0514-38647* E02-0514-38648 EO2-0514-38648* E02-0514-38649 E02-0514-38649* E02-0514-38650 EO2-0514-38650* Exygen ID 0203933 0203933 Dup Inj 0203934 0203934Dup h j 0203935 0203935Dup Inj 0203936 0203936Dup Inj 0203937 0203937 Dup Inj 0203938 0203938Dup Inj 0203939 0203939 Dup Inj 0203940 0203940 Dup Inj 020394 1 0203941 Dup Inj 0203942 0203942Dup Inj 0203943 0203943 Dup Inj 0203944 0203944 Dup Inj 0203945 0203945 Dup Inj 0203946 0203946Dup h j 0203947 0203947 Dup Inj 0203948 0203948 Dup Inj 0203949 0203949 Dup hj 0203950 0203950 Dup Inj 0203951 0203951 Dup Inj 0203952 0203952Dup Inj 0203953 0203953 Dup Inj 0203954 0203954 Dup Inj 0203955 0203955 Dup Inj 0203956 0203956Dup Inj 0203957 0203957 Dup Inj 0203958 0203958 Dup Inj 0203959 0203959 Dup Inj 0203960 0203960 Dup Inj 0203961 0203961 Dup Inj 0203962 0203962 Dup Inj Sample Description B-418-028-A (29MarO2) 0m g / d 1 of 12T B-418-028-A (29MarO2)0 m&nL 1 of 12T B-418-028-A (29MarO2) 0 mg/mL 2 of 12T B-418-028-A (29MarO2)0 mg/mL 2 of 12T B-418428-A (29Mar02) 0mg/mL 5 of 12M B-418-028-A (29MarO2) 0 mg/mL 5 of 12M B-418-028-A (29MarO2) 0 mg/mL, 6 of 12M B 418-028-A(29MarO2) 0 mg/mL 6 of 12M B-418-028-A (29Mar02) 0 mg/mL 9 of 12B B-418-028-A(29MarO2) 0 mg/mL 9 of 12B B-418-028-A (29MarO2) 0 mg/mL 10 of 12B B-418-028-A (29-2) 0 mglmL 10 of 12B B-418-028-B (29MarO2) 0.03mg/mL 1 of 12T B-418-028-B (29MarO2) 0.03 mg/mL 1 of 12T B-418-028-B (29-2) 0.03mg/mL2 of 12T B-418-028-B (29MarO2)0.03 mg/mL2 of 12T B418O28-B (29-2) 0.03 m g / d 5 of 12M B-418-028-B (29-2) 0.03 mg/mL 5 of 12M B-418-028-B (29MarO2) 0.03mg/mL 6 of 12M B-418-028-B ( 2 9 W 2 ) 0.03 mg/mL 6 of 12M B-418-028-B (29Mar02) 0.03 mg/mL 9 of 12B B-418-028-B (29Mar02) 0.03 mg/mL 9 of 12B B-418-028-B (29MarO2) 0.03 mg/mL 10 of 12B 8-418-028-B (29MarO2)0.03 m g / d 10of 128 B-418-028-C (29MarO2) 0.1 mg/mL 1 of 12T B 418-028-C(29Mar02) 0.1 mg/mL 1 of 12T B-418-0284 (29MarO2) 0.1 mg/mL 2 of 12T B-418-0284 (29MarO2)0.1 mg/mL 2 of 12T B418-028-C (29MarO2) 0.1 mg/& 5 of 12M B-418-028-C (29MarO2) 0.1 mg/mL 5 of 12M B-418-02842 (29MarO2) 0.1 mg/mL 6 of 12M B-418-028-C (29MarO2) 0.1 mg/mL 6 of 12M B-418-028-C (29MarO2) 0.1 mg/mL 9 of 12B B-4 18-028-C (29MarO2) 0.1 mg/mL 9 of 12B B-418-028-C (29MarO2) 0.1 mg/mL 10 of 12B B-418-028-C (29-2) 0.1 mg/mL 10 of 12B B-418-028-D (29MarO2) 0.3 mg/mL 1 of 12T B-418-028-D (29Mato2) 0.3 mg/mL 1 of 12T B-418-028-D (29Mar02) 0.3 mg/mL 2 of 12T B-418-028-D (29MarO2) 0.3 mg/mL 2 of 12T B-418-028-D (29MarO2) 0.3 mg/mL 5 of 12M B-418-028-D (29Mar02) 0.3 mg/mL. 5 of 12M B-418-028-D (29Md2) 0.3mg/mL 6 of 12M B-418-028-D (29MarO2) 0.3 mg/mL 6 of 12M B-418-028-D (29MarO2) 0.3 mg/mL 9 of 12B B-418-028-D(29MarO2)0.3 mg/mL 9 of 12B B-418-028-D (29MarO2)0.3mg/mL 10 of 12B B418-028-D (29Mar02) 0.3 mglmL 10 of 12B B-418-028-E (29MarO2) 1 mg/mL 1 of 12T B-418-028-E (29MarO2) 1 mg/mL 1 of 12T B-418-028-E (29MarO2) 1 mg/mL 2 of 12T B-418-0284 (29Mar02) 1 mg/mL2 of 12T B418-028-E (29Md2) 1mghnL 5 Of 12M B-418-028-E (29MarO2) 1 mg/mL 5 of 12M B-418-028-E (29Mar02) 1 mg/mL 6 of 12M B-418-028-E (29MarO2) 1 mg/mL 6 of 12M B-418-028-E (29MarO2) 1 mg/mL 9 of 12B B-418-028-E (29Mar02) 1 mg/mL 9 of 12B B-418-028-E (29MarO2) 1 mg/mL 10 of 12B B-418-028-E (29MarO2) 1 mg/mL 10 of 12B Set Number 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 110702A 1 10702A 110702A 110702A 110702AR 110702AR 110702A 110702A 1 10702A 110702A 110702AR 110702AR 110702A 110702A 110702A 110702A 110702B 110702B 110702B 110702B 110702B 110702B 110702B 110702B 110702B 110702B 110702B 110702B 110702B 110702B 110702B 1 10702B 110702B 110702B 110702B 110702B PFHS Known Cone. (n-) 0 0 0 0 0 0 0 0 0 0 0 0 30000 30000 30000 30000 30000 30000 30000 30000 30000 30000 30000 30000 100000 100000 100000 100000 100000 100000 100000 100000 100000 100000 100000 100000 300000 300000 300000 300000 300000 300000 300000 300000 300000 300000 300000 300000 1000000 1000000 1000000 1000000 1000000 1000000 1000000 1000000 1000000 1000000 1000000 1000000 PFHS Found (ng/mL) ND ND ND ND ND ND ND ND ND ND ND ND 24400 23700 26900 25800 25500 26100 25800 25800 24200 24400 24800 24500 75400 73800 76300 74700 93300 95100 69800 72300 84400 83700 95700 92800 270000 268000 270000 279000 254000 266000 24 1000 239000 255000 250000 280000 293000 1020000 1090000 987000 980000 1 160000 1170000 1040000 994000 1090000 110ooM) 1 1 10000 1070000 PFHS Reeoverr (%) 81 79 90 86 85 87 86 86 81 81 83 82 75 74 76 75 93 95 70 72 84 84 96 93 90 89 90 93 85 89 80 80 85 83 93 98 102 109 99 98 116 I17 1 04 99 109 110 111 107 * Duplicate Injection M> = Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 42 of 153 418-028:PAGE G-43 Exygen StudyNo.:023-072 FIGURES Exygen Research Page 43 of 153 418-028:PAGE G-44 Exygen Study No.: 023-072 Figure 1. Typical Calibration Curve for PFHS Compound 1 name: PFHS Coefficient of Determination: 0.999484 Calibration curve: 2633.68 * x + 44.4695 Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: lix, Axis trans: None Exygen Research Page 44 of 153 418-028:PAGE G-45 Exygen Study No.: 023-072 Figure 2. Chromatogram Representing a 0.1 ng/mL standard for PFHS CO82002-6.0.1 nglmL Standard 0919028-2012 Sm (Mn.2x3) 1001 21Sep-2002 1034:30 LCmnSlMS#? MRM of 1 Channel ES- 8.& 399>80 292 4.21e3 Area % G 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 Time )O 10.00 11.00 12.00 Exygen Research Page 45 of 153 418-028:PAGE G-46 Exygen Study No.:023-072 Figure 3. Chromatogram Representing a Control Rat Plasma Sample for PFHS (Exygen ID 0202913 Control, Data Set: 091602B) 0202913 Control 0916028-207 Sm (Mn. 2x3) 3.66 b '1 0.17 1 I 17Sep-200222:5259 LClMSlMS #7 MRM of 1 Channel ES 399 > 8( 386 13 ?0.21 Exygen Research Page 46 of 153 418-028:PAGE G-47 Exygen StudyNo.:023-072 Figure 4. Chromatogram Representing a Control Rat Serum Sample for PFHS (Exygen ID: 0202987 Control, Data Set: 091902B) 0202987Control A 0919026-2008 Srn (Mn. 2x3) "3 21Sep-200209:07:34 LCIMSMS w7 MRM of 1 Channel ES- 10,18 399>80 289 f 9 f - p 10.00 11.00 Time 12.00 Exygen Research Page 47 of 153 418-028:PAGE G-48 Exygen Study No.: 023-072 Figure 5. Chromatogram Representing a Control Rat Liver Sample for PPHS, (Exygen ID: 0202877 Control Data Set: 100202A) 0202877 Control 100202A-208 SIT(IMn. 2x3) 1 om 03O~t-200208:12:19 LCIIJISMS 67 MRM of 1 Channel ES- 399>80 z.ooe3 Area % a Exygen Research Page 48 of 153 418-028:PAGE G-49 Exygen StudyNo.:023-072 Figure 6. Chromatogram Representing Control Rat Plasma Sample Fortified with 10 ppb of PFHS (Exygen ID: 0202913 Spk A, Data Set: 091602B) 0202913 Spk A. 10 ppb 0916028-208 Sm (Mn, 2x3) 1 oc 1 9.08 937 1 174ep-20022314:44 LClMSlMS #7 MRM of1 ChannelES 399 > 8C 8.25e2 Area % Exygen Research Page 49 of 153 418-028:PAGE G-50 Exygen Study No.:023-072 Figure 7. Chromatogram Representing Control Rat Serum Sample Fortified with 10 ppb of PFHS (Exygen ID: 0202987 Spk A, Data Set: 091902B) 0202987 Spk A, 10 ppb 0919028-2009 Sm (Mn, 2x3) "'1 21-~ep-200209:2914 LCIMSIMS #7 MRM of 1 ChannelES399>80 1.we4 AIW Exygen Research Page 50 of 153 418-028:PAGE G-51 Exygen Study No.:023-072 Figure 8. Chromatogram Representing Control Rat Liver Sample Fortified with 10 ppb of PFHS (Exygen ID: 0202877 Spk A, Data Set: 100202A) 0202877 Spk A, 10 ppb 100202A-209Sm (Mn.2x3) 100 03Qct-2002 0833:57 LCMSlMS #7 MRM of 1 Channel ES- 399>80 I.08& Area % 0 Time 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 Exygen Research Page 51 of 153 418-028:PAGE G-52 Exygen Study No.:023-072 Figure 9. Chromatogram Representing Rat Plasma Sample Analyzed for PFHS (Exygen ID: 0201816, Sponsor ID: 19176 GROUP I, Data Set: 091602B) 0201816 0916028-212 Srn (Mn. 2x3) 100 9.07 27721 18-Sep-2002 00:41:24 LCIMSrnS #7 MRM of 1 Channel ES399>80 2.16e4 Area 9c Exygen Research Page 52 of 153 418-028:PAGE G-53 Exygen Study No.:023-072 Figure 10. Chromatogram Representing Rat Serum Sample Analyzed for PFHS, DF=lO (Exygen ID: 0201863, Sponsor ID:19079 GROUP 11, Data Set: 091902BR) 0201863, DF-10 091902BR-308 Sm (Mn, 2x3) 100 8.80 24887 24Sep-2002 02:5015 L C i M S l S #7 MRM of 1Channel E S 3~ 5 ao 3.91 a4 0 Exygen Research Page 53 of 153 418-028:PAGE G-54 Exygen Study No.: 023-072 Figure 11. Chromatogram Representing Rat Liver Sample Analyzed for PFHS (Exygen ID: 0202050, Sponsor ID: 19018 GROUP I1 Male 1, Data Set: 100202A) 03.0ct-2002 10:00:52 LCrrmSINlS #7 MRM of 1 Channel E S 100- 0.28 399s80 1.19e5 Area %- 0 ,?, , , , , , , , . . . , I I . . . I , , . , , I I . . I . . . . I . . . . , .. .,I . . I , , . . , I ~ . . . , I . . . . ( . , , , , , , , . , , , , , ,, , , , , , , , , , , , , ,,Time Exygen Research Page 54 of 153 418-028:PAGE G-55 Exygen Study No.: 023-072 APPENDIX A Study Protocol 418-028 (Exygen Study No. 023-072) and Amendments, Deviation and Note to File Exygen Research Page 55 of 153 418-028:PAGE G-56 Exygen StudyNO.:023-072 PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 STUDY TITLE: Oral (Gavage) CombinedRepeated Dose ToxicityStudyof T-7706 with the ReproductiorUDevelopmental Toxicity Screening Test PURPOSE: c TESTING FACIm The purpose of this study is to provide iaformaton on the possible health hazards that may result firomrepeated exposureof Crl:CD@(SD)IGSBR VAF/Plus@male and femalerats to a test subsiance beginning before cohabitation, through mating and continuing for at least 42 days (male rats) or throughparturitionuntil day 21 oflactation (femalerats). This repeateddose study incorporatesa reproduction/developmentaltoxicity screeningtest thatCM be used to provideinitial informationon possible effectson male and female reproductivepdoxmance(e.g., gonadal fimctioqmatingbehavior, conception, developmentof the conceptus and parturition). The study also places emphasis on neurological cffocts as a specific endpoint and should identie the neurotoxic potential of a test substance, which may warrant further in-depthinvestigation. Because of the selectivity of the endpoints and the short duration of the study, the d g test wiIl notprovide evidence for definitiveclaims of no reproductioddevelopmentaleffects. In particular, it offas ody limited means of detecting postnatal manifestations of prenatal exposure or effects that may be induced duringpostnatal exposure. Argus Research 905 SheehyDrive, BuildingA Horsham, Pennsylvania 19044-1297 Telephone: (215) 443-8710 Telefax: (215) 443-8587 STUDY DIRECTOR RaymondG. York,Ph.D.,DABT Associate Director of Research Email: rapond.york@criver..com Address 89 cited above for Testing Facility SPONSOR ..- 3M Corporate Toxicology 3M Center, Building 220-23-02 St. Paul, Minnesota 55144-1000 Exygen Research c Page 56 of 153 418-028:PAGE G-57 Exygen Study No.: 023-072 - STUDY MONtTOR John ButenhoE Ph.D., DABT, CIH 3M Corporate Toxicology 3M Medical Depzatment Telephone: (651) 733-1962 Telefax: (651) 733-1773 Email: jlbutenho ff@mmm.com Rotocol4 18-028 Page 2 REGULATORY CITATIONS: Organisationfor Economic Co-operationand Development (1996). OECD Guidelinefor Testing of Chemicab. Section4, No. 422: Combined RepeatedDoseToxicity Studywith the Repmduction/DevelopmentalToxicityScreeningTest, adopted22 March 1996. Organkition forEconomicCoopemtion andDevelopment (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Fhal]. U.S.Food and Drug Administration. Good LaboratoryPracticeRegulations;FinalRule. 21 CFR Part 58. Japanese Minishy of Health and Welfare (1997). CodLaboratoryPractice Standardfor Safety Sfudieson Drugs, MHW OrdinanceNumber21, March 26,1997. c REGULATORY COMPLIANCE: This studywill be conducted in compliancewith the GoodLaboratoryPractice(GLP) regulationscited above. All changes or revisions of this protocol shall be documented, signed by the Study Director and the Sponsor, dated and maintainedwith the protocol. The Testing Facility's Quality Assu~anceUnit (QAU) will audit the protocol,the raw data and the report,and will inspectcriticalphases of those portions ofthe studyconductedat the Testing Facility in accordance with the StandanlOperatingProceduresof the Testing Facility. The iinal report will include a compliance statementsignedby the StudyDirectorthat &c report accurately reflsts the raw data obtained duringthe performanceof the stndy and that all applicable GLP regulationswere followedin the conduct of the study. Shouldsignificant deviations fiom GLP regulationsoccur, each will be described in detail, togetherwith howthe deviation might affect the quality or integrity of the study. Should any portion of the study be conducted by a subcontractoror by the Sponsor,the Study Directorwill ensure that a qualifiedPrincipalInvestigatoris identifiedby the facilityconducting that portion of the study. The QAU forthis facility will conduct critical phase inspectionsand F audit respective results and reportsfor that studyportion accordingto the SOPSof that facility. such critical phase inspectionreports and report audits will be submitted by the facilityto the principal Investigator and the Study Director. The dates of the inspectionsand report submissionswill be incorporatedinto aQAU Statementgeneratedby that facilityandprovided Exygen Research Page 57 of 153 418-028:PAGE G-58 Exygen Study No.:023-072 - Protocol418-028 Page 3 to the TestingFacility for inclusion in the final report. In addition,thisfacilitywillprovide a statement of GLP compliance, as described above, signed by theprincipal Investigator for inclusion in the final report, SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT 1 to the protocol. TEST SUBSTANCE AND VEHICLE: 'Jest Substance: T-7706lperfluorohexane SuUonate Potassium Salt (PFHS)] Lot identificationwillbe documented in the raw data The Sponsor will provideto the Testing Facilitydocumentation or certificationof the identity, composition, method ofsynthesis,strength and activity/purityof the test substance, This documentationwill be included in the final report. Vehicle: Aqueous 0.5%carboxynethlycellulose(CMC) (medium viscosity) prepared using reverse osmosismcmbmneprocessed deionizedwater (RO. deionized water). Lot identificationand Supplier will be docurnented in the raw data Neither the Sponsor nor the StudyDirector is aware of any potentialcontaminantslikely to be present in the vehiclethatwould interfereWith the resultsof this study. Therefore, no analyses otherthan those mentioned in this protocolwill be conducted. Safetv Precautions: Gloves, dust-mistlHEPA-filtered mask,appropriate eye protection and unifodab coat to be worn during formulationpreparation and dosage. The MaterialSafetyData Sheet (MSDS)is attached to the protocol (A'JTACHMENT 2). Storape: Bulk Test Substance: Bulk Vehicle Components: prepared Test Substance and Vehicle Formulations: R w m temperature. Room temperature. Reliigerated(2Tto 8C). A All test substance shipmentsshouldbe addressedto the attentionofJulian Gdbinaki,Managerof Formulation Laboratory, at the previously cited TestingFacility address and telephone number. ... .. . Exygen Research Page 58 of 153 418-028:PAGEG-59 Exygen Study No.:023-072 - Protocol 418-028 Page 4 Shipmentsshould include infomationconcerningstorageconditions and shipping cartons should be labeled appropriately. The recipient should be notified in advanceof shipment. FORMULATION Freauencv of Prenaration: Formulations (suspensions)will be preparedweekly at the TestingFacility. Detailed preparationprocedures will be attachedta thisprotocol (ATTACHMENT3). Adiustment for hri* Thetest substancewill be considered 100%pure fixthe purpose of dosage calculations. Testing F a c W Reserve Samnles: The Testing Facilitywill reserve a sample of each lot of bulk test substance (approximately 1 g) and bulk vehicle components (approximately 1 g or 5 mlL) used during the come ofthe study. Samples wilt be stored under the previously cited conditions. ANALYSES: .- Results of requiredanalyseswill beprovidedto the TestingFacility for inclusion in the study report. Samples additionalto those described below may be taken if deemed necessaryduringthe course ofthe study. Additional analyses, if required, will be documented by protoool amendment. Bulk Test Substance SamDling: A sample of approximately 1 g of the test substance will be taken on the last day of treatment and sent (ambient conditions) to: Principal Investigator: Lisa Clemm 3M EnvirOnrnentalTechtaoIogyand Safety Services Buildiag 2-3E-09 St. Paul, M i ~ e ~ o5t5a133-3331 Telephone: (65 1) 778-5568 Telefax: (651) 778-6176 Email: laclemm@mmtn.com The recipientwill be notifiedin advance of sample shipmmt. Exygen Research Page 59 of 153 418-028:PAGE G-60 Exygen StudyNo.:023-072 Analvses of PreDared Formulations: Protocol 418-028 Page 5 Concentrationand homogeneity of the prepared fomulationswill be verified duringthe course of thisstudy. Quadruplicatesamples(2mL each) will be taken fbm the top, middle and bottom of each conccnttatouon the firstday of prcpm~on.Two samples&om each quadruplicateset Will be shipped for analysiq ther e W g samples will be retained at the TestingFacility as backup samples. Quadruplicatesampleswill be taken&om each concentration on the last dayof preparation. Two samples firom each quadruplicate set will be shippedfor analysis;the remaitlingsampleswill be retained as backup samples. Backup sampleswill be stored under the previouslycited conditionsand discardedat the TestingFacilityupontherequest of the Sponsor. Stabilityof the prepared formulations will be documentedduring thisstudy. Two sets of duplicate samples (2mL each) Born each concentrationwill be talcen on the h t day of preparation. One sampleof each duplicateset will be shipped on the day of preparation. These samples willbe analyzed at the followingtime points: as soonafter preparationaspossibIeand ten days after the first analysis. The remaining sampleswill be retained at the Testing Facility as backup samples. Backup sampleswill be stored under the pmviouslycited conditionsand -- . discarded at the Testing Facility upon the request of the Sponsor. Shirraine hstructions: Samples to be analyzedwill be shipped(refiigerated) to: Principal Investigator: Lisa Clemen 3M EnvironmentalTechnologyand Safety Services Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone: (651) 778-5568 Telefax: (651) 778-6176 Emaif: laclemen-.corn The recipient willbe notifiedinadvance of sample shipment. DXSPOSXTION: Prepared formulationswill be discardedat the Testing Facility. AU remainingbulk test substancewill be returned to: - Dan Hakes 3M EHSR Auto & C h a Grp c 3M Center, Building 236-1B-10 St. Paul, Mirmesota 55144-1000 Telephone: (651) 733-2392 Exygen Research Page 60of 153 418-028:PAGE G-61 Exygen Study No.:023-072 TEST SYSTEM: Rotacol418-028 Page 6 The Crl:CD@(SD)IGSBR VAF/Plus@rat was selected as the Test Systembecause: I) it is one mammalian species accepted for use in toxicity studies and it has been widely used throughout m d w , 2) this strainofrat has beendemonstratcdto be sensitiveto reproductive and developmental toxins; and 3) historicid data and experience exist at the Testing Initial population acclimated: lo0male and 100virgin fernalerats. Population selected for main study: 75 male and 75 virgin f d e rats (15 per sex per dosage group). Populationselected for toxicokineticstudy: 15 male and 15 femalerats (three per sex per dosage group). Bodv W e i d t and Ape: Malerats will be orderedto weigh h m 275 g to 300 g each at receipt,at which t h e they will be - expected to be at least 60 daysof age. Female rats will be orderedto weigh fnnn200 g to 225 g each at receipt, at which time they will be expected to be at least 56 days of age. Actual body weights will be recordedthe day afterreceiptand will be documented in the raw data. Tbe weight rangeswill be includedin the finalreport. At studyinitiation,the weight variation of the rat13will not exceed*2O% of the mean weight of each sex. Both male and femalerats will be evaluated. Source: CharlesRiver Laboratories,Inc. The rats will be shipped in filtered cartonsby air &eightand/or truck fromCharlesRiver Laboratories, hc., to the Testing Facility. Rats arepermanentlyidentifiedusing MonelUBstif-piercingear tags (Gey Band and Tag Co., hc., No. MSPT 20101). Male and ftmderats are assignedtemporarynumbersat receiptand given unique permanent identification numbers when assigned to the study before administration of the first dosage. Pups will notbe individually identified during lactation;all parameterswill be evaluated in tern of the litter. c Exygen Research Page 61 of 153 418-028:PAGE G-62 Exygen StudyNo.:023-072 hWc01418-028 Page 7 ANIMALHUSBANDRY All cage sues and housing conditions are itl compliancewith the Guidefor the Care and Useof LaboratoryAnimals(4).Argus Researchis an AAALAC-accredited facility. Housing: Fo generationratswill be individually housed in stainless steelwire-bottomedcagesexcept during the cohabitation and postpartum periods. During cohabitation, each pair of rats will be housedinthe male rat's cage. Beginningno later than day 20 of presumedgestation,Fo generationfemale ratswill be individuallyhoused in ncstingboxes. Each dam and delivered litterwill be housed in a commonnestingbox duringthepostparhunperid Nestingmaterial (bed-o'cobse)will be provided. Bedding will be changed as often as necess@ to keep thed u d s dry and clean. Analysesfor possible contaminationare conducted semi-atmually and documentedin the raw data Room Air. Temperatureand Humidity: - The animalmom is independentlysuppliedwith at least ten changesper hour of 100% h s h air that has been passed through99.97% HEPA filters. Room temperahuewill be maintain4at 64F to 79'F (18'C to 26OC) and monitorcd constantly. Room humidity will also be monitored constantly and maintainedat 30% to 70%. An automaticallycontrolled 12-hourlight:12-hourdark fluorescentlight cycle will be maintained. Each dark period will begin at 1900hours EST. The light cyclemay be adjustedby the StudyDirectoror designee if deemed necessary to accommodate scheduled laboratory activities. Any such adjustment will be docummtcd in the raw data. Rats will be given CertifiedRodent Diet@#002 (PMI Nutrition International) availablead libitam h r n individual feeders. Feed will be removedthe eveningprior to the scheduled sacrifice. - Water: - Water will be available ad fi6itm h m individual bottles attached to the cagesor h m an automaticwatering accesssystem. All water will be fiom a local s o m e and passedthrough a rewerse osmosis membrane before use. Chlorine will be added to the processed water as a backtriostat;processed water is expectedto contain no more than 1.2 ppm chlorineat the time of analysis. Water is analyzed monthly for possiblebacterialcontamhation andtwice annually for possible chemical contamination. . Exygen Research Page 62 of 153 4 18-028:PAGE G-63 Exygen Study No.: 023-072 h t o d 4 18-028 Page 8 Contaminants: Neither the Sponsornor the Study Directoris aware of any potential contamhuntslikelytobe present in thc certifieddiet, in the chinking water or in the nestingmaterials at levelsthatw d d interferewith theresultsof this study. Therefore,no analyses other than those routinely performedby the feed supplieror those mentioned in thisprotocolwillbe conducted. DAY NUMBERING SYSTEM Gestationday 0 is defined as the day spermatozoa are observed in a smearof the vaginal contents and/or a copulato~yplug observed in sia. - The day of birth is designated lactationday 0 (postpartumday 0) in the Health Eff`ects Test Guidelines- Reproduction andFertility Effects (Officeof F'rcvention, Pesticides and Toxic Substances870.3800, August, 1998) and in the OECD Guideline for the Testingof Chemicals Combined Repeated Dose ToxicityStudywith the RqroductionlDeveloprnentalToxicity ScreeningTest (Section 4, No.422,22 March 1966). This same day is designatad day 1 postpartum (day 1 of lactation)in the StandardOperating Proceduresof the TestingFacility, Throughout this protocol, the day ofbirth willbe designatedday 1postpartum (day 1 of lactation) and all subsequent ages of the F1 generationrats and days of the lactation period will - be d e t d e d and cited accordingly. RANMlMIzATIoN AND COHABITATION: Upon arrival, ratswill be assignedto individualhousing on the basisofcomputer-generated random units. During m acclimationperiod of at least five days, male and female ratswill be selected for study on thebasis of physical appearanceand body weights recorded during acclimation. The rats will be assigned to dosage groupsbased on computer-generated(weightordered) randomizrrtion procedures. Within each dosage group,consecutiveorderwill be used to assign rats to cohebitation, w e male rat per fmahrat. The cohabitation period Will consist ofa maximum of 14 days. Femalerats with spermatozoaobserved in a smear of the vaginal contentsand/or a copulatoryplug observed in siiu will be considered tobe at day 0 of presumed gestationandassigned to individual housing. Female ratsnot mated within the first seven days of cohabitationwill be assigned alternate male rats that have mated (same dosage group) and will remain in cohabitation for a maximum of seven additionaldays. Day 1 oflactation(postpartum) is defined as the day of birth and is alsothe hTst day on which all pups in a litter areindividuallyweighed(pup body weightswill be recordedafter ail pups in a - litter are delivered and groomed by the dam). Litters will not bc d l e d during the laciationperiod, because random selection ofpups for culling could r d t in potentialbiases in pup viabilitiesand body weight gainsoverthispaiod. Exygen Research Page 63 of 153 418-028:PAGE G-64 Exygen Study No.: 023-072, - prDtoCol418-028 Page 9 Within each dosage group,consecutiveorderwill be used to assign the first 10male and the &st 10female ratsto a functional observationalbatteryFOB) and motor activityassessment, blood sample collection for clinical chemistryand hemtology(CC&H), and histological evaluations. On day 22 postpartum, a table of random units will be used to select five male and five female pups per litter for blood sample and liver collection; these pups will only be selected from the ten dams selected for FOB, motor activity, CC$H and histologicalevaluation. ADMINISTRATION: Route and Reason for Choice: The oral (gavage) mutewas selectedfor use because: 1) in comparisonwith the dietary route, the exact dosage can be accuratelyadministered;and 2) it is one ofthe possibleroutes for environmental exposure. Method and Freuuency: Dosageswill be adjusteddaily for body weight changes and given at approximatelythe same time each day. The first day of dosage is designated as day 1 ofstudy. - Male ratswill be giventhe test substance once daily beginning 14daysbefore a cohabitation period that consistsof a maximum 14 days. Dosage will continue throughthe daybefore sacrifice,aftercompletion of the cohabitationperiod, after aminimumof42 daysof admiuistration. Fernalcrats will be giventhe test substanceonce dailybeginning 14 days before a cohabitation period that consistsofa maximum of14days. Dosagewill continuethrough the day befbn scheduledsacrihce (day 21 of lactation). Rationale or Dosaee Selection: Dosageswefe selectedby the Sponsorbased onprevious studies conduotdwith the test substance,taking into accountpossible differencesin sensitivitybetweenpregnant and nonpregnantrats. The bighest dosagewill be expected to cause toxic effectsbut notmortalityor obvious suffering. The descendingsequence of the lowerdosage levels will be selected farthe purpose of demonstratingany dosage-relatedresponse, withno adverseeffects expectedat the lowest level. Exygen Research Page 64 of 153 - Dosage Levels. Concentrationsand Volumes: 418-028:PAGE G-65 Exygen Study No.: 023-072 Pmtw014 I 8-028 Page 10 TESTS. ANALYSES AND - MEASUREMENTS FOGENERATION: ViabiW - Male and Female Rae: All Periods: At least twice daily. - Clinical Observations andor GeneralAtwarawe Male and Female Rats: Accltnation Period. Weekly. Dosage Period: Daily before dosage. On the first day of dosage, postdosageobservationswill be recorded at appmxiznately hourly intervals fbrthe first four hours and at the end ofthe n o d workingday. Subsequentpostdosageobservations will be ~ c ~ ~ r adt eindtervals deemed appropriatebythe Study Direotorordesignee after determinationof peak toxicologic effects. Maternal Behavior: Days 1,5,8,15 and22 postparhun Observedabnormal behavior recorded daily. Clinid observationsmay be recordedmore hquentlythan citedabove, if deerned appropriate by the Study Director andor Study Monitor. Data collectedfor rats assignedto toxicokinetic samplecollectionwill not be summarizedor aualyzed statistically. - Detailed ClinicalObservations Male and Female Rats: Once before the first dosage and at least once weekly thereafter, detailed clinical observations will be conducted for allmale and f a d e rats. These obsewationswill be made outside the a g e - in a standardarena at the same time eachdayof conduct. Effort will be made to ensurethat variationsin the test conditionsare minimal and that observationsare conducted by observers unawareof treatment groups. Signsnoted shouldinclude, but not be limitedto: changesin skin, fiu, eyes, mucousmembranes,occmmce of secretioxls and excretionsand autonomic activity (e.g., lacrimation,piloendon, pupil size, unusualrespiratorypattern). Changes in gait, postwe Exygen Research Page 65 of 153 418-028:PAGE G-66 Exyga StudyNO.:023-072 M i l 418-028 Page I t and response to handling as well as the presence of clonic or tonic movements, sterotypic behavior (e.g., excessivegmoming, repetitive circling), difficultorprolonged pathuitionor bizarrebehavior (e.g., self-mutilation, walkingbackwards) should also be recorded. Bodv Weights - Male and Female Rats: Acclimation Period: weekly. Dosage Period: Daily. Sacrifice: Terminal weight. Feed ConsumDtion Values - Male Rats (recorded and tabulated): Dosage Period: Weekly. - Feed Consamdon Values FemaleRats (recorded and tabulated): Dosage Period Weekly to cohabitation. Gestation Period. Days 0,7,10,12, lS, 18,20 and 25 (if necessary). Postpartum Ptriod: Days 1,5,8 and 15. Feed consumption not tabulated after day 15 poslpartum, when it is expected that pups will begin to consume maternal feed. - Feed ConsumtionValues Male and FemaleRab: Feed consumption values may be recoded more frequently than cited above if it is necessary to replenishthe feed. During cohabitation,when two ratsoccupythe same cage with one feedjar, replenishmentof the feedjars will be documented. Individual vaIueswill not be recordedor tabulated. Toxicokinetic Samule Collection: On day 14 and 42 of study, blood samples(approximately 1 mL each) will be collected h m eachmalerat assignedto thetcrXicolcinetic sample collectionportion of the study (3 p a group). On day 14 of study and day 21 of presumed gestation,blood samples (approximately 1 mCeach) will be collectedfhrn each fmale rat assignedto the toxicokinetic sample collectionportion of the study (3 per group). Sampleswill be collectedprior to dosage on day 14 of study. The time of each blood collection will be recorded in the raw data. Exygen Research Page 66 of 153 418-028:PAGE G-67 Exygen Study No.:023-072 pnOtOCOl418-028 Page 12 Blood will be collected ikom the orbital sinus. Ifnecessary, blood may be collected &om an alternate site; if so,the alternate site will be documentedin the raw data.) The samples will be transferredinto EDTA-coated(purple top) tubes and spunin a centrifuge. Theresulting serum will be transfmed into polypropylenetubes labeledwith the protocol number,Sponsorstudy number, animalnumber, sex,p u p number,dosage Iwel, day ofstudy, collectionintend, date of collection, species,generationand storage conditions. AU sampleswill be immediately frozen on dry ice and maintainedbzcn (S-7OoC) until shipmentfor aualysis. After the lastblood sarnplecollection, ratswill be sacrificed and samplesofthe liverwill be collected for analysis. Samples to be analyzedwillbe shippedon dry ice to: Rincipal Investigator: Lisa Clemen 3M EnvironmentalTecbnologyandSafety Services Building 2-3B-09 St. Paul, Minnesota 55 133-3331 Telephone: (651) 778-5568 Telek: (651) 778-6176 Email: laclemen-.corn The recipient will be notified in advance of sample shipment. Estrous cyclingwill be evaluatedby examination of vaginal cytologyb e g b h g withtheday afterthe firstadministration and then until spermatozoaarc observedin a smear of the vaginal contents andor a copulatoryplug is observed insitu during the cohabitationperiod. - Caesarean-SectioainP Toxicokinetic Studv: On day 21 of presumed gestation, blood and liver samples will be collected &om aI1 femalerats designated for toxicokinetic sample collection. Blood sampleswill be collectedh m the rats as previously described. After sacrifice,the liver of each rat will be excised and the liver weight will be recorded. The median liver iobewill be h z e n and stored (I-20C) until shipment for possible analysis. The fetuseswill be removed h m the uterus and blood sampleswill be collectedfromeach fetus via decapitation. Blood will be placed into tubes, pooled per litter, allowed to clot and spun in a centrifuge;the resulting serum will be transferred intolabeled polypropylene tubes. All samples will be immediately h z e n on dry ice and maintainedhzen &?O"C) until shipmentfor e analysis. Exygen Research Page 67 of 153 418-028:PAGE G-68 Exygen StudyNo.:023-072 - RO~OCOI 418-028 Page 13 The liver &omeach fetuswill be collected,pooled per litter and placed into labeled tubes. Thc sampleswill be hzen and stored (520C) until shipment for analysis. Sampleswill be shippedon dry iceto Lisa Clernen at the previouslycited address, Natural Deliverv: Female rats will be evaluated for: Adverse Clinical Sips Observed DuringPartmition. Duration of Gestation(day 0 ofpresumed g d o n tothe time the firstpup isobserved), LitterSize (defined as all pups delivered). Pup Viability at Birth. Functional Observational Battery: On one occasion duringthe come of the study, a functional observationalbattcry will - be conducted on 10male and 10femaleratsper group. Formale rats,this assessmentwillbe conducted shortly before scheduled sacrifice. but prior to blood sample collection for hematology and clinical chemistryevaluations. Femalerats shouldbe tested during the lactation period, shortlybefore scheduledsacrifice. The FOB,to be conducted by an observerunawareof tbe group assignmentof the rat,will assess the following parameters: 1. Lacrimation,salivation,palpebral clomue, prominence of the eye. pupillary reaction to light, piloerection,respiration,and urinationand defecation (autonomic functions). 2. Sensorimotorresponsesto visual, auditory, tactile and painlid stimuli (reactivity and sensitivity). 3. Reactions to handling and behavior in the open field (excitability). 4. Gait pattern in the open field, severityof gait abnormalities, air rightingreaction, and landing foot splay (gait and sensorimotor coordination). 5. Fortlimband hindlimb grip strength. 6. Abnormal clinicalsigns including but not limited to convulsions, CremOrs and other m d behavior, hypotonia orhypextonia, emaciation,dehydration, c uukempt appearanceand deposits mund the eyes, nose or mouth. Exygen Research Page 68 of 153 418-028:PAGE G-69 Exygen Study No.:023-072 - Protocol 418-028 Page 14 Evidenceof the ability of thisbattery to detect the effectsof positivecontrol substances will be provided (TestingFacilityPositiveControl Data). Data will also be provided to document intembserverreliability if morethan one observeris involvedin the testing. Motor Activitv Test: Motor activitywill be evaluatedon 10male and 10 fmale ratsper group once during the course of the study. This assessment will be conducted shortly befose scheduled sacrifice, but prior to blood sample collection. The movementsofeach rat willbe monitoredby apassive infraredsensormounted outside a stainless-steel wirt-bottamed cage (40.6 x 25.4 x 17.8 cm). Eachtest sessionwill be 1.5 hours in duration with the number of movements and time spent in movement tabulated at each fiveminuteinterval. The apparatus will monitor a rack of up to 32cagesand sensorsduringeach session, with each rat tested in the saxhe looation on the rackmoss test sessions. Groupswill be comterbalmceci across testing sessions and cages. Data will be provided to demonstratethat the test system iscapableof &tecting increasesin activityproduced by positive controlsubstances(Testing Facility PositiveControl Data). HEMATOLOGYAND CLINICALCHEMISTRY: ..-. At scheduled sacrifice, the rats (fasted) assigned to hematology and clinical chemistry (H&CC) samplecollection will be exsanguinatedh m the inferiorm a cava fbllowing sacrificeby carbon dioxideasphyxiation. Approximately5 mL ofblood will be collectedand processed as describedbelow. Determinations additionalto those describedbelow may be conductedrfthe knownpropertiesofthe test substaucemay, or are suspectedto, a&ct related metabolic profiles (e.g., calcium, phosphate, fasting tiglycerides and fasting glucose, specific hormonea, and cholinamase). Hematology: Approximately 1 mL of blood will be collected into EDTAcoated tubes and maintained on wet ice or refrigerateduntil shipmentfor analysisof the ibllowinghematologicparameters: Erythrocyte Count (RBC) Hcmatocrit (HCT) Hemoglobin (HGB) Mean Corpuscular Hemoglobin (MCH) Mean Corpuscular Hanoglobin Concentration (MCHC) Mean Corpuscular Volume (MCV) LeukocyteCount, Total (WBC) Leukocyte Count, Differential Platelet Count (PLAT) Mean Platelet Volume (MPV) Cell Morphology Two blood smear slides wiU be prepared at the TestingFacilityfor each samplefor measurementsofdifferential leukocytecount. All samples(on wet ice) and slides (ambient conditions) will be shipped to Redfield Laboratories at the following address. Approximately 1.8 mL ofblood will be added to a tubecontaining0.2 mLofsodium citrate (0.129 M). The contentswill be mixedand maintainedon wet ice until the tubes are ccnaifugtd Exygen Research Page 69 of 153 418-028:PAGE G-70 Exygen StudyNo.:023-072 - prat0~01418-028 Page 15 (within30 minutesof the collection time). The resultingplasma will be trausfmed 2.0mL polypropylene tubeslabeledwith studynumber,Sponsor's studynumber,rat number,dosage level, day of study, collectioninterval, date of collection, species, generationand storage. All sampleswill be hzen on dry ice and maintained l?ozen (57OOC) until shipmenton dry ice by ovemightcourier for measurement ofprothrombintime(PT) and activatedpartial thromboplastintime (APT"). Clinical Chemistry: Approximately 2 mL of blood will be collectedinto serum separatortubes and centrifuged. The resulting sera sampleswill Ba immediatelyfrozenon dry ice and lnaiatainedhzea (S7OoC) until shipment for analysisof the following parametem Total Protein (TP) Triglycerides crru) Albumin (A) Globulin (G) Albumin/Globulin Ratio (NG) Glucose (GLU) Cholesterol (CHOL) Total Bilirubin (TBILQ ' Urea Nitrogen (BUN) Creatinine (CREAT) CreatinineKinase (CK) Alanine Aminofransferase (&T) Aspartate Amhotransferase (AST) Alkaline Phosphatase (ALK) Calcium (CA) Phosphorus (PHOS) Sodium (NA) Potassium (K) Chloride (CL) Sampleswill be shipped(ondry ice) to RedfieldLaboratories at the following address. S M D U ~IPnstructions: Sampleswill be shipped to anive on Monday throughFridayaccording to the conditions described above to: Principal Investigator: Ms. Phyllis Powell Redfield Laboratories A Division of CRL-DDS 100 East Boone Street P.O.Box 308 Redfield, Arkansas 72132 Telephone: (501) 397-2540 Telefax: (501) 397-2002 The recipient will be notified in advance of sample shipment. URINALYSIS: h Urinalysiswill not be conducted unless indicatedbasedon expectedor observed toxicityof the test substance. Exygen Research Page 70 of 153 418-028:PAGE G-71 Exygen Study No.: 023-072 rrr METHOD OF SACRIFICE: Protocol 418-028 Page 16 Fo generationrats willbe sacrificedby carbondioxide asphyxiation. - GROSS NECROPSY AND HISTOPATHOLOGY FOGENERATION RATS: ScheduledSacrifice- ToxicokineticStudy: Scheduledsacrificeof male rats will be conducted on day42 of study. Scheduledsacrifice of female rats will be conductedon day 21 ofpresumed gestation. Blood sampleswill be collectedfbm the rats as previously described. After sacrifice,the liver of each rat will be excised and the liver weight will be recorded. The median liver lobe will be frozen and stored (S-2OoC) Until shipment for analysis. Fetal samples will be collected as previously described. Carcasses wiU be discarded without Mer evaluation. Sampleswill be shippedon dry ice to LisaClemcnat the previously cited address. - ScheduledSacrifEce Mnin Study: Scheduled sacrificeofmale rats will be conducted on the day followingthe last dosage administration, aftera minimum of42 days of dosage. Soheddedsacrificeof f d e rats will be conducted on day 22 of lactation. Gross necropsy ofall male and female rats will include an initial physicalexaminationof external surfkces and all orifices,as well as the cranial, thoracic and abdominalcavities and their contents. Special attentionwill be paid to the organsof the reproductive system. The number of implantation sites and corporalutea will be recorded. Male and female rats will be examined for &rots lesions. Gross lesions will be retained in neutral buffered 10%formalin and examinedhistologically. Tissue trimming and histopathologywill be performed under the supervision of or by a Board-CertifiedVeterinary Pathologist. The ovaries and the uterus with cervix of each female rat will be weighed, and ovaries, uterus, vagina and a mammary gIand will be retained in neutral buffered 10% formalin. Uteri of apparently nonpregnantrats will be examinedafterbeingpressedbetweenglass plates to coDfirm the absence of implantationsites, and retainedin neutral buffced 10% fomdin. Exygen Research Page 71 of 153 418-028:PAGE G-72 Exygen Study No.: 023-072 Protocol 418-028 Page 17 A S u m Eval~u~tionosf Male Rats: To assess the potential toxicity of the test article on the male reproductive system, the endpoints listed below will be evaluated fromthe first 10male rats in each dosagegroup. OrganWeights The followingorganswill be individuallyweighed righttestis,left testis, left epididymis(whole and cauda), right epididymis, seminal vesicles (with and without fluid) and prostate. Spenn Evaluations: Spermconcentrationand motilitywill be evaluated wingcomputerassistedspennanalysis (CASA). Motilitywill be evaluated by the HamiltonT h m e NOS by collectionof a sample h m the left vas deferens. A homogtnate will be prepared h m the left caudaepididymisfor evaluationby the Hamilton T h m e NOS to det&m sperm concentration(spermper gram oftissue weight). The remainingportion of the left cauda cpididymiswill be used to manually evaluate sperm morphology. Spenn morphology ewduationswill include the fbllowing: 1) determinationof the percentage of no& spermin a sampleofat least 200; and 2) quaIitativeevaluationof abnomalsperm,including such categories as abnormalhead, abnormaltail, and abnormal head and tail. See ATTACHMENT 4 for additionaltissues to be weighed andretainedfrom the ten ratspersex per p u p assignedto histological sample colIeetionand wduatioi~ AII other tissues will be discarded. Scheduled Sacrifice of Female Rats that Do Not DelPver Litters: Rats that do not deliver a litter will be sacrificedon day 25 of presumedgestation. Gross nempsy, examinationand tissueretentionwill be conducted as describcdpreviouslyfor rats at scheduled sacrifice. Damswithno survivingpups will be sacrificed afterthe last pup is found dead, missingor presumed cannibalized. Gross necropsy, examination and tissueretention will be conducted as describedabove for rats at scheduid sacrifice. Exygen Research Page 72 of 153 418-028:PAGE G-73 Exygen Study No.:023-072 Rats Faund Dead or Moribund PromcOl4 18-028 Page 18 Rats that die or are sacrificed because of moribund condition,abortion orpremature deiiverywiU be exatnined for the cause ofdeath or moribund conditionon the day the observationis made. The rats will be examined for gross lesions. Testes and epididymides of male rats will be excised and paired organ weights will be recorded. The epididymides will be retained in neutral buffered 10%formalin. The testeswill be fixed inBouin's solutionfor 48 to 96 hoursand then retained in neutral buffered 10%formalin. Pregnancy status and uterine contents of femalerats will be recorded. Aborted &tuses and/ordelivered pups will be examinedto the extent possible. Uteri of apparentlynonpregnimt ratswill be examined afterbeing pressed between glassplates to c o d m the absence of implantationsites. Ovaries and uteri will be retained hneutral buffered 10% formalin. TESTS. ANALYSES AND MEASUREMENTS- F1 GENERATION Viabilify Preweaning Period Litters will be observed for dead pups at least twice daily. The pups in each littm will be counted once daily. Clmical Observationsand/or General Amentame: .- Preweaning Period Once daily. Pups will be observed if they are warm and clean, for evidenceof a nest and if pups are grouped together and nursing or have milk in stomach. Each pup will be examined for generalshape of the head, trunk,limbs, tail and presence of anus. Clinicalobservations may be reconded more fresuently thancited above, if deemed appropriate by the StudyDirectorand/or the Study Monitor. Body Weights: Preweaning Period: Days 1(birth), 8,15 and 22 postpartunz Sacrifice: Terminal weight. Feed Consumution Values (recorded and tabulated): pI.eweaning Period: Not recorded. - METHOD OF SACRIFICE F1 GENERATION PUPS: F1 generation pups will be sacrificedby carbon dioxide asphyxiation. Exygen Research Page 73 of 153 418-028:PAGE G-74 Exygen Study No.: 023-072 Protocol 418-028 Page 19 - c NECROPSY F1 GENERATION Grosslesionswillberetained inneutral M e r e d 10% Fonnalin forpossible futureevaluation. Unless specificallycited below, all other tissues will be discarded. PUDSFound Dead om Dav 1 PostDartum: Pups that die before examination of the litter for pup viability will be evaluated for vital status at biah. The lungs will be removedand immersed in water. Pups with lungs that sink willbe identified as stillborn; pups with lungs that float will be identified as Liveborn, and to have died shortly after birth. Pups with gross lesions will be preserved in Bouin's solutionfor pckble fiture evaluation. i m : Pups found dead or sacrificedbecauseofmoribunditywill be examined forgross lesions and for the came of deathor the moribund condition. Pups with gross lesions will be preserved in Bouin's solution for possible futureevaluation. Scheduled Sacrifice: - Onday 22 postpartum, pups will be will be sacrificed and examined for gross lesions; gross lesionswill be preservedin neutral buffered 10% formalin. Necropsy will include a single crosssection ofthehead at the level of the fiontal-parietal suture and examinationofthe cross- sectioned brain for apparent hydrocephaly. Blood samples will be collected from each selected pup (5 per sex per litter h m the 10f d e s per group selectedfor FOB and motor activityassessment, blood sample collection for H&CC, and histologicalevaluations)&om the vena cava The bloodwill be placed into tubes, pooled per litter, allowedto clot and spun in acentrifuge; the resulting serum will be tra~~fkreindto labeled polypropylene tubes. All sampleswill be immediatelyfrozen on dry ice and maintained hzen (470C) until shipment foranalysis. The liver h m each selectedpup will be collectedexcised and the organweight recorded. The median lobe will be fiozen and stored &2OoC) until sbipment for possible analysis. Fmzen samples will be shipped to Lisa Clemm at the previously cited address. The remainingportion ofeach liverwillbe retainedin nwtral bufkcd 10% formalin for possible histological evaluation. The livers will be pmcessed and tvaluated histologicallyas described for the Fo generation ratsin HistologicalEvaluation in A'ITACHMENT 4. c Exygen Research Page 74 of 153 418-028:PAGE G-75 Exygen Study No.: 023-072 PROPOSED STATISTICAL TESTS(*"? The followingschematicrepresentsstatisticalanatysesofthe data. ProtDcoI 418-028 Page 20 1. Parametric A BarUWsTmtc Tvoe of Testa II. Nanoarametriob 111. Test for Prauortion Data Variance Test for Homogeneily of the Binomial Distibution * --------------I- a Statisticallysignificantprobabilitiesare reportedaseitherp5 0.05 orp I0.01. b. Proportiondata are not included in thiscategory. c. Test for homogeneityof variance. Exygen Research Page 75 of 153 418-028:PAGE G-76 Exygen StudyNo.:023-072 PrOtOCd418-028 Page 2 1 c Test items in the FOB using interval scales, such 85 the grip-strmgthtests and the lauding faot splay test, as well as body weight data and feed consumptionvalueswill be analyzedas described under the Parametricheading of the schematic. Bartlett's Test of Homogeneity of Variances@w) ill be used to estimatetheprobabilitythat the groups had diffaent variances. A nonsignificant result @>0.001)will indicatethat an assumptionof homogeneity ofvariance is not inappropriate, and the data will be compared using the Analysis of Variance Test("). If that test is significant (pS.OS), the groups exposed to the test articldsubstancewill be comparedwith the control group using Dunnett's Test("). IfBartlett's Test is significant(,pS.OOl), the Analysis of Variance Test is not apprupriate, and the datawill be analyzed as describedunder the Nonnarametric heading of the schematic. When 75%or fewer of the scores in all the groups are tied, the Kruskal-WallisTest"') will bc used to analyzethc data, and in the event ofa significant result @9.05), Dunn's TestcLnwiIl be used to comparethe goups exposed to the test articldsubstancewith the control group. When more than75%of the scoresin any group are tied, Fisher's Exact Test('4)will be used to comparethe proportion ofties in the groups. Data from the motor activity test, with repeated measurementswithina session,will be analpmi using anAnalysis ofVariance with Repeated Measures('5),as described under that heading in the schematic. A Significanteffect(pg.05) in that test can appear as effect of Concentration(a differencebetweenp u p s in the total across all measurementsin a session) or as an mteraction betweenConcentrationand Block (adifferencebetween groups at specificmeasurement c periods). If the Concentrationeffect is significant,the totals for the control group and the groups giventhe test articldsubstancewill be compared using Dmett's Test. If the Concentrationx Block interaction is significant, an Analysisof VarianceTest will be usedto Bvaluafethe data at each measurementperiod, and a significant result (p40.05) will be followedby a comparisonof the p u p s using Dunnett's Test. Test items in the FOB having graded or count scores will be analyzed using the procedures describedunderthe NomaaamCtricheadingof the schematic. Clinical observation incidencedata, as well as the descriptive andquantal data from the FOB, as contingencytablesusing the Variance Test for Homogeneityof the Binomial Alternate or additionalstatistical evaluationsmay be performed if deemed necessaryor appropriate. Exygen Research Page 76 of 153 418-028:PAGE G-77 Exygen Study No.: 023-072 Aotocol418-028 Page 22 DATA ACOUISITION. VERTJ?ICATIONAND STORAGE: Data generated during the course of this study will be recorded either by hand or using the Argus Automated Data Collectionand Management System, the VivariumTemperatureand Relative HumidityMonitoring System, the Coutbourn Inrtnunents PassiveInfinredMotor Activity System, the CoulbournInstrumentsAuditoryStartieastern,the CoulboumInstnunentsSptial Delayed Alternation System, andor the passive avoidance software. All data will be tabulated, summasized and/or statisticallyanalyzed using theArgrrsAutomatedData Coitectionand Management System, the Yivnrium Temperatureand Retative Humidity MonitoringSystem, Microsoft Excel [part of MimsofiOffice97 (version SR-2)] and/orTheSASSystem (version 6.12). Recordswill be reviewed by the StudyDirectorand/or appropriatemanagementpersonnelwithin 21 days after generation. All originalrecordswill be stored in the archives of the Testing Facility. AX originaldatawill be bound and indexed. A copy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the TestingFacilityat no charge for one year after mailingofthe draftfinal report, after which time the Sponsorwill be contactedto determine the dispositionof these materials. ExecutiveDirectorof Research MiIdre+dS. Christian, Ph.D.,Fellow,ATS Directorof Research Alan M.Hobeman, Ph.D.,DABT AssociateDirectorofResearch and Study Director: Raymond G. York, Ph.D.,DABT Directorof Operations and Compliance: BarbaraJ. Patterson, B.A. Directorof Laboratory Operations: JohnF. Barnett,B.S. Directorof StudyManagement: ValerieA. Sharper,M.S. Manager of Animal Operations: Dena C. bbo, V.M.D. Chairperson, Institutional h i m d Care and Use Committee: DouglasB. Learn, Ph.D. ConsuItant,VeterinaryPathology: W. Ray Brown, D.V.M.,Ph.D., ACVP c Exygen Research Page 77 of 153 418-028:PAGE G-78 Exygen StudyNo.: 023-072 L RECORDS TO BE MAlNTAINED: Protocol 418-028 Page 23 Protocol and Amendments. Test Substance, Vehicle andor Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment(ifprescribedby StaffVeterinarian). General Comments. ClinicalObservationsa d o r GeneralAppearance. Body Weights. Feed Consumption Values. Natural Delivery Observations. FOB and MotorActivityObsemtions. Blood SampleCollection, Processingand Shipment. Gross Necropsy Obstrvations. organ weights. Photographs (if required). Study Maintenance(roomand enviromental records). Feed and Water Aadyses. Packing andor Shipment Lists. .c FINAL REPORT: The Study Directat will provide periodic updates of study progress to the Sponsor. Draft summary tablesofunaudited computer-mordeddatamay accompany these updates. Statistical analyseswill not be performed on these interim data A comprehensivedraft find reportwill be prepared on completion of the studyand will be finalizedfollowing consultationwith the Sponsor. The reportwill include the following: Summary and Conclusion. Experimental Design and Method Evaluation of Test Results. Appendices: Figures, Summary and IndividualTablesSurdmarizingthe Above Data, Protocol and Associated Amendments and Deviations, Study Director's GLP Compliance Statement,Reports of Supporting Data (if appropriate) and QAU Statment. The Sponsorwill receive one copy of the draft report and two copiesof the final report. Data will be hand- andor computer-recorded. Records will be reviewed by the Study Director and/orappropriate managementpersonnel within 21 days after generation. All original records will be stored in the archives of the Testing Facility. All original datawill be boundand indexed. A copy of all raw data will be supplied to the Sponsorupon request. Prescwd tissues will be stored at the TestingFacility at no charge for one year aftermailing of the dtaR final ,- report, after which time the Sponsorwill be contacted to deteiminethe disposition of these materials. Exygen Research Page 78 of 153 418-028:PAGE G-79 Exygen Study No.: 023-072 Protocol 418-028 Page 24 n INSTITUTIONAL, A",C mAND USE COMMITTEESTATEMENT: The procedures describedin thisprotocol havebeen reViewedby the Testing Facility's Institutional AnimaI Care and Use Committee. AU procedures described m this protocol that invoivestudy animals wit1be conducted in a mannerto avoid or minimizediscomforgdistressor pain to the auimals. The Sponsor's signaturebelow documentsthe fact that informationconcerningthe necessityfor conducting this study and the fact that this is not an unaecessarilyduplicativestudymay be obtainedBorn the Sponsor. No alternative(in vilro)procedures were availableformeetingthe stated purposes of the study. 1. Christian, M.S. and Voytek, P.E.(1982). In V i wReprodudve and Mutugenicity Tests. EnvironmentalProtectionAgency, Washington, D.C. NationalTechnical Idomation Service, U.S.Departmentof Commerce, SpringGeld,VA 22161. 2. Christian, M.S.(1984). Reproductivetoxicity and tmatology evaluationsof naltrexone (ProceedingsofNdtrexone Symposium,New Yo& Ademy of Sciences, November 7, 19831,J. C l i i Psychiat. 45(9):7-10. 3. h g , P.L.(1988). EmbryoandFetal Deveiopmntul Toxiciv (Teratology)ControlData I in the CharlesRiver Crl:CD@BRRat. CharlesRiver Laboratories, Inc., Wilmington, MA 01887-0630. (Database providedby Argus Research Laboratories,Inc.) 4. Institute of LaboratoryAnimal Resources (1996). Guidefor the Care and Useof LaboratoryAnimals. National Academy Press,Washington, D.C. 5. Haggerty,G.C. (1989). Developmentof Tier I neurobehavioraltestingcapabilitiesfor incorporation into pivotal rodent saf'ety assessmentstudies. J. h e r . Col. Toxiwl. 853- 70. 6. S. (1968). Comprehensiveobservational assessment: Ia A systemic quantitative procedure for assessing the behavioral and physiologic state of the mouse. Psychopharmacologia (Berlin)13:222-257. 7. Moser?V.C.(1989). Screeningapproachesto neurotoxicity: A functionalobservational battery. J. h e r . Col. Toxicol. 8:85-94. 8. O'Donoghue, J.L. (1989). Screeikg for neurotoxicityusing a neurologicallybased examinationand neumpathology. J. h e r . Coi. Toxicol. 8:97-116. 9. Sokal, R.R.and Rohlf, F.J. (1969). Bartlett's test ofhomogeneity ofvariances. Biometry, W.H. Freeman and Co., San Francisco, pp. 370-371. Exygen Research Page 79 of 153 418-028:PAGE G-80 Exygen StudyNo.:023-072 Protocol 418-028 Page 25 rcI, 10. Snedecor,G.W. and Cochran, W.G. (1967). Analysis of Variance.StatirZicul Methods, 6th Edition, Iowa Sate UniversityPress,Ames, p ~2.58-275. 11. Dunnett, C.W.(1955). A multiplecomparisonprocedureforcomparing several treatments with a control. J. h e r . Stat. Assoc. 501096-1121. 12. Sokal,ICR.and Rohlf, F.J. (1969). Kruskal-WailisTest. Biometry, W.H. Freemanand CO., Sm Francisco, pp. 388-389. 13. Dunu, O.J. (1964). Multiple comparisons using rank sums. Tcchnometrics 6(3):241-252. 14. Siegel, S. (1956). Nonparametric Sfatisticsfor the Behavioral Sciences, McGraw-Hili, New York, pp. 96-104. 15. SAS Institute. hc. (1 988). Repeated measures analysisof variance. SAUSTAP User's Guide,Release 6.03 Edition, Cary,NC,pp. 602-609. 16. Snedecor,G.W. and Cochran, W.G. (1967). Variance test for homogeneityof the binomial distribution. Stdstical Methods,6th Edition,IowaStateUniversityPress, Ames, pp. 240-241. Exygen Research Page 80of 153 - PROTOCOL APPROVAL: FOR THE TESTING FACIL,ETY Alan M.Hobennan, PhD., DABT Director of Research 418-028:PAGE G-81 Exygen Study No.: 023-072 Protocol 418-028 Page 26 Date FOR TEE SPONSOR John Butdoff:PhD., DABT, CIH Study Monitor and Sponsor's Rqresmtative c Exygen Research Page 81 of 153 418-028:PAGE G-82 Exygen Study No.: 023-072 ATTACHMFJIT 1 SCHEMATICOF STUDYDESIGN AND STUDY SCBEDULE c Exygen Research Page 82 of 153 418-028:PAGE G-83 Exygen StudyNo.:023-072 AlTACHMENT 1 STUDY SCHEMATIC COMBINEDREPEAT DOSE AND REPRODUCTIVElDEVELOPMENTALTOXICITY SCREEN" Male Rats1 Remating 2W~kS cohabitaton 2W& Nahnal SUbSkUlCC Dosage Dosageperiod a. For additional detailssee "Tests,Analyses and Measurements"sectionof the protocol. b. FOB and motor activity evaluations conducted on ten malesper p u p . c. Male rats sacrificedaRercompletionofat least 42 days ofdosage; necropsy and retention of male reproductive organs. Hematology, clinical biochemistry and histological samples collected fiom ten male rats per group. d. Ten femaleratsper group assignedto FOB evaluationand motor activity evaluation. e. Ten female rats per groupand their litters sacrificed on day 22postparhun;necropsy and A retention of &male reproductiveorgans. Hmatology,clinicalbiochemistry aud histological samples collected. Remainingfemale rats sacrificedonday 22 postpartum and discarded. Exygen Research Page 83 of 153 418-028:PAGE G-84 Exygen Study No.:023-072 ATTACHMENT 1 Protocol 418-028 Page 2 Of 3 SCHEDULE' 26 MAR 02 OlAPRM 01 APR02- 12JUN02 - AnimalReceipt Acclimation Begins. Start of Dosage Period - Male Rats (14daysbefore cohabitation and through a 14-day cohabitation period until the day before sacrifice after at least 42 days of dosage). - DosagePeriod Female Rats (14daysbefore cohabitationthrough Day 22 of lactation). 14APROZPM-21APR02AM 21 APR 02 PM - 28 APR 02 AM Cohabitation Period (Maximum of 14 days), Malel(7days) Male 2 (7 days) 14 APR 02 Day 14 Toxicokinetic Sample Collection ..A 15APR02 First Possible Day 0 of Presumed Gestation. 28 APR 02 Last Possible Day 0 of Presumed Gestatim 06MAY02 19 MAY 02 06 MAY 02 - 07 MAY 02 First Possible Day 21 of Presumed Gestaiion ToxicokineticSampleCollectionand Sacrifice- ToxicokineticStudyFemale Rats - Last PossibleDay 21 of Presumed Gestation Toxicokinetic Sample Collection and Sacrifice ToxicokindcStudyFemale Rats - FOB and Motor Activity Evaluation 10Male Rats per Group a. Theastartdate of the study is the day the StudyDirector signs the protocol. b. Throughout this schedule,the day ofbirth is designated day 1 postpartum (day 1 of lactation)and all subsequent ages of the Fl generationrats and days of the lactation c period will be determined and cited accordingly, as described above the protocol section, "DayNumbering System." Exygen Research Page 84 of 153 418-028:PAGE G-85 Exygen Study No.: 023-072 ATTACHME" 1 06 MAY 02 23 MAY 02 10 MAY 02 23 MAY 02 12 MAY 02 13 MAY 02 23 MAY 02 - 09 J"02 c 27 MAY 02 - 13 Ill"02 24 SEP 02 ProtocoI 4 18-028 Page 3 Of 3 First Possible Delivery (Day 21 of presumed gatation). Last Possible Delivery (Day 25 of presumed gestatiim). First Possible Day 25 of Presumed Gestation Female Sacrifict. Last Possible Day 25 of Presumed Gestation Female Sacrifice. - Day 42 ToxicokheticSample Collectionand ScheduledSacrifice ToxicokineticStudyMale Rats - ScheduledSacrifice Main StudyMale Rats (Earliestpossibledate). Hematology,Clinical Biochemistry and Histological Sample Collection of SelectedMale Rats. FOB and Motor Activity Evaluation - 10 Female Rats per Group. - Day 22 Postpartum SacrificeFemale Rats and Pups. Hematology, Clinical Chemistry and HistoIogical Sample Collectionof SelectedFemale Rats and Pups. Draft Find Report Exygen Research Page 85 of 153 418-028:PAGE G-86 Exygen Study No.: 023-072 ATTACEMENT 2 MATERIAL SAFETY DATA SHEET F Exygen Research Page 86 of 153 418-028:PAGE G-87 Exygen Study No.: 023-072 h MATERIAL SAFEPY DATA SHEET (Experimental) 3N 3H Center S t . Paul, Minnesota 55144-1000 1-800-364-3577 or (651) 737-6501 (24 hours) Copyright, 1999, Minnesota Mining and Manufacturing Company. All r i g h t s reserved. Copying and/or downloading a t this information f o r the purpoae o f properly u t i l i x i n a 3?4products is alloned provided that: 1) the information is copied i n full n l t h no ohonges unless p r i o r agreement is obtalncd from 3M, and 2) neither t h e copy nor the o r i g i n a l is resold or otherwise distributed with the intention o f earning 8 profit thereon. DIVZSfON: 3EI SPECIALN WTERWS MTERIAL: L-9051 OWELOPMENTAL PRWUM ISWED: O8eember 07, 1890 SUPERSEDES: May 17, 1999 DOCUMENT: 04-5470-2 ... POTASSIUM PERFLWROHEXANE SULFONATE.. ...... .c. RESIOUAL ORGANIC FLUOROCHEMICALS.. 3871 -99-6 Mixture 100.0 Unknwn T h i s material i s n o t l i s t e d on the TSCA inventory and should be used f o r research and devtlopment purposes only under the direct supervision of a technically qualified individual. ............... ................ BOILIW POINT:. VAPOR PRESSURE:. ................. VAPOR DEMSITY: MIA Negligible MIA EVAWRATION RATE:.............. Negligible ...... SOLUBILIrY I# WATER:........,,. SPECIFIC GRAVITY:........ sllfpht ca. 1.0 W8ter-1 (Bulk) ............... pPHER: C.E.N..T.V.OL.A.TI.LE.:................................ Negligible, N/A VISCDSIM:..................... NIP MELTING POINT:.. NIQ APPEARANCE AN0 MWR: O f f -white crystalline s o l i d , sharp odor. Exygen Research Page 87 of 153 418-028:PAGE G-88 Exygen Study No.: 023-072 F MSDS: L-9OS1 DEVELOPMENTAL PAOOUCT Oecember 07, 1999 ---------------------.---.------.------------------.-----------.. -3..-F-IR-E-A.ND--EX-P.LO-S-IO-N -HA-ZA-R.D-DA-TA-.--..-----------------------------.---.-..--- .................. FLASH POINT:. * 212 F Setaflesh Set8tla8h Closed Cup FLAMHABLE LIMITS LEL:........ N/A - ..... FUWMLE Lmrs UEL:.. Iv/A ... AUTOIGNITION TEMPERATURE:... N/D EXTINOUISHINO MEOIA: Water, Carbon d i o x i d e , Dry Ch9RiCal, Fom SPECIAL FIRE FIOMINO PROCEDURES: Hear f u l l protective clothing, including hslaet, self-contained, positive pressure o r pressure damand braatthing apparatus, bunker coat and pants, bands amund oms, -1st and legs, Taco mask, a d protective covering for expoled ereas of the head. UNUSUAL FIRE AND EXPLOSION HAu\RDs: See Hazardous Decomposition seetion for products of omburtion. STABILITY: Stable . INCOMPATIBILIN None knawn - MATERIALS/CONDITIOHS TO AVOID: HAUAWUS POLYMERIZATION: Harordour polymerization w i l l not occur. HAZARDOUS DECCMPOSITIaN PROWCTS: Hay produce fluorocarbon gases it eXpO8ed t o very high tmp8ratures (over 300 C). SPILL RESPONSE: Observe precautions from other sections. Collect spilled aaterial. Use wet sweeping compound or water t a avoid dusting. Clean up residue. Place i n a closed container. AECOWIENDED DISPOWL : Incinerate i n an i n d u s t r i a l or commercial f a c i l i t y i n the presence o f a combustible m a t e r i a l . Combustion products will include HF. Oisposal alternative: Ofspase of Wste product i n a t a c i l i t y permitted t o accept chemical wste. Exygen Research Page 88 of 153 418-028:PAGE G-89 Exygen Study No.: 023-072 LA HSDS: L-9051 DEVELOPMENTAL PRODUCT Oeceaber 07, 1999 PAGE 3 ENVIRONMENTAL MTA: Not determined. REUlUTORY INFORMATION: Volatile Organic Corpaundr: NIb. - VOC Lesr H20 8 Exempt Solvents: MID. Since regulations vary, consult applicable regulations o r authorities before disposal. U.S. EPA Hazardous Worts Nu8ber None (Not U.S. PA Hazardous). OTHER ElQVIR0"TAL INFOMTZOM: . T h i s product may contain one or more organic fluorocheaiaals that have t h e potential t 0 resist degradation M d persist in the environment EPCRA HAUAD CLASS: FIRE HAZARD: No PRESSURE: No REACTIVITY: Na ACUTE: Yas CHRONIC: No EYE CONTACT: Immediately flush e y e s with large amounts of water. Qet immediate medical attention. S K I N CIINTACT: Flush skin with large amounts o f M t w . Ifi r r i t a t i o n persiats, get medical sttentfon. INHALATION: If signsisynptors QCDUC, remove person t o fmsh air. If signslsyaptoas continue, call a physician. IF SWALLOWED: I? swallowd, call a physician irmediatsly. Only induce vomiting a t the instruction of a physician. Never give anything by mouth t o an unconscious person. EYE PROTECTTON: Avoid eye contact. Avoid eye contact uith vapor, spray, or sfot. The following should br worn alone or i n colbination, as appropriate, t o prevent eye contact: Wear vented goggles. I - A_tab_rev_kti_ons.: _N/D_-_No_t O_ctsr_min_ed_NI_A -_No_t A_ppl-ic_abl_e C~A -A-p-pr-ox-im-at-ely- - - - - - - - - - - - - - - - . - - - - - - . - - Exygen Research Page 89 of 153 418-028:PAGE G-90 Exygen StudyNo.:023-072 A MSDS: L-9051 OEVELOPHENTAL PAOOUCT December 07, 1999 ma 4 SKIN PROTECTION: Avoid skin contact. mar appropriate ptoves when handling t h i s material. A p a i r o f gloves made froa the tollowfng i a t a r k l ( s ) are recommended: butyl rubber, polyethylene/polyvinylidenechloride (Saranex). Use one o r a w e o f thr following personal protection iteas as necessary t o prevent skin contact: head cowring, coveralls. Proteetive garments (other than glover) should be made o f . either o f the follOUing chloride (Saranex) materials: polyethylantfpolyvinylidene RECOWENDED VENTILATION: Use n i t h appropriate local rxhrust ventilation. ProvLd. appropriate local exhaust v e n t i l s t i o n a t tranafer points. Use i n a will- ventilated area. Provide suff i c i a n t ventilation t o maintain saisaions b e l w recoamcnded sxporure li.its. If cxhrust v e n t i l a t i o n is not adequate, use mppropriate respiratory protection. Provide , v e n t i l a t i o n adequate t o control vapor concentration8 below recommended exposure l i m i t s rndlor c o n t r o l spray o r r i s t . - Local exhaust v e n t i l a t i o n is recoarcnded where the met airborne. material beco RESPIRATORY PROTECTION: Avoid breathing of airborne material. Select one o f the following NIOSH approved respirators based on airborne concentration of contaminants and i n accordance with OSHA regulations: h8lf-aask supplied a i r respirator, full-faCO supplied air respirator. PREVENTION OF ACCIDE'KTAL INGESTION: Do not eat, drink o r smoke when using t h i s product. Ha6h exposed areas thoreuphly n i t h soap and water. Wash hands a f t e r handling and before eating. RECUMMENDED STORAGE : Store a t room temperature. closed uhen not i n use. Keep container dry. Keep container FIRE AND E%PLOSIUUAVOIDANCE: Nut determined. Exygen Research Page 90 of 153 418-028:PAGE G-91 Exygen Study No.:023-072 - MSDS: L-QD51 DEVELOPMENTAL PROWlCT Oecesmbtr 07, 1999 PAQE 5 EXPOSURE LIMITS (continued) ---'--'---------------------.---.----.-..----.------------.----- XNGREDXENT VALUE UNIT WPE M H SKIN. the p o t e n t i a l c o n t r i b u t i o n t o the overall exposure by the cutaneous route i n c l u d i n g mcous meabrano and aye, e i t h e r by airborne or, aore p8rticulorly, by d i r e c t oontact with t h e substance. Vehicles oan a l t e r s k i n absorption. -SOURCE 9M: OF EXPOSURE LIMIT DATA: 313 Recommended Exposure Guidelines EYE CONTACT: No i n f o n a t i o n &as found regarding effects from eye contact. Single exposure 8ay cause: M i l d Eye frritation: rignslrymptomr can include redness, s i d l i n g , pain, and tearing. SKIN CONTACT? No information was found regarding e f f e c t s rrom 'skin contrct. Hay be absorbed through t h e s k i n and p e r s i s t in the body f o r an extended time. Single exposure may cause: Moderate Skin I r r i t a t i o n : signslrymptoms tan include rednei8, anelling, i t c h i n g , 8nd dryness. INHALATION : No information was found regarding effects fron inhalrtion exposure. Nay be absorbed by inhalation and p e r s i s t fn the body for an extended tide. Single overexposure, abovm recommended guidelines, MY cause: I r r i t a t i o n (upper r e s p i r a t o r y ) : signstsymptoms can include soreness o f t h e nose and throat, coughing and sneezing. I F SWALLOWED: I n g e s t i o n is n o t a l i k e l y route of exposure t o t h i s product. No infomation was found regarding affects fmn S H a l l O W h g . Animal studies canducted on OrgrniC fUJOrOGheRiGal9 rhich nay be present i n t h i s product indicate effects including liver disturbances, u c i g h t lass, lass o f appetite, lethargy, and neurological, pancreatic, ...----------*..- F _a.d.re.n_al -an_d _hem_at_olo-g_lc -ef-fe-c-ts_. T_he-re*a-re-.no-*kn.ow-n-h-un.a.n.h-ea-l-th-. -.--. **---.-.------* - - - Abbreviations: N I Q Not D e t r r l l i n r d NlA Not Applicable CA Approximately Exygen Research Page 91 of 153 418-028:PAGE G-92 Exygen StudyNo.:023-072 I . - HsaS: L-9051 DEVELOPMENTAL PRoouct December 0 7 , 1999 PAGE 6 effects from o n t i c i p t t i d exposure t o there organic fluorochcaicrls when used as intended and instructed. OTHER HEALTH HAZARD INFOAHATION: This product ray contain one or . a r m organic fluarochmrloalr t h 8 t have the potential t o be absorbed and remain i n t h e body tor lona periods of time, e i t h e r as the parent molecrule o r as metabolites, and ray acoumulstc with repeated exposures. There are no known huaan health e f f e c t s fron anticipated exposure to these organic fluorochemicals nhen used as intended and instructed. The presence of organic fluorochemlorls i n t h e blood of t h e g e n e r a population and &UbpapUl!atlon8, such as warkers, has been published dating back t o t h e 1970's. 3M'r epidemiologic81 rtudy o f its ow workers indiGate8 no adverse effects. _-I_- Abb - r -_L___ eviati ___I_ ons: _ -_ N/ -- D - -- - . ----.. Not - --.------------ Determined - - . --. Nf -- A - -- - ---. No - t - ------ App - l - i ---- cab l e CA - Approxirately --------------------;-----r-------.--l----.--.----.--.--*-----.-.*------.----..------- The information i n tnir Material Safety Data Sheet (HSDS) is believed t o be correct as af the date issued. 3n MKS NO HAWWTIES, UCPRESSED bR IMPLIED, INCLUDING, But NOT LIHZTED TO, ANY IsWLIEDIWIWITY OF HERCHANTABILITY OR FITNESS FOR A PARTIGUUR PURPOSE OR COURSE OF PERFORHANCE OR USAGE OF TRADE. User i s responsible foF deterainlng whether t h e 3H product is f i t f o r a plrticulor purpo8e and suitable f o r user's method of use o r applicatian. Given the variety o f factors that can a f f e c t the use and application of a 3M product, some o f uhich are uniquely within t h e user's knowledge and control, it is e s s e n t i a l that the user evaluate the 3M procluct t o determine whether it is f i t f o r 8 particular purpose and suitable f o r user's rethod o f use o r application. .a 31.1provides information i n electronic fern as a service t o i t s cvstolers. Due t o the remote p o s s i b i l i t y t h a t electronic transfer ray haye resulted in erFors, omissions o r rltenltionrr i n this infom8tion, 3M makes no representations as t o its completeness o f clccuracy. I n addition, information obtained from a database nay not be a s current as t h e information i n the HSDS available directly from 3M. Exygen Research Page 92 of 153 418-028:PAGE G-93 Exygen Study No.: 023-072 ATTACHMENT 3 TEST SUBSTANCE PREPARATION PROCEI)URE .- Exygen Research Page 93 of 153 418-028:PAGE G-94 Exygen StudyNo.:023-072 n AlTACHMENT 3 Pmtocol418428 Version: 418-028(19 MAR 021 Page 1 af2 TEST SUBSTANCE PREPARATION PROCEDURE Test Substance: T-7706 Vehicle: Aqueous 0.5% CMC (medium viscosity) A. Purpose: The purpose of this procedureis to pmvide a method forthe preparation of dosage suspensionsof T-7706 for oral (Savage) administrationto rats on Argus Research Study number 418-028. 6. General Information: 1. All suspensioncontainerswill be labeledand color-coded.Each label will specify the protocol number, test substance identifmhn, Argus batch number, concentration,dosage level, preparation date,expiration date and storage conditions. c 2. Suspensions will be prepared: - Daily X Weekly - Approximately every ten days - For- days of use - Bysponsor 3. Suspensionswill be administeredat a finaldosage volume of a rnUkg. 4. Safety: -X Gloves, uniformnabcoat, goggles or safety glasses with side shields & Dust-misWEPA-filtered Mask - Half-Face Respiratorif not used in a chemicalfume hood - Full-FaceRespirator/PositivePressureHood __ Tyvek Suit or tyvek apron and sleeves 6. Dosage suspensions adjusted for % Activit\/lPurity or Correction Factor. - Yes X No (Calculations based on 100%) - - - %Activity % Purity Correction Factor 6. Sampling requirements:Cied in protocol 7. Storage: Cited in protocol Exygen Research Page 94 of 153 418-028:PAGE G-95 Exygen StudyNo.:023-072 - AlTACHMENT 3 Protocol 418-028 Version: 416-026M Q MAR 02) Page 2 of 2 TEST SUBSTANCE PREPARATION PROCEDURE NOTE: Priorto test substance preparation accurately measure the required amount of the appropriatevehicle (R.O. deionizedwater should be used for calibration purposes) in a graduated cylinder, pour the required amount of vehide into a beaker. Carefully mark each beaker at the meniscus. This markwill be used during the preparationto bringthe test substance slurry up to volume. C. Dosage Suspension Preparation: 1. Weigh the required amount of test substance on a piece of weigh paper or into an appropriately sized mortar (see PREPARATION CALCULATIONS). 2. If weigh paper is used,transferthe test substanceto an appropriately sized mortar. If necessary, grind the test substance into a fine powder. Slowly add a small amount of vehicle and grind. Continue to add vehicle slowly and grindthe vehicle and the test substancetogether to form a fine .c slurry. Transfer the vehideltest substance slurry to a marked beaker. 3. Rinse the mortar and pestle with additional vehde to remove any remaining test substance. Transfer rinse to beaker. 4. Add additionalvehicle to the beakerto bring volume up to the mark. Place on magnetic stir plate and agitate prior to and during sampling, aliquotting andlot administration. 5. Repeat steps (1) through (4) for each concentration. Date: W P ~ Q L attached clarificationform] InitiaVDate : Exygen Research Page 95 of 153 4 18-028:PAGEG-96 Exygen Study No.: 023-072 ATTACHMENT 4 TISSUES TO BE WEIGHED, RETAINED AND EXAMINED HISTOLOGXCALLY Exygen Research Page 96 of 153 418-028:PAGE G-97 Exygen Study No.:023-072 ATTACHMENT 4 Protocol 418-028 Page 1 of 2 r.4 TISSUES TO WEIGDZDAND RETAINED FOR POSSIBLE EXAMINATION FROM TEN RATS PER SEX PER GROUP The tenrats per sex pet group assigned to functional observationalbattery andmotor activity tests will be assigned to hematology,clinicalbiochemistryand histological evaluations. Tissues to be Weiehed: The following organswillbe excised, trimmed and individually weighed as soonaspossible after excision to avoid drying. liver spleen kidneys brain adrcnals heart thyIllus ovaries testes uterus (with cenrix) right epididyrnis prostate left epididymis(whole and cauda) seminal vesicles (With and without fluid) Tissues to be Retained .-- The followingtissues or representativesampleswill be retainedin neutralbuffered 10% formalin.. brain (representative regions including cerebrum, cerebellum, pons) smalland large intestines(includingPeyer'spatches) lungs(perfirsed withneutral buffered 10% formalin) lymph nodes (submandiblar and mediastinal) peripheralnerve (sciatic ortibial) gross lesions spinal cord (cervical, thoracic and lumbar) stomach liver kidneys adrenals spleen heart thymus thyroidparathymid trachea uterus urinarybladder bone marrow (sternum) testes* ovaries prostate uterus seminalvesicles (with coagulatinggland) vagina mammary gland (female rats only) * Testeswill be fixed in Bouin's solutionfor 48 to 96 hoursbefore beingrebind inneutral buffmed 10% fomalin. LL. Additionally, the remainingportion of the left epididymis(corpus and caput), as well as theright epididyrniswill be fixed in neutral buffered 10%fonnalh. Exygen Research Page 97 of 153 418-028:PAGE G-98 Exygen Study No.: 023-072 ATTACHMENT 4 - Histolodeal Examination: Rotoco14 18-028 Page 2 of 2 Histologicalexamination of retained tissues, includingreproductiveorgans,will be couducted forthe assignedten rats per sex from the control and highdosage ~ O U a~ndSfrom the F1 generationpups (livers) fromthe control and high dosage pups. If lesions attributed to the test substanceare observedin the rats exposedto the high test substanceconcentration,the game tissues will be examinedfiom the assignedtentats per sex exposed to the lower test substance concentrations. Shouldfesultswarrant examination of the lower dosagegraups and conductof quantitativeevaluation,scheduled report date andprices will be adjustedaccordingly. The postlactional ovary should contain primordial and growing foJIiclcs as well as the large corpora lutea of lactation. Histopathological examination may detect qualitative depletion of the primOniial folliclepopulation, A quantitativeevaluationofprimordialfollicles will be conducted for Fo generation fmalerats;the number of rats, ovariansectionselectionand section sample sizewill be statisticallyappmpriate for the evaluation procedureused. Examinationwill includeenumerationof the number of primordial follicles,which can be combinedwith small growingfollicles,for comparisonof ovaries of rats assigned to treatedand control groups. Tissuesto be examinedhistologicallywill be shipped (ambient conditions)to: .c Rhcipal Investigator: W. Ray Brown,D.V.M., Ph.D., ACW Veterinary Pathologist Research PathologyS d c c s , Inc. 438 E.Butler Avenue New Bntah, Pennsylvania 18901 Telephone: (215) 345-7070 Telek: (215) 345-4326 Email: WRBRPS@conccntric.net The recipient will be notifiedin advanceof sample shipment. Exygen Research Page 98 of 153 418-028:PAGE G-99 Exygen Study No.: 023-072 PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706WITH THEREPRODUCTION/DEVELOPMENTAL , TOXtCITY SCREENINGTEST SPONSOR'S STUDY NUMBER: T-7706.1 Amnzdment 1 - 17 April 2002 1. Detailed CIinicaI - Observations Male and Female Rats (page 10 oftheprotocol): [EffectiveDate: 1April 20021 Detailed clinicalobscnraticmswill not be recorded for male and female rats assigned to toxicokiueticsampling. Reason for Chan--g-e: This changewas made becausesufficicnt data for evaluationof detailedclinical observationswill be availableh m the rats assignedto the main study. 2. Feed ConsumutionValues - Male Rats and Female Rats (page 11 of theprotocol): [EffectiveDate: 1 April 20021 Feed ccmsumptionvalues will not bc recordedor tabulated farmate and f d e rats assignedtotoxicokineticsampling. Reason for Change: This chaugewas madebecause sufficientdata for evaluationof f e d consumption will be available h m the rats assignedto the main study. C. Any revisions made to this fmpiiztd amendment must be made by subsequent amendment. Exygen Research Page 99 of 153 418-028:PAGE G-100 Exygen Study No.: 023-072 Protocol 418-028 Amendment 1 Page 2 .- 3. Estrous Cvclinn and Mati% (page 12of the protocol): [EffectiveDate: 1April 20021 For the rats assignedto toxkokheticsampling, estmuscyclingwill be evaluated duringthe cohabitationperiod Until spumatozoa are observed in a smear of the vaginal contents and/or a copulatory plug is observedin sihc, but not during the dosagepuiod, prior to cohabitation. Reason for Chanpe: This change was made because sufficient data for evaluation of estrous cycling will be available h m the rats assignedto themain study. 4. Scheduled Sacri5ce - ToxicokineticStudy (page 16 of the protocol): (EffectiveDate: 5 April 20021 Thenumber of hplautationsitasand corpora lutea will be recorded. Carcasses willbe discardtdwithout fiathcrevaluation. Reason of chance: This change was made in orderto provide moreinformation about possible toxicity of the test substance m pregnant rats. 5. ScheduledSacrifice - Main Study (page 16 of the protocol): [EffectiveDate: 27 March 20023 The number of implantationsiteswill be recorded, rather than the number of implantationsitesand corpora luteawili be recorded. Reason'of Change: The number of corpora lutea will not be recorded because corpora lutea regress at a rapid rate and are not counted on studies at weaning. 6. ScheduledSacrifice(page 19 ofthe protocol): PffeCtiveDate: 4 April 2002J The liver&om each selectedpup will be excised and the organ weight recorded, xather thanthe liver h m each selected pup will be collected excised and the organ weight recorded Any revisions made ta this fh?lded amendmentmust be made by subsequmt amendment. Exygen Research Page 100 of 153 418-028:PAGE G-101 Exygen Study No.: 023-072 Reason for Chanec: This change clarifies the protocol by removiag an extraneous word. * t Alan M.Hobtnnao,Ph.D., DABT Director of Research Date Associate Directoro X a e a r ~ h Study Director Theresa H. Woodard, D.V.M. Member,InstitutionalAnimal Care and Use Committee Date John BuWo& PhD, DABT, CIHDate Study Monitor and SponsOis Rqresenmtive -. Any revisions d e to this ftnaiizedunendment mustbe mmdeby subrquentrmcndment. Exygen Research Page 101of 153 4 18-028:PAGE G-102 Exygen Study No.: 023-072 PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH TNE REPRODUCTIONLDEVELOPMENTAL TOXICITY SCREENING TEST SPONSORSSTUDYNUMBER T-7706.1 Amendment 2 - 15 July 2002 1. Safetv Precautions(page 3 of the protocol and Attachment 3 page 1 of the protocol): [Effitive Date: 29 April 20021 A halffacerespiratorwill be worn in additionto gloves, appropriateeye protection and a uniformflabcoat during formulation preparation of the bulk test substance. Reason for Chmw: This change was made to match the bulk test substancetext with the text locattd within the Material Safety Data Sheet. 2. Studv Schedule (Attachment 1page2 of the protocol): [EffectiveDate: 29 April 20021 The datesfor FOB and motor activity - evaluations havebeen extended to four days (06 MAY 02 09 MAY 02) rather - than two days (06 MAY 02 07 MAY 02). Any revisions to this finalized amendment must be made by subsequent amendmeut. Exygen Research Page 102 of 153 4 18-028:PAGE G- 103 Exygen Study No.:023-072 Protocol 418-028 Amendment 2 Page 2 Reason for Change: This change was made because m m timeis needed to evaluate animalsassigned to FOB and motor activity evaluations. U Study Director Member, Institutional Animal Care and Use Committee Study Monitor and SponS0r"s Representative ,.- Any revisbns to this fialized amendmentmust be made by subsequentamendment. Exygen Research Page 103 of 153 418-028:PAGE G-104 Exygen Study No.: 023-072 -/- ARGUS RESEARCH churfes aver f.c?borotories Dlsrovcy and Dcvolopmmt Sewices PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706WITH THE REPRODUCTION/DEVELOPMENTAL TOMCITY SCREBNING TEST SPONSOR'S STUDY NUMBER: T-7706.1 Ameneent 3 - 17 July 2002 - 1. Buk Test SubstanceSamDlinq, ShioDine Instructions, ShiDDinEInstructions, Caesarean-Sectionim - ToxicokineticStud%ScheduledSacrifice Toxicokinetic Study3and Scheduled Sacrifice, (pages 4,5,12,13,16 and 19, respectivelyof the protocol) c [Effictive Date: 29 M a y 20021 Samples for perfluorohexanesulfonate (PFHS) analysis shipped to Lisa Clemen at 3M Environmental Technology and Safety Services will be reshipped and remaining samples, that were to be shipped to Lisa * Clemenand have not yet been shipped,will be shipped to: John Flaherty,Ph.D. (Principal Investigator) Exygen Research 3058 Research Drive State College, Pennsylvania 16801 Telephone: (814) 272-1039, ext. 122 Telefax: (814) 231-1580 Email: john.flaherty@exygen.com Any revisionsto this finalized amendment must be made by subsequent amendment. _--I_-.. .. . . . . . -. Exygen Research Page 104 of 153 4 18-028:PAGE G-105 Exygen Study No.: 023-072 Protocol 418-028 Amendment 3 Page 2 These samples include: Bulk test substance sampling (page 4 of the protocol) Concentration and homogeneity @age 5 of the protocol) Stability@age 5 of the protocol) Plasma toxicokineticsamples(page 11to 12 and page 16 of the protocol) Plasma samples, rather than serum samples, were retained. Liver toxicokineticsamples(pages 12 and 16 of the protocol) Pooled fetal serumtoxicokineticsamples (pages 12 and 16 of the protocol) Pooled fetal liver taxicokineticsamples (pages 13 and 16 ofthe protocol) Pooled pup serum toxicokineticsamples (page 19of the protocol) Pup livertoxicokineticsamples (page 19of the protocol) The analyses will be subcontractedto Exygenby tbe Sponsorand the Quality Assurance Unit forExygenResearch will conduct criticalphase inspectionsand audit the respective results and reports according to the Standard Operating procedures of that facility. Suchcritical.phaseinspectionreports and audit reports will be submitted by that facilityto the Study Director, Raymond G. York. The date of the inspectionsand report submissionswillbe incorporated into a QAU statement generated by ExygenResearch for inclusion in the final report for Protocol 418-028. n Reason for Channe: This change was made at the request of the Sponsorbecause 3M is not able to complete the formulationanalysiswith their currentstaffing. Directorof Research Study Director Member,Institutional Animal Care and Use Committee Study Monitor and Sponsor's Representative I) n Any revisionsto this finalized amendment must be madeby subsequent amendment. Exygen Research Page 105 of 153 . . ., .. ._._....._.._..._..._...-... . . . 8 418-028:PAGE G-106 Exygen Study No.: 023-072 .- - r I ! . I i ' I i i I i: . i ! i i j I I I 1 3. PROTOCOL 418-028 ORAL (GAVAGE) COMBXNBDRHPBATEDDOSE TOXCWY STUDY OF T-7706WTl'H R A L TOMclTy SCREENINGTEST SPONSOR'S STUDY NUMBER: T-7706.1 - hendmcnt 4 20 March2003 - - 1. BdkTest SubstanceSarwlin&ShiwinpInstmcticms, S h b i u E~nstnrctionS, Caesarean-Sectiibnin~Toxiwkinetic Study, ScheduledSacrifice Toxicokhetic Study, and ScheduledSacrifice,@ages 4,5,12,13,16 and 19,respcctiVtly ofthe protoad) and Amendment 3. Item 1: Bulk Teat Substance Samnlinp;, Shi- - JUS~KU~OZSISh, i W g In~cructionsC. aesaregnSectionin~ T O X ~ C O ~S~tUud&y,C - ScheduledSacrifice ToxicakineticStudy, and SchechredS d c e @age I of Amen-& 3) mfbtiveDate: 27 February 20031 Tbe analysesp&md by Fkygen Research willbedoneacwr~gtoExygenMethadExM-023-MlRevision 1,entitled "Method of Analysis for the Determinationof P e f l u o m h ~ a u E ~ a(Pt eFfHS), * PerflUOMOCtanesuLfon~(PFOS) andPentadecafluoroooicAcid (PFQA) in Rat Liva, Serum and Urine Revision 1". Any revisionstotbisfinalizedamendmentmustbe madeby subsequentamendment. Exygen Research Page 106 of 153 418-028:PAGE G-107 Exygen Study No.:023-072 Reason for Change: Thischange was made at the requestofthe Exygen Resemh in order to clarify I the method of analysis. I ~ h & ,Iastitutiond~nimactare and Use C d t t e e ------ a * -e3 John Baenhoff, Ph.D.,DABT.CIHDab Study Monitor and Sponsor's Representative i b y revisionstothishalized ammdmentmustbe madeby subsequentammdment. ... ! .* ._... ., . ~ .. - I - - Exygen Research Page 107 of 153 418-028:PAGE G-108 Exygen Study No.: 023-072 PROTOCOL DEVIATION deviation Number: 1 Date of Occurrence: l o / 14/02 Page J- of 2 Exygenwdy Number: 023-072 Protocol Number: 418-028 A k recovery 62%for PFHS i s DESCRIPTIONOF DEVIATION being accepted for the 5000 ppb spike (0202119 Spk C) in data set Deviation Issued. ACTIONS TAKFN lor examde-deviationissued. SOP reviston. ete, Recorded By: Date: wlIs103 'IMPACT ON STUDY The 10 ppb and 50 ppb control matrix s p i k includedwith the set gave acceptable recoveries. Study Monitor . .- Exygen QAU ,Review _____.______.-_.._______.__._ . .-. .-..... . . Exygen Research Page 108 of 153 4 18-028:PAGE G- 109 Exygen Study No.:023-072 - E RESEARCH 3058 Research Drive State College, PA 16801 Phone: 874-272-1039 Fw: 814-231-1580 NOTE TO FILE Date: 01/14/03 Exygen Research Study # 023-072 PROTOCOL # 41 8-028 NOTE The PFHS stock solution was inadvertentlynot corrected for salt content when prepared, The stock solution was used to make the fortification and calibration standards usedfor the study. Due to the Large amount of data associated m'th this study, the concentrations of fortification and calibration solutions listed throughout the raw data were left uncorrected for the salt content. The final PFHS values recorded were corrected to reflect the difference i n PMS concentration due to the salt content. The correctionwas performed by multiplyingthe final ppb value found by 0.91, I Date: OlIl4I03 F Exygen Research July 31,2001/0 Page 109 of 153 418-028:PAGE G-110 Exygen StudyNo.:023-072 APPENDIX B Analytical Method: Method of Analysis for the Determination of Perfluorohexanesulfonate (PFHS), Perfluorooctanesulfonate (PFOS) and Pentadecafluorooctanoic Acid (PFOA) in Rat Liver, Serum and Urine Revision 1 (Exygen Method No. ExM-023-071 Revision 1) Exygen Research Page 110 of 153 418-028:PAGE G-111 Exygen Study No.: 023-072 TITLE Method of Analysis for the Determinationof Peduomhexanesulfonate(PFHS), Perfluomoctanesulfonate (PFOS)and Pentadecafluorooctanoic Acid ( W A )in Rat Liver, S e m and Urine Revision 1 AUTHORS John Flaherty, Karen Risha, and Emily Decker DATE ISSUED . November 20,2002 3M Medical DepartmentCorporateToxicology 3M Center, Building 220-2E-02 St. Paul, MN 55144-1000 PERFORMING LABORATORY Exygen Research 3058 Research Drive State College,FA 16801 METHOD NUMBER ExM-023-071Revision 1 TOTAL NUMBER OF PAGES 43 .. ..._ Exygen Research Page 1 1 1 of 153 418-028:PAGE G-112 Exygen Study No.: 023-072 ExygenMethod NO:ExM-023-071Revision 1 MAMAGElVENT APPROVAL ' Laboratory Manager Exygen Research John L.Butenhoff Date Sponsor Representative 3M Medical Department Corporate Toxicology pygen Research .-....,-. Exygen Research Page 2 of 43 Page 112 of 153 . 418-028:PAGEG-113 Exygen StudyN o. 023-072 ExygcnMethod No:ExM-023-071Revision 1 TABLE OF CONTENTS TMITALNEAG...E...M....E..N...T...A...P..P..R...O...V...A...L.....................................................................................................................................................................................2.1 TABLE OF CONTENTS...................................................................................................3. LISTOFTAEWS ............................................................................................................4.. LISTOF FIGURES.............................................................................................................5 1. SUMMARY ..................................................................................................................7 32.. EXPEWMENTALCOMPOUNDS.............................................................................. 8 CHEMICALSAND SUPPLIES...................................................................................9 3.1. CHEMICALS...........................................................................................................9 3.2. STANDARDS.........................................................................................................9. 3.3. EQUIPMENATM)SUPPLJES.................................................................................... 9 3.4. S O L ~ ~..N....S.................................................................................................... 10 3.5. PREPARATIOONFSTANDARD SOLUTION..S......................................................... 10 3.5.1. Stock solution......................................................................................... 11 3.5.2. Fortification Solutions ............................................................................ 11 3.5.3. CalibrationStandards.............................................................................11 4. METHOD..................................................................................................................1..2 4.1. FLOWDWGR.A...M............................................................................................1..2 4.2. SAhfFLEPROCESSING..........................................................................................12 4.3. BATCHSETU...P.................................................................................................. 13 4.4. S,uwr.~EXTRACTIO..N.......................................................................................13 4.4.1. LiverBxtraction...................................................................................... 13 4.4.2. Serum andUrineExtraction...................................................................13 4.4.3. SPEColumnConditioning.....................................................................14 4.5. QUANTITATIO...N................................................................................................ 14 4.5.1. Lc/Ms/MS System andO p t i n g Conditions......................................14 4.5.2. CalibrationCurveProccdutes................................................................ 15 4.5.3. Sample Analysis ..................................................................................... 16 4.6. ACCEPTANCCERITER.I..A..................................................................................... 17 4.7. PBRFORMANCCREITERI..A................................................................................... 17 4.8. TIME FOR ANALYSIS........................................................................... 18 5. CALCULATIONS...................................................................................................... 18 6. SAFETY ...................................................................................................................... 19 Exygen Research ...... Exygen Research Page 3 of 43 Page 113 of 153 418-028:PAGE G-114 Exygen Study No.: 023-072 ExygenMethod No:ExM-023-071Revision 1 LIST OF TABLES Table 1. Recovery Summary of- in Rat Liver and Serum.............................,.,....20 Table 2. Recovery Summary of PFOS in Rat Liver, Serum and Urine ........................... 21 Table 3. RecoverySummary of PFOA in Rat Liver, SerumandUrine...........................22 Exygen Research . .-.. ._- Exygen Research ... . . - .~ Page 4 of43 Page 114 of 153 418-028:PAGE G-115 Exygen Study No.: 023-072 Exygen MethodNo:ExM-023-071Revision 1 LIST OF F'IGURES Figure 1. Calibration Curve forPFHS............................................................................ Figure 2. CalibrationCurveforPFOS............................................................................ 24 Figure 3. Calibration CurveforPFOA............................................................_....._2..5.... Figure 4. Figure 5. Figure 6. Figure 7. Figu? 8. . RCeopnrteasiennintagtPivFeHCSh..r.o..m..a..t.o..g...r.a..m...o..f...a...0....1...ng../..m....L....S...t..a..n...d...a..r..d............ ...............26 ... Representative chromatogramofa 0.1 n&nL StandardContaining PFOS 26 Representative Chromatogramof a 0.1 nglmLStandardContaining PFOA..27 Representative Chromatogramof a 0.5 ng/mLStandardContaining PFHS...27 Representative Chromatogramof a 0.5 n g / d StandardContaining PFOS...28 ... Figure9. Representative Chromatogramof a 0.5 n g h L Standard Containing WOA..28 Figure 10. Representative Chromatogramof a 5.0 ng/mL StandardContaining PFHS 29 Figure 11. Representative Chromatogramof a 5.0 ng/rnL StandardContaining PFOS...29 Figure 12. RepresentativeChromatogram of a 5.0 ng/mL StandardContaining PFOA..30 Figure 13. Representative Chromatogramof a ReagentBlank SampleAnalyzed for PFNS ............................................................................................................... 30 Figure 14. Reprcsentative Chromatogramof a Reagent Blank SampleAnalyzed for PFOS ...............................................................................................................31 Figure 15. Representative Chromatogram of PFOA.................................................. a Re.a...g..e..n.t..B...l.a.-n..k...S..a..m...p...l.e..A...n...a..ly..z.e..d...f.o..r.......31 Figure 16. Representative Chromatogram for PFHS........................................ of a,.C...o..n..t..r.o..l..L...i.v..e..r...S..a..m...p.l.e..A...n..a..l.y..z.e..d...............32 Figure 17. RfoerpPreFsOenSt.a...t.i.v...e...C....h...r.o...m....a..t..o.gram...o..f..a....C....o..n...t.r..o...l..L....i.v...e...r..S..ampl.e..A...n..a..l.y..z..e.d...............32 Figure 18. Representative Chromatogramof a ControlLiver SampleAnalyzed forPFOA......................................................................................................... 33 Figure 19. RfoerpPreFsHenSt.a..t.i.v..e...C..h..r..o..m..a..t..o..g..r...a..m..o..f...a..C...o..n..t.r.o..t..S..e..r..u..m.....S..a...m....p..l..e..A....n..a...l.y..z...e..d...... 33 Figure 20. Representative Chromatogramof a for PFOS............................................... Con,.t.r.o.l..S.e..r.u..m...Sampl.e....A....n...a...l.y...z...e...d. .... 34 Figure21. RfoerpPreFsOenAta..t.i.v..e..C...h..r..o..m...a..t.o..g.,r..a.m....o..f..a...C...o..n..t.r.o..l..S...e..r.u..m....S..a..m...p..l.e...A...n..a.l.y..z..e..d...,.,.......34 Figure 22. RfoerpPreFsOenSt.a..t.i.v..e...C..h..r..o..m...a..t.o..g..r..a_m...o..f..a...C..o.n..t..r..o.l...U..r..i.n..e..S...a..m...p..l.e...A..n..a..l.y..z..e..d.....,........35 Figurt 23. fRoerpPrFesOenAta..t.i.v..e...C...h..r..o.m....a...t..o...g...r..a...m.. of a..C....o...n...t.r..o...l...U....r..i.n...e. Sam.p..l..e...A....n...a..l..y..z...e..d........ 35 Figure 24. Representative Chromatogram of a Control Liver Sample Fortified at 10 ng/g with PFHS ...................................................................................... 36 Exygen Research ...___..._. . ...-_ - . ..__--_. , . . ..... Page 5 of 43 Exygen Research Page 115 of 153 418-028:PAGE G-116 Exygen Study No.: 023-072 Exygen Method No:EXM-023-071Revision 1 LIST OF FIGURES (continued) Figure 25. Representative Chromatogram of a Convol Liver Sample Fortified at 10nglg with PFOS ......................................................................,.....,.........36 P i p 26. Representative Chromatogramof a ControlLiver SampleFortified at 10nglg withPFOA........,............................................................................. 37 Figure 27. Representative Chromatogram of a Control Liver Sample Fortified at 50 ng/g with PFHS ......................................................................................37 F i g m 28. Representative at 50 ng/g with CPFhrOoSm.a..t.o..g.r.a..m...,o..f..a...C...o..n..t.r.o..l..L...i.v..e..r..S..a...m...p..l...e...F...o...r.t..i.f..i.e...d......... 38 Figure 29. Representative at 50 ng/g with CPhFrOoAm.a..t.o.g..r.a..m....o.f...a..C...o.n..t.r.o..l..L..i.v.-e.r..S..a..m...p..l.e..F..o..r.t.i.f..i.e.d..-............3..8...... Figure30. RatelpOrensgen/mtaLtivweiCthhProFmHaSt.o..g..r.a..m....o..f...a..C...o..n..r.r.o..l..S...e..r.u..m....S...a..m...p...l.e..F...or,.t.i.f.i.e..d..............39 Figure 31. RateLprOesnegnlmtatLivweiCthhProFmOaSto...g..r..a..m...o..f..a...C..o..n..t.r.o..l..S..e.r.u.m Sam,,.p..l.e...F..o..r..t.i.f.i.e..d..............39 Figure32. Rate1p0rensge/nmtaLtiwvietChhProFmOAato..g...r.a..m...o..f..a...C...o..n...t.r.o.l..S...e..r..u..m....S.a..m...p..l.e...F..o..r.t..i,f.i..e.d,.........,...40 Figure 33. Representative Chromatogram of a at 50nglmLwith PFHS........................ Cont..r..o..l...S..e..r...u..m...S..a..m....p.le..F...o..r..t.i.f.i.e..d..............40 Figure34. Rate5p0rensge/nmtaLtivweitChhPrFoOmSat.o.,g..r..a.m....o..f..a...C...o..n..t.r.o..l..S...e.r..u..m....S..a..m...p..l.e...F..o..r..t.i.f.i.e..d..............41 Figure35. Rate5p0rensge/nmtaLtivwietChhProFmOaAto..g..r..a..m...o..f...a...C...o..n..t.r..o..l..S..e..r..u..m....S...a..m...p..l.e...F...o..r.t..i.f..i.e..d......... 41 Figure 36. Representative Chromatognun of at 10n g / dwith PFOS........... a C.o..n..t.r.o..l..U....r.i..n..e...S...a..m....p..l..e..F...o...r.t..i.f..i.e..d... ...... 42 Figure 37. Representative Chromatogram of at 10 ng/mL with PFOA .......... a C.o..n..t.r.o..l..U..r..i..n.e...S...a.m....p..l.e..F..o.,.r.t.i.f..i.e..d...............42 Figure 38. RepresentativeChromatogram of a Control Urine SampleFortified at 50 ng/mLwith PFOS.......................................................,...........................43 Figure 39. Rate5p0rensgejnmtaLtivwietChhPrFomOAato..g..r.a..m,...o.f...a..C...o..n..t.r.o..l..U..r.i...n.e...S...a.m....p..l.e..F...o..r.t..if..i.e..d...............43 Exygen Research ......-.. - .- Exygen Research Page 6 of 43 Page 116 of 153 418-028:PAGE G-117 Exygen Study No.: 023-072 ExygenMethad No: ExM-023-071 Revision I 1. SUMMARY This report details a method of analysis for residues of Peffluorohexanesulfonate (PFHS), Perflwrooctanesulfonate (PJXlS)and PentadecafluorooctmoinoicAcid (PFOA)in Rat Liver, Serum and Urine. Residues of PFHS, PFOS and PFOA are extracted from each matrix with acetonitrile. The acetonitrile extract is added to water and loaded onto a conditioned C18 solid phase extraction (SPE)cartridge. Analyte residues are eluted with 2 mL of methanol. Quantification of PFHS,PFOS and PFOA is accomplished by liquid chromatography/tandemmass spectrometry(L,C/MS/MS) analysis using multiple reaction monitoring (MRM). The proposed limit ofquantitation W Q ;the lowest fortification specified by the method which gives adequate recovery according to EPA guidelines) for this method is 10ng/g (parts-per-billion)each for PFHS,PFOS and PFOA. The theoretical limit of detection &+OD)will be based on the signal to noise ratio and will be at least greater than 3 times the level of noise, based on the instrumentation system used. For all analytes, the lowest analytical standard correspondsto 0.1ng/mL. This method was developed using rat liver, serum and urine. Typical percent recoveries f standard deviations (at 10 and 50 ng/g) are shown below: ' Fortification Level (ndg) 10 50 PFNS Recovery in Rat Liver 115% f 9.9% (-3) 98% f 3.5% (-3) Fortification Level (nglmL) 10 50 PFHS Recovery in Rat SCnUn 108%& 4.7% ( ~ 3 ) 111% f9.6% (-3) - :k: - Fortification 10 pFosRacovery inRatLiver 96% k 8.5% (n=3) 50 88% f 1.5% (nd) Fortification 10 50 PFOS Recovery in Rat Serum 88% f 9.8% (n=3) 120%f 2.1%(w3) PFOS Recovery in Rat Urine 93%rt: 4.7% (-3) 79% f 1.296 (n=3) , hdg) , 10 50 PFOA Recovery F o inRat Liver (ndmL) 98% 2 3.1% ( ~ 3 ) 10 94%f25% (n=3) 50 PFOA~Recovery~in Rat Serum 117% 1.5% (n=3 111% f 4.0% (m3) PFO~ARecover~y m Rat Urine 89%f25%(1133) 87% f 2.1% (1133) Exygen Research -.- -_... . .- __.I Page 7 of 43 Exygen Research Page 117 of 153 418-028:PAGE G-118 Exygen Study No.: 023-072 Exygen MethodNo:EXM-023-071Revision 1 2. EXPERTMENTAL COMPOUNDS The structutes for PFHS, FTOS and PFOA are given below. PFHS Chemical Name = Molecularweight = Perfluorohexanesulfonate 399,as shown Pl?HS.is suppliedas the potassium salt (C&iSO;K+), molecular weight =438 PFOS Chemical Name = Molecular weight = Perfluorooctanesulfonate 499, as shown F-\/\/\/\/ S 03- F FFF PFOS is supplied as the potassium salt (CEFWSO~Km")olecularweight =538 PFOA Chemical Name = Molecular weight = Pentadecafluorooctanoic Acid 413, as shown F-H"' 0' pygcn Research Page 8 of 43 Exygen Research Page 118of 153 418-028:PAGE G-119 Exygen StudyNo.:023-072 Exygen McthodNo:EM-023-071Revision 1 3. CHEMICALSA N D SUPPLIES 3.1. CHEMICALS Chemical Grade Source Catalog No. Methanol (MeOH) HPLC EM Science MXO47S-1 Ammonium Acetate Reagent JT Baker 0596-01 Water Acetonitrile Type I HPLC hygen EMScience NA AXO145-1 Type I water = electrical resistivity, minimum of 16.67 W c m at 25 OC, from a Labconco Waterprom workstation. 3.2. STANDARDS ~ Standard Peffluorohexanesulfonatc (PFHS) Perflumooctanesulfonate (PFOS) Pentadecafluorooctanoic Acid (PFOA) TCR Number SE-036 SD-018 Lot No: 08316DO Purity (%) 99.99 all isomers, 84.36 straight chain 86.9 96 Source 3M 3M Aidrich Chem 3.3. EQUIPMENATND SUPPLIES Equipment Balance, analytical (display at least 0.0001g) 125-mL LDPE narrow mouth bottles Disposable glass micropipets (50-100& 100-200pL) Tissumizer Wrist action shaker Sorvall RC 5C plus Centrifuge 50 mL disposable polypropylene centrifuge tubes 15 mL disposable polypropylene centrifuge tubes Visiprep vacuum manifold Sep Pak Vac 6cc (lg) tC18 cartridges(part # WAT 036795 2-mL clear HPK vial Kit (cat # 5181-3400) ClassA pipets and volumetricflasks Standard lab equipment (graduated cylinders, disposable tubes etc.) Stand-alone drop-in guard cartridge holder (part #844017-400) Hypercarbdrop-in guard column (4 mm) (part # 844017-400) HPLC Pump (Lc-10AD) Lc/Ms/MS and HPLC systems Supplier Mettler Nalgene Drummond (VWR) Tekmar Burrell Scientific Dupont VWR VWR Supelco Waters Hewlett-Packard various suppliers various suppliers Keystone Scientific Keystone Scientific Shimadzu As described in section 4.5. Exygen Restarch -. . .. .. ...-.____ ...._.. . Page 9 of 43 Exygen Research Page 119 of 153 418-028:PAGE G-120 Exygen Study No.: 023-072 ExygenMethod No:Em-023-071Revision 1 Notes: 1. In order to avoid contamination,the use of disposablelabware (containers, tubes,pipettes, etc.) is highlyrecommended. 2. Teflon or Teflon-linedcontainersorequipmentshould not be used. 3. It m a y be necessary to check the solvents (acetonitrile,methanol) for the presence of contaminants (esFially PFOA) by LC/MS/MS before use. (Ilertainlot numbers have been found to be unsuitablefor use. 4. Use disposable micropipettes or pipettes to aliquot standard solutions when pt.eparingstandardsand samplesfor extraction. 5. Equivalentmaterialsmay be substitutedfor those specifiedin this method. 3.4. SOLUTIONS (1) . 2 mM ammonium acetate in water is pnpared by weighing 0.154 g of ammonium acetate anddissolvingin 1L of water. (2) Hypercarb filtered type I water is prepared by filtering type I water through a Hypercarb guard column using a HPLC pump at -2-3 mUmin. - Before use, wash the guard cartridge with -25 mL of HPLC grade acetonitrile, then 25 mL of type I water, then begin collecting the filtered type I water eluate for use in the extraction. Rqeat the wash after 5ltering -2Lof water. Note:The aforementionedexampleis provided for guidance, alternativevolumes may be prepared as long as the appropriate ratios of the solvent to solute are maintained. 3.5. PREPARATIONOF STANDARD SOLUnoNS Analytical standards are used for three purposes: 1. CalibrationStandards-These standardsace pteparedin methanol and are used to calibratethe response of the detectorused in the analysis. 2. Laboratory Control Spikes - These fortifications are prepared at concentrations correspondingto the LOQ and 5x LOQ and are used to determine analytical recovery. Laboratory control spikes are prepared in control matrix. 3. Matrix Spikes - These fortifications are prepared by spiking into the field samples at a known concentration. Matrix spikes are used to evduate the effect of the sample matrix on analytical recovery and are prepared at the client's request. The analyst m a y vary the absolute volumes of the standardsas long as the correct proportions of solute to solvent are maintained. Exygcn Restarch Page 10 of 43 Exygen Research Page 120 of 153 4 18-028:PAGE G- 121 Exygen Study No.: 023-072 ExygenMethodNo:ExM-023-Ml Revision 1 3.5.I . Stocksolution Prepare individual stock solutions at 100 ~g/rnLfor PPHS, PFOS and PFOA by weighing out 10 mg of each anslytical standard (corrected for purity and if necessary, salt content) and dilute to 100 mL with methanol in separate 100-mt volumetric flasks. The stock solutions (in 125-mL LDPE bottles) are to be stored in a refrigerator at 2OC to 6OC and m stable for a maximum period of one year from the date of preparation. 3.5.2. Fortification Solrrtianr a. Prepare a mixed fortification standad at 1.0 pg/mL (1000 ng/mL) of PFHS, PFOS and PFOA by adding 1.0mL of each of the 100 p g / d stock solutions into a 100-mL volumetric flask and bring up to volume with methanol. . b. Prepare a mixed fortification standard at 0.1 pg/mL (100 ng/mL,) of PFHS,PFOS and PFOA by diluting 10.0 mL of the 1.0 p@mLmixed fortification solution to 100 mL with methanol in a volumetric flask. ExampIe: one hundred microliters of the 0.1 pg/mL solution spiked into 1 g of liver or 1mt of s e d u r i n e is equivalent to a 10 ppb (10 ng/mL or ndg) fortification. Store all fortificationstandardsolutionsin a refrigerator (in 125-mLLDPE bottles) at 2OC to 6C for a maximum periodof one year fromthe date of preparation. Note also that additionalconcentrationsmay be prepared if necessary. 3.5.3. Calibration Standards LC/MS/MS calibmtion standards containing PFHS, PFOS and PFOA tm prepared at 0.1,0.2,0.5, 1.0.2.0 and 5.0 ng/mL in methanol via dilutionof the 0.1 pg/mLmixedfortificationsolution (section 3.5.2.b). The following is a typical example; additional concentrations may be prepared as needed. Initial Conc. (nm) 100 100 100 5.0 2.0 1.o Volume (mt) 5 .O 2.0 1.o 10.0 10.0 10.0 Diluted to 100 100 100 100 100 100 Final Conc. (nghL) 5 .O 2.0 1.o 0.5 0.2 0.1 The standards may be used for a period of one year (in 125-mLLDPE bottles) when stored refrigerated (at 2C to 6C). Exygcn Research Page I 1 of 43 Exygen Research Page 121 of 153 418-028:PAGE G-122 Exygen Study No.: 023-072 Exygen Method No:ExM-023-071Revision I 4. METJ3OD 4.1. FLOWDIAGRAM The flow diagram of the method is given below, followed by a detailed descriptionof each step. Method How Diamam Weigh apprqniate mount ofliver or measure appropriatevolume of serumor urine (fortify samples designated as matrix spikes and laboratory control spikes) 1 Add water to liver for a final volume of 10mL, add waterto serum and urine ora final volume of 20 d,homogenize J Remove 1 mL and add 5 mL of ACN, shake .L Centrifuge 3. Decant supernatant into 35 mL of water L Load onto conditioned SPE L Elute with 2 mL methanol J. LCIMSMS Analysis 4.2. SAMPLEPROCESSING For liver samples, place frozen samples in a food processor and homogenizewith dry ice. Then place samples in containers and leave open in frozen storage overnight to allow for COZsublimation. Seal and piace the samples in frozen storage below -10C until time of extraction. Alternately, if there is an insufficient amount of sample (- less than 5 g), then no processing is necessary and the sample can be used as supplied. No sample processing is needed for serum and urine samples. However, frozen serum and urine samples must be allowed to completely thaw to room temperature before use. Exygen Research Page 12 of 43 Exygen Research Page 122 of 153 4 18-028:PAGE G- 123 Exygen Study No.: 023-072 ExygenMethod No:ExM-023-071Revision 1 4.3. BATCHSETUP a. A batch of samples should not contain more than 20 field samples. b. Each batch of samples analyzed must include at least one control (method blank using control matrix) and two matrix controls fortified at known concentrations (typicalIy 10 and 50 ng/g for liver or ng/mL for serum and urine) to verify procedural recovery for the batch. c. At least one field sampIe in each batch must also be separately fortified at a known concentration and carried through the procedure to verify recovery. Additional samples in the batch may also be fortified if desired d. All samples require duplicateinjections. 4.4. SAMPLEEX~~UCTION 4.4.1. Liver Ercrmction a. Weigh I g of liver sample into a 50 mL disposable centrifuge tube and fortify, if appropriate. Note that alternate weights of liver may be measud dependingon the sample size availablefor use. b. Add water to the sample for a final volume of 10 mL. Cap tightly. c. Homogenize sample using a tissuemizer for -1 minute. d. Transfer 1 mL of the sample using a disposable pipette into 15 mL disposable centrifuge tubes. Add 5 mL of ACN and shake for -20 - minutes on a wrist action shaker. e. Centrifuge tubes at -3000 rpm for 5 minutes. Cmfillly decant supernatant into a 50 mL disposable centrifuge tube and add 35 mL. of water. f. Load the sample onto a conditioned SPE column (for conditioning details, see section 4.4.3.). Discard the eluate. Any analyte residues will be trapped on the SPEcolumn at thispoint. g. Elute with 2 mL of methanol. Collect 2 mL of elute into a graduated 15 mL centrifuge tube. h. Analyze samplesusing electrospray LC/MSMS. 4.4.2. Serum and Urine Ertraction a. Measure 1 mL of serum or urine sample into a 50 mL disposable centrifuge tube and fortify, if appropriate. Note that alternate volumes of serum and urine may be measured depending on sample size available for use. e y g e n Research Page 13 of 43 Exygen Research Page 123 of 153 418-028:PAGE G-124 Exygen StudyNo.:023-072 FlxygenMethod No:ExM-023-071Revision 1 b. Add water to sample for a final volume of 20 mL. Cap tightly and vortex for -1 minute. Then continue with steps d-h in section 4.4.1. 4.4.3. SPE Column Conditioning Place the unconditioned SPE columns on the vacuum manifold. Condition the - SPE columns by passing 10 mL of methanol through the column fallowed -by 5 mL of water. The washes m a y be pulled through the SPE column using vacuum at a flow rate of -1 drop/sec or may be allowedto pass through the columnunaided. Discardall washes. Do not allow the columnto dcy. 4.5. QUANTFFATION 4.5.1. L C M W S System and Operating ConditiDns Mass Spec: Interface: Computer: Software: Micromass Quattro Ultima (Micromass) Electtospray (Micromass) Harvard infusion pump (Harvard Instruments), for tuning COMPAQ ProfessionalWorkstationAP200 Windows NT, Masslynx 3.3 HPLC Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven Note: A 4 x 10mm hypercarb drop in guard cartridgeis attachedon-line after the purge valve and before the sampleinjectorport to trap any residue contaminants that may be in the mobile phase and/or HPLC system. HPLC Column: Genesis C g (Jones Chromatography),2.1 mrn x 50 mm, 4p Column Temperature:3 5 O C Injection Volume: 15 pL Mobile Phase (A): 2 mMAmmonium Acetate in Type I water Mobile Phase (B): Mtthanol Time - % A - % B Flow Rate ~ W m i n ) 0.0 90 10 0.3 2.0 90 10 0.3 5.0 10 90 0.3 9.0 10 90 0.3 9.5 0 100 0.3 14.0 0 100 0.3 14.5 90 10 0.3 20.0 90 10 0.3' FxygenResearch Page 14 of43 Exygen Research Page 124 of 153 418-028:PAGE G-125 Exygen Study No.:023-072 Exypn Method No:ExM-023-071Rdvision 1 It may be necessary to adjust the IlpLC gradient in order to optimize instrument performance. Columns with differentdimensions(e.g. 2.1 x 30)and also columns fiom different manufacturers (Keystone Bemil Cla etc.) could be used, providcd equivalent chromatographyis obtained. Ions monitored: Analyte PFHS PFOS PFOA Mode Negative Negative Negative Transition Monitorcd 399 3 80 499 3 99 413 3 369 Approximate Retention Time -8.2 min. -8.8 min -8.6 min The retention times may vary, on a day to day basis, depending on the batch of mobile phase etc. Drift in retention times (up to f 4 96) is acceptable within an analytical run, as long as the drift continues through the entire analysis and the standards are included at the beginning and end of the analytical run. Note:An alternative K/MS/MS system may be used once demonstrated to be equivalent. - The mass sptrometer is tuned for each analyte by infusing a 1.0 p g / d standard solution (at 10 pUrnin, using an infusion pump) via a "T" into a stream of mobile phase containing 50% methanol and 50% 2mM ammonium acetate in water at 0.2 a m i n flow rate. Each analyte is initially tuned for the parent ion and then tuned for the product ion. Once the instrument is tuned, the optimized parameters are saved as a tune file. This tune file is then used during routine analysis. 4.5.2. Calibration Curve Procedures a. Inject the same aliquot (between 10 to 50 pL) of each calibration standard . (ranging from the lowest level standard to the highest level prepared), into the LC/MS/MS. b. Use weighted linear standard curves for quantitation. Linear standard curves are generated for each analyte by linear regression using Ilx weighting of peak area versus calibration standard concentration using Masslpx (or equivalent) software system. Any calibrationstandardfoundto be a statistical outlier by using an appropriate outlier test, may be excluded from the calculation of the calibration curve. However, the total number of calibration standards that may be excluded must not exceed 20% of the total number of standards injected, c. The correlation coefficient (R) for calibration curves generated must be 20.9925 (R2 10.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standardsor the relevant set of samples should be reanalyzed. Fxygen Rcsearch Page 15 of 43 Exygen Research Page 125 of 153 418-028:PAGE G-126 Exygen Study No.:023-072 ExygenMethod No:ExM-023-071Revision 1 Typical calibration curves for PFHS, PFOS and PFOA can be found in Figures 1- 3. 4.5.3 Sample Analysis a. Inject the same aliquot (between I0 to 50 pL) of each standard, sample, recovery, control, etc. into the LC/MS/MS system. b. Standards comsponding to at least five or mom concentrationlevels (starting with the LOQ level or below) must bc included in an analytical set. c. An entire set of calibration standards should be injected at the beginning of a set followed by calibration standards interspersed approximately every 5-10 simples (to account for a second set of extracted standards). As an alternative, an entire set of calibration standards may be included at the beginning and at the end of a sample set. In either case, calibration standards must be the first and last injection in a sample set. d. The concentration of each sampk/fortification/controJ is determined from the standard curve, based on the peak area of each analyte. The standard responses should bracket responses of the residue found in each sample set. Results may be quantitated up to 10% outside the e w e by extrapolation. If necessary, dilute the samples to give a response within the standard curve range. e. Fortification recoveries falling within 70 to 130% are considered acceptable. f, Samples must be stored refrigerated between 2C to 6C until analysis. g. Samples in which either no peaks are detected or peaks less than the lowest concentration of the calibration standarch are detected at the comsponding analyte retention time will be reported as ND (not detected). Samples in which peaks are detected at the corresponding analyte retention time that ace lessthan the LOQ and greater than or qua1to the lowest concentration of the calibration standards will be reported as NQ (not quantifiable). The analysisperformed during the method development included fortifications at 10 and 50 ng/g of PFHS in rat liver, 10 and 50 ng/mL ofPFHS in serum,10 and 50 ng/g of PFOS and PFOA in rat liver and 10 and 50 ng/mL of PFOS and PFOA in serumand urine. Typical chromatogramscan be found in Figures 4-39. Exygen -arch Page 16 of 43 Exygen Research Page 126 of 153 4 1 8-028:PAGE G- 127 Exygen Study No.:023-072 Exygeo MetbodNo:ExM-023471 Revision 1 4.6. ACCEPT~CECRITERU The following criteria must be met to ensure the presence of PPHS,PFOS and PFOA 1. The chtomatogram must show a peak of a daughter ion at 80 amu froma pmnt of 399 m u for PFHS,a daughter ion at 99 m u froma pmnt of 499 m u for PFOS, and a daughter ion at 369 m u from a parent of 413 m u for PFOA. 2. Method blanks must not contain analytc at levels greater than the LOQ. If a blank contains the analyte at levels greater than 10 ng/mL, then a new blank sample must be obtained and the entire set must be m-extracted. 3. Recoveries of control spikes and matrix spikes (if any) must be between 70-130% of their known values. If a control spike falls outside the ' acceptable limits, the entire set of samples should be re-extracted. Any matrix spike outside 70-13096 should be evaluated by the analyst to determineif reextraction is warranted. 4. Any calibration standard found to be a statisticaloutlierby using the Huge Error Test, may be excluded from the calculation of the calibration curve. However, the total number of calibration standards that could be excluded must not exceed 20% of the total number of standardsinjected. 5. The correlation coefficient (R) for calibration curves generated must be 10.9925 (R* 29.985). if calibraion results fall outside these lirnita, then appropriate steps must be taken to adjust instrument operation, and the standardsor the relevant set of samplesshould be reanalyzed. 6. Retention times between standards and samples must not drift more than f 4 % within M analytical run. If retention time dtift exceeds this limjt within an analytical run then the set must be reanalyzed. 4.7. PERFoRMANcEcRITERlA The following two criteria must be performed once as a system suitability test, before the commencement of analysis, when using an instrumentation set-up that has not been used for this method. First Crite&Sx Run a standard solution on tc/Ms/Ms corresponding to the estimated LOQ (10 ng/mt) in matrix and obtain a signal to noise ratio for the analyte transition of at least 91, compared to a reagent blank. Ifthis criterion cannot be met, optimize and change instrument operating parameters (or increase the injection volume, if appropriate). Exygen Research Page I7 of 43 Exygen Research Page 127 of 153 418-028:PAGE G-128 Exygen StudyNo.:023-072 ExygenMethod No:ExM-023-071Revision 1 second Criterion: Run a set of standards of five or mom concentration levels, from at or below the LOQ, up to the highest concentration level to be included in the analysis. Generate a calibration curve for the analyte and obtain a linear regression with a coefficient of determination (R') of at least 0.985 for the analyte. Once this criterionis met, samplesm a y be analyzedwith standardsinterspersed. 4.8. TIME REQUIREDFOR ANALYSIS One person can take a set of 20 samples through the sample preparation procedure in approximately 4 hours. The Lc/Ms/MS analysis of the set (containing20 field samples, 1 matrix blank, 2 laboratory control spikes, 1 matrix spike and 12 standardinjections)will take approximately 14 hours. 5. CALCUT?ONS a. Use Equation 1 to calculate the amount of a n a l 9 found (in ng/mL, based on peak area) using the standard curve (llx weighted linear regression parameters)generated by the MassIynx softwareprogram. Eauation 1: - Andyte found (ng/mL) = Ibeak m a intercerq) x DF x aliquot factor slope DF = factor by which the final volume was diluted, if necessary. Aliquotfactor=10for liver, 20 orserum and urine b. For samples fortified with known amounts of analyte prior to extraction, use Equation 2 to calculatethe percent recovery. Eauation 2 Recovery (9%)= [ total analytefound (ng/mL) - analytefoundin control (ng/mL)] xloo analyteadded (ng/mL) Note: Subtract analytc found in control (nglmL) from analyte found (ng/mL), if ng/mL, in control is greater than LOQ. Exygen Research -. . .-- Page 18 of 43 Exygen Research Page 128 of 153 418-028:PAGE G-129 Exygen Study No.:023-072 ExygenMethod No:ExM-023-071Revision 1 c, Use Equation 3 to calculatethe amount of analyte found (in ppb) Eauation 3: Analyte found (ndg or ng/mL)= Wytefound Cn&&\ x M CmL] sample weight (9) or sample volume (mL) FV = final volume For reporting purposes, samples in which either no peaks rn detected or peaks less than the lowest concentration of the calibration stantlards are detected at the comsponding analyte retention time will be reported as ND (not detected). Samples in which peaks are detected at the corresponding analyte retention time that are less than the LOQ and greater than or equal to the lowest concentratiou of the calibrationstandardswill be reposed as NQ (not quantifiable). 6. SAEETY The analyst should read the material safety data sheets for all standards and reagents before performing this method. Use universal precautions when handling standards and reagents, including working in fume hoods and wearing laboratorycoats, safety glasses, and gloves. Exygen Research Exygen Research Page 19 of 43 . . Page 129 of 153 418-028:PAGE G-130 Exygen Study No.: 023-072 ExygenMethd No:ExM-023-071Revision 1 Table 1. Recovery Summary of PFHS in Rat Liver and Serum Recovery Summaryof PFHS In Rat Liver Sample ID 0201684Spk A 0201684 Spk B Analyte Added (ndg) 10 10 10 Average: Standard Devbtlon: Percent Recovery (9b) 108 110 126 I 115 9.9 Sample ID 0201684 Spk D 0201684 Spk E 0201684 Spk F Analyte Added (ndg) 50 50 50 Average: Standard Deviation: Percent Recovery (%) i 101 98 94 98 3.5 Recovery Summary of PFHS in Rat Serum Sample ID Analyte Addbd (nglmL) Percent Recovery (%) 0201682Spk10A 1i2 r n 0201692Spk 8 10 1 03 0201682 Spk c 10 110 108 Standard Devlatlon: 4.7 Sample ID 0201682 Sok E 0201682Spk F Analyte Added (ndmL) 5.0. 50 Average: Standard Deviatlon: Percent Recovery (%) 107 104 122 i 111 9.6 Exygen Research Page 20 of 43 Page 130 of 153 4 18-028:PAGEG- 131 Exygen Study No.: 023-072 ExygenMethod No:ExM-023-071Revision I Table 2. Recovery Summary of PFOS in Rat Liver, Serum and Urine Recovery Summary of PFOS In Rat Uver Sample ID 0201684 Spk A 0201684 Spk B A n a l p Added wg) 10 10 Percent Remvery (%J88 ae Sample ID 0201684 Spk D 0201684Spk E 0201684 Spk F Standard Deviation: Analyte Ad 50 AVOrpg.: Standard Oevkffon: 8.5 am 87 c 8_1_1 1.5 Recovery Summary of PFOS In Rat Serum Sample ID Analyk Added(npmL) Percent Recovery (%) 0201682 Spk A 10 80 0201682 Spk B l.a- 0e9e AWHIO.: 0201682&k C 10 89 88 Standard DeviaUon: 9.8 0201682 Spk E 0201682 Spk F - 50 50 Average: Standard Devlatlon: 118 122 121 120 d 2.1 RecoverySummary of PFOS In Rat Urine Sample ID 0201682 Sok B 2201682 Spk C Analyte Added (nghL 1i.0n- lo Standard Daviati& Percent Recovery(%) i 9..,8. 4.7 Sample ID 0201682 S p 0201682Spk E 0201682 Spk F Analyte Added (ng/mL) v 50 50 50 Avense: Standard Devlatlgn: Percent Recovery- 80 78 78 7_a - 1.2 - - -.--- Exygen Research - I _ _ - Page21 of43 Exygen Research Page 131 of 153 418-028:PAGE G-132 Exygen Study No.: 023-072 ExygenMethod No:ExM-023-07I Revision 1 Table 3. Recovery Summary of PFOA in Rat Liver, Serum and Urine Recovery Summary of PFOA In Rat Liver 101 ~ Sample ID - 020 0201684 Spk E 0201684 Spk F Standard Devlatlon: Analyte Added (nglg) 50 50 50 Average: Standard Devhtlon: 3.1 - 97 92 94 94 2.5 Recovery Summary of PFOA In Rat Serum 118 0201662 Spk E 10 117 5 0201882 Spk C 10 115 Standard Deviation: 11137 0201682 Spk E 0201682 Spk F --c -7 107 50 112 60 Avenge: 116 Standard Deviatlon: 4.0 Recovery Summary of PFOA in Rat Urine Sample ID 0201682 Spk A 0201682 Spk B 0201682 Spk C 10 10 10 Average: Stendad Dsvlatlon: Percent Recovery(%) 88 91 89 as 2.5 Sample ID 0201682 Spk D 0201682 Spk E 0201682 Spk F Analyts Added (ndmL) 50 50 50 Average: Standard Omviati&: Percent R e w v e r y L 89 EB 88-75_ 2.1 .fiygen Research -.---- ... . .._ Page 22 of 43 Exygen Research Page 132 of 153 4 18-028:PAGE G-133 Exygen Study No.: 023-072 ExygenMethadNo:ExM-023-071Revision I Figure 1. Calibration Curve for PFaS Mpound 1 name:PFHS CW4llkbrahtlatnoc(uD~vco&:1s9im16i.m28lm* x-0+.89W.1O41S34 Rasponrelype:Memal Sfd,Ares Cumtype: Unear,O@I: Erelude,We&- 1s tnns:None 9.W- f Exygen Research Exygen Research Page 23 of 43 Page 133 of 153 418-028:PAGE G-134 Exygen StudyNo.:023-072 ExygenMethad No:ExM-023-071Revision 1 Figure 2. Calibration Curve for PFOS Compound 2 name: PFOS Coeffient of Determination:0.997009 Calibration curve: 3211.30 x + 48.9285 Response type: External Std, Area C u m type: Linear, Origin: Exclude, Weighting: lk h i s trans: None 1.62e4- Response- Exygen Research -I_--.. Exygen Research Page 24 of 43 Page 134 of 153 4 18-028:PAGE G- 135 Exygen Study No.:023-072 ExygcnMethadNo:ExM-023-071Revision 1 3. CalibrationCurve for PFOA Compound 1 name: PFOA Coefficientof Determination:0.996045 Calibration curve: 31270.9 x + 3675.36 Response type: Wernal Std, Area Curve type: Linear, Origin: Emlude, Weighting: lk Ms trans: None Exygcn Research .. - Exygen Research Page 25 of 43 Page 135 of 153 418-028:PAGE G-136 Exygen StudyNo.:023-072 ExygenMethod No:ExM-023-071Revision 1 F'igure 4. Representative Chromatogram of a 0.1 ng/mL Standard Containing PF'HS Figure 5. Representative Chromatogram of a 0.1 ng/mL Standard Containing PFOS Exygen Research Page 26 of 43 Page 136 of 153 418-028:PAGE G-137 Exygen Study No.: 023-072 ExygenMethod No:ExM-022071Revision I Figure 6. Representative Chromatogram of a 0.1 ng(mL Standard Containing PFOA COPSard 0.1 nohLPFOA an8 PFOS 0 7 Q ~ 0Sm2(Mn. 2r2) ldo ouuI-3wIUrtR4a LVMSIIISli7 am MRM d 2 ChmW ES Wa- 4 143-. WArmS I Figure 7. Representative Chromatogram of a 0.5 ng/mL Standard Containing PFHS Exygen Research +-. . ._. Exygen Research Page 27 of 43 Page 137 of 153 418-028:PAGE G-138 Exygen StudyNo.:023-072 ExygenMethodNo:ExM-023-071Revision I Figure 8. Representative Chromatogram of 8 0.5 ng/mL Standard Containing PFOS GO52502403npm* WOA ~ nPdFOS WuMDW 19:W:tT LcmWMsn YRMdZ chmuI*1p.4E>18m54 h . i . . . Figure 9. Representative Chromatogramof a 0.5 ng/mL Standard Containing PFOA Exygen Research Exygen Research Page 28 of 43 Page 138 of 153 4 18-028:PAGE G- 139 Exygen Study No.:023-072 Exygm Method No:ExM-023-071Revision 1 Figure 10. Representative Chromatogram of a 5.0 n g / d Standard Containing PFXS 0062lOZ-1, MnplmlPFHS oQ1ouI.IoDan m,=) L17 Bt10-4 Z U U M O m 1%61:21 LmSMSn Wdluun3*ImE*Ilm. 1.A1W N Figure 11. Representative Chromatogramof a 5.0 ng/& Containing PFOS COC1SOO-1,5.0nghnL PFOAudPFOS Standard Exygen Research Page 29 of 43 Page 139 of 153 418-028:PAGE G-140 Exygen Study No.:023-072 ExygenMethod No:ExM-023-071Revision 1 Figure 12. RepresentativeChromatogramof a 5.0 nghL Standard Containing PFOA Figure 13. Representative Chromatogram of a Reagent Blank Sample Analyzed for PFHS R4lg.nt BlMk A `i` Exygen Research _- . _.- Exygen Research Page 30 of 43 Page 140 of 153 418-028:PAGEG-141 Exygen Study No.:023-072 ExygenMethodNo:ExM-023-071Revision 1 Figure 14. Representative Chromatogram of a Reagent Blank Sample Analyzed for PFOS Figure 15. Representative Chromatogramof a Reagent Blank Sample Analyzed for PFOA ExygenResearch . .. ...._. . ...-. _-.. Page 3 1 of 43 Exygen Research Page 141of 153 418-028:PAGE G-142 Exygen Study No.:023-072 ExygenMethod No:Exh4-023-071Revision 1 Figure 16. Representative Chromatogram of a Control Liver Sample Analyzed for PFHS figure 17. RepresentativeChromatogram of a Control Liver SampIe Analyzed for PFOS Exygen Research Exygen Research Page 32 of 43 .. Page 142of 153 418-028:PAGE G-143 Exygen Study No.:023-072 Exygen Method No: ExM-(n3-071 Revision I Figure 18. Representative Chromatogram of a Control Liver Sample Analyzed for PFOA F'igure 19. Representative Chromatogram of a Control Serum Sample Analyzed for PFHS I? T Exygcn Research Exygen Research Page 33 of43 Page 143 of 153 4 18-028:PAGE G- 144 Exygen Study No.: 023-072 ExygenMethod No:ExM-023-071Revision 1 Figure 20. Representative Chromatogram of a Control Serum Sample Analyzed for PFOS Figure 21. Representative Chromatogramof a Control Serum Sample Analyzed for PFOA Fxygen Research - ...... ..-- -..... . ... .... ... ..._ Exygen Research PagC 34 Of 43 -... Page 144 of 153 4 18-028:PAGE G-145 Exygen Study No.: 023-072 ExygenMethod No:ExM-023-071Revision 1 Figure 22. Representative Chromatogram of a Control Urine Sample Analyzed for PFOS Urlnslink A OT#IOu.l;W Sm(MI, Z@ '1 oQJul-#=* LCWSWSH MRM d 2 C h n n l h E6 499rW 1-. ku Figure 23. Representative Chromatogram of a Control Urine Sample Analyzed for PFOA Exygen Research Page 145 of 153 4 18-028:PAGE G- 146 Exygen Study No.: 023-072 ExygenMethod No: ExM423-071Revision 1 Figure 24. Representative Chromatogram of a Control Liver Sample Fortinedat 10 ng/g with PFHS =muuu8f8pk h 10ppb Figure 25. Representative Chromatogram of a Control Liver Sample Fortifiedat 10nglg with PFOS 1 ExygenResearch Exygen Research Page 36 of 43 ... . - __I__ Page 146 of 153 4 18-028:PAGE G- 147 Exygen StudyNo.:023-072 ExygenMethodNo:ExM-023-071Revision 1 Figure 26. Representative Chromatogram of a Control Liver Sample Fortitied at 10ng/g with PFOA Figure 27. Representative Chromatogram of a Control Liver Sample FortEed at SO ng/g with PFHS '9 Exygen Research ...-_. - . . . . . . . I . . . . . . . . . . . . . ..-- ...- .--., Page 37 of 43 ...... .--__.. -. . Exygen Research Page 147 of 153 418-028:PAGE G-148 Exygen Study No.: 023-072 ExygenMethod No:ExM-023-071Revision 1 Figure 28. Representative Chromatogram of a Control Liver Sample Fortifiedat 50 ng/g with PFOS Figure 29. Representative Chromatogram of a Control Liver Sample Fortified at 50 ng/g with PFOA Exygen Research ....--. -_..-.--- Exygen Research Page 148 of 153 418-028:PAGE G-149 Exygm StudyNO.:023-072 ExygenMethod No:ExM-023-071Revision 1 Figure 30. Representative Chromatogram of a Control Serum Sample Fortified at 10 n&/mLwith PFHS Figure 31. Representative Chromatogram of a Control Serum SampIe Fortified at 10 ng/mL with PFOS Exygen Research .-- .. - . .-. ...-_ .... Page 39 of 43 ...--.-... - . . Page 149 of 153 418-028:PAGEG-150 Exygen Study No.: 023-072 ExygcnMethodNo:ExM-023-071Revision 1 Figure 32. Representative Chromatogramof a Control SerumSample FortiFIed at 10 ng/mC with PFOA Figure 33. Representative Chromatogram of a Control Serum Sample Fortifiedat 50 ng/mL with PFHS Exygen Research Page 150 of 153 418-028:PAGE G-151 Exygen Study No.:023-072 ExypnMethod No:ExM-023.071Revision 1 Figure 34. RepresentativeChromatogrmof a ControlSerum Sample FortifiedatSO n#mL with PF'OS Figure 35. Representative Chromatogramof a Control Serum Sample Fortiffedat 50 n g / d with PFOA Exygen Research - ~ -- Page41 of 43 -- Exygin Research Page 151 of 153 4 18-028:PAGEG- 152 Exygen StudyNo.:023-072 Exygen Method No:EM-023-071Revision 1 F'igure 36. Representative Chromatogra3nof a Control UrineSample RortBedat 10ng/mL with PMlS Figure 37. Representative Chromatogramof a Control UrineSample Fortified at 10ng/mL with PF`OA Exygen Research .- .. .. _--- - - _..- Page42 of43 -. Exygen Research Page 152 of 153 4 18-028:PAGE G- 153 Exygen Study No.: 023-072 ExygenMethodNo:ExM-023471Revision 1 Figure 38. Representative Chromatogram of a Control Urine Sample FoMied at 50 ng/mL with PFOS Figure 39. Representative Chromatogram of a Control Urine Sample Fartifiedat 50 ng/mL with PFOA Exygen Research .--.- .... . . .... -..-_ ... I Page43 of43 -.- -- - Exygen Research Page 153 of 153 APPENDIX H TEMPERATURE AND RELATIVE HUMIDITY REPORTS ARGUS 418-028:PAGE H-1 Temperature and Relative Humidity Report Location: Room 05 Protocol Number: 418-028 Range of Dates: 26-Mar12002 1420 to IOJun-2002 O W 9 Target Range: Species: Rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Mean (fSD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low rh): Temperature 64F to 79F 77 181 8.25 1817 69.2 71 .O 69.2 66.0 (A 0.7) Relative Humidity 30% to 70% 1 1 1 77 1 18 1 8.25 4817 11 54.6 (k4.4) 1, 1 64.5 55.3 36.4 1817 0 0 (100.0) (0.0) (0.0) Report Generated: 10-Jun-2002at 1326 COMMENTS: APPENDIX I POSITIVE CONTROL DATA 4 18-028:PAGE I- 1 Historical Control Data This Functional Observation Battery Standard Operating Procedure and Studies conducted to document the training and competency of the technical staff and Motor Activity Negative Control Data and Positive Control Data are available at the Testing Facility. Page 1 418-028:PAGE 1-2 Summary Information for Functional Observati Study Number - Title 012-006-Validationof Fmctional ObservationalBattery and MotorActivity Measure Using Positive Test Substances 012-014-NeurotoxicityEvaluation of Positive Control Substances in Crl:CD@BRVAFIPlusB Rats 012-015 - Neurotoxi&y Evaluationof DDT in Crl:CD@BR VAF/Plus@ Rats In-Life start 12/89 919 1 3/92 Test Substance acrylamide physostigmiae DDT acryhnide IDPN carbaryl DDT triadiinefon DDT Dosage Information mg/kg Number of mUkg Dosages 50 1 7 0.75 I .5 1 75 1 1 40 1 9 200 1 3 75 5 1 75 5 1 200 5 1 75 1 1 012-017 -NeurotoxicityEvaluation of PositiveControl Substancesin Crl:CD@BR VAF/PlusB Rats 5/92 acrylamide 40 1 9 JDPN 200 1 3. cfirbaryl 40 5 1 DDT 75 1 1 d-amphetamine 4.0 1 1 012-022- Neuotoxiuty Evaluation of Carbaryl hCrl:CD@BRVAF/plus@Rats 10192 carbaryl 40,200 5 1 012-03 1 - NeurotoxiciQEvaluationof PositiveControl Substancesin Crl:CD@BRVAFiPlus@ Rats 7/93 acrylamide 45 1 10 IDPN 250 1 4 carbaryl 40 5 1 DDT 75 1 1 d-amphetamine 4 1 1 Page 2 012-056-Neurotoxicity Evaluation of Positive Control Substances in Crl:CD@BRVAF/Plus@Rats 012-075 -Neurotoxicity Evaluation of Positive Control Substances in Crl:CD@BRV A F ~ l u s BRats 012-081 - Neurotoxicity Evaluation of Positive Control Substances in Crl:CD@BR VAF/plusBRats 418-028:PAGE 1-3 11/95 acrylamide 45 1 10 DPN 250 1 5 carbaryl 40 1 1 DDT 75 1 1 d-amphetamine 4.0 1 1 3/98 acsylamide 30 1 17 trimethyltin 8 1 1 MK-80I 0.3 1 1 CarbaI-JIl 100 4 1 DM 100 2 1 11/01 acrylamide 30 1 10 IDPN 250 1 1 d-amphetamine 40 1 1 CaTbaryl 100 4 1 DDT 100 2 5 Page 3 418-028:PAGE 1-4 Summary Informationfor Motor Activity Study - Title In-Life Start 012-011 - The Assessmentof Motor Activity in Neonatal and Adult Rodents using Passiveh h r e d Sensors 519 1 012414- NeurotoxiCityEvaluation of PositiveControlSubstancesin CrlCD@BRVAF/Plus@Rats 919 1 012-016-Motor Activity Evaluation in CrlCDOBR VAF/Plm@ Rats AdministeredChlorpromazineand dAmphetamine (PositiveControl Study) 012-058 - Neurotoxiuty Evaluation of Positive Control Substances in Crl:CD@BR VAFiPlusB Rats 4/96 Test Substance Dosage Wormation Number of mgkg mUkg Dosages d-amphetmhe 0.75, 1.5,4 1 I chlorpromazine 1,294 1 1 acrylamide 40 1 9 IDPN ' 200 1 3 carbaryl 75 5 1 DDT 75 5 1 triadimefon 200 5 1 d-hetamine 0.5, 1,4 1 1 chlorpromazhe 1,294 1 1 acrylamide 45 1 10 d-amphetamhe 0.75 1 1 trimeulyltin 8 1 1 MK-80 1 10 1 1 Page 4 APPENDIX J HISTOPATHOLOGY REPORT 418-028:PAGE J-1 RESEARCH PATHOLOGY SERVICES. INC. 438 East Butler Avenue, New Britain, PA 18901 Phone: 215-345-7070 Fax: 215-345-4326 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCEENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706. I HISTOPATHOLOGY REPORT SUBMITTED TO: Raymond G. York, Ph.D., D.A.B.T. Argus Research 905 Sheehy Drive Horsham, PA 19044 SUBMITTED BY: W. day Brown, D.V.M., PdD.' Veterinary PathoIogist July 3, 2003 418-028:PAGE J-2 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTlON/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT TABLE OF CONTENTS REPORT Page Method ....................................................................................................................... I Results ....................................................................................................................... 3 Summary.................................................................................................................... 5 Quality Assurance Unit Statement ...............................................,............,................6 Good Laboratory Practice Compliance Statement .....................................................7 TABLE 1. Incidenceand Degree of Severity of Histomorphologic Observations.................... 8 APPENDICES I. Histomorphologic Observations ...............................................................1-1to 1-12 Key to Histomorphologic Observations.............................................................,.I-1 Tables 1-1. Histomorphologic Observations - Group I Male Rats ............................... 1-2 1-2. HistomorphologicObservations - Group II Male Rats .............................. 1-4 1-3. HistomorphologicObservations - Group 111 Male Rats ............................. 1-5 1-4. Histomorphologic Observations - Group IV Male Rats............................. 1-6 1-5. Histomorphologic Observations - Group V Male Rats.............................. 1-7 1-6. HistomorphologicObservations - Group I Female Rats........................... 1-9 1-7. Histomorphologic Observations - Group V Female Rats........................ 1-1I II. IndividualAnimal Gross and Histomorphology Data.............................. 11-1 to 11-70 Ill. Histomorphologic Observationsin the Liver - F I Generation Pups .......111-1to 111-2 Tables 111-1. Histomorphologic Observations in the Liver - Group I - F1 Generation Pups............................................................................... 111-2 111-2. Histomorphologic Observations in the Liver - Group V - F I Generation Pups ............................................................................... 111-3 418-028:PAGE J-3 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTlON/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT METHOD Microscopic examination was made of the specified tissues from 50 adult male and 20 adult female Crl:CD@(SD)IGSBR VAF/PIUS@rats from five groups in an oral (gavage) combined repeated dose toxicity study of T-7706 (PFHS) with the reproduction/developmentaltoxicity screening test. In addition, microscopic examination was made of the liver of I 0 pups in each of the control and high dosage groups of the F I Generation. A brief outline of the study design showing the dose group identification, number of rats per sex per group, and dosage levels of the control and test substances are shown below. DOSAGE GROUP I II Ill IV V NUMBER OF RATS PER SEXa 15 + 3' 15 + 3' 15 + 3' 15 + 3' 15 + 3' DOSAGE (mg/kg/day) 0 0.3 1 3 10 CONCENTRATION^ (mg/mL) 0 0.03 0.1 0.3 1 DOSE VOLUME (mUkg) 10 10 10 10 I0 Male rats were given the test substance once daily beginning 14 days before a cohabitation period that consisted of a maximum of 14 days. Dosing continued through the day before sacrifice, after completion of a cohabitation period, after a minimum of 42 days of administration. Female rats were given the test substance once daily beginning 14 days before a cohabitation period that consisted of a maximum of 14 days. Dosing continued through the day before scheduled sacrifice (Day 22 of lactation). Dosages were adjusted daily for body weight changes and were given at approximately the same time each day. The first day of dosing was desig- Research Pathology Services, Inc. -1 418-028:PAGE 5-4 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT nated as Day 1 of the study. Pups were sacrificed and necropsied on Day 22 postpartum. All rats were necropsied and the specified tissues were collected and placed in 10% neutral buffered formalin for fixation. The testes were fixed in Bouin's solution for 48 to 96 hours and then retained in 10% neutral buffered formalin. The in-life portion of the study, necropsies, and recording of the gross necropsy observations were performed by the staff of Argus Research, Horsham, PA. The tissue processing, microscopic slide preparation and histopathologic evaluation were performed by Research Pathology Services, Inc. The tissues specified for microscopic evaluation from 10 male and 10 female rats of Groups I and V included: brain, duodenum, jejunum, ileum, cecum, colon, rectum, Peyer's patch, lung, submandibular and mediastinal lymph nodes, sciatic nerve, stomach, kidneys, spleen, thymus, trachea, urinary bladder, testes, epididymides, seminal vesicles, coagulating gland, prostate, spinal cord (cervical, lumbar and thoracic), liver, adrenal glands, heart, thyroid, parathyroid, uterus, bone marrow (sternum), ovaries, uterus, vagina, mammary gland (female rats) and all other tissues with gross changes. In addition, the liver and thyroid of 10 male and 10 female rats in each of the intermediate dosage groups were examined. The liver of 9 or I 0 pups F1 generation pups from each of the I 0 selected control and high dosage female rats were also examined. Representative samples of these tissues were routinely processed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin for microscopic evaluation. In addition, sections of the testes from the control and high dosage group male rats were stained with periodic acid-Schiff (PAS) reaction and examined. The study was initiated on June 24, 2002 and completed on July 3, 2003. Upon completion of the project, all raw data (remaining wet tissue, paraffin blocks, microscopic slides and histology records) will be returned to Argus Research for archiving. Research Pathology Services, Inc. -2 418-028:PAGE J-5 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTlON/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT RESULTS The type, incidence and degree of severity of the histomorphologic changes in the specified tissues for the male and female rats are presented in Table I.The microscopic observations in each rat are summarized in tabular form in Appendix I (Tables 1-1 to 1-7). A key to the histomorphologic observations precedes Table 1-1. The gross necropsy observations, detailed descriptions of the microscopic observations, and a correlation of the microscopic findings with the gross changes in these rats, when applicable, are contained in Appendix II. The histomorphologic observations in the liver of the F1 Generation pups of the control and high dosage groups are summarized in tabular form in Appendix 111 (Tables 111-1 and 111-2). A key to the histomorphologic observations is included on Tables 111-1 and 111-2. No treatment-related microscopic changes were observed in any of the male rats given 0.3 or 1 mg/kg/day of the test substance or in female rats given I 0 mg/kg/day of T-7706. Treatment-related microscopic changes were observed in the liver and thyroid gland of male rats of the 3 and 10 mg/kg/day dosage groups. The treatment-related microscopic change in the liver consisted of minimal to moderate enlargement (hypertrophy) of centrilobular hepatocytes (Table 1). The affected hepatocytes were enlarged with an increased amount of dense eosinophilic granular cytoplasm. The treatment-related effect in the thyroid gland consisted of an increased incidence of male rats of the 3 and 10 mg/kg/day dosage groups with hypertrophy (enlargement) of follicular cells and hyperplasia (increased follicular cells and small follicles) in male rats (Table I ) . Although the incidence in Group IV was minimally increased over the controls, the hypertrophy and hyperplasia in this group of rats could have been associated with the liver-cell changes. These microscopic changes in the liver and thyroid are consistent with the known effects of compounds that cause microsomal enzyme induction where the hepatocellular hypertrophy results in a compensatory hypertrophy and hyperplasia of Research Pathology Services, Inc. -3 418-028:PAGE J-6 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT the thyroid due to increased plasma turnover of thyroxine and an associated stimulation of thyroid-stimulating hormone in rats.' There were no compound-related microscopic changes observed in the liver of the F1 generation pups from the dams given I 0 mg/kg/day of the test substance. There were a few other microscopic changes observed in the various organs and tissues which were considered to have occurred spontaneously and to be incidental and unrelated to compound administration. The type, incidence and severity of these changes were not influenced by compound administration. These changes also are listed in the attached histomorphology tables. 1 Sanders, J.E., Eigenberg, D.A., Bracht, L..J., Wang, W.R., and van Zwieten, M.J., Thyroid and Liver Trophic Changes in Rats Secondary to Liver Microsomal Enzyme Induction Caused by an Experimental Leukotriene Antagonist (L-649,923), Toxicology and Pharmacology 95, 378-387 (1988) Research Pathology Services, Inc. -4 418-028:PAGE J-7 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTlON/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REP0RT SUMMARY Microscopic examination was made of the specified tissues from five groups of male and female Crl:CD@(SD)IGSBR VAF/PIUS@rats used in an oral (gavage) combined repeated dose toxicity study of T-7706 with the reproduction/developmental toxicity screening test. The five groups of rats had been given the vehicle (aqueous 0.5% carboxymethylcellulose), or 0.3, 1, 3 or I 0 mg/kg/day of T-7706, orally by gavage, once daily for the protocol-specified number of days. In addition, the liver was examined microscopically from 9 or I 0 pups of the F I generation from 10 dams each of the control and high dosage groups. No treatment-related microscopic changes were observed in female rats given I 0 mg/kg/day of T-7706 or in male rats given 0.3 or Img/kg/day of T-7706. Treatment-related microscopic changes were observed in the liver and thyroid of male rats of the 3 and I 0 mg/kg/day dosage groups. The treatment-related change in the liver consisted of minimal to moderate hypertrophy of centrilobular hepatocytes and the effect in the thyroid was an increased incidence of male rats with a compensatory hypertrophy and hyperplasia of the follicular epithelium in these groups of male rats. No treatment-related microscopic changes were observed in the liver of the F I generation pups from dams given 10 mg/kg/day of T-7706. All other microscopic changes were considered to be spontaneous in origin and not treatment-related. The type, incidence or severity of these changes were not considered to be influenced by administration of the test substance. Research Pathology Services, Inc. -5 418-028:PAGE J-8 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATH0LOGY REPORT QUALITY ASSURANCE UNIT STATEMENT All aspects of the tissue processing, microscopic slide preparation, histopathologic evaluation and report preparation for the study listed above have been performed according to the Standard Operating Procedures of Research Pathology Services, Inc. and were audited in accordance with the procedures established by the Quality Assurance Unit of Research Pathology Services, Inc. in compliance with the Good Laboratory Practice regulations specified in the protocol. Organisationfor Economic Cooperation and Development (1996). OECD Guideline for Testingof Chemicals.Section 4, No. 422: Combined Repeated Dose Toxicity Study with the ReproductionlDevelopmentalToxicity Screening Test, adopted 22 March 1996. Organisationfor EconomicCooperation and Development (1998). The RevisedOECD Principlesof Good Laboratory Practices [C(97)186/Final]. US Food and Drug Administration. Good Laboratory PracticeRegulations:Final Rule.21 CFR Part 58. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26,1997. Quality assurance unit study-basedinspectionswere performed as shown below. There were no devia- tions from the protocol, standard operating procedures and/or appropriate good laboratory practice regulations noted during the conduct of the study. The summary report of QA inspectionsis includedin the final report sub- mitted to the study director on July 3, 2003. Dates of Dates Reported to Date Reported Inspection Study Phase Management to Study Director 12/16/02 12/16/02 01/07/03 01/09/03 01/11/03 01/ I3/03 01/13/03 01/3 1/03 01/31/03 02121/03 02/24/03 02/26/03 02/27/03 02/28/03 07/03/03 Master Schedule Pre-initiation Trimming Embedding Microtomy Staining Organization& Review Histopathology Data Entry Data Verification Data Processing Report Preparation Pre-submissionAudit Draft report Final report I2/31/02 12131/02 01/30/03 01/31/03 01/31103 01131/03 01/27/03 01131/03 02/27/03 02127/03 02/27/03 02/27/03 02/27/03 02/28/03 07/03/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 02/28/03 07/03/03 Research Pathology Services, Inc. Kared W. Harkins, BS Quality Assurance Unit Date -6 418-028:PAGE J-9 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT All aspects of the above-referencedstudy performed by Research Pathology Services, Inc. were conducted according to: Organisation for EconomicCooperationand Development (1996). OECD Guidelinefor Tesfingof Chemicals.Section 4, No. 422: Combined Repeated Dose Toxicity Study with the ReproductionlDevelopmental Toxicity Screening Test, adopted 22 March 1996. Organisation for Economic Cooperation and Development (1998). The Revised OECD Principlesof Good Laboratory Practices [C(97)186/Final]. US Food and Drug Administration. Good Laboratory Practice Regulations: Final Rule. 21 CFR Part 58. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26,1997. No deviations were noted that had any significant impact on the validity of the study. Research Pathology Services, Inc. -7 418-028:PAGE J-10 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418 - 028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1 Incidence and Degree o f Severity o f H i stomorphologic Observations Dose Group: Sex: Number o f Animal s/Group: I I 1 I11 I V v MMMMM 10 10 10 10 10 ADRENAL GLANDS : NO. EXAMINED NO. NORMAL 10 0 0 0 10 70009 -degeneration, c y s t i c , cortex, mu1tifocal mild COl [Ol COl LO1 COl 00000 - hypertrophy/vacuolation, cortex, mu1ti focal mild r01 COI COl COl COl 00000 - necrosis , cortex, focal mild [Ol COl COI COI COl 00000 -vacuolation, cortex, diffuse m inimal mild 131 Col C01 Col L-11 20000 10001 - vacuol a t ion, zona g lomerulosa mild COl COl COl COl COl 00000 BONE MARROW (STERNUM) : NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 NO. NORMAL CECUM: NO. EXAMINED NO. NORMAL -inflammation, mucosa, chronic mild moderate COAGULATING GLAND: NO. EXAMINED NO. NORMAL COLON : NO. EXAMINED NO. NORMAL -inflammation, mucosa, chronic mild DUODENUM: NO. EXAMINED NO. NORMAL EPIDIDYMIDES : NO. EXAMINED NO. NORMAL I: 3 = Total incidence of specified lesion, a l l grades. 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 80009 121 COI COl COl C11 00001 20000 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 900010 C11 COl COl COl COl 10000 10 0 0 0 10 10 0 0 0 10 10 0 0 1 10 60009 I V FF 10 10 10 10 87 [Ol C11 01 COl C11 01 COl 111 01 [OI COl 00 00 L-21 COl 20 10 10 10 10 10 10 10 10 10 10 8 10 C21 COl 10 10 10 10 10 10 COl COl 00 10 10 10 10 -8 418-028:PAGE J-11 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING.TES~- PROTOCOL NUMBER 418- 028 SPONSOR'S STUDY NUMBER T - 7706.1 TABLE 1 (Continued) Incidence and Degree o f Severity o f H istomorphologic Observations Dose Group: Sex: Number o f Anima1s/Group : I I1 I11 I V v MMMMM 10 10 10 10 10 EPIDIDYMIDES (Continued) : - exfol iated spermatogeni c c e l l s moderate C01 COl COl C11 COI 00010 - hypospermia marked COl COl C01 C11 COI 0 0 0 10 - i n f i l t r a t i o n , mononuclear-cell, focal minimal c41 coi coi coi c11 40001 HEART : NO. EXAMINED NO. NORMAL 10 0 0 0 10 80009 - in f l ammati on, chronic, focal mini ma1 mild P I COl COl COl C11 10001 10000 ILEUM : NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 JEJUNUM : NO. EXAMINED NO. NORMAL - d iv e r t icul um 10 0 0 0 10 100 0 0 9 00001 NO. NORMAL -basophilia/degeneration, cortical tubules, focal /mu1 tifocal m inimal -cyst(s), medulla - d i1atation, pel v i s m inimal mild - i n f i l t r a t i o n , mononuclear-cell, focal/mul tifocal m inimal -mineralization, mu1tifocal m i n ima1 10 3 0 0 10 71007 COl C01 COl c01 Ell 00001 10002 01000 C21 C01 C01 COl c21 20002 COI COI 0 0 I V FF 10 10 10 10 10 10 COl COl 00 00 10 10 10 10 10 10 10 10 00 10 10 98 COI COl 00 00 00 [Ol COl 00 [ 1 = Total incidence of specified lesion, a l l grades. -9 418-028:PAGE J-12 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418 - 028 SPONSOR S STUDY NUMBER T-7706.1 TABLE 1 (Continued) Incidence and Degree o f Severity o f H i stomorphologic Observations Dose Group: Sex: Number o f Animal s/Group: I I 1 I11 I V v MMMMM 10 10 10 10 10 KIDNEYS (Continued) : - nephritis, chronic, focal m i n ima1 COl c01 101 COl C11 0 0 0 0 1 LIVER: NO. EXAMINED NO. NORMAL 10 10 10 10 10 232 0 0 -hypertrophy, hepatocellular m i n ima1 mild moderate c e n t r i1obul a r -inflammation, chron c, foca /mu1 tifocal m i n ima1 mild - 1ip idosi s, tension, focal -necrosis , focal m inimal COl COl COl C9I ClOl 0 0 0 84 0 0 0 15 0 0 0 0 1 c C81 C61 5 4 3 2 71 C6l ~ 5 1 7 6 5 0 0 0 0 2 0 0 0 COl COl COI COl COl 0 0 0 0 0 - vacuol a t i on, hepatocellu la r , m idzonal mild COl COl C11 COl COl 0 0 10 0 -vacuolation, hepatocellular, multifocal m inimal mild COl C11 c11 COl c01 0 10 0 0 0 0 10 0 NO. NORMAL - i n f i l t r a t i o n , polymorphonuclear, perivascul ar/peri bronchial mild -inflammation, i n t e r s t i t i a l , focal minimal -macrophages, a1veol i, focal m i n ima1 LYMPH NODE, MEDIASTINAL: NO. EXAMINED NO. NORMAL -congestion m i n ima1 10 0 0 0 10 10 0 0 0 10 c01 COI C01 COl COl 0 0 0 0 0 COl LO1 COl COl COl 0 0 0 0 0 COl COl COl COl [Ol 0 0 0 0 0 8 0 0 0 10 7 0 0 0 8 COl c01 [Ol [Ol [Ol 0 0 0 0 0 I V FF 10 10 C11 C11 1 1 10 10 8 7 COI COl 0 0 0 0 0 0 C21 c 11 2 1 0 0 0 0 COl C21 0 2 COl COl 0 0 101 COl 0 0 0 0 10 10 9 9 C11 COl 1 0 C O l [11 0 1 COl C11 0 1 10 10 7 10 t-21 c01 2 0 I: 1 = Total incidence o f specified lesion, a l l grades. - 10 418-028:PAGEJ- 13 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1 (Continued) Incidence and Degree o f Severity o f H istomorphologi c Observations Dose Group: Sex: Number o f Animal s/Group: I I 1 I11 I V v MMMMM 10 10 10 10 10 LYMPH NODE, MEDIASTINAL (Continued) : - hyperplasia, lymphocytic/plasmacytic mild COI COl COl COl COl 0 0 0 0 0 -mastocytosis m inimal c11 COl COl COl C2l 10 0 0 2 LYMPH NODE, SUBMANDIBULAR: NO. EXAMINED NO. NORMAL 10 0 0 0 10 50 0 0 4 - hyperpl asi a, 1ymphocytic/pl asmacytic m inimal mild moderate ~ 5 1 COl COl COl C6l 10 0 0 2 2 0 0 0 0 2 0 0 0 4 MAMMARY GLAND : NO. EXAMINED NO. NORMAL -necrosis , focal m inimal NO. NORMAL OVARIES: NO. EXAMINED NO. NORMAL -cyst(s), intraovarian PARATHYROID : NO. EXAMINED NO. NORMAL PEYER 'S PATCH: NO. EXAMINED NO. NORMAL -mineralization, focal m inimal 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 9 COl COl COl COI r.11 0 0 0 0 1 NO. NORMAL 10 0 0 0 10 6 0 0 0 6 I V FF 10 10 111 [Ol 10 C11 COl 10 10 9 43 C6l 161 10 35 21 10 10 10 9 COl r11 01 10 9 10 9 10 10 9 10 10 10 10 10 10 10 10 10 10 COl COI 00 [ ] = Total incidence o f specified lesion, a l l grades. - 11 418-028:PAGE J- 14 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418.028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1 (Continued) Incidence and Degree o f Severity o f H istomorphologic Observations Dose Group: Sex: Number o f Animal s/Group: I I1 I11 I V v MMMMM 10 10 10 10 10 PROSTATE (Continued) : -atrophy, focal /mu1 tifocal m i n ima1 moderate c11 COl L-01 COl L-21 1 0001 00 0 0 1 -inflammation, chronic, focal/multifocal m in ima1 c31 coi c01 COl C31 30003 NO. NORMAL -inflammation, mucosa, chronic m i n ima1 mild SEMINAL VESICLES: NO. EXAMINED NO. NORMAL SKIN (GROSS LESION) : NO. EXAMINED NO. NORMAL -dermatitis, chronic, focal m i n ima1 mild SPINAL CORD, CERVICAL: NO. EXAMINED NO. NORMAL SPINAL CORD, LUMBAR : NO. EXAMINED NO. NORMAL SPINAL CORD, THORACIC: NO. EXAMINED NO. NORMAL 10 0 0 0 10 90009 c11 c01 c01 c01 c11 00001 1 00 00 10 0 0 0 10 10 0 0 0 10 00000 00 0 0 0 L-01 c01 L-01 L-01 L-01 00 0 0 0 00 0 0 0 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 NO. NORMAL -atrophy mild STOMACH : NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 L-01 co1 co1 [OI COl 00000 10 0 0 0 10 80 0 0 9 [ 3 = Total incidence o f specified lesion, a l l grades. I V FF 10 10 10 10 10 10 c01 c01 0 0 0 0 1 1 0 0 1-11 c11 0 1 1 0 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 9 101 e11 0 1 10 10 7 6 - 12 418-028:PAGE J-15 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1 (Continued) Incidence and Degree o f Severity of H i stomorphologic Observations Dose Group: Sex: Number o f Animal s/Group: I I1 I11 I V v MMMMM 10 10 10 10 10 STOMACH (Continued) : - d i1atation, mucosal glands m i n ima1 c11 c01 c01 r.01 co1 1 0000 -edema/infl ammation, submucosa, forestomacti mild el1 COl c01 COI c11 1 000 1 - edema/i n f lammation, submucosa, g l andul a r area m in ima1 mild moderate c21 COl I?] 1 00 00 0 1 00 c01 COl 0 0 0 0 0 0 -erosion(s), glandular mucosa moderate COl COl COl c01 c01 00000 TESTIS (H&E) : NO. EXAMINED NO. NORMAL 10 0 0 1 10 10 0 0 0 10 -degeneration, mult foca mild co1 COl c01 c11 c01 0 0 0 10 TESTIS (PAS) : NO. EXAMINED NO. NORMAL 10 0 0 1 10 10 0 0 0 10 -degeneration, mult foca mild co1 COl c01 [ll COl 0 0 0 10 THYMUS: NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 -atrophy moderate marked c01 c01 COl c01 c01 00000 00000 THYROID : NO. EXAMINED NO. NORMAL 10 10 10 10 10 53553 - hypertrophy/hyperplasia, f o l l i c u l a r epithelium minimal mild moderate C21 C31 c21 c41 c71 0 1 12 0 2 2 12 3 00004 - u ltimobranchial body/cyst 3442 2 TRACHEA : NO. EXAMINED NO. NORMAL 10 0 0 0 10 60009 [ ] = Total incidence o f specified lesion, a l l grades. I V FF 10 10 c11 c11 1 1 c01 c11 0 1 C21 C31 0 0 1 3 1 0 COl c11 0 1 10 10 9 9 c11 c11 1 0 0 1 10 10 5 5 c01 c01 0 0 0 0 0 0 5 5 10 10 10 9 - 13 418-028:PAGE J-16 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVE LOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418- 028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1 (Continued) Incidence and Degree o f Severity o f H istomorphologi c Observations Dose Group: Sex: Number o f Animal s/Group: I I 1 I11 I V v MMMMM 10 10 10 10 10 TRACHEA (Continued) : -inflammation, chronic, focal m i n ima1 mild C41 Col [OI Col C11 20000 20001 URINARY BLADDER: NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 -inflammation, chronic, focal minimal COl COl COl COl COl 00000 UTERUS : NO. EXAMINED NO. NORMAL - d i s t e n t i on, 1umen mild moderate - macrophages, pigmented m inimal mild moderate marked VAGINA: NO. EXAMINED NO. NORMAL I V FF 10 10 COl C11 01 00 9 10 a 10 C11 LO1 10 10 10 01 COI C21 01 01 ClOl C81 10 22 75 01 10 10 10 10 [ 3 = Total incidence o f specified lesion, a l l grades. - 14 418-028:PAGE J-17 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT APPENDIX I HISTOMORPHOLOGIC OBSERVATIONS 418-028:PAGE J-18 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTlON/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATH0LOGY REPORT KEY TO HISTOMORPHOLOGIC OBSERVATIONS No change (not remarkable, within normal histologic limits or indicated change not present). * Tissue not available (specified tissue missing, insufficient tissue in plane of section, artifact precludes evaluation, or specified tissue not present in section). <> Microscopic finding(s) in tissue(s) with gross observation(s). <-> Within normal limits [no microscopic change(s) to correlate with the gross observation(s)]. P Indicated change or lesion present I Minimal degree or amount of indicated change or lesion. 2 Mild degree or amount of indicated change or lesion. 3 Moderate degree or amount of indicated change or lesion. 4 Marked degree or amount of indicated change or lesion. ss Scheduled Sacrifice 1-1 418-028:PAGE J-19 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1-1 Histomorphologic Observations Dose Group: Animal Number: Sex : Death Type: ADRENAL GLANDS: -vacuolation, cortex, diffuse 1 1 1 1 1 1 1 1 1 1 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 M M M M M M M M M M ss ss ss ss ss ss ss ss ss ss BONE MARROW (STERNUM): BRAIN: CECUM: -inflammation, mucosa, chronic COAGULATING GLAND: - COLON : -inflammation, mucosa, chronic DUODENUM : EPIDIDYMIDES: - i n f i l t r a t i o n , mononuclear-cel , foca -HEART : inf 1ammation, chronic , focal ILEUM: JEJUNUM : KIDNEYS: -cyst (s) , medulla -in fi1t r a t ion, mononucl ear-cel 1 , f o c a l /mu1 t i focal LIVER: -inflammation, chronic, focal/mul tifocal 1 LYMPH NODE, MEDIASTINAL: -mastocytosis LYMPH NODE. SUBMANDIBULAR: -hyperpl asia, lymphocyti c/plasmacytic NERVE, SCIATIC: PARATHYROID : PEYER'S PATCH: PROSTATE : -atrophy, focal/mul tifocal 1 1-2 418-028:PAGE 5-20 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1-1 (Continued) H i stomorphol ogi c Observat ions Dose Group: Animal Number: Sex : Death Type: 1 1 1 1 1 1 1 1 1 1 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 M M M M M M M M M M ss ss ss ss ss ss ss ss ss ss PROSTATE (Continued): -inf 1ammation, chroni c , f o c a l /mu1 ti f o c a l 1- RECTUM : -inflammation, mucosa, chronic SEMINAL VESICLES: SPINAL CORD, CERVICAL: SPINAL CORD, LUMBAR: SPINAL CORD, THORACIC: SPLEEN : STOMACH : -di 1a t a t i on, mucosal -edema/inflammation, -ederna/i n f l ammation, area glands submucosa, submucosa , forestomach glandular 12- - 3 1- TESTIS (H&E) : T E S T I S (PAS): THYMUS : THYROID : -hypertrophy/hyperplasia, f o l l i c u l a r epi the1iurn -ul timobranchi a1 bodylcyst TRACHEA : -in f l ammation, chroni c , f o c a l - 2 - - 1 - 2 - 1- URINARY BLADDER: 1-3 4 18-028:PAGE J-2 1 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1-2 Hi stomorphologi c Observat i ons Dose Group: Animal Number: Sex : Death Type: KIDNEYS : -di 1 atation, pel vi s LIVER: -inflammation, chronic, focal/multifocal -1 ipi dosi s, tension , focal -vacuolati on, hepatocell ul ar , mu1 t i focal I1 I1 I1 I1 I1 I1 I1 I1 I1 I1 19102 19106 19108 19110 19113 19120 19129 19136 19138 19139 M M M M M M M M M M ss ss ss ss ss ss ss ss ss ss <-> <2> <1> 2 - 2 - 1 1 P - - - 1 P 1 - - - - - - - - - - - - -1- - THYROID : -hypertrophy/hyperpl asi a, foll icul ar epi the1 i um -ul timobranchi a1 body/cyst ---2- - 1 - - 2 PP-----PP- 1-4 4 18-028:PAGE 5-22 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1-3 H i stomorphol ogi c Observat i ons Dose Group: Animal Number: Sex : Death Tme: LIVER: -inflammation, chronic, focal/multifocal -vacuol at i on, hepatocell ul ar , mi dzonal -vacuolat i on, hepatocell ul ar , mu1 ti focal I11 I11 I11 I11 I11 I11 I11 I11 I11 I11 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 M M M M M M M M M M ss ss ss ss ss ss ss ss ss ss - - 1 1 1 - - - 1 - 1 1 - 1 - - - - 2 -----2- --- THYROID : -hypertrophy/hyperpl asi a, foll i cul ar epi the1 ium -ul timobranchi a1 body/cyst 1-5 418-028:PAGE 3-23 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1-4 H i stomorphol ogi c Observations Dose Group: Animal Number: Sex : Death Type: I V I V I V I V IV I V I V I V I V I V 19100 19103 19104 19118 19133 19141 19142 19144 19148 19150 M M M M M M M M M M ss ss ss ss ss ss ss ss ss ss EPIDIDYMIDES: -exfol iated spermatogeni c c e l l s -hypospermia LIVER: - - -hypertrophy, hepatocellular, c e n t r i l o b u l a r 1 1 2 1 1 1 -inflammation, chronic, focal/multifocal 1 1 1 - -11 11- T E S T I S (H&E): -degenerat ion, mu1t i focal T E S T I S (PAS): -degeneration, mu1ti focal THYROID : -hypertrophy/hyperplasia, f o l l i c u l a r epi the1 iurn -ul timobranchi a1 body/cyst 1-- -1- ----PP 1-6 418-028:PAGE J-24 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/OEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 TABLE 1-5 Dose Group: An i ma1 Number : Sex : Death Tme: ADRENAL GLANDS: -vacuolati on, cortex, di ffuse H i stomorphologic Observati ons vvvvvvvvvv 19111 19117 19124 19126 1 9 1 2 7 1 9 1 2 8 19140 19145 19149 1 9 1 5 2 MMMMMMMMMM ss ss ss ss ss ss ss ss ss ss BONE MARROW (STERNUM): CECUM : -inflammation, mucosa, chronic COAGULATING GLAND: EPIDIDYMIDES: -infiltration, mononuclear-cell, focal HEART : -i nflammation, chronic, focal --1- - ----- - - - - - - - -1 JEJUNUM : -divert icul um <p> - - - KIDNEYS: -basophi1 i a/degeneration, cortical tubules. focal/mu1 ti focal 1- - - - - - - - - -cyst(s), medulla P - P - - - - - - - -i nfi 1tration, mononuclear-cel1 , focal/mu1 ti focal 1- - - - - -1- - -nephritis, chronic, focal --1- ------ LIVER: -hypertrophy, hepatocellular, centrilobular 2 1 2 2 3 2 1 2 1 1 -i nf1ammation, chronic, focal/mu1tifocal 1 - - 1 - 1 1 - - 1 LYMPH NODE, MEDIASTINAL: -mastocytosi s LYMPH NODE, SUBMANDIBULAR: -hyperplasia, 1ymphocyti c/pl asmacyti c 1 3 - - 1 3 - 3 - 3 PEYER'S PATCH: -mineral i tati on, focal 1- - - - - - - - 1-7 418-028:PAGE J-25 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1-5 (Continued) Dose Group: Animal Number: Sex : Death Type: H i stomorphologic Observations vvvvvvvvvv 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 M M M M M M M M M M ss ss ss ss ss ss ss ss ss ss PROSTATE : -atrophy, focal/mul tifocal -inflammation, chronic, focal/multifocal RECTUM: -inflammation, mucosa, chronic -1 -- -- 1- -- 3- -- -- -- 11 SPINAL CORD, CERVICAL: S P I N A L CORD. LUMBAR: SPINAL CORD, THORACIC: - STOMACH : -edema/inflammation, submucosa, forestomach THYMUS : THYROID : -hypertrophy/hyperplasia, fol1 i c u l a r epi the1iurn -ul t imobranchia1 body/cyst TRACHEA : -inflammation, chronic, focal URINARY BLADDER : 3 P - 2- -- 2- 3- 3- - 3 P 2 - 1-8 418-028:PAGE 5-26 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING T S T PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 TABLE 1-6 Histomorphologic Observations Dose Group: Animal Number: Sex : Death Type: ADRENAL GLANDS: -vacuolati on, zona glomerulosa 1111111111 19012 19019 1 9 0 2 1 1 9 0 2 3 19041 19042 19044 19050 19053 19065 F F F F F F F F F F ss ss ss ss ss ss ss ss ss ss BONE MARROW (STERNUM): BRA1N : - CECUM: -inflammation, mucosa, chronic COLON: DUODENUM : HEART : - ILEUM : JEJUNUM: K IDNEYS : -nephritis , chronic, focal LIVER: -inflammation, chronic, focal/mu ti focal - LUNG : -i nfi1trati on, polymorphonuclear peri vascular/peri bronchia1 - 1 2 - LYMPH NODE, MEDIASTINAL: -congesti on -hyperplasia, lymphocytic/plasmacytic -mastocytosi s LYMPH NODE, SUBMANDIBULAR: -hyperplasia, 1ymphocyti c/pl asmacyti c - 3 1 2 2 MAMMARY GLAND: NERVE, SCIATIC: OVAR IES : -cyst(s), intraovarian PARATHYROID: PEYER'S PATCH: S K I N (GROSS LESION): -dermatitis, chronic, focal 1-9 418-028:PAGE J-27 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1-6 (Continued) H i stomorphol ogi c Observat ions Dose Group: Anima1 Number : Sex : Death Type: SPINAL CORD, CERVICAL: 1 1 1 1 1 1 1 1 1 1 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 F F F F F F F F F F ss ss ss ss ss ss ss ss ss ss - - - - - - - - - - SPINAL CORD, THORACIC: - - - - - - - - - - STOMACH : -di 1a t a t ion, mucosal g l ands 1- - - - - - - - - -edema/i nflammation, submucosa, glandular area - - - 2 3 - - - - - THYMUS : -atrophy - 3 - THYROID: -ul timobranchi a1 body/cyst TRACHEA : URINARY BLADDER: -inflammation, chronic, focal UTERUS : -macrophages , pigmented VAGINA : 1-10 418-028:PAGE J-28 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 4 1 8 - 0 2 8 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 TABLE 1-7 H i stomorphologi c Observations Dose Group: Ani ma1 Number : Sex: Death T y p e : ADRENAL GLANDS: -degenerati on, cysti c, cortex, mu1tifocal -hypertrophylvacuolation, cortex, mu1 ti focal -necrosis, cortex, focal vvvvvvvvvv 19001 19006 19011 19020 19022 19025 1 9 0 2 7 1 9 0 2 8 19030 19031 FFFFFFFFFF ss ss ss ss ss ss ss ss ss ss BONE MARROW (STERNUM) : BRAIN: CECUM : COLON : DUODENUM: HEART: ILEUM: JEJUNUM : KIDNEYS: -mineral i zati on, mu1ti focal -nephritis , chronic, focal LIVER: -i nf1ammation, chroni c, focal/mu1tifocal -necrosis, focal - LUNG : -inflammati on, i ntersti tia1 , focal 1 -macrophages, a1 veol i , focal 1 LYMPH NODE. MEDIASTINAL: LYMPH NODE, SUBMANDIBULAR: -hyperplasia, 1ymphocyti c/pl asmacytic 2- 22-23*2 MAMMARY GLAND : -necrosis, focal NERVE, SCIATIC: OVARI ES : PARATHYROID: PEY ER 'S PATCH: RECTUM : 1-11 418-028:PAGE J-29 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TABLE 1-7 (Cont inued) H i stomorphol ogic Observations Dose Group: Animal Number: Sex : Death Type: vvvvvvvvvv 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 FFFFFFFFFF ss ss ss ss ss ss ss ss ss ss S K I N (GROSS LESION): -dermat it i s, chronic, focal SPINAL CORD, CERVICAL: - - - - - - - - - - SPINAL CORD, THORACIC: SPLEEN: -atrophy - - - <2> - - - - - - STOMACH : -di 1a t a t ion, mucosal g l ands -------1- - -edema/i nflammation, submucosa, forestomach - - - <2> - - - - - - - -edema/inflammation, submucosa, glandular area - - -<2>- -erosi on( s ) , g l andul a r mucosa - - - <3> 2 2 - - - - - - - THYMUS : -atrophy - <4> - - - - - THYROID : -ul timobranchi a1 body/cyst - P - P P - P - P - TRACHEA: -inflammation, chronic, focal UTERUS: - d i s t e n t ion, 1umen -macrophages , pigmented 2- - - - - - - -3 3 2 3 4 2 3 - 3 3 - 1-12 418-028PAGE J-30 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT APPENDIX II INDIVIDUALANIMAL GROSS AND HISTOMORPHOLOGYDATA 4 18-028:PAGE 5-3 1 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l o g y D a t a HISTOMORPHOLOGIC OBSERVATIONS: LYMPH NODE, MEDIASTINAL: mastocytosis (minimal ) LYMPH NODE, SUBMANDIBULAR: hyperplasia, l y m p h o c y t i c / p l a s m a c y t i c (moderate) PROSTATE : inf 1ammat ion, chronic , focal /mu1 t ifocal ( m i n ima1 ) T.H.Y..R.O.I.D.:....................................h.y.p.e.r.t.r.o.p.h.y./.h.y.p.e.r.p.l.a.s.i.a.,..f.o..l l.i.c.u..l a..r..e.p.i.t .h.e.l.i u..m..(.m..i l.d.)...... THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN COAGULATING GLAND COLON DUODENUM HEART ILEUM JEJUNUM LIVER LUNG NERVE, SCIATIC PEYER S PATCH S P I N A L CORD, LUMBAR RECTUM S P I N A L CORD, THORACIC SEMINAL VESICLES SPLEEN TESTIS (H&E) TESTIS (PAS) THYMUS .U.R.I N. A. R. .Y. .B.L.A.D.D.E.R. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . E n d o f R e c o r d - 19109 CECUM EPIDIDYMIDES KIDNEYS PARATHYROID S P I N A L CORD, STOMACH TRACHEA CERVICAL 11-1 418-028:PAGE J-32 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i d u a l Animal Gross and H i stornorphol ogy Data GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o.t..a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: ADRENAL GLANDS: EPIDIDYMIDES: HEART : LIVER: THYROID : TRACHEA: vacuolation, cortex, diffuse (minimal) i n f i l t r a t i o n , mononuclear-cell, focal (minimal) in f l ammation, chroni c , f o c a l (mi nima1) inflammation, chronic, focal/multifocal (minimal) u l t imobranchi a1 bodylcyst inflammation, chronic, focal (mild) THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL L I M I T S : BONE MARROW (STERNUM) COLON KIDNEYS NERVE, S C I A T I C RECTUM SPINAL CORD, THORACIC T..E.S.T.I.S..(.P.A..S.)............ End o f Record- 19115 BRAIN CECUM COAGULATING GLAND DUODENUM ILEUM JEJUNUM LUNG LYMPH NODE, MEDIASTINAL LYMPH NODE, SUBMANDIBULAR PARATHYRO ID PEY ER S PATCH PROSTATE SEMINAL VESICLES SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPLEEN STOMACH TESTIS (H&E) . T.H.Y.M. .U.S. . . . . . . . . . . . . . . . . . . . .U.R. I.N.A.R.Y. .B.L.A.D.D.E.R. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-2 418-028:PAGE 5-33 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Individual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): .K.ID.N.E..Y.S.: ..M.o..t t.l.e.d..t.a.n..a.n.d..r.e.d.,..b..i l.a..t e..r.a.l...................N.o..m..i c.r.o.s..c .o.p.i.c.c.h.a..n.g.e..t .o..c.o.r.r.e.l.a.t.e........... HISTOMORPHOLOGIC OBSERVATIONS: KIDNEYS : No m i croscopi c change t o c o r r e l a t e LIVER: in f1ammat ion, chronic , f o c a l /mu1 ti f o c a l ( m i nima1 ) .LY.M..P.H..N.O.D.E., ..S.U.B.M.A.N.D.I.B.U.L.A.R.:.................h..y.p.e.r.p.la..s.i a..,..1.y .m.p.h.o.c.y.t.i.c./.p.l.a.s.m.a.c.y.t.i.c..(.m.i.1.d.)................. THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COAGULATING GLAND HEART LUNG PEYER 'S PATCH S P I N A L CORD, CERVICAL STOMACH T..H.Y.R.O..I.D.............. E n d o f R e c o r d - 19116 BONE MARROW (STERNUM) COLON ILEUM LYMPH NODE, MEDIASTINAL PROSTATE S P I N A L CORD, LUMBAR TESTIS (H&E) TRACHEA BRAIN DUODENUM JEJUNUM NERVE, SCIATIC RECTUM SPINAL CORD, THORACIC TESTIS (PAS) URINARY BLADDER CECUM E P ID IDYMIDES KIDNEYS PARATHYROID SEMINAL VESICLES SPLEEN THYMUS --------------------______________c_____--------------------------------------- 11-3 418-028:PAGE 5-34 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphol ogy Data HISTOMORPHOLOGIC OBSERVATIONS: ADRENAL GLANDS: vacuol ation, cortex, d i f f u s e (minimal ) CECUM : COLON : KIDNEYS : inflammation, mucosa, chronic (moderate) inflammation , mucosa, chronic (mi I d ) i n f i l t r a t i o n , mononuclear-cell , focal/multifocal (minimal) LIVER: inflammation, chronic, focal/multifocal (mild) R. .EC. .TU. M. .:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .i n.f.l a. .m.m. .a.t.i o.n. ,. . m. .u.c.o.s.a., . .c.h.r.o.n.i.c. .(.m. .i l.d.). . . . . . . . . . . . . . . . . . . . . . . . THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL LIMITS: BONE MARROW (STERNUM) BRAIN COAGULATING GLAND DUODENUM EPIDIDYMIDES HEART ILEUM JEJUNUM LUNG LYMPH NODE, MEDIASTINAL LYMPH NODE, SUBMANDIBULAR NERVE, S C I A T I C PARATHYROID PEYER S PATCH PROSTATE SEMINAL VESICLES SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPINAL CORD, THORACIC SPLEEN STOMACH TESTIS (H&E) TESTIS (PAS) THYMUS T.H. Y. .R.O.I.D. . . . . . . . . . . . . . . . . . . .T.R.A.C.H.E.A. . . . . . . . . . . . . . . . . . . . .U.R.IN. A. R. .Y. .B.LA. D. .D.E.R. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19119 11-4 418-028:PAGE 3-35 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Individual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N..o.t .a.p.p..l i.c.a.b.1.e................................ H ISTOMORPHOLOGIC OBSERVATIONS : E P ID IDYMIDES : infiltration, mononuclear-cell, focal (minimal) LIVER: inflammation, chronic, f o c a l / m u l t i f o c a l (minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia, l y m p h o c y t i c / p l a s m a c y t i c (minimal ) T.R.A.C..H.E.A.:...................................i.n.f.l.a.m.m..a.t.i o..n., ..c.h.r.o.n.i.c.,..f.o.c.a.l..(.m.i.n.i.m.a.l.)...................... THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN CECUM COAGULATING GLAND COLON DUODENUM HEART ILEUM JEJUNUM KIDNEYS LUNG NERVE, SCIATIC PARATHYROID PEYER S PATCH PROSTATE RECTUM SEMINAL VESICLES S P I N A L CORD, CERVICAL S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC SPLEEN STOMACH TESTIS (H&E) T..E.S.T.I.S..(P..A.S.) ...............T.H.Y.M.U.S......................T.H.Y.R.O.I.D.....................U.R.I.N.A.R.Y..B.L.A.D.D.E.R........ T I S S U E ( S 1 NOT A V A I L A B L E FOR EVALUATION: .LY.M..P.H..N.O.D.E., ..M.E.D.I.A.S.T.I.N.A.L................................................................................ End of Record- 19122 11-5 418-028:PAGE J-36 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 4 1 8 - 0 2 8 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Indi v i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.:..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N..o.t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: CECUM : LIVER: i n f l a m m a t i o n , mucosa, chronic (moderate) in f1ammat ion, chronic , f o c a l /mu1 t i focal ( m i n ima1 ) LYMPH NODE, SUBMANDIBULAR: hyperpl a s i a , 1y m p h o c y t i c/pl a s m a c y t i c ( m o d e r a t e ) PROSTATE : atrophy, focal /mu1 ti focal ( m i n i m a l ) T.H.Y..R.O.I.D.:....................................u.l.t .i m..o.b.r.a.n.c.h.i.a.1..b.o..d.y.lc..y.s.t.................................. THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COLON ILEUM BONE MARROW (STERNUM) DUODENUM JEJUNUM BRAIN EPID IDYMIDES KIDNEYS COAGULATING GLAND HEART LUNG LYMPH NODE, MEDIASTINAL RECTUM NERVE, SCIATIC SEMINAL VESICLES PARATHYROID SPINAL CORD, CERVICAL PEYER I S PATCH S P I N A L CORD, LUMBAR SPINAL CORD, THORACIC SPLEEN STOMACH TESTIS (H&E) T..E.S.T.I.S..(P..A.S.) ...............T.H.Y.M.U.S......................T.R.A.C.H.E.A.....................U.R.I.N.A.R.Y..B.L.A.D.D.E.R........ E n d o f R e c o r d - 19125 11-6 4 18-028:PAGE J -37 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WXTH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 Individual Animal Gross and H i stomorphology Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: ADRENAL GLANDS: EPIDIDYMIDES: vacuolation, cortex, diffuse (mild) i n f i l t r a t i o n , mononuclear-cell , focal (minimal) HEART: inflammation, chronic, focal (mild) LIVER: inflammation, chronic, focal /mu1 t i focal (mi 1d) LYMPH NODE, SUBMANDIBULAR: hyperplasi a, 1ymphocyt ic/pl asmacyt ic (mi I d ) PROSTATE : inflammation, chronic, focal/multi focal (minimal) T.R..A.C.H.E.A.: ....................................in..fl.a.m..m.a..ti.o.n.,..c.h.r.o..n.ic.., .f.o.c.a..l ..(m..i.I.d.)........................ THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL L I M I T S : BONE MARROW (STERNUM) BRAIN CECUM COAGULATING GLAND COLON KIDNEYS DUODENUM LUNG ILEUM JEJUNUM LYMPH NODE, MEDIASTINAL NERVE, S C I A T I C PARAT HYROID SPINAL CORD, CERVICAL PEY ER ' S PATCH SPINAL CORD, LUMBAR RECTUM SPINAL CORD, THORACIC SEMINAL VESICLES SPLEEN STOMACH TESTIS (H&E) TESTIS (PAS) THYMUS T. H. .Y.R.O.I.D. . . . . . . . . . . . . . . . . . . .U. R. .IN.A.R. Y. . .B.LA.D.D. E. R. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19131 11-7 418-028:PAGE J-38 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Individual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): H ISTOMORPHOLOGIC OBSERVATION (S 1: HISTOMORPHOLOGIC OBSERVATIONS: LIVER: STOMACH : i n f l a m m a t i o n , chronic, f o c a l / m u l t if o c a l ( m i 1d) e d e m a / i n f1ammat ion, s u b m u c o s a , g l andul a r area ( m o d e r a t e ) T. H. .Y.R.O.I D. .:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . u. .l t. .i m. .o.b.r.a.n.c.h.i .a.1. .b.o.d.y./.c.y.s.t. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN CECUM COAGULATING GLAND COLON DUODENUM E P I D I D Y M IDES HEART ILEUM JEJUNUM KIDNEYS LUNG LYMPH NODE, SUBMANDIBULAR NERVE, S C I A T I C PARATHYROID PEYER 'S PATCH PROSTATE RECTUM SEMINAL VESICLES S P I N A L CORD, CERVICAL S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC SPLEEN TESTIS (H&E) TESTIS (PAS) THYMUS TRACHEA U. .R.I N. A. R. .Y. .B.L.A.D.D.E.R. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . T I S S U E ( S ) NOT A V A I L A B L E FOR EVALUATION: .LY.M..P.H..N.O.D.E., ..M.E.D.I.A.S.T.I.N.A.L................................................................................ E n d o f R e c o r d - 19132 11-8 418-028:PAGE J-39 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGI C OBSERVATION( S1: G. .E.N.E.R.A.L.: . .N.o. .g.r.o.s.s. .c.h.a.n.g.e.s... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .N.o.t. .a.p.p.l.i.c.a.b.1.e. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . HISTOMORPHOLOGIC OBSERVATIONS: EPI D I DYMIDES : KIDNEYS : STOMACH : i n f i l t r a t i o n , mononuclear-cell , focal (minimal) cyst (s) , medulla d i l a t a t i o n , mucosal glands (minimal) edema/inflammation, submucosa, glandular area (minimal) edema/inflammation, submucosa, forestomach (mi I d ) hypertrophy/hyperplasia, f o l l i c u l a r epithelium (mild) ..in.f.l.a.m..m.a.t.i.o.n.,..c.h..ro..n.i.c., ..f.o.c.a.l..(.m..in..im..a.l.)............... THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL LIMITS: ADRENAL GLANDS COAGULATING GLAND ILEUM LYMPH NODE, MEDIASTINAL PEYER `S PATCH SPINAL CORD, CERVICAL T..E.S.T.I.S...(H..&.E.)............ End o f Record- 19134 BONE MARROW (STERNUM) BRAIN CECUM COLON DUODENUM HEART JEJUNUM LIVER LUNG LYMPH NODE, SUBMANDIBULAR NERVE, S C I A T I C PARATHYROID PROSTATE RECTUM SEMINAL VESICLES SPINAL CORD, LUMBAR SPINAL CORD, THORACIC SPLEEN . T. .E.S.T.I.S. .(P. .A.S.). . . . . . . . . . . . . . .T.H.Y.M.U.S. . . . . . . . . . . . . . . . . . . . . .U.R.I.N.A.R.Y. .B.L.A.D.D.E.R. . . . . . . . . 11-9 418-028:PAGE J-40 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 4 1 8 - 0 2 8 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o..t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: KIDNEYS: in f i 1t r a t i on, m o n o n u c l e a r - c e l l , focal /mu1 ti focal ( m i n i m a l ) LIVER: i n f l a m m a t i o n , chronic, f o c a l / r n u l t i f o c a l ( m i n i m a l ) .P.R.O.S.T.A.T.E.:..................................i.n..f1.a.m..m.a.t.i.o.n., ..c.h.r.o.n.i.c.,..f.o.c.a.l./m..u1..t.i.f.o.c.a.l..(.m.i.n..im.a.1..) .......... THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN CECUM COAGULATING GLAND COLON DUODENUM E P IDIDYMIDES HEART ILEUM JEJUNUM LUNG LYMPH NODE, MEDIASTINAL LYMPH NODE, SUBMANDIBULAR NERVE, S C I A T I C PARATHYROID PEYER 'S PATCH RECTUM SEMINAL VESICLES S P I N A L CORD, CERVICAL S P I N A L CORD, LUMBAR TESTIS (H&E) S P I N A L CORD, THORACIC TESTIS (PAS) SPLEEN THYMUS STOMACH THYROID T.R. A. C. H. .E.A. . . . . . . . . . . . . . . . . . . .U.R.I.N.A.R.Y. .B.L.A.D.D.E.R. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f R e c o r d - 19135 11-10 418-028:PAGE 5-41 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i d u a l Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): K..ID.N.E.Y..S.: ..B.i.l.a.t.e.r.a.l.-..m..o.tt.l.e.d................................N.o..m.i.c.r.o.s.c.o.p.i.c..c.h.a.n.g.e..t.o..c.o..rr.e..la..te............ H ISTOMORPHOLOGIC OBSERVATIONS : KIDNEYS : No microscopic change t o c o r r e l a t e LIVER: inf 1annnation, chroni c , f o c a l /mu1 t i f o c a l (mi 1d) T.H.Y..R.O.I.D.:...................................u..lt.i.m.o.b..ra.n..c.h.i .a.1..b.o.d.y./.c.y.s.t................................... 11-11 418-028:PAGE J-42 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING T E S T PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i d u a l Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.:..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o..t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: LIVER: inf 1ammation , chronic , foca1/mu1t if o c a l (mi 1d) T.H. Y. .RO. .I.D.:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .u.l.t i.m. .o.b.r.a.n.c. h. .i a. l. .b.o. d. .y./c.y.s.t. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19106 11-12 418-028:PAGE J-43 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I I I n d iv i dual Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: LIVER: lipidosis, tension, focal ............................................in.f.l.a.m..m.a.t.i.o.n.,..c.h.r.o.n..ic.,..f.o.c..a.l/.m..u.l.t.if.o.c.a..l ..(.m.i.n.i.m.a.l.}.......... THE FOLLOWING TISSUE(S1 WERE W I T H I N NORMAL LIMITS: T.H. Y. .R.O.I.D. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19108 11-13 418-028:PAGE J-44 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphology Data GROSS OBSERVATION(S): H ISTOMORPHOLOGIC OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATIONS: LIVER: in f1ammation, chroni c , focal /mu1 t i focal (min ima1) T. H. .Y.R.O.I D. .:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .h.y. p. e. r. t. r. o. .p.h.y./.h.y.p.e.r.p.l.a.s.i.a.,. .f .o.l.l.i.c.u.l.a. r. .e.p. .i t.h.e. l.i.u.m. . .( m. .i.l d. .) . . . . . . End o f Record- 19110 11-14 418-028:PAGE J-45 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 4 1 8 - 0 2 8 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Indi v i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATIONS: K..ID.N.E.Y..S.:...................................d..i l.a.t.a.t.i.o.n.., ..p.e.l.v.i.s..(.m.i.l .d.) ................................... 11-15 418-028:PAGE J-46 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR S STUDY NUMBER T-7706.1 Appendix I1 I n d iv i dual Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S) : HISTOMQRPHOLOGIC OBSERVATION(S): H ISTOMORPHOLOGIC 06sERVAT IONS : LIVER: in f1ammation I chroni c I f o c a l /mu1 t i focal (mi n ima1) ...........................................l.i.p.i.d.o.s..i s.,...te.n..s.io.n.., ..fo..c.a.l................................... THE FOLLOWING TISSUE(S) WERE WITHIN NORMAL LIMITS: 11-16 418-028:PAGE 3-47 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODlJCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphol ogy Data HISTOMORPWOLOGIC OBSERVATIONS: LIVER: THYROI D : End o f Record- 19129 inflammation, chronic, focal/mul tifocal (minimal) hypertrophy/hyperplasia, f o l l i c u l a r epithelium (minimal) 11-17 418-028:PAGE 5-48 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF 1-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I1 Individual Animal Gross and Hi stomorphology Data ANIMAL NI IBER: 19 SEX: M GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): .K.I.D.N.E.Y.S.:. .B.i.l.a.t.e.r.a.l.-. .m.o.t.t.l.e. d. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .d.i.1.a.t.a.t. i.o. n. .,. p.e.l. .vi. .s. . . . . . . . . . . . . . . . . . . . . . . . . . . HISTOMORPHOLOGIC OBSERVATIONS: KIDNEYS: dilatation, pelvis (minimal) LIVER: vacuolation, hepatocellular, mu1 tifocal (minimal ) T. H. .Y.R.O.I.D.:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . u.l.t. i. m. o. b. r. a. .n.c.h.i.a1. . .b.o.d.y./.c.y.s.t. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19136 11-18 418-028:PAGE 3-49 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Indiv i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S) : HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o.t..a.p.p.l.i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: T.H.Y..R.O.ID..:....................................ul..t i.m..o.b.r.a.n.c.h.i.a.1..b.o.d..y./c.y.s.t................................... THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : 11-19 4 18-028:PAGE J-50 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix 11 Indi vi dual Animal Gross and Hi stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: .T.H.Y.R.O.I.D.:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .h. y. p. e. r. t. r. o. p. h. y. /. h. y. p. e. r. p. l. a. s. i. a. ,. . .f.o.l.l.i.c.u.l.a.r. .e.p.i.t.h.e.l.i.u.m. . .(.m.i.l.d.). . . . . . THE FOLLOWING TISSUEfS) WERE WITHIN NORMAL LIMITS: L. .I .V.E.R. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19139 11-20 418-028:PAGE J-51 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTIONIDEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N..o.t .a.p.p..l i.c.a.b.1.e................................ THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : 11-21 418-028:PAGE 5-52 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Indiv i dual A n i m a l Gross and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : 11-22 418-028:PAGE J-53 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I1 Indiv i dual Animal Gross and Histomorphology Data GROSS OBSERVATION(S) : HISTOMORPHOLOGIC OBSERVATION(S) : G. .E.N.E.R.A.L.:. .N.o. .g.r.o.s.s. .c.h.a.n.g.e. s. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .N.o.t. .a.p.p.l. i.c.a.b.1. e. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . HISTOMORPHOLOGIC OBSERVATIONS: I 1-23 418-028:PAGE J-54 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTIONJDEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphology Data GROSS OBSERVATION(S) : HISTOMORPHOLOGIC OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATIONS: LIVER: in f1ammation, chroni c , focal /mu1 t i focal (mi n ima1) T. H. .Y.R.O.I D. .:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .u. .l t. i.m. .o.b.r.a.n.c.h.i .a1. . .b.o.d.y.l.c.y.s.t. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19112 11-24 418-028:PAGE J-55 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i d u a l Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.:..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o..t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: LIVER: inflammation, chronic, focal/mul t if o c a l (mi njmal) T.H.Y..R.O.I.D.:....................................u.l.t.i.m.o.b.r.a.n.c.h.i.a.1..b.o.d..y.lc.y..s.t .................................. End o f Record- 19114 11-25 418-028:PAGE 5-56 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I1 I n d i v i d u a l Animal Gross and H i stomorphology Data GROSS OBSERVATION(S): H ISTOMORPHOLQGIC OBSERVATION (S 1: G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o.t..a.p.p.l.i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: LIVER: inflammation, chronic, focal/mul tifocal (minimal) vacuolation, hepatocellular, multifocal (mild) T. H. .Y.R.O.I.D.:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .u.l .t i.m. .o.b.r.a.n.c.h.i.a.1. .b.o. d. .y./ c.y. s. t. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19121 11-26 418-028:PAGE 5-57 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i d u a l Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G. .E.N.E.R.A.L.:. .N.o. .g.r.o.s.s. .c.h.a.n.g.e.s... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .N.o.t. .a.p.p.l.i.c.a.b.1.e. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . H ISTOMORPHOLOGIC OBSERVATIONS : .L.IV.E..R.: .....................................i.n.f.la.m..m..a.t.io.n.,...c.h.r.o.n.i.c., ..f.o.c.a.l./m..u.1.t.i.f.o.c.a.l..(.m..in.i.m..a.l .) .......... THE FOLLOWING TISSUE(S1 WERE WITHIN NORMAL LIMITS: 11-27 418-028:PAGE 3-58 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i d u a l Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: LIVER: inflammation, chronic, focal/mul tifocal (minimal ) T.H..Y.R.O.ID..:...................................h.y.p.e.r.t.r.o.p.h.y./.h.y.p.e.r.p.l.a.s.i.a.,...f.o.l.l.i c..u.l.a.r..e.p..it.h.e.l.i.u.m...(m..i.ld.)....... End o f Record- 19130 11-28 418-028:PAGE J-59 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 4 1 8 - 0 2 8 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS O!SERVATION(S) : HISTOMORPHOLOGIC OBSERVATION(S): THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : .L .I V.E. .R. . . . . . . . . . . . . . . . . . . . . .T.H.Y.R.O.I.D. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End of Record- 19137 11-29 418-028:PAGE 3-60 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Indiv i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: .LI.V.E.R..: .....................................v.a.c.u..o.l a..t.io.n.., ..h.e.p.a.t.o.c.e..ll.u.l.a.r.,..m.i.d.t.o.n.a.l..(.m.i.1.d.)................ THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : T. H. .Y.R.O.I.D. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . E n d o f Record- 19146 11-30 418-028:PAGE J-6 I ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTIONfDEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S1: H ISTOMORPHOLOGIC OBSERVATION[S 1: G. .E.N.E.R.A.L.: . .N.o. .g.r.o.s.s. .c.h.a.n.g.e.s... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . N. .o.t. .a.p.p.l .i.c.a.b.1.e. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . HISTOMORPHOLOGIC OBSERVATIONS: LIVER: THYROID : End of Record- 19100 hypertrophy, hepatocell u la r , cent ri1obul a r (mi n ima1) inflammation, chronic, focal/multifocal (minimal ) hypertrophy/hyperplasia, f o l l i c u l a r epithelium (minimal) 11-31 418-028:PAGE 5-62 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I1 I n d iv i dual Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o..t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: .L.IV.E.R..:.....................................h..y.p.e.r.tr.o.p.h.y.., ..h.e.p.a.t.o.c.e..ll.u.l.a.r., ..c.e.n..tr.i.l.o.b.u.l.a.r..(.m..in..im..a.l)........ THE FOLLOWING TISSUE(S1 WERE WITHIN NORMAL L I M I T S : T. H. .Y.R.O.I D. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19103 11-32 418-028:PAGE J-63 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I1 I n d iv i dual Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATIONS: LIVER: inflammation, chronic, focal/multifocal (minimal) ............................................h.y.p.e.r.t.ro..p.h.y.,..h.e.p..a.t.o.c.e.l.lu..l a..r,..c..e.n.t.r.i.1.o.b.u.l.a.r..(.m.i.1.d.).......... THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL LIMITS: T.H. .Y.R.O.I D. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End of Record- 19104 I1-33 418-028:PAGE J-64 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i d u a l Animal Gross and H istomorphology Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S1: G. .E.N.E.R.A.L.: . .N.o. .g.r.o.s.s. .c.h.a.n.g.e.s... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .N.o.t. .a.p.p.l.i.c.a.b.1.e. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . THE FOLLOWING TISSUE(S1 WERE WITHIN NORMAL LIMITS: T. H. .Y.R.O.I D. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19118 11-34 4 18-028:PAGE J-65 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPNENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR ' S STUDY NUMBER T-7706.1 Appendix I 1 Individual Animal Gross and H i stomorphology Data GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S1: G..E.N.E.R.A.L.:..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N..o.t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: LIVER: hypertrophy, hepatocellular, c e n t r i l o b u l a r (minimal ) THYROID : hypertrophy/hyperplasia, f o l l i c u l a r epithelium (minimal) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .u. .l t. .im. .o.b.r.a.n.c.h.i .a1. . .b.o.d.y./.c.y.s.t . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19133 11-35 418-028:PAGE J-66 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphology Data GROSS OBSERVATION(S1: H ISTOMORPHOLOGIC OBSERVATION (S1: G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N..o.t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS : LIVER: hypertrophy, hepatocellular, centri lobular (minimal ) T.H..Y.R.O.I.D.:...................................u..lt.i.m.o.b..ra.n..c.h.i .a.1..b.o.d.y.f.c.y.s.t................................... End o f Record- 19141 11-36 418-028:PAGE J-67 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i d u a l Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): EPIDIDYMIDES: B i l a t e r a l - small ; cauda epididymis, hypospermi a l e f t , small. e x f o l iated spermatogenic c e l l s TESTES: B i 1a t e r a l - small . degeneration, m u l t i f o c a l , TESTIS (PAS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . d. .e.g.e.n.e. r.a.t.io. .n., . .m. u. .l t.i.f .o.c.a.l.,. .T.E.S. T. .I S. . (.H.&. .E.). . . . . . . HISTOMORPHOLOGIC OBSERVATIONS: EPIDIDYMIDES: LIVER: TESTIS (H&E) : TESTIS (PAS) : hypospermi a (marked) e x f o l iated spermatogenic c e l l s (moderate) inflammation, chronic, focal /mu1 ti focal (minimal ) hypertrophy, hepatocell u l a r , centri lobular (minimal) degenerat ion, mu1t i focal (mi 1d) . d. .e.g.e.n.e. .ra. t.i.o.n.,. .m. .u.l.t.i .f o. .c.a.l. .(.m. .i l.d.). . . . . . . . . . . . . . . . . . . . . . . . . . . . . THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL LIMITS: T. H. .Y.R.O.I.D. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19142 11-37 4 18-028:PAGE J-68 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Individual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): H ISTOMORPHOLOGIC OBSERVATION( S1: HISTOMORPHOLOGIC OBSERVATIONS: L.I.V.E.R..: .....................................i.n..f1.a.m.m..at..io..n., ..c.h.r.o.n.i.c.,..fo..c.a.l./m..u1..t.i.f.o.c.a.l..(.m.i.n.i.m.a.1.)........... THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : 11-38 418-028:PAGE J-69 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i dual Animal Gross and H i stomorphology Data GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: LIVER: inflammation , chronic , f o c a l /mu1 t i f o c a l (minimal ) hypertrophy, hepatocell u l a r , centri lobular (minimal) T.H..Y.R.O.ID..:...................................h.y.p.e.r.t.r.o.p.h.y./.h.y.p.e.r.p.l.a.s.i.a.,...f .o.l.l.i.c.u.l.a.r..e.p.i.t.h.e.l.iu..m..(.m..i.ld.)....... End o f Record- 19148 11-39 418-028:PAGE J-70 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I1 Individual Animal Gross and H i stomorphology Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S1: G. .E.N.E.R.A.L.: . .N.o. .g.r.o.s.s. .c.h.a.n.g.e.s... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .N.o.t. .a.p.p.l.i.c.a.b.1.e. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . HISTOMORPHOLOGIC OBSERVATIONS: LIVER: hypertrophy, hepatocell ular, centrilobular (minimal ) T.H..Y.R.O.ID.:....................................h..y.p.e.r.tr.o.p.h..y./h..y.p.e.r.p.l.a.s.i.a.,..f.o.l.1.i.c.u.l.a.r..e.p.i.t.h.e.1.i.u.m..(.m..i 1.d..) ...... End o f Record- 19150 11-40 4 18-028:PAGE J-7 1 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I 1 Indivi dual Animal Gross and H i stomorphology Data HISTOMORPHOLOGIC OBSERVATIONS: JEJUNUM : KIDNEYS: LIVER: LYMPH NODE, SUBMANDIBULAR: PEYER s PATCH: PROSTATE : THYROID : di vert i cul um basophi1i a/degeneration, cortical tubules, focal/mu1tifocal (minima1 ) cyst( s ) , medulla infiltration, mononuclear-cell , focal/multifocal (minimal) hypertrophy, hepatocellular, centrilobular (mild) inflammation, chronic, focal/multifocal (minimal) hyperplasia, lymphocytic/plasmacytic (minimal) mineralization, focal (minimal) i nf1ammati on, chronic, focal/mu1 tifocal (minimal ) ul timobranchia1 body/cyst . h. .y.p.e.r.t.r.o.p.h.y./.h.y.p.e. r. p. .l.a.si. .a.,. .f.o.l.l.i.c.u.l.a. r. . .e.pi. .t.h.e.1.i.u.m. . (. .m.o.d.e.r.a.t.e.). THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COAGULATING GLAND HEART NERVE, S C I A T I C S P I N A L CORD, CERVICAL STOMACH TRACHEA -___-_-------------__I_ End of Record- 19111 BONE MARROW (STERNUM) COLON ILEUM PARATHYROID SPINAL CORD, LUMBAR TESTIS (H&E) URINARY BLADDER BRAIN DUODENUM LUNG RECTUM S P I N A L CORD, TESTIS (PAS) THORACIC CECUM EPIDIDYMIDES LYMPH NODE, MEDIASTINAL SEMINAL VESICLES SPLEEN THYMUS 11-41 4 18-028PAGE J-72 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: LIVER: hypertrophy, hepatocellu l a r , c e n t r i 1obul a r ( m i n i m a l ) LYMPH NODE, MEDIASTINAL: mastocytosis (minimal ) LYMPH NODE, SUBMANDIBULAR: hyperplasia, l y m p h o c y t i c / p l a s m a c y t i c (moderate) .ST.O.M..A.C.H.:....................................e.d.e.m.a./.i .n.f.l.a.m.m..a.t.i o.n.,..s.u.b.m..u.c.o.sa.,...f o.r.e.s.t.o.m..a.c.h..(.m.i.1.d.).......... THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COAGULATING GLAND HEART LUNG PROSTAT E S P I N A L CORD, LUMBAR TESTIS (PAS) U..R.IN..A.R.Y..B..L.A.D.D.E.R......... E n d o f R e c o r d - 19117 BONE MARROW (STERNUM) COLON ILEUM NERVE, SCIATIC RECTUM SPINAL CORD, THORACIC THYMUS BRAIN DUODENUM JEJUNUM PARATHYROID SEMINAL VESICLES SPLEEN THYROID CECUM EPIDIDYMIDES KIDNEYS PEYER 'S PATCH S P I N A L CORD, CERVICAL TESTIS (H&E) TRACHEA 11-42 4 I8-028:PAGE 3-73 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION( S 1: HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: EPIDIDYMIDES: KIDNEYS : LIVER: LYMPH NODE, MEDIASTINAL: THYROI D : infiltration, mononuclear-cell, focal (minimal) nephritis , chronic, focal ( m i n i m a l ) cyst (s), m e d u l l a hypertrophy, hepatocellu l a r , c e n t r i 1obul a r ( m i 1d) mastocytosis (minimal) hypertrophy/hyperplasia, f o l l i c u l a r e p i t h e l i u m ( m i l d ) THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COAGULATING GLAND ILEUM NERVE, S C I A T I C RECTUM S P I N A L CORD, THORACIC T..E.S.T.I.S...(P..A.S.)............. E n d o f R e c o r d - 19124 BONE MARROW (STERNUM) BRAIN COLON DUODENUM JEJUNUM LUNG PARATHYROID PEY ER 'S PATCH SEMINAL VESICLES SPINAL CORD, CERVICAL SPLEEN STOMACH THYMUS TRACHEA .----------------------------------------------------------------------------- CECUM HEART LYMPH NODE, SUBMANDIBULAR PROSTATE S P I N A L CORD, LUMBAR TESTIS (H&E) URINARY BLADDER 11-43 418-028:PAGE 3-74 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 4 1 8 - 0 2 8 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: LIVER: i n f l a m m a t i o n , chronic, focal /mu1 tifocal (minimal ) hypertrophy, hepatocellul a r , centri 1obul a r ( m i 1d) .P.R.O.S.T.A.T.E.:...................................i.n.f.1.a.m.m.a.t.i.o.n.,..c.h.r.o.n..ic..,.f.o.c.a..l /.m.u.1..t.i.fo..c.a.l..(.m.i.n.i.m.a.1.).......... THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COAGULATING GLAND HEART LUNG PARATHYROID S P I N A L CORD, CERVICAL STOMACH THYROID BONE MARROW (STERNUM] COLON ILEUM LYMPH NODE, MEDIASTINAL PEYER 'S PATCH S P I N A L CORD, LUMBAR TESTIS (H&E) TRACHEA BRA1N DUODENUM JEJUNUM LYMPH NODE, SUBMANDIBULAR RECTUM S P I N A L CORD, THORACIC TESTIS (PAS) URINARY BLADDER CECUM E P ID IDYMIDES KIDNEYS NERVE, SCIATIC SEMINAL VESICLES SPLEEN THYMUS 11-44 418-028:PAGE 5-75 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 5 . 1 Appendix I1 Individual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: ADRENAL GLANDS: LIVER: LYMPH NODE, SUBMANDIBULAR: THYROID : vacuolation, cortex, diffuse ( m i l d ) hypertrophy, hepatocellul a r , c e n t r i 1obul a r ( m o d e r a t e ) hyperplasi a, 1y m p h o c y t ic/pl a s m a c y t ic ( m i n ima1 ) hypertrophy/hyperplasia, f o l l i c u l a r e p i t h e l i u m ( m i l d ) --c-----------------___________c________--------------------------------------------------------------- THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : BONE MARROW (STERNUM) COLON BRAIN DUODENUM CECUM EPIDIDYMIDES ILEUM JEJUNUM KIDNEYS LYMPH NODE, MEDIASTINAL PROSTAT E SPINAL CORD, LUMBAR NERVE, SCIATIC RECTUM S P I N A L CORD, THORACIC PARATHYROID SEMINAL VESICLES SPLEEN TESTIS (H&E) TESTIS (PAS) THYMUS U. .R.I N. A. R. .Y. .B.L.A.D.D.E.R. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . E n d o f R e c o r d - 19127 COAGULATING GLAND HEART LUNG PEY ER 'S PATCH S P I N A L CORD, CERVICAL STOMACH TRACHEA 11-45 418-028:PAGE J-76 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION($): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: CECUM: LIVER: LYMPH NODE, SUBMANDIBULAR: PROSTATE : THYROID : inflammation, mucosa, chronic ( m i l d ) in f 1amnation , chronic , focal /mu 1t ifocal (mi nima1) hypertrophy, hepatocellular, centri lobular (mild) hyperpl asi a, 1ymphocyt ic / p l asmacyt ic (moderate) atrophy, focal /mu1 t i f o c a l (moderate) hypertrophy/hyperpl asi a , f o l 1ic u l a r epi the1 iurn (moderate) .----__---__---____------------------------------------------------- THE FOLLOWING TISSUE(S) WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN COAGULATING GLAND COLON DUODENUM EPIDIDYMIDES HEART ILEUM JEJUNUM KIDNEYS LUNG LYMPH NODE, MEDIASTINAL NERVE, S C I A T I C PARATHYROID PEYER S PATCH RECTUM SEMINAL VESICLES SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPINAL CORD, THORACIC SPLEEN STOMACH TESTIS (H&E) T. .E.S.T.I .S. .(P. .A.S.). . . . . . . . . . . . . . .T.H.Y.M.U.S. . . . . . . . . . . . . . . . . . . . . .TR. .A.C.H.E.A. . . . . . . . . . . . . . . . . . . .U.R. I.N.A.R.Y. .B.L.A.D.D.E.R. . . . . . . . . End o f Record- 19128 11-46 418-028:PAGE J-77 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 A p p e n d i x 11 Individual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATIONS: LIVER: i n f l a m m a t i o n , chronic, f o c a l / m u l t i f o c a l (minimal) hypertrophy, hepatocellu l a r , c e n t r i 1obul a r ( m i n i m a l ) T.H.Y..R.O.I.D.:....................................h.y.p.e.r.t.r.o.p.h.y./.h.y.p.e.r.p.l.a.s.i.a.,..f.o..l.l .i c..u.l.a.r..e.p.i.th..e.1.i.u.m..(.m.o..d.e.r a.t.e.).. THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) COAGULATING GLAND HEART COLON ILEUM LUNG LYMPH NODE, MEDIASTINAL PARATHYROID PEYER ' S PATCH SEMINAL VESICLES S P I N A L CORD, CERVICAL SPLEEN STOMACH T.H. Y. M. .U.S. . . . . . . . . . . . . . . . . . . . . .T.R.A.C.H.E.A. . . . . . . . . . . . . . . . . E n d o f R e c o r d - 19140 BRAIN DUODENUM J E3 UNUM LYMPH NODE, SUBMANDIBULAR PROSTATE S P I N A L CORD, LUMBAR TESTIS (H&E) URINARY BLADDER CECUM EPIDIDYMIDES KIDNEYS NERVE, SCIATIC RECTUM S P I N A L CORD, THORACIC TESTIS (PAS) 11-47 418-028:PAGE J-78 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o..t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: KIDNEYS : infiltration, mononuclear-cell, focal/multifocal (minimal) LIVER: hypertrophy, hepatocellular, centri lobular (mi I d ) LYMPH NODE, SUBMANDIBULAR: RECTUM : hyperpl asia , 1y m p h o c y tic/pl a s m a c y t ic ( m o d e r a t e ) in f lammat ion, m u c o s a , chroni c ( m i n ima1 ) T.R.A.C..H.E.A.:....................................i .n.f l.a.m..m.a.t.i.o.n.,..c.h.r.o.n..ic.,..f.o.c..a.l ..( .m.i.l d..) ........................ THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COAGULATING GLAND HEART LYMPH NODE, MEDIASTINAL PROSTATE S P I N A L CORD, THORACIC T..E.S.T.I.S...(P..A.S.)........... E n d o f R e c o r d - 19145 BONE MARROW (STERNUM) BRAIN COLON ILEUM DUODENUM JEJUNUM NERVE, SCIATIC PARATHYROID SEMINAL VESICLES S P I N A L CORD, SPLEEN THYMUS STOMACH THYROID ,----------------------------------------------------------------------------- CERVICAL CECUM EPID IDYMIDES LUNG PEYER S PATCH S P I N A L CORD, LUMBAR TESTIS (H&E) URINARY BLADDER 11-48 4 18-028:PAGE 5-79 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMEIER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Indiv i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S1: H ISTOMORPHOLOGI C OBSERVATION ( S 1: G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s.................................N..o.t..a.p.p.l.i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: LIVER: hypertrophy, hepatocellular, centrilobular ( m i n i m a l ) THYROID : u l t i m o b r a n c h i a1 body/cyst ............................................h.y.p.e.r.t.r.o.p.h.y./.h.y.p.e.r.p.l.a.s.i.a.,..f.o..l.l .i c..u.l.a.r..e.p.i.t.h.e.l.i.u.m..(.m.o..d.e.r a.t.e.).. THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COAGULATING GLAND HEART LUNG PARATHY RO ID SEMINAL VESICLES SPLEEN T.H..Y.M.U.S................. E n d o f R e c o r d - 19149 BONE MARROW (STERNUM) COLON ILEUM LYMPH NODE, MEDIASTINAL PEYER'S PATCH S P I N A L CORD, CERVICAL STOMACH TRACHEA BRAIN CECUM DUODENUM EPIDIDYMIDES JEJUNUM KIDNEYS LYMPH NODE, SUBMANDIBULAR NERVE, S C I A T I C PROST A T E RECTUM S P I N A L CORD, LUMBAR SPINAL CORD, THORAC IC TESTIS (H&E) TESTIS (PAS) URINARY BLADDER .------------------_--------------------------------- 11-49 4 18-028;PAGE 5-80 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 Individual Animal Gross and H i stomorphology Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: HEART: inflammation, chronic, focal (minimal ) LIVER: inflammation, chronic, focal /mu1 t i focal (minimal ) hypertrophy, hepatocellular, centrilobular (minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia, 1ymphocytic/pl asmacytic (moderate) PROSTATE : in f1ammation , chroni c , focal /mu1 ti focal (mi n ima1) atrophy, focal/mul tifocal (minimal) T. H. .Y.R.O.I D. .:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . h. y.p.e.r.t.r.o.p.h.y./.h.y.p.e.r.p.l.a.s.i.a.,. . .f.o.l.l.i.c.u.l.a. r. .e.p. .i t.h.e. l.i.u.m. . .( m. .i.l d. .) . . . . . . THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COAGULATING GLAND ILEUM LYMPH NODE, MEDIASTINAL RECTUM SPINAL CORD, THORACIC T..E.S.T.I.S...(P..A.S.)............ End o f Record- 19152 BONE MARROW (STERNUM) BRAIN COLON DUODENUM JEJUNUM KIDNEYS NERVE, SCIATIC PARATHYROID SEMINAL VESICLES SPINAL CORD, SPLEEN STOMACH THYMUS TRACHEA .------------------------------------------------- CERVICAL CECUM E P IDID Y M IDES LUNG PEYER I S PATCH SPINAL CORD, LUMBAR TESTIS (H&E) URINARY BLADDER 11-50 418-028:PAGE 5-8 1 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: STOMACH : dilatation, m u c o s a l glands ( m i n i m a l ) THYROID : ult i m o b r a n c h i a l body/cyst U..TE.R..U.S.:....................................m..a.c.r o.p..h.a.g.e.s., ..p.i.g.m.e.n.t.e.d..(.m.o.d.e.r.a.t.e.) ........................... THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN COLON DUODENUM HEART JEJUNUM KIDNEYS LIVER LYMPH NODE, MEDIASTINAL OVARIES LYMPH NODE, SUBMANDIBULAR MAMMARY GLAND PARATHYROID PEYER S PATCH S P I N A L CORD, CERVICAL S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC T.H.Y.M. .U.S. . . . . . . . . . . . . . . . . . . . . .T.R.A.C.H.E.A. . . . . . . . . . . . . . . . . . . . .U.R. I.N.A.R.Y. . B. .LA. D. .D.E.R. . . End o f Record- 19012 CECUM ILEUM LUNG NERVE, RECTUM SPLEEN VAGI NA SCIATIC 11-51 4 18-028:PAGE 5-82 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 4 1 8 - 0 2 8 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o..t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: LYMPH NODE, SUBMANDIBULAR: hyperplasia, l y m p h o c y t i c / p l a s m a c y t i c ( m i l d ) .U.T.E.R.U.S.:.....................................m.a.c.r.o.p.h.a.g.e.s.,..p.i.g.m.e..n.t e.d...(m..i I.d..).............................. THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COLON JEJUNUM LYMPH NODE, MEDIASTINAL PARATHYROID S P I N A L CORD, LUMBAR THYMUS V..A.G.I.N.A.................. E n d o f R e c o r d - 19019 BONE MARROW (STERNUM) DUODENUM KIDNEYS MAMMARY GLAND PEYER'S PATCH S P I N A L CORD, THORACIC THY RO ID BRAIN HEART LIVER NERVE, SCIATIC RECTUM SPLEEN TRACHEA CECUM ILEUM LUNG OVAR IES S P I N A L CORD, CERVICAL STOMACH URINARY BLADDER 11-52 4 18-028:PAGE 5-83 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Indi v i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S1: - H..E.A.D.: ..H.e.a.d.-..s.c.a.b.,..0...2.cm..x.0..4.c.m...............................d.e.r.m..a.t.i .t i.s.,..c.h..r.o.n.i.c., ..f o..c.a.l.,..S.K.I.N..(.G.R.O.S.S..L.E.S.I.ON) HISTOMORPHOLOGIC OBSERVATIONS: LIVER: i n f l a m m a t i o n , chronic, focal/mul tifocal (minimal) LYMPH NODE, MEDIASTINAL: congestion ( m i n i m a l ) S K I N (GROSS LESION) : d e r m a t i t i s , chronic, focal ( m i l d ) THYROID : u l t i m o b r a n c h i a1 body/cyst U..TE.R..U.S.:....................................m..a.c.r o.p.h..a.g.e.s.,..p.i g.m..e.n.t.e.d..(.m.o.d.e.r.a.t.e.)........................... THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) COLON DUODENUM JEJUNUM KIDNEYS MAMMARY GLAND NERVE, SCIATIC PEY ER S PATCH RECTUM S P I N A L CORD, THORACIC SPLEEN T.R. A. .C.H.E.A. . . . . . . . . . . . . . . . . . . . .V.A.G.I.N. A. . . . . . . . . . . . . . BRAIN HEART LUNG OVARIES SPINAL CORD, STOMACH CERVICAL CECUM ILEUM LYMPH NODE, SUBMANDIBULAR PARATHYRO ID S P I N A L CORD, LUMBAR THYMUS T I S S U E ( S 1 NOT A V A I L A B L E FOR EVALUATION: 11-53 418-028:PAGE J-84 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S) : HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: LUNG : in fi1t r a t ion, polymorphonucl ear, p e r i vascul a r / p e r i bronchi a1 (mild) LYMPH NODE, MEDIASTINAL: mastocytosis (minimal) STOMACH: edema/i n f l ammation, submucosa, g l andul a r area (mi 1d) U..TE.R..U.S.:....................................m..a.c.ro.p.h.a.g..e.s., .p..ig.m..e.n.t.e.d..(.m.i.n.im..a.l.)............................ THE FOLLOWING TISSUE(S) WERE WITHIN NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) COLON JEJUNUM DUODENUM KIDNEYS MAMMARY GLAND PEY ER S PATCH NERVE, SCIATIC RECTUM SPINAL CORD, THORACIC SPLEEN T.R. A. C. .H.E.A. . . . . . . . . . . . . . . . . . . . .U.R. I.N.A.R. Y. . .B.LA.D. .D.E.R. . . . End o f Record- 19023 BRAIN CECUM HEART ILEUM LIVER LYMPH NODE, SUBMANDIBULAR OVAR IES PARATHYROID SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR THYMUS THYROID . V. .A.G.I.N.A. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-54 418-028:PAGE 5-85 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Indiv i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATIONS: LYMPH NODE, SUBMANDIBULAR: hyperpl a s i a, 1y m p h o c y t ic / p l a s m a c y t ic ( m o d e r a t e ) STOMACH : edema/i n f lammati on, submucosa, g landul a r area ( m o d e r a t e ) THYROID : u l t i m o b r a n c h i a1 body/cyst U. .TE.R.U. .S.:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .m. .a.c.ro.p.h.a.g.e.s.,. .p. .ig.m. .e.n.t.e.d. .(.m.o.d.e.r.a.t.e.) . . . . . . . . . . . . . . . . . . . . . . . . . . . THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COLON BONE MARROW (STERNUM) DUODENUM JEJUNUM LYMPH NODE, MEDIASTINAL PARATHYROID S P I N A L CORD, LUMBAR KIDNEYS MAMMARY GLAND PEYER 'S PATCH S P I N A L CORD, THORACIC T.R. A. C. .H.E.A. . . . . . . . . . . . . . . . . . . . .U.R.I.N.A.R.Y. .B.L.A.D.D.E.R. . . . . End o f Record- 19041 BRA1N HEART LIVER NERVE, S C I A T I C RECTUM SPLEEN -V--A-G-IN-A---------- CECUM ILEUM LUNG OVARIES S P I N A L CORD, THYMUS CERVICAL 11-55 418-028:PAGE 5-86 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMNTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l o g y D a t a GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o.t..a.p.p.l.i.c.a.b.1.e................................ THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COLON JEJUNUM LYMPH NODE, MEDIASTINAL PARATHYROID BONE MARROW (STERNUM) BRAIN DUODENUM HEART KIDNEYS LIVER LYMPH NODE, SUBMANDIBULAR MAMMARY PEYER'S PATCH RECTUM S P I N A L CORD, LUMBAR THYMUS S P I N A L CORD, THORACIC TRACHEA SPLEEN URINARY ------------------c------------------------------------------------- GLAND BLADDER End of Record- 19042 CECUM ILEUM LUNG NERVE, SCIATIC S P I N A L CORD, CERVICAL STOMACH VAG INA .--------------------------- 11-56 4 18-028:PAGE 5-87 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l o g y D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o..t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: KIDNEYS: nephritis, chronic, focal (minimal) LYMPH NODE, MEDIASTINAL: congestion ( m i n i m a l ) hyperpl a s i a, 1y m p h o c y tic / p l a s m a c y t ic ( m i 1d) LYMPH NODE, SUBMANDIBULAR: hyperpl a s i a, 1ymphocyt ic/pl a s m a c y t ic ( m o d e r a t e ) U..TE.R..U.S.: ....................................m..a.c.r o.p.h..a.g.e.s., ..p.i g.m..e.n.t.e.d..(.m.o.d.e.r.a.t.e.)........................... THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COLON JEJUNUM NERVE, S C I A T I C RECTUM SPLEEN T..R.A.C.H.E.A.................. End o f R e c o r d - 19044 BONE MARROW (STERNUM) DUODENUM LIVER OVAR IES S P I N A L CORD, CERVICAL STOMACH ..U.R.I.N.A.R..Y..B.L.A.D.D.E..R..... BRAIN CECUM HEART ILEUM LUNG MAMMARY GLAND PARATHYROID PEYER 'S PATCH S P I N A L CORD, LUMBAR SPINAL CORD, THORACIC THYMUS THYROID . .V.AG. .I.N.A. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-57 418-028:PAGE 5-88 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTIONJDEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Individual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: ADRENAL GLANDS: vacuol a t i on, zona g l o m e r u l osa ( m i 1d) LYMPH NODE, SUBMANDIBULAR: hyperplasia, l y m p h o c y t i c l p l a s m a c y t i c ( m i n i m a l ) U..T.E.R.U.S.: .....................................m.a.c.r.o.p.h.a.g.e.s.,..p.i.g.m.e..n.t e..d..( m..o.d.e.r.a.t e.)........................... THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : BONE MARROW (STERNUM) BRAIN CECUM DUODENUM HEART ILEUM KIDNEYS LIVER LUNG MAMMARY GLAND PEYER 'S PATCH NERVE, SCIATIC RECTUM OVAR IES S P I N A L CORD, CERVICAL S P I N A L CORD, THORACIC SPLEEN STOMACH T. H. .Y.R.O.I.D. . . . . . . . . . . . . . . . . . . . .T.R.A.C.H.E.A. . . . . . . . . . . . . . . . . . . . .U.R.I.N.A.R.Y. . B. L.A. D. .D.E.R. . . . . E n d o f R e c o r d - 19050 COLON JEJUNUM LYMPH NODE, MEDIASTINAL PARATHYROID S P I N A L CORD, LUMBAR THYMUS VAG INA 11-58 418-028:PAGE 5-89 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPME"AL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphol ogy Data GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: ADRENAL GLANDS: vacuol a t ion I zona g l omerulosa (mi 1d) CECUM: inflammation, mucosa, chronic (moderate) LYMPH NODE, SUBMANDIBULAR: hyperpl a s i a I 1ymphocyt ic / p l asmacyt ic (mi 1d) THYMUS : atrophy (moderate) THYROID : u l timobranchi a1 body/cyst U..TE.R..U.S.:....................................m..a.c.ro.p.h.a..g.e.s., ..p.ig.m..e.n.t.e.d..(.m.o.d.e..ra.t.e.)........................... THE FOLLOWING TISSUE(S1 WERE WITHIN NORMAL LIMITS: BONE MARROW (STERNUM) BRAIN COLON DUODENUM HEART ILEUM JEJUNUM KIDNEYS LIVER NERVE, SCIATIC LUNG OVARIES LYMPH NODE, MEDIASTINAL MAMMARY GLAND PARATHYROI 0 PEY ER 'S PATCH RECTUM SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPINAL CORD, THORACIC SPLEEN STOMACH TRACHEA URINARY BLADDER V. A. G. .I.N.A. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . End o f Record- 19053 11-59 4 18-028{PAGEJ-90 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: CECUM : i n f l a m m a t i o n , mucosa, chronic ( m i l d ) LIVER: LYMPH NODE, SUBMANDIBULAR: in f1ammat ion , chroni c , focal /mu1 t i f o c a l ( m i n ima1 ) hyperpla s i a , 1ymphocyt ic/pl a s m a c y t ic ( m i 1d) URINARY BLADDER: i n f l a m m a t i o n , chronic, focal ( m i n i m a l ) .U.T.E.R.U.S.:.....................................m.a.c.r.o.p.h.a.g.e.s.,..p.i.g.m..e.n.t e.d...(m..i 1.d..).............................. THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS DUODENUM KIDNEYS NERVE, S C I A T I C RECTUM SPLEEN T.R..A.C.H.E.A.................. E n d o f R e c o r d - 19065 BONE MARROW (STERNUM) BRAIN COLON HEART ILEUM JEJUNUM LUNG LYMPH MODE, MEDIASTINAL MAMMARY GLAND OVARI ES PARATHYR O I D PEYER S PATCH S P I N A L CORD, CERVICAL S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC STOMACH THYMUS THYROID . V.A.G. .I.N.A. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-60 418-028:PAGE J-91 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL T O X I C I T Y SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR S STUDY NUMBER T-7706.1 Appendix I 1 I n d iv i dual Animal Gross and H i stomorphol ogy Data HISTOMORPHOLOGIC OBSERVATIONS: LUNG : inflammation, i n t e r s t i t i a l , focal (minimal) macrophages, a1veol i, focal (mi nimal ) TRACHEA : inf 1ammation, chronic , focal (mi ni ma1) UTERUS : macrophages, pigmented (moderate) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .d. .i s.t.e.n. .t .io.n.,. .1.u.m. .e.n. .(m. .i.1.d.). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . THE FOLLOWING TISSUE(S) WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN CECUM COLON DUODENUM HEART ILEUM JEJUNUM KIDNEYS LIVER LYMPH NODE, MEDIASTINAL LYMPH NODE, SUBMANDIBULAR MAMMARY GLAND NERVE, SCIATIC O V A R I ES PARATHYROID PEYER'S PATCH RECTUM SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPINAL CORD, THORACIC SPLEEN STOMACH T.H.Y.M. U. .S. . . . . . . . . . . . . . . . . . . . .T.H.Y.R.O. .I D. . . . . . . . . . . . . . . . . . . . .U.R.IN. A. R. .Y. .B.LA. D. .D.E.R. . . . . . . . . . . .V.A.G.I.N.A. . . . . . . . . . . . . . . . . . End o f Record- 19001 11-61 418-028:PAGE J-92 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 4 1 8 - 0 2 8 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l o g y D a t a GROSS OBSERVATION(S) : HISTOMORPHOLOGIC OBSERVATION(S) : HISTOMORPHOLOGIC OBSERVATIONS: LYMPH NODE, SUBMANDIBULAR: hyperpl a s i a, 1ymphocyt ic / p l a s m a c y t i c ( m i 1d) THYROID : u l t i m o b r a n c h ia1 body/cyst U..TE.R..U.S.:....................................m..a.c.r o.p..h.a.g.e.s., ..p.i g.m..e.n.t.e.d..(.m.i.1.d.)............................... THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) COLON J EJUNUM DUODENUM KIDNEYS LYMPH NODE, MEDIASTINAL MAMMARY GLAND PARATHYROID PEYER 'S PATCH S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC T.H.Y.M..U.S......................T.R..A.C.H.E.A............. E n d o f R e c o r d - 19006 BRAIN CECUM HEART ILEUM LIVER LUNG NERVE, SCIATIC OVAR IES RECTUM SPINAL CORD, CERVICAL SPLEEN STOMACH . . . .U.R.I N. .A.R.Y. .B.L.A.D.D.E.R. . . . . . . . . . . . .V.A.G.I.N.A. . . . . . . . . . . . . . . 11-62 418-028;PAGE 5-93 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 4 1 8 - 0 2 8 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Individual A n i m a l G r o s s and H i s t o m o r p h o l o g y D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATIONS: ADRENAL GLANDS : necrosis, cortex, focal (mi Id) LIVER: Not remarkable U..TE.R..U.S.: ....................................m..a.c.r o.p..h.a.g.e.s., ..p.ig.m..e.n.t.e.d..(.m.o.d.e.r.a.t.e.) ........................... THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : BONE MARROW (STERNUM) BRAIN CECUM COLON DUODENUM KIDNEYS HEART LUNG ILEUM LYMPH NODE, MEDIASTINAL JEJUNUM LYMPH NODE, SUBMANDIBULAR MAMMARY GLAND PEYER S PATCH NERVE, SCIATIC RECTUM O V A R I ES S P I N A L CORD, CERVICAL PARATHYROI D S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC SPLEEN STOMACH THYMUS T. H. .Y.R.O.I .D. . . . . . . . . . . . . . . . . . . .T.R.A.C. H. E. A. . . . . . . . . . . . . . . . . . . . .U.R.I.N.A.R.Y. .B.L.A.D.D.E.R. . . . . . . . . . . . .V.A.G.I.N.A. . . . . . . . . . . . . . . . . E n d o f R e c o r d - 19011 11-63 4 18-028:PAGE 5-94 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Appendix I 1 I n d i v i d u a l Animal Gross and H i stomorphology Data GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S): ADRENAL GLANDS: Bi 1a t e r a l - 1arge. GASTROINTESTINAL TRACT: Abdominal d i s t e n t i o n , l a r g e and small i n t e s t i n e s distended w i t h gas. SPLEEN: Small . STOMACH: Fundic mucosa surface contains approximately 12 black areas ranging i n size from p i n p o i n t t o 0.3cm i n diameter. T.H. Y. M. .U.S.: . .S.m. .a.l.l.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . degeneration, c y s t ic , cortex, mu1t i focal No microscopic change t o c o r r e l a t e , DUODENUM No microscopic change t o c o r r e l ate, JEJUNUM No mi croscopi c change t o c o r r e l ate, ILEUM No m i croscopi c change t o c o r r e l ate, COLON No m i croscopi c change t o c o r r e l ate, CECUM No m i croscopi c change t o c o r r e l ate, RECTUM atrophy edema/i n f la m a t ion, submucosa , g l andul a r area edema/inflammation, submucosa, forestomach erosi on( s), g l andul a r mucosa . . . . . . . . .a.t.r .o.p.h.y. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . HISTOMORPHOLOGIC OBSERVATIONS: ADRENAL GLANDS : degeneration, cystic, cortex, multifocal (mild) CECUM: No microscopic change t o c o r r e l a t e COLON: No m i croscopi c change t o c o r r e l a t e DUODENUM : No microscopic change t o c o r r e l a t e ILEUM: No m i croscopi c change t o c o r r e l a t e JEJUNUM: No m i croscopi c change t o c o r r e l a t e LIVER: necrosis, focal (minimal ) LYMPH NODE, SUBMANDIBULAR: hyperpl a s i a, 1ymphocyt ic / p l asmacyt ic (mi1d) RECTUM: No microscopic change t o c o r r e l a t e SPLEEN: atrophy (mild) STOMACH: erosion( s) , glandular mucosa (moderate) edema/inflammation, submucosa, glandular area (mi I d ) edema/inflammation, submucosa, forestomach (mi I d ) THYMUS: atrophy (marked) THYROID : u l t imobranchi a1 body/cyst U. .T.E.R.U.S.:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .m. .a.c.ro. p. .h.a.g.e.s.,. .p.ig.m. .e.n.t.e.d. .(.m.a.r.k.e.d.). . . . . . . . . . . . . . . . . . . . . . . . . . . . . THE FOLLOWING TISSUE(S1 WERE WITHIN NORMAL LIMITS: BONE MARROW (STERNUM) DUODENUM KIDNEYS NERVE, S C I A T I C RECTUM T.R..A.C.H.E.A.............. End o f Record- 19020 BRAIN CECUM HEART ILEUM LUNG LYMPH NODE, MEDIASTINAL OVARIES PARATHYROID SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR ....U.R.I.N.A.R.Y..B.L.A.D.D.E.R.............V.A.G..I.N.A................ COLON JEJUNUM MAMMARY GLAND PEY ER s PATCH SPINAL CORD, THORACIC 11-64 418-028;PAGE J-95 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTIONIDEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l o g y D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N..o.t .a.p.p..l i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: LIVER: i n f l a m m a t i o n , chronic, f o c a l / m u l tifocal ( m i n i m a l ) LYMPH NODE, SUBMANDIBULAR: hyperpl a s i a, 1ymphocyt ic/pl a s m a c y t ic ( m i 1d) MAMMARY GLAND: necrosi s , focal ( m i n i m a l ) THYROID : u lt i m o b r a n c h i a l body/cyst U..TE.R.U..S.: ....................................m..a.c.r o.p.h..a.g.e.s., ..p.ig.m..e.n.t.e.d..(.m.i.1.d.)............................... THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN CECUM COLON DUODENUM HEART ILEUM JEJUNUM KIDNEYS LUNG LYMPH NODE, MEDIASTINAL NERVE, SCIATIC OVARI ES PARATHYROID PEYER 'S PATCH RECTUM S P I N A L CORD, CERVICAL S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC SPLEEN STOMACH THYMUS TRACHEA .U.R.I N. A. R. .Y. .B.L.A.D.D.E.R. . . . . . . . . . . .V.A.G. I. N. A. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . E n d o f R e c o r d - 19022 11-65 418-028:PAGE J-96 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 A p p e n d i x 11 Indiv i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S1: HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: ADRENAL GLANDS: hypertrophy/vacuolation, cortex, mu1ti focal ( m i l d ) KIDNEYS : nephritis , chroni c, focal (mi n i m a l ) STOMACH : edema/inflammation, submucosa, glandular area (mi Id) U..TE.R..U.S.:....................................m..a.c.ro.p.h.a.g.e.s.,...p.i g.m..e.n.t.e.d..(.m.o.d.e.r.a.t.e.)........................... THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : BONE MARROW (STERNUM) DUODENUM BRAIN HEART CECUM ILEUM COLON JEJUNUM LIVER LUNG LYMPH NODE, MEDIASTINAL LYMPH NODE, SUBMANDIBULAR MAMMARY GLAND NERVE, SCIATIC OVARIES PARATHYROI D PEYER 'S PATCH RECTUM S P I N A L CORD, CERVICAL S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC SPLEEN THYMUS THYROID T.R. A. C. H. .E.A. . . . . . . . . . . . . . . . . . . .U.R.I.N.A.R.Y. .B.L.A.D.D.E.R. . . . . . . . . . . . .VA. G. .I.N.A. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . E n d o f R e c o r d - 19025 11-66 4 18-028:PAGE 5-97 OR 9VA DMBINED R ATED X TY TUDY F T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Individual A n i m a l G r o s s and H i s t o m o r p h o l o g y D a t a GROSS OBSERVATION(S1 : HISTOMORPHOLOGIC OBSERVATION(S1: G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o.t..a.p.p.l.i.c.a.b.1.e................................ H ISTOMORPHOLOGIC OBSERVATIONS : THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COLON JEJUNUM MAMMARY GLAND RECTUM SPLEEN .U.T.E.R.U.S................. BONE MARROW (STERNUM) BRAIN DUODENUM HEART LIVER LUNG OVARIES PARATHYROID S P I N A L CORD, CERVICAL S P I N A L CORD, THYMUS TRACHEA VAG INA .---------------_--------------------------------- LUMBAR T I S S U E ( S 1 NOT AVAILABLE FOR EVALUATION: CECUM ILEUM LYMPH NODE, MEDIASTINAL PEYER ' S PATCH S P I N A L CORD, THORACIC URINARY BLADDER I1-67 418-028:PAGE 5-98 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Indiv i dual A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S) : H ISTOMORPHOLOGIC OBSERVATION(S1: G..E.N.E.R.A.L.: ..N.o..g.r.o.s.s..c.h.a.n.g.e.s..................................N.o.t..a.p.p.l.i.c.a.b.1.e................................ HISTOMORPHOLOGIC OBSERVATIONS: LYMPH NODE, SUBMANDIBULAR: hyperpl a s i a, 1y m p h o c y t ic / p l a s m a c y t ic ( m o d e r a t e ) STOMACH: dilatation, m u c o s a l glands ( m i n i m a l ) U..TE.R..U.S.:....................................m..a.c.r o.p.h..a.g.e.s., ..p.i.g.m.e.n.t.e.d..(.m.o.d.e.r.a.t.e.)........................... THE FOLLOWING TISSUE(S) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN CECUM COLON DUODENUM HEART ILEUM JEJUNUM KIDNEYS LIVER LUNG LYMPH NODE, MEDIASTINAL MAMMARY GLAND NERVE, S C I A T I C OVARIES PARATHYROID PEYER S PATCH RECTUM S P I N A L CORD, CERVICAL S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC SPLEEN THYMUS T. H. .Y.R.O.I D. . . . . . . . . . . . . . . . . . . . .T.R.A.C.H.E.A. . . . . . . . . . . . . . . . . . . . .U.R.I N. A. R. .Y. .B.L.A.D.D.E.R. . . . . . . . . . . .V.A.G. I.N.A. . . . . . . . . . . . . . . E n d o f R e c o r d - 19028 11-68 418-028:PAGE J-99 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 Individual A n i m a l G r o s s and H i s t o m o r p h o l o g y D a t a GROSS OBSERVATIONIS): HISTOMORPHOLOGIC OBSERVATION(S): - H..E.A.D.: ..H.e.a.d.-..s.c.a.b.,..0...3.c.m..x..0...3.c.m............................d.e.r.m.a..t i.t.i.s., ..c.h.r.o.n.i.c.,..f.o.c.a..l ,..S.K..I N...(G.R.O.S.S...L.E.S.IO.N) HISTOMORPHOLQGIC OBSERVATIONS: LIVER: Not remarkable S K I N (GROSS LESION) : d e r m a t i t i s , chronic, focal ( m i n i m a l ) THYROID : u l t i m o b r a n c h i a1 body/cyst U..TE.R..U.S.:....................................m..a.c.r o.p..h.a.g.e.s.,..p.i g.m..e.n.t.e.d..(.m.o.d.e.r.a.t.e.)........................... THE FOLLOWING T I S S U E ( S ) WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS BONE MARROW (STERNUM) BRAIN CECUM COLON DUODENUM HEART ILEUM JEJUNUM KIDNEYS LUNG LYMPH NODE, MEDIASTINAL MAMMARY GLAND NERVE, S C I A T I C OVARI ES PARATHYROID PEYER'S PATCH RECTUM S P I N A L CORD, CERVICAL S P I N A L CORD, LUMBAR S P I N A L CORD, THORACIC SPLEEN STOMACH THYMUS T. R. .A.C.H.E.A. . . . . . . . . . . . . . . . . . . . .U.R.I N. .A.R.Y. .B.L.A.D.D.E.R. . . . . . . . . . . . .V.A.G.I.N.A. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . T I S S U E ( S ) NOT A V A I L A B L E FOR EVALUATION: L.Y.M. .P.H. .N.O.D.E.,. .S.U.B.M.A.N.D.I.B.U.L.A.R. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . E n d o f R e c o r d - 19030 11-69 418-028:PAGE J-100 ORAL (GAVAGE) COMBINED REPEATED DOSE T O X I C I T Y STUDY OF T - 7 7 0 6 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR S STUDY NUMBER T - 7 7 0 6 . 1 Appendix I1 I n d i v i d u a l A n i m a l G r o s s and H i s t o m o r p h o l ogy D a t a GROSS OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATION(S): HISTOMORPHOLOGIC OBSERVATIONS: LIVER: necrosi s , focal (mi nimal ) LYMPH NODE, SUBMANDIBULAR: hyperplasia, l y m p h o c y t i c l p l a s m a c y t i c (mi Id) U..TE.R..U.S.:....................................d..i s..te..n.t.i .on..,..1.u.m.e.n..(.m.o.d.e.r.a.t.e.) ................................ THE FOLLOWING T I S S U E ( S 1 WERE W I T H I N NORMAL L I M I T S : ADRENAL GLANDS COLON JEJUNUM MAMMARY GLAND PEYER S PATCH S P I N A L CORD, THORACIC T..H.Y.R.O..ID............... E n d o f R e c o r d - 19031 BONE MARROW (STERNUM) BRAIN DUODENUM HEART KIDNEYS LUNG NERVE, SCIATIC OVARIES RECTUM SPINAL CORD, CERVICAL SPLEEN TRACHEA STOMACH URINARY BLADDER ,------------------------------------------------------------------------------- CECUM ILEUM LYMPH NODE, MEDIASTINAL PARATHYROID S P I N A L CORD, LUMBAR THYMUS VAG INA 11-70 418-028:PAGEJ- 101 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTlON/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT APPENDIX Ill HISTOMORPHOLOGIC OBSERVATIONS IN THE LIVER F I GENERATION PUPS 418-028:PAGE J-102 ORAL (GAVAGE) COMBINED REPEATED DOSE STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Table 111-1 HistomorphologicObservationsin the Liver - Group I - F1 Generation Pups Dose Group: Dam Number: Pup Number: I I I I I I I I I I 19012 19012 19012 19012 19012 19012 19012 19012 19012 19012 1 2 3 4 5 7 8 141516 LIVER: Dose Group: Dam Number: Pup Number: LIVER: I I I I I I I I I I 19019 19019 19019 19019 19019 19019 19019 19019 19019 19019 1 2 3 7 8 11 12 13 14 15 Dose Group: Dam Number: Pup Number: LIVER: I I I I I I I I I I 19021 19021 19021 19021 19021 19021 19021 19021 19021 19021 1 2 6 8 9 10 11 12 13 14 Dose Group: Dam Number: Pup Number: LIVER: I I I I I I I I I I 19023 19023 19023 19023 19023 19023 19023 19023 19023 19023 1 2 3 6 7 8 9 101112 Dose Group: Dam Number: Pup Number: LIVER: I I I I I I I I I I 19041 19041 19041 19041 19041 19041 19041 19041 19041 19041 1 2 3 4 5 8 9 151617 111-1 418-028;PAGE J-103 ORAL (GAVAGE) COMBINED REPEATED DOSE STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Table Ill-I (Continued) HistomorphologicObservationsin the Liver- Group I - F1 Generation Pups Dose Group: Dam Number: Pup Number: I I I I I I I I I I 19042 19042 19042 19042 19042 19042 19042 19042 19042 19042 1 2 4 5 6 7 8 I01213 LIVER: Dose Group: Dam Number: Pup Number: LIVER: I I I I I I I I I I 19044 19044 19044 19044 19044 19044 19044 19044 19044 19044 1 2 5 6 7 9 11 12 13 14 Dose Group: Dam Number: Pup Number: LIVER: I I I I I I I I I I 19050 19050 19050 19050 19050 19050 19050 19050 19050 19050 1 2 3 4 5 6 7 8 1314 Dose Group: Dam Number: Pup Number: LIVER: Dose Group: Dam Number: Pup Number: LIVER: I I I I I I I I I I 19053 19053 19053 19053 19053 19053 19053 19053 19053 19053 1 3 4 5 6 10 11 12 13 14 I I I I I I I I I I 19065 19065 19065 19065 19065 19065 19065 19065 19065 19065 3 4 5 7 8 10 11 14 15 17 I 11-2 418-028:PAGE J-104 ORAL (GAVAGE) COMBINED REPEATED DOSE STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Dose Group: Dam Number: Pup Number: LIVER : Table 111-2 HistomorphologicObservationsin the Liver- GroupV - F1 GenerationPups vvvvvvvvv 19001 19001 19001 19001 19001 19001 19001 19001 19001 1 234567911 Dose Group: Dam Number: Pup Number: LIVER : vvvvvvvvvv 19006 19006 19006 19006 19006 19006 19006 19006 19006 19006 1 2 3 4 5 8 10 11 13 18 Dose Group: Dam Number: Pup Number: LIVER : vvvvvvvvvv 19011 19011 19011 19011 19011 19011 19011 19011 19011 19011 2 3 4 5 6 9 10111213 Dose Group: Dam Number: Pup Number: LIVER : vvvvvvvvvv 19020 19020 19020 19020 19020 19020 19020 19020 19020 19020 2 4 5 7 8 9 10111213 Dose Group: Dam Number: Pup Number: LIVER : vvvvvvvvvv 19022 19022 19022 19022 19022 19022 19022 19022 19022 19022 1 4 5 7 8 11 12 13 14 15 I 11-3 418-028:PAGE J- 105 ORAL (GAVAGE) COMBINED REPEATED DOSE STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Dose Group: Dam Number: Pup Number: LIVER : Table 111-2 (Continued) HistomorphologicObservationsin the Liver - Group V - F1 Generation Pups vvvvvvvvvv 19025 19025 19025 19025 19025 19025 19025 19025 19025 19025 1 2 7 8 13 14 15 18 19 20 Dose Group: Dam Number: Pup Number: LIVER : vvvvvvvvv 19027 19027 19027 19027 19027 19027 19027 19027 19027 1 2 3 4 5 6 7 1314 Dose Group: Dam Number: Pup Number: LIVER : vvvvvvvvvv 19028 19028 19028 19028 19028 19028 19028 19028 19028 19028 1 2 3 4 5 6 7 101112 Dose Group: Dam Number: Pup Number: LIVER : vvvvvvvvvv 19030 19030 19030 19030 19030 19030 19030 19030 19030 19030 1 2 4 5 6 8 9 111213 Dose Group: Dam Number: Pup Number: LIVER : vvvvvvvvvv 19031 19031 19031 19031 19031 19031 19031 19031 19031 19031 1 2 3 5 6 9 11 12 14 15 I 11-4 APPENDIX K HEMATOLOGY AND CLINICAL CHEMISTRY REPORTS Study Report f o r Hematology 418-028:PAGE K-1 GROUP: 11 91 09 19115 19116 19479 I9122 19134 13151 13.5 15.9 15.1 12.7 16.4 17.6 47. 7 "9*9 45.0 14.3 -- 7.10 8.58 6.95 7.16 6.77 7.61 7.37 7.45 7.43 8.71 -- 15.4 19.0 16.3 16.2 15.7 16.2 16.3 16.3 15.6 18.2 -- 40.6 50.0 42.6 42.1 40.6 41 *4 41 -6 43.3 42.7 49.7 -- 57.2 58.3 61.3 58.8 59.9 54.4 56.5 58.1 57.5 57.1 21.7 22.1 23.5 22.6 23.2 21.3 22.1 21.9 21 .o 20.9 -- cc 37.9 38.0 38.3 38.5 38.7 39.1 39.2 37.6 36.5 36.6 -- 1219 1072 964 1354 1 074 1322 1 049 1264 7 269 1203 MEAN SD N 15.3 1.63 10 7.51 0.648 10 16.5 1.16 10 43-5 3.415 10 57.9 1.89 10 22.0 0.87 10 38.0 0.93 10 1179 10 418-028:PAGE K-2 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Animal ID WBC RBC M MILL/CU MIM GROUP: 2-H 19102 19106 19108 19140 19143 19120 19129 19147 19153 40.9 17.0 12.7 13.1 15.7 18.4 14.7 10.5 17.2 19.0 -- 7.55 6.93 7.02 8.20 6.80 7.42 7.29 7.32 7.53 7.20 -- HGB GRAWS/DL 16.2 '15.2 16.1 15.9 16.1 16.4 IS.? 15.4 16.4 15.5 -- HCT MCV MCH % CU MICRONS PIC0 GRAMS 43.6 39.6 40.3 43.9 42.0 44.4 43.2 41 - 3 42.2 41.7 -- 57.7 57.1 57.4 53.5 61.8 59.8 59.3 56.4 56.0 57.9 -- 21.5 21 .? 22.9 19.4 23.7 22.1 21.8 21 .o 21.8 21.5 -- SEX: MALE MCHC % PLT THSN/CU MM 37.2 38.4 40.0 36.2 38.3 36.9 36.8 37.3 38.9 37.2 -- 926 970 959 1468 1170 1382 1184 1183 1069 994 -- MEAN SD N 14.9 3.03 10 7.33 0.396 10 15.9 0.42 10 42.2 1.57 10 57.7 2-27 10 21. 1.13 10 37.7 1. I 5 I0 1131 183.7 10 LABCAT HE4.43 418-028:PAGE K-3 HEAN SD N 13.8 2-84 10 7.32 0.441 10 15.7 0.59 10 42.0 2.22 10 57.4 1.78 10 21.5 0.75 10 37.5 0.79 10 1115 149.2 10 (--I - Data Unavailable LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-4 GROUP: 4-H 19100 19103 19104 19118 19133 19141 19142 19144 19148 19150 19156 19160 19162 49163 19164 MEAN SD N 12.6 21 .o 18.1 12.0 22.0 17.4 12.8 19.6 15.6 11 .I 16.2 3.97 10 6.59 6.73 6.75 6.83 7.50 7.a7 6.87 7.37 6.97 6.Q -- I - 6.93 0.305 10 14.6 15.0 15.6 14.8 16.8 15.5 15.0 16.1 15.6 15.0 -- -- 15.4 0.67 10 38.2 39.0 39.3 38.4 45.5 40.3 33.9 42.4 40.6 - 38.4 57.9 58.0 58.2 56.2 60.7 57.0 58.1 57.5 58.2 57.6 -- I 40.2 2.29 I0 57.9 1.15 10 22.2 22.3 23.1 21 -7 22.4 21.9 21.8 21- 8 22.4 22.7 38.2 38.5 39.7 38.5 36.9 38.5 37.6 38.0 38.4 39-.4- 22.2 0.45 10 38.4 0.80 10 1072 1144 1029 1274 1124 1125 998 962 1155 11-6-1 -- 1104 91- 4 10 LABCAT HE4. 43 27-JUN-2002 Study Report for Hematology STUDY NO: 060-069 SEX: MALE GROUP: 5-H 19111 19117 19124 13126 19127 19128 19140 4 91 45 1914 19152 19154 19158 19168 19169 19170 SD N 15.7 14.3 10.8 12.5 11.o 14.2 15.2 20.7 18.3 13.9 -- ..,- 14.7 3.07 10 7.20 6.72 6.68 8.07 6.61 7.09 7.05 7.13 6.66 6.68 -- 6.99 0.442 10 16.1 15.7 15.0 17.3 14.7 15.5 16.2 15.7 14.8 14.8 c- 15.6 0.81 10 41.7 39.8 39.0 44.4 38.1 41 .I 41.9 41.4 39.8 39.5 57.9 59.2 58.4 55 .O 57.6 58.0 59.5 58.1 59.8 59.2 ^C 22.4 23.4 22.5 21-4 22.2 21.9 23.0 22.0 22.2 22.2 38.6 39.4 38.5 39.0 38.6 37.7 38.7 37.9 37.2 37.5 1013 1181 1249 1142 1040 1001 1059 4 523 1234 1041 40.7 1.81 10 58.3 1-37 10 22.3 0.56 10 38.3 0.70 10 I148 159.9 10 LABCAT HE4 - 43 27-JUN-2002 418-028:PAGE K-6 L__IC______-III__CCI_I.-----------C-------------------------------------------------------------------~-~------~---------- Animal I D WBC THSN/CU MM RBC MILL/CU GROUP: 1-F 19010 1 901 2 19019 19021 19023 19042 1 9044 19050 79068 19074 19075 -- 15.9 6.6 7.4 7.3 8.0 6.8 10.6 8.2 6.6 12.5 -- ---- -- 5.60 6.95 7.10 6.62 6.86 6.45 6.09 6. 6.08 6.67 --- -- -a 9.0 6.51 SD 3.09 0.461 N 10 10 HGB -- 74.0 16.4 16.7 16.3 16.2 15.1 14.3 16.0 14.7 15.7 I -- I - -- 15.5 0.95 10 HCT -- 35.2 42.5 44.7 42.4 42.4 40.2 37.6 42.2 37.5 40.2 -- -- -- -.". 70 MCV ICRONS CM P I C 0 GRAMS -- 62.9 61 .I 63.0 44.1 61.8 62.3 61.7 63.6 61.6 60.2 -- ---- 62.2 1.19 10 -- 25.0 23.6 23.5 24.6 23.6 23.4 23.5 24.1 24.2 23.5 -- -- -- -- 23.9 0.55 10 RCHC % PLT THSN/CU RH -- 39.8 38.6 37.4 38.4 38.2 37.6 38.0 37.9 39.2 39.1 -- --- . . . c 38.4 0.76 10 -- 1421 1344 1199 1352 1529 1225 1343 '1629 1524 1623 ---- -- 1419 153.0 10 27-JUN-2002 418-028:PAGE M-7 GROUP: 2-F 19004 1WO9 19016 I9018 19026 19037 19043 19047 19048 19052 19055 19861 19067 19071 SD N 7.8 14.6 47.0 13.3 9.1 18.0 26.3 13.5 11 .I 7.1 -- 13.8 5.73 10 7.07 5.90 6.32 5.61 6.36 6.30 6.23 5.83 5.93 6.60 -- c- 6.22 0.423 10 16.6 14.8 14.3 14.1 15.5 14.5 14.8 14.7 14.5 16.0 -- 15.0 0.77 10 43.8 38.3 38.5 35.3 41.6 38.9 37.8 38.3 37.5 43.4 -- 62.0 64.9 60.9 63.0 65.4 61.7 60.6 65.7 63.3 65.8 -- 38.3 2-72 10 63.3 2.01 10 23.5 25.1 23.6 25.1 24.4 23.9 23.8 25.2 24.5 24.2 -- 24.2 0.76 18 37.9 38.6 38.7 39.9 37.3 37.3 39.2 38.4 38.7 36.9 1540 1157 1529 1057 1443 1166 1257 942 1041 1264 -- -- 38.3 0.94 10 - I 1240 208.1 10 27-JUN-2002 418-028:PAGE K-8 11-1 6.40 15.2 39.5 61.8 23.3 38.7 1274 SD 2.98 0.554 0.91 3.24 1.50 0.88 1.29 226.3 N 10 10 10 I0 10 10 10 10 LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-9 GROUP: 4- F 19002 19005 19035 19039 19040 1 9045 19046 19051 19054 19058 7 9062 19063 19066 19069 1 9073 MEAN SD N -- 9.5 9.6 12.8 10.0 9.7 -- -- 14.9 15.7 6.2 12.9 7.1 -- 10.8 3.15 10 -- 6.51 6.81 6.48 6.48 5.77 --- 7.33 5.94 6.86 5.68 6.01 -- 6.39 0 532 10 -- 15.8 16.2 14.8 15.8 - 13.7 -- 15.7 14.2 15.7 14.0 15.7 -- 15.2 0.90 10 -- 42.0 42.4 38.9 41. 36.7 -- -- 44.1 37.2 43.3 35.2 40.3 4o.l 3-02 10 -- 64.5 62.3 60.1 63.8 63.6 -- -- 60.2 62.6 63.1 61 .9 67.0 -- 62.9 2.04 10 -- 24.3 23.8 24.4 23.7 --- 21.4 23.9 22.9 24.6 26.1 -- 23.8 1.25 '10 -- 37.6 38.2 38.0 38.3 37.3 --- 35.6 38.2 36.3 39.8 39.0 -- 37.8 1.22 10 -- 1479 IS27 1066 1333 1148 -- c- 1497 1094 1399 999 1506 --- 130s 207.2 10 Study Report for Hematology 418-028:PAGE K-10 MEAN SD N 8.9 3.48 10 6.42 0.538 10 15.3 0.77 10 4Q.4 3.15 10 63.0 1.60 10 23.8 1.05 10 37.8 1.37 10 1355 176.8 10 27-JUN-2002 418-028:PAGE K-ll MEAN SD N 8.1 0.52 10 43.4 0.24 10 21 .o 3.09 10 0 11.9 2.2 0.0 0.8 0.0 1.72 0.73 0.00 0.56 10 10 10 10 10 418-028:PAGE K-12 GROUP: 2-M 19102 19106 19108 19110 19113 19120 19129 13136 19138 13439 1914? 19153 19164 19165 19171 MEAN SD N 7.5 7.6 8.4 8.4 7.2 7.6 7.2 8.8 8.1 7.8 -- 7.3 0.54 10 14.0 14.0 14.6 14.2 14.4 14.4 14.3 14.2 13.7 14.5 -- 14.2 0.27 10 22.1 22.0 24.6 26.2 20.0 17.5 20.8 19.3 16.6 31.4 -- 22.1 4.41 10 0 6.5 2.7 0.0 I.o 0 12.1 2.9 0.0 1.4 0 10.9 1.4 0.0 0.1 0 10.0 2.0 0.0 0.7 0 11- 1 3.6 0.0 0.8 0 13.4 3.1 0.0 1.5 8 9.4 4.0 0.0 0.7 0 8.6 1.4 0.0 0.3 0 11.4 5.2 0.0 0.2 0 16.3 2.5 0.0 0.2 -- -- -- -- -- 8 11.o 2.9 0.0 0.7 0.0 2.68 1.18 0.00 0.50 10 10 10 10 10 27-JUN-2002 418-028:PAGE K-13 MEAN SD N 8.2 0.72 10 13.6 0.23 10 0.6 2.38 10 0 10.8 2.0 0.0 0.4 0.0 2.02 1.06 0.03 0-51 10 10 10 10 10 418-028:PAGE I(-I4 STUDY I D : ARGUS 418-028 STUDY NO: 040-069 Animal ID MPV PT seconds GROUP: 4-H 19100 7.1 19103 8.2 19104 7.9 8.3 7.4 19141 8.4 19142 7.5 19144 9.3 19148 8.7 19150 9.3 19156 13.5 14.6 14.0 13.9 44.0 13.6 13.5 13.9 73.8 13.1 APTT seconds 19.8 26.3 23.0 21.4 19.1 21.5 20.8 21.5 19.7 18.6 C COUNT Lymphocyte THSN/CU MM Segment THSN/CU THSN/CU MM THSN/CU MM 0 7.7 4.0 0.0 0.6 0 16.4 2.3 0.0 1-7 0 12.1 3.1 0.0 2.2 0 9.4 1.4 0.0 0.5 0 17.4 2.9 0-0 0.7 0 12.7 3 .? 0.0 0.5 0 8.6 2.8 0.1 1.0 0 15.9 2.7 0.0 0.6 0 12.3 3.1 0.0 0.0 0 8.3 2.4 -- 0.0 -- 0.2 -- MEAN SD N 8.2 0.76 10 13.8 0.40 10 21- 2 2.24 10 0 12.1 2.8 0.0 0.8 0.0 3.57 0.73 0.03 0.67 10 10 10 10 10 LABCAT HE4.43 27- JUN-2002 418-028:PAGE K-15 GROUP: 519111 191 17 1912a 1 94 27 191 28 497 40 191 45 191 49 19452 19154 19158 19168 191 69 194 70 MEAN SD N 7.7 7.5 7.3 9.0 8.8 8.6 8.0 7.5 7.8 8.2 8.0 0.59 40 13.9 15.1 13.8 13.7 14.1 13.5 13.9 14.3 13.5 13.7 23.6 23.3 20.8 21.8 20.7 21.6 19.6 21 .9 19.5 20.0 14.0 0.47' 10 21.3 1.4 10 0 13.0 1.4 0 11 -3 1.7 0 10-2 0.3 0 10.1 1 .1 0 8.0 2. 0 12.4 1.4 0 12.2 2.1 0 17.2 2.5 0 16.4 2.2 0 13.1 0.4 -- c- 0 12.4 1.6 0.0 2.75 0.78 10 10 10 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 -- 0.0 0.00 10 0.6 0.9 0.2 0.9 0.2 0.3 0.2 0.8 0.0 0.4 -- 0.5 0.33 10 27'-JUN-2002 418-028:PAGE K-16 GROUP: 1-F 1904 0 19012 19019 19021 19023 19041 19042 19044 1 9850 1905 19865 19068 19072 19074 19075 MEAN SD N 7.7 8.9 7.8 7.9 9.1 8.4 8.4 8.2 8.1 8.9 8.3 0.49 10 -- 13.9 13.0 13.8 12.9 13.4 12.8 12.8 13.1 13.3 12.8 -- 0.41 10 26.6 20.4 24.2 21.9 79.3 21- 8 18.5 20.5 22.5 21.6 21.7 2.36 I0 -- -- -- 0 12.6 2.7 0.0 0.5 0 4.4 2.0 0.0 0.2 0 5.7 1.4 0.0 0.0 0 6.1 0.3 0.0 0.1 0 5.0 2.9 0.0 0.1 0 5.3 1.5 0.0 0-0 0 7.3 3.1 0.0 0.1 0 6.1 1.8 0.0 0.2 0 4.7 1.8 0.0 0.0 0 8.4 3.8 0.0 0.1 -- -- -- 0 6.6 2.2 0.0 0. I 0.0 2.44 0.90 0.00 0.15 10 10 10 10 10 CABCAT HE4.43 27-JUN-2002 418-028:PAGE K-17 MEAN SD N 8.0 0.254 10 13.4 0.29 10 23 - 3.19 10 0 9.8 0.0 3.44 10 10 0.0 0.3 0-00 0.49 10 10 10 27-JUN-2002 418-028:PAGE K-18 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ____I_ ____________---c------------------------------~----------------------------------------------------- SEX: FEMALE Animal ID ________ MPV CU MICRON PT seconds APTT seconds C Lymphocyte Segmented Bands Monocytes T THSN/CU MM THSN/CU W THSN/CU M M THSN/CU HM ___1_3-__--_--_-------------------------------------------------------------------------------------- GROUP: 3-F 19003 7.2 19007 7.3 19008 9.3 1901 3 8.5 1 901 4 -- 1901 5 9.0 1901 7 7.8 19024 8.8 19829 7.4 19034 19838 8-.1- 19056 9.3 19057 -- 19060 -- 190 -- 12.9 12.9 12.8 13.7 -- 13.6 13.3 12.9 13.3 12.7 -- 13.6 --- -- 22.7 21 -7 22.6 22.-7- 27.6 21 - 9 22.1 24.4 21-.2- 27.6 -... -- -- 0 6.5 4.0 0.0 0.3 0 5.7 2.0 0.0 0.2 0 5.0 1 .? 0.0 0.0 -0- 12.3 .,.- 3-. 8 0.0 -- 0.3 -- 0 5.7 1.6 0.0 0.1 0 9.8 2.6 0.0 0.1 0 7.4 2.6 0-0 0.2 0 10.3 2.1 0.0 0.0 -0- 10-.9- 1 .o -- 0-. 0- 0.1 -- ----0 10.2 2.2 0.0 0.4 c- -- -- -- -- -. I - -- -- -- -- -- -- MEAN SD N 8.3 0.82 10 13.2 0.37 10 23.5 2.35 10 0 8.4 2.4 0.0 0.2 0.0 2.60 0.93 0.00 0.13 10 10 10 10 10 _____-________---------------------------------------------------------------------------------------------- (--I - Data U n a v a i l a b l e 418-028:PAGE K-19 NEAN SD N 8.0 0.63 I0 13.5 0.42 10 25.3 2.45 10 0 8.2 2.4 0.0 0.2 0.0 2.52 0.96 0.09 0.11 10 I0 I0 10 10 LAECAT HE4.43 27- J UN-2002 418-028:PAGE K-20 STUDY ID: ARGUS 4 STUDY NO: 060-069 Animal I D MPV CU MICRON GROUP: 5-F 001 6.9 19006 9.2 19011 7.4 19020 7.0 19022 7.6 19025 8.3 19027 7.3 8.? 7.9 19031 6.8 1903 -- 1903 19049 PT seconds 13.8 12.8 12.9 16.8 13.7 13.5 13-2 13.6 13.7 13.5 APTT seconds 30.4 22.1 22.3 27.6 21' 6 25.3 27.5 22.2 31.5 21.6 RBC Lymphocyte Segmented M THSN/CU MM THSN/ 0 10.7 4.2 0.0 0 4.6 2.7 0.0 0 4.4 1.8 0.0 2 2.2 1.9 0.0 0 3.1 2.0 0.0 0 5.2 2.0 0.0 0 10.2 2.0 0.0 0 7.1 2.8 0.0 0 5.2 2.8 0.0 0 8.2 3.7 0.0 -- -- -- SEX: FEMALE 0.5 0.2 0.7 0.1 0.2 0.1 0.1 0.3 0.1 0.0 -- 7.7 13.8 25.2 0 6.0 0.0 0.2 0.74 1.12 3.81 0.6 2.77 0.00 0.14 N 10 10 10 10 10 10 10 10 428-028:PAGE K-21 GROUP: 1-M 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 19143 19151 19157 19161 19167 MEAN SD N 0.7 0.2 0.2 0.1 0.2 0.5 0.2 0.0 0.2 0.3 0.3 0.20 10 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0,OO 10 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.5 0.0 0.4 0.0 0.2 0.0 0.3 0.0 0.0 0.0 0.0 0.8 0.1 0.20 10 0.0 0.00 10 7-JUN-2002 Study Report f o r Hematology 418-028:PAGE K-22, MEAN SD N 0.3 0.21 10 0-0 0.00 10 0.1 0.74 10 0.0 0.00 10 T HE4.43 27-JUN-2002 418-028:PAGE M-23 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Animal ID Eosinaphil Basophils Abnormal L THSN/CU MM Other THSN/CU M GROUP: 3-M 19101 0.3 0.0 0.1 0.0 19105 0.0 0.0 0.1 0.0 19107 0.5 0.0 0.0 0.0 19112 0.3 0.0 0.1 0.0 19114 0.1 0.0 0.0 0.0 19121 0.4 0.0 0.2 0.0 19123 0.0 0.0 0.1 0.0 19130 0.1 0.0 0.0 0.0 19137 0.2 0.0 0.0 0.0 19146 19155 0.6 0.0 0.0 0.0 -- -- -- 19159 MEAN SD N 0.3 0.21 10 0.0 0.00 10 0.1 0.07 10 0.0 0.00 10 SEX: MALE LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-24 STUDY NO: 060-069 Animal JD Eosinophil THSN/CU GROUP: 4-M 19100 0.1 79103 0.0 19104 0.4 19118 0.2 0.9 0.3 19142 0.1 19144 0.4 19148 0.2 19150 19156 0.1 -- 19160 79162 1916 1916 c- MEAN SD N 0.3 0.26 10 Basophils THSN/CU MM Other THSN/CU 0.0 0.1 0.0 0-0 0.6 0.0 0.0 0.4 0.0 0.0 0.5 0-0 0.0 0.2 0.0 0.0 0.2 0.0 0.0 0.1 0.0 0.0 0.0 Q.0 0.0 0.0 0.0 0.0 0.0 0.0 -- -- 0.0 0.00 10 0.2 0.22 10 0.0 0.00 10 SEX: MALE 27-JUN-2002 418-028:PAGE M-25 Animal ID Eosinophi 1 Basophi 1s THSN/CU MM Other THSN/CU GROUP: 5-M 19117 0.2 0.0 0.5 0.0 19117 0.4 0.0 0.0 0.0 1912s 0.1 0.0 0.0 0.0 19126 0.4 0.0 0.0 0.0 19127 0.1 0.0 0.2 0.0 19128 0.1 0.0 0.0 0.0 19140 0.3 0.0 0.5 0.0 19145 0.2 0.0 0.0 0.0 19149 0.0 0.0 0.0 0.0 19152 19154 19158 0.0 0.0 0.0 0.0 -- -- -- -- - I -- 19168 I - MEAN SD N 0.2 0.15 10 0.0 0. 10 0.1 0.21 10 0.0 0.00 10 SEX: MALE Study Report for Hematology 418-028:PAGE If-26 STUDY NO: 060-069 GROUP: I - F 19010 19012 19019 19021 19023 19041 13042 19044 19050 19053 19065 19068 19072 1 9074 19075 MEAN SD N -- 0.2 0.0 0.3 0.1 0.0 0.0 0.1 0.1 0.0 0.1 -- 0.1 0.10 10 bnormal b HSN/CU MM 0th THSN/CU -- -- 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.1 0.0 0-0 0.0 0-0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.8 0.0 0.1 0.0 0.0 0.1 0.0 -- 0.0 0.00 10 0.0 0.05 10 0.0 0.00 10 27-JUN-2002 418-028:PAGE M-27 Animal I D Eosinophi1 Basophi 1s THSN/CU HM Other THSN/CU _I____1-____c1------_----------c--------------~------------------------------------------------------------------ I GROUP: 2-F 1 9004 0.0 0.0 0.0 0.0 19009 0.1 0.0 0.0 0.0 19016 0.3 0.0 0.0 0.0 19018 0.1 0.0 0.0 0.6 19026 0.Q 0.0 0.0 0.0 19036 0.4 0.O 0.0 0.0 1903-P 0.0 0.0 0.0 0.0 1 9043 0.1 0.0 0.0 0.0 1 904? 0.1 0.0 0.0 0.0 19048 13052 19055 19061 1 9067 1 9071 -0 . 1 - 0.0 -- 0.0 -- 0-.0- -- -- -- -- -- -- -- -c -- -- I - -- -- -.... -- MEAN SD N 0.1 0.13 10 0.0 0.00 10 0.0 0.00 10 0.0 0.00 10 LABCAT HE4.43 7- JUN-2002 418-028:PAGE K-28 MEAN SD N 0.1 0.13 10 0.0 0. 10 0.7 0.07 10 0.0 0.00 10 LABCAT HE4.43 27-JUN-2002 Study Report f o r Hematology 418-028:PAGE K-29 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 GROUP: 4-F 19002 1 9005 19035 19039 19040 19045 19046 1905 1 19054 19058 19062 19063 19066 19069 190 MEAN SD N -- 0.1 0.0 0.1 0.0 0.0 0.0 0.2 0.0 0.3 0.0 -- 0.1 0.11 10 bnormal L HSN/CU MM Qth THSN/CU -- -- 0.0 0.0 0.0 0.0 0.1 0.0 0.0 0.0 0.0 0,o 0.0 0.0 0.0 0.0 0.0 -- -- -- 0.0 0.3 0.0 0.0 0.0 0-0 0.0 0.0 0.0 0.0 0.0 0.0 -- 0.0 0. I 0.0 -._ -."." 6.0 0.00 10 0.1 0.10 10 0.0 0.00 10 SEX: FEFIALE 418-028:PAGE K-30 MEAN SD N 0.1 0.15 10 0.0 0.00 70 0.0 0.05 10 0.0 0.00 10 27-JUN-2002 418-028:PAGE K-31 Group: 1-M SD N Group: 2-M SD N Group: 3-M MEAN SD N Group: 4-M SD N Group: 5-M MEAN SD N 15.3 1.63 10 7.51 0,648 10 16.5 1.16 10 43.5 3.413 10 14.9 -03 10 7.33 0 - 396 10 15.9 0.42 10 42.2 1.57 10 13 2. 10 7-32 0.441 10 15.7 0.59 10 42.0 2.22 10 16.2 3.97 10 6.93* 0.305 10 15.4 0.67 10 40.2 2.23 10 14.7 3.07 10 6.90.442 10 15*6* 0.81 10 40.7 1.87 10 57.9 1.89 10 57.7 2.27 10 57.4 1.78 10 57.9 1.15 10 58.3 1.37 10 2.0 0.87 10 21 .% 1.13 10 21.5 0.75 10 22.2 0.45 10 22.3 0.56 10 38.0 0.93 10 1179 130.9 10 37.7 1.15 10 1131 183.7 10 37.5 0.79 10 1115 149.2 10 8.4 0"80 10 1104 91 - 4 10 38.3 0-70 10 1148 159.9 10 8. 1 0.52 10 7.9 0.54 70 8.2 0.72 10 8.2 0.76 10 8.0 0.59 10 7-JUN-2002 418-028:PAGE K-32 Group: I - M NfAN SD N Group: 2-M N SD N SD N Group: 4-M N SD N Group: 5-M MEAN SD N 13.4 0.24 10 21 .o 3.09 10 14.2* 0.27 I0 22.1 4.41 10 13.6 0.23 10 20.6 2.38 10 13.8* 0.40 10 21.2 2.24 10 14.0* 0.47 10 21 - 3 1.43 10 0 14 -9 2.2 0.0 0.8 0.3 0.0 0.0 1.72 0.73 0.00 0.56 0.20 0.00 10 10 10 10 40 10 10 0 11 .o 2.9 0.0 0.7 0.3 0.0 0.0 2.613 1 .I8 0.00 0.50 0.21 0.00 10 I0 10 10 10 10 10 0 10. 2.0 0.0 0.6 0.3 0.0 0.0 2.02 1.06 0.03 0.51 0.21 0.00 10 10 10 10 10 10 10 0 12.1 2.8 0.0 0.8 0.3 0.0 0.0 3.57 0.73 0.03 0.67 0.26 0.00 10 10 10 10 10 10 40 0 12.4 1.6 0.0 0.5 0.2 0.0 0.0 2.75 0.78 0.00 0.33 0.15 0.00 10 10 10 10 10 10 10 LABCAT HE4.43 7-JUN-2002 STUDY NO: 060-069 Study Report for Hematolo 418-028:PAGE I(-33 Y DUNNETT'S PROCEDURE SEX: MALE SD N Group: 2MEAN SD N Group : MEAN SD N Group: BEAN SD N SD N 0.1 0.20 40 0.1 10 8.1 0.0'7 40 0.2 0.22 10 8.1 0.21 10 0.0 0.00 10 0.0 0.00 10 0.0 0.00 10 0.0 0.00 10 0.0 0.00 10 418-028:PAGE K-34 F Ratio = Coeff. Var. % = 0.90 20.065 I F ' table values Dunnett's 'T' table values F.01 = P. 3.78 3.12 F.05 = P.05 = 2.58 2.51 F Ratio = Caeff. Var. X = 3-71 4.874 I F ' table values Dunnett's I T ' table values F.01 = P.01 = 3-12 P.05 = 2.58 2.54 27- JUN-2 418-028:PAGE K-35 F Ratio = Coeff. Var. % = 1.77 3.593 I F ' table values Dunnett's IT' table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 LABCAT HE4.43 27- J UN-2002 418-028:PAGE K-36 F Ratio = Coeff. War. % = 0.56 7.823 I F ' table values Dunnett's IT' t a b l e values F.01 = P.01 = 3-78 3.12 F-05 = P.05 = 2.58 2.51 MBCAT ME4.43 27- JUN-2002 418-028:PAGE K-37 Y DUNNETT'S PROCEDURE 1-F.4 0.24 TREATMENTS 4 4.13 1-03 2-M 10 142.3 14.2 ERROR 45 5.13 0.11 3-M 10 136.3 13.6 ------c------------_cf_c____c___c_ 4- 10 437.9 13.8 0.40 2.71 13.0 13.8* 12.9 43.9 TOTAL 49 9.26 5-x 10 139.5 14.0 13.0 13.8* 12.9 13. 1- 10 209. 21 .o 3.09 TREATMENTS 4 11.75 2. 2- 10 220.5 22.1 4.41 0.83 17-7 24.2 17.0 25.0 ERROR 45 374.92 8. 3-M 16 205.7 20.6 4- 40 211.7 21.2 4.2 47.0 25.0 TOTAL 49 386.66 5-M 10 212.8 21.3 1.43 0.23 17.7 24.2 17.0 25.0 F Ratio = Coeff. Var, X = 0.35 13. 09 ' F ' table values Dunnett's ' T ' tab1 F.01 = 3-78 F-05 = 2.58 3.12 P.05 = 2.57 F Raleio = Coeff. Var. X = 0.00 0.000 I F ' table values Dunnett's I T ' table values F.01 = P,O1 = 3.78 3.12 F.05 = P.05 = 2.58 2.51 27- JUN- 4 18-028:PA 418-028:PAGE K-39 F Ratio = Coeff. War. % = 0.93 78.222 I F ' table values Dunnett's 'T' t a b l values F.01 = P.01 = 3.78 3.12 F.05 = P.05 = 2-58 2.51 1-M 234-# 5-t4 10 2.6 0.3 0.20 10 3.0 0.3 0.21 0 . 4 10 2.5 0.3 0.21 0.11 10 2.7 0.3 0.26 0.41 10 1 .8 0.2 0.0 0.5 0.0 0.5 0.0 0.5 0.0 0.5 0.0 0.0 0.0 0.0 0.5 0.5 0.5 0.5 TREATMENTS 4 0.08 ERROR 45 1.93 -c--------------------------------~--- TOTAL 49 2.00 0.02 0.04 F Ratio = Coeff. War. % = 0.00 0.000 I F ' table values Dunnett's ' T ' table values F.01 = P.01 = 3.78 3.12 F.05 = P.05 = 2.58 2.51 Source-sour ce o f War ia t ian T HE4.4 Treatments-between groups 7- JUN-2002 418-028:PAGE M-40 F Ratio = Coeff. Var. % = 8-95 137. 8 ' F ' table values Dunnett's 'T' tabl F-01 = 3.78 3.72 F.05 = P.05 = 2.58 2.51 1-M 10 0.0 0.0 0.00 TREATMENTS 4 0.00000 0. 2-M 18 0.0 0.0 0.00 0.00 0.8 0.0 0.0 0.0 ERROR 45 0.00000 0 . 3-w 10 0.0 0.0 0.00 0.00 0.0 0.0 4- I0 0.0 0.0 0.00 0.00 0.0 .o 0.0 0.0 TOTAL 49 O.OOOO0 5-PI 10 0.0 0.0 0.00 0. 0 - 0 0.0 0.0 0.0 SourceSource of Variation tments-between groups 418-028:PAGE K-41 Group: l - F MEAN SD N Group: 2-F MEAN SD N Group: 3-F MEAN SD N Group: 4-F MEAN SD N Group: 5-F MEAN SD N 9.0 3.09 10 6.51 0.461 10 13.8* 5.73 10 6.22 0.423 10 11 .I 2.98 10 6.40 0.554 10 10.8 3.15 10 6.39 0.532 10 8.9 3.48 10 6.42 0.538 10 15.5 0.95 10 15.0 0.77 10 15.2 0.91 10 15.2 0.90 10 15.3 0.77 10 40.5 2.93 10 39.3 2.72 10 39.5 3.24 10 40.1 3.02 10 40.4 3.15 10 62.2 1. I 9 10 63.3 2.01 10 61.8 1.50 10 62.9 2.04 10 63.0 1.60 10 23.9 0.55 10 24.2 0.76 10 23.9 0.88 10 23.8 1.25 10 23.8 1.05 10 38.4 0.76 10 1419 153.0 10 38.3 0.94 10 1240 208.1 10 38.7 I.29 10 1274 226.3 10 37.8 1.22 10 1305 207.2 10 37.8 1.37 10 1355 176.8 10 8.3 0.49 10 8.0 0.84 10 8.3 0.82 10 8.0 0.63 10 7.7 0.74 10 *-Significant Difference from Control P C .05 LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-42 Group: I - F MEAN SD N Group: 2-F MEAN SD N Group: 3-F MEAN SD N Group: 4-F MEAN SD N Group: 5-F MEAN SD N 13.2 0.41 10 13.4 0.29 10 13.2 0.37 10 13.5 0.42 I0 13.8 1.12 10 21.7 2.36 10 23.9 3.19 10 23.5 2.35 10 25.3 2.45 10 25.2 3.81 10 0 6.6 2.2 0.0 0.1 0.1 0.0 0.0 2.44 0.90 0.00 0.15 0.10 0.00 10 10 10 10 10 10 10 0 9.8* 3.6 0.0 0.3 0.1 0.0 0.0 3.44 3.69 0.00 0.49 0.13 0.00 10 I0 10 10 I0 10 10 0 8.4 2.4 0.0 0.2 0.1 0.0 0.0 2.60 0.93 0.00 0.13 0.13 0.00 10 10 10 10 10 10 10 0 8.2 2.4 0.0 0.2 0.1 0.0 0.0 2.52 0.96 0.09 0.11 0.11 0.00 10 10 I0 10 10 10 10 0 6.0 2.6 0.0 0.2 0.1 0.0 0.6 2.77 0.82 0.00 0.14 0.15 0.00 10 10 10 10 10 10 10 *-Significant Difference from Control P < -05 LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-43 Group: 2-F MEAN SD N Group: 3-F MEAN SD N Group: 4-F MEAN SD N Group: 5-F MEAN SD N 0.0 0.05 10 0.0 0.00 10 0.1 0.07 10 0.1 0.10 10 0.0 0.05 10 0.0 0.00 10 0 .o 0.00 I0 0.0 0.00 10 0.0 0.00 10 0.0 0.00 10 418-028:PAGE K-44 STUDY ID: ARGUS 418-028 SEX: FEMALE STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE ------------------------------------------------------------------------------------------------ __ White Blood Count _ _ _G r o u p S t d . DUNNETT'S DUNNETT'S RANGES Degree Sum of Mean _N___T_o t_a _l ----M-e-a-n-------D-e-v-.------' t--' -------L-O----9-5-%----H--I -----L-O---9-9-%----H--I-------S-o-u-r-c-e-----F-dm-------S-q-u-a-r-e-s-----S-q-uare I-F 10 89.9 9.0 3.09 TREATMENTS 4 158.42 39.61 2- F 3- F 10 137.8 10 110.7 13.8 11.1 5.73 2.80 2.98 1.22 4.7 13.3* 4.7 13.3 3.6 3.6 14.3 14.3 -----E-R-R-O-R----4-5-------6-5-9-.-1-2-------1-4-.-6-5- 4- F 10 108.4 10.8 3.15 1.08 4.7 13.3 3.6 14.3 TOTAL 49 817.54 5- F 10 88.9 8.9 3.48 0.06 4.7 13.3 3.6 14.3 * - S i g n i f i c a n t D i f f e r e n c e from Control E r r o r - w i t h i n groups P < -05 LABCAT HE4.43 Source-Source of Variation Treatments-between groups 27-JUN-2002 418-028:PAGE K-45 I-F 10 239.0 23.9 0.55 2- F 10 242.4 24.2 0.76 0.82 22.9 24.9 3-F 10 238.9 23.9 0.88 0.02 22.9 24.9 4- F 10 237.9 23.8 1.25 0.26 22.9 24.9 5-F 10 238.3 23.8 1.05 0.17 22.9 24.9 F Ratio = Coeff. Var. % = 0.37 3.886 I F ' table values Dunnett's ' T I table values F.01 = P.01 = ~~-- TREATMENTS 4 I.28 0.32 22.6 22.6 25.2 25.2 -----E-R-R-O-R----4-5--------3-8-.9-2--------0--.8-6- 22.6 25.2 TOTAL 49 40.21 22.6 25.2 3.78 F.05 = 3.12 P.05 = 2.58 2.51 Error-within groups Source-Source o f Variation LABCAT HE4.43 Treatments-between groups 27-JUN-2002 418-028:PAGE K-46 __---------------------------------- --- Study Report for Hematology ------u_------------------------------------------------- SUMMARY REPORT ________ PERIOD: TERMINAL _____--------------------------------------------------------------------------------------------- STUDY I D : ARGUS 418-028 SEX: FEMALE STUDY NO: 060-069 ________ ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE -----------___--------------------------------------------------------------------------------- _ _ _ _ _ _ Mean Corpuscular Hem- C O ~ C . Group N Total Mean Std. DUNNETT'S Dev . It' DUNNETT'S RANGES LO -95%- H I LO -99%- ______--------------____c_______________----------------------------------------------------------------- HI Degree Source Fdm Sum o f Squares Mean Square __ I-F 10 384.2 38.4 0.76 TREATMENTS 4 5.34 1.34 2- F 3- F 10 382.9 38.3 10 386.6 38.7 0.94 0.26 1.29 0.47 37.1 39.7 37.1 39.7 36.8 36.8 40.0 40.0 -----E-R-R-O-R----4-5--------5-8-.2-5---------1-.2--9- 4- F 10 378.3 37.8 1.22 1.16 37.1 39.7 36.8 40.0 TOTAL 49 63.60 5-F 10 378.4 37.8 1.37 1.74 37.1 39.7 36.8 40.0 F Ratio = 1.03 ' F ' table values F.01 = 3.78 F.05 = 2.58 -Coeff. Var. % = 2.978 Dunnett's ' T I t a b l e values P.01 = ---_--_-------------__________L_________----------------------------------------------------------------------- 3.12 P.05 = 2.51 __ Platelets ___________-_--------------------------------------~------------------~---------------------------~--------.----------- I-F 10 14189 1419 153.0 TREATMENTS 4 198797 49699 2- F 3- F 10 12396 10 12735 1240 1274 208.1 226.3 2.05 1.66 1199 1799 1639 1639 1145 1145 1692 1692 -----E-R-R-O-R----4-5------1-7-2-9-2-1-6------3-8-4-2-7-- 4- F I 0 13048 1305 207.2 1.30 3 1 9 9 1639 1145 1692 TOTAL 49 1928012 5-F 10 13554 1355 176.8 0.72 1199 1639 1145 1692 ______ I-F 10 2-F 10 3- F 10 4- F 10 5- F 10 83.4 80.0 82.7 80.3 76.5 ___________--___ 8.3 0.49 8.0 0.84 1.06 8.3 0.82 0.22 8.0 0.63 0.97 7.7 0.74 2.15 7.5 9.1 7.5 9.1 7.5 9.1 7.5 9.1 TREATMENTS 4 2.95 0.74 7.3 9.3 ERROR 45 23.11 0.51 7.3 9.3 ---------- 7.3 9.3 7.3 9.3 TOTAL 49 26.06 ___ ____________------------------------------------------------------------------------------------------------- Error-wi t h i n groups Treatments-between groups Source-Source o f Variation LABCAT HE4.43 27-JUN-2002 Study Report f o r Hematology 418-028:PAGE K-47 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: FEMALE Prothrombin Group N Total Mean - Std DUNNETT' S Dev . It' DUNNETT'S RANGES LO -95%- H I LO -99%- HI Degree Source Fdm Sum of Squares Mean Square I-F 10 131.8 13.2 0.41 TREATMENTS 4 2.32 0.58 2- F 3- F 10 133.5 13.4 10 131.7 13.2 0.29 0.63 0.37 0.04 12.5 13.9 12.5 13.9 12.3 12.3 14.0 14.0 ------ER-R-O-R----4-5--------1-6-.4-8--------0--.3-7-- 4- F 10 134.8 13.5 0.42 1.11 12.5 13.9 12.3 14.0 TOTAL 49 18.80 5-F 10 137.5 13.8 1.12 2.11 12.5 13.9 12.3 14.0 F Ratio = 1.58 ' F ' t a b l e values F.01 = 3.78 F.05 = 2.58 coeff. Var. % = 4.521 Dunnett's ' T I table values P.01 = 3.12 P.05 = 2.51 F Ratio = Coeff. Var. X = 2.53 12.081 I F ' table values Dunnett's ' T I table values --------------------------------------------------------------------------------------------------------------------,-------- Nucleated Red Cells ________~--~~-~ I-F 10 0 0 0.0 2- F 10 0 0 0.0 0.00 3-F 10 0 0 0.0 0.00 4-F 10 0 0 0.0 0.00 5-F 10 2 0 0.6 1.58 F.01 = P.01 = 0 0 0 0 0 0 0 0 F Ratio = Coeff. Var. % = 1.00 707.107 ' F ' table values Dunnett's I T ' table values F.01 = P.01 = 3.78 F.05 = 2.58 3.12 P.05 = 2.51 0 0 0 0 3.78 3.12 TREATMENTS 4 0 ERROR 45 0 ---------------L---------------------- 0 TOTAL 49 0 F.05 = 2.58 P.05 = 2.51 0.32 3.60 3.92 ~ ~_____ 0.08 0.08 LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-48 *-Significant Difference from Control P C .Os Error-within groups LABCAT HE4.43 Source-Source o f Variation Treatments-between groups 418-028:PAGE K-49 ______ -------------S--t--u-d--y---R--e--p--o--r-t---f--o--r--H--e--m-a--t--o-l--o--g-y------------------------------ __ SUMMARY REPORT PERIOD: TERMINAL ---------------____________------------------------------------------------------------------------------------- - STUDY I D : ARGUS 418-028 SEX: FEMALE STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE _____ ________y___________------------------------------------------------------------------------------ Monocytes Group Std. DUNNETT'S DUNNETT'S RANGES Degree Sum o f Mean N Total Mean . Dev It' LO -95%- H I LO -99%- H I Source Fdm Squares Square _____________-_-------------------------------------------------------------~--------------------- I-F 10 1.3 0.1 0.15 TREATMENTS 4 0.11 0.03 2- F 3- F 10 2.7 0.3 0.49 1.26 10 1.7 0.2 0.13 0.36 -.I 0.4 -.I 0.4 -.2 -.2 0.5 0.5 -----E-R-R-O-R----4-5--------2-.-7-9--------0-.-0-6-- 4- F 10 1.9 0.2 0.11 0.54 -.I 0.4 -.2 0.5 TOTAL 49 2.90 5- F 10 1.7 0.2 0.14 0.36 -.I 0.4 -.2 0.5 F Ratio = 0.43 I F ' tabLe values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 133.942 Dunnett's I T ' t a b l e values P.01 = 3.12 P.05 = 2.57 I-F 10 0.9 0.1 0.10 TREATMENTS 4 0.014 0.004 2- F 3- F 10 1.2 0.1 0.13 0.54 I0 0.9 0.1 0.13 0.00 0.0 0.2 0.0 0.2 -.I 0.3 -.I 0.3 ERROR 45 0.691 ---------------L---------------------- 0.015 4- F I0 0.7 0.1 0.11 0.36 0.0 0.2 -.I 0.3 TOTAL 49 0.705 5-F 10 0.8 0.1 0.15 0.18 0.0 0.2 -.I 0.3 F Ratio = Coeff. Var. % = 0.23 137.686 I F ' table values Dunnett's ' T I table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 - Basophi1s -_-----------------------------------------------------------~----------------------- I-F 10 0.0 0.0 0.00 TREATMENTS 4 0.00000 0.00000 2- F 3- F 10 0.0 0.0 10 0.0 0.0 0.00 0.00 0.00 0.00 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 ------ER-R-O-R----4-5------0-.-0-0-0-0-0-----0-.0-0-0-o-O 4- F 70 0.0 0.0 0.00 0.00 0.0 0.0 0.0 0.0 TOTAL 49 0.M)OOO 5- F 10 0.0 0.0 0.00 0.00 0.0 0.0 0.0 0.0 LABCAT HE4.43 27- JUN-2002 Study Report f o r Hematology 418-028:PAGE K-50 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: FEMALE Abnormal Lymphocytes Group N Total Mean . Std. DUNNETT' S Dev It' I-F 10 0.3 0.0 0.05 2-F 10 0.0 0.0 0.00 1-09 3-F 10 0.6 0.1 0.07 1.09 4-F 10 0.5 0.1 0.10 0.73 5-F 10 0.3 0.0 0.05 0.00 DUNNETT'S RANGES LO -95%- H I LO -99%- H I 0.0 0.1 0.0 0.1 0.0 0.1 0.0 0.1 -.I 0.1 -.I 0.1 -.I 0.1 -.I 0.1 D e g r e e Sum of Mean Source Fdm __~Sq_ua-r-e-s----S-q-u-a-r-e----~~ TREATMENTS 4 0.021 O.QO5 -----E-R-R-O-R----4-5--------0-.-1-7-1-------0-.-0-0-4- TOTAL 49 0.192 _ _ ~ ~ ~ ~ I-F 10 0.0 0.0 2- F 10 0.0 0.0 3- F 10 0.0 0.0 4- F 10 0.0 0.0 5-F 10 0.0 0.0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 ~-----_--_------- TREATMENTS 4 O.OO000 0.00000 0.0 0.0 ERROR 45 O.OOOO0 0.00000 0.0 0.0 ...................................... 0.0 0.0 0.0 0.0 TOTAL 49 0.00000 E r r o r - w i t h i n groups Source-Source of Variation LABCAT HE4.43 Treatments-between groups 27- JUN-2002 418-028:PAGE K-51 19109 19115 19116 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi1s Basophi 1s Abnormal Lymphocytes Other WBC 0 72 9.7 15 2.0 0 0.0 8 1 .I 5 0.7 0 0.0 0 0.0 0 0.0 13.5 0 72 11.4 17 2.7 0 0.0 10 1.6 1 0.2 0 0.0 0 0.0 0 0.0 15.9 0 n 11.6 20 3.0 0 0.0 2 0.3 I 0.2 0 0.0 0 0.0 0 0.0 15.1 418-028:PAGE K-52 19119 1 91 22 191 25 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocy tes Segmented Neutrophi 1s Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 72 9.1 17 2.2 0 0.0 10 1.3 1 0.1 0 0.0 0 0.0 0 0.0 12.7 0 86 14.1 9 1.5 0 0.0 1 0.2 1 0.2 0 0.0 3 0.5 0 0.0 16.4 0 81 14.3 a 1.4 0 0.0 6 1 .I 3 0.5 0 0.0 2 0.4 0 0.0 17.6 418-028:PAGE K-53 19131 19132 19134 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocy tes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 74 13.1 1s 2.7 0 0.0 9 1.6 1 0.2 0 0.0 1 0.2 0 0.0 17.7 0 85 12.7 10 1.5 0 0.0 3 0.4 0 0.0 0 0.0 2 0.3 0 0.0 14.9 0 73 11.o 23 3.5 0 0.0 3 0.5 I 0.2 0 0.0 0 0.0 0 0.0 15.0 418-028:PAGE K-54 19135 Nucleated Red Cells 0 Lymphocytes 85 Segmented Neutrophils 11 Bands 0 Monocyt es 2 Eosinophi 1s 2 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19143 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19151 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-55 19157 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19161 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Ecrsinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19167 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocyt es 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE I(-56 19102 19106 19108 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi Cs Abnormal Lymphocytes Other WBC 0 60 6.5 25 2.7 0 0.0 9 1.o 3 0.3 0 0.0 3 0.3 0 0.0 70.9 0 71 12.1 17 2.9 0 0.0 8 1.4 3 0.5 0 0.0 1 0.2 0 0.0 17.0 0 86 10.9 1'1 1.4 0 0.0 7 0.1 0 0.0 0 0.0 2 0.3 0 0.0 12.7 4 18-028:PAGE K-57 19110 19113 191 20 Nucleated Red Cells Lyrnphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 76 10.0 15 2.0 0 0.0 5 0.7 2 0.3 0 0.0 2 0.3 0 0.0 13.1 0 71 11 .I 23 3.6 0 0.0 5 0.8 1 0.2 0 0.0 0 0.0 0 0.0 15.7 0 73 13.4 17 3.1 0 0.0 8 1.5 2 0.4 0 0.0 0 0.0 0 0.0 13.4 418-028:PAGE K-58 19129 Nucleated Red Cells Lymphocytes . Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 19136 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 19138 Nucleated Red Cells Lymphocytes Segmented Nwtrophi 1s Bands Monocyt es Eosinophi 1s Basophi1s Abnormal Lymphocytes Other WBC 0 64 9.4 27 4.0 0 0.0 5 0.7 4 0.6 0 0.0 0 0.0 0 0.0 14.7 0 82 8.6 13 1.4 0 0.0 3 0.3 2 0.2 0 0.0 0 0.0 0 0.0 10.5 0 66 11.4 30 5.2 0 0.0 'I 0.2 3 0.5 0 0.0 0 0.0 0 0.0 17.2 4 18-028:PAGE K-59 19139 Nucleated Red Cells 0 Lymphocytes 86 Segmented Neutrophi 1s 13 Bands 0 Monocytes 1 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19747 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19153 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC LABCAT HE4.43 27- JUN-2002 418-028:PAGE K-60 19164 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocyt es 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19165 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other `0 WBC 19171 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27- JUN-2002 418-028:PAGE K-61 19101 19105 19107 Nucleated R e d Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophils Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocytes Segmented Neutrophi 1s Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other UBC 0 77 10.1 16 2.1 0 0.0 4 0.5 2 0.3 0 0.0 1 0.1 0 0.0 13.1 0 80 8.8 17 1.9 0 0.0 2 0.2 0 0.0 0 0.0 1 0.1 0 0.0 11.o 0 78 9.8 5 0.6 1 0.1 12 1. s 4 0.5 0 0.0 0 0.0 0 0.0 12.5 418-028:PAGE K-62 19112 ,19114 191 21 Nucleated R e d Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s BasophiI s Abnormal Lymphocytes Other WBC 0 83 11 .o 10 1.3 0 0.0 4 0.5 2 0.3 0 0.0 1 0.1 0 0.0 13.3 0 70 7.8 24 2.7 0 0.0 5 0.6 1 0.1 0 0.0 0 0.0 0 0.0 11.2 0 71 14.8 19 4.0 0 0.0 7 1.5 2 0.4 0 0.0 1 0.2 0 0.0 20.9 418-028:PAGE K-63 19130 19137 Nucleated Red Cells Lymphocytes Segmented Neutrophiks Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 84 11.7 12 1.7 0 0.0 3 0.4 1 0.1 0 0.0 0 0.0 0 0.0 13.9 0 87 10.6 9 1 .I 0 0.0 2 0.2 2 0.2 0 0.0 0 0.0 0 0.0 12.2 418-028:PAGE K-64 19146 Nucleated Red Cells 0 Lymphocytes 85 Segmented Neutrophi 1s 9 Bands 0 Monocytes 2 Eosinophi I s 4 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19155 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi I s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19159 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi I s 0 Basophi I s 0 Abnormal Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-65 19172 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19173 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19174 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 UBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-66 19103 19104 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 78 16.4 11 2.3 0 0.0 8 1.7 0 0.0 0 0.0 3 0.6 0 0.0 21 .o 0 67 12.1 17 3.1 0 0.0 12 2.2 2 0.4 0 0.0 2 0.4 0 0.0 18.1 418-028:PAGE K-67 19118 19133 19141 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi I s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 78 9.4 12 1.4 0 0.0 4 0.5 2 0.2 0 0.0 4 0.5 0 0.0 12.0 0 79 17.4 13 2.9 0 0.0 3 0.7 4 0.9 0 0.0 I 0.2 0 0.0 22.0 0 73 12.7 21 3.7 0 0.0 3 0.5 2 0.3 0 0.0 1 0.2 0 0.0 17.4 418-028:PAGE K-68 I9142 19144 19148 Nucleated R e d Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 67 8.6 22 2.8 1 0.1 8 1.o 1 0.1 0 0.0 1 0.1 0 0.0 12.8 0 81 15.9 14 2.7 0 0.0 3 0.6 2 0.4 0 0.0 0 0.0 0 0.0 19.6 0 79 12.3 20 3.1 0 0.0 0 0.0 1 0.2 0 0.0 0 0.0 0 0.0 15.6 418-028:PAGE K-69 19150 Nucleated Red Cells 0 Lymphocytes 75 Segmented Neutrophils 22 Bands 0 Monocytes 2 Eosinophi 1s 1 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19156 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19160 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-70 19162 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 79163 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19166 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC (-1 - Data Unavailable LABCAT HE4.43 ~~~________________ 27-JUN-2002 418-028:PAGE K-71 19117 19124 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocy tes Segmented Neutrophi 1s Bands Monocytes Eosinophi Ls Basophi 1s Abnormal Lymphocytes Other WBC 0 79 11.3 12 1.7 0 0.0 6 0.9 3 0.4 0 0.0 0 0.0 0 0.0 14.3 0 94 10.2 3 0.3 0 0.0 2 0.2 1 0.1 0 0.0 0 0.0 0 0.0 10.8 418-028:PAGE K-72 19126 19127 19128 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnorma 1 Lymphocytes Other WBC 0 81 10.1 9 1 .I 0 0.0 7 0.9 3 0.4 0 0.0 0 0.0 0 0.0 12.5 0 73 8.0 22 2.4 0 0.0 2 0.2 1 0.1 0 0.0 2 0.2 0 0.0 11.o 0 87 12.4 10 1.4 0 0.0 2 0.3 1 0.1 0 0.0 0 0.0 0 0.0 14.2 418-028:PAGE K-73 19145 191 49 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 83 17.2 12 2.5 0 0.0 4 0.8 1 0.2 0 0.0 0 0.0 0 0.0 20.7 0 88 16.1 12 2.2 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 18.3 418-028:PAGE K-74 191 52 Nucleated Red Cells 0 Lymphocytes 94 Segmented Neutrophils 3 Bands 0 Monocytes 3 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19154 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 191 58 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 AbnormaI Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-75 19168 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19169 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19170 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocyt es 0 Eosinophi I s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27- J UN-2002 418-028:PAGE K-76 19010 19012 19019 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- -- 0 79 12.6 17 2.7 0 0.0 3 0.5 1 0.2 0 0.0 0 0.0 0 0.0 15.9 0 66 4.4 31 2.0 0 0.0 3 0.2 0 0.0 0 0.0 0 0.0 0 0.0 6.6 418-028:PAGE K-77 19021 19023 19041 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnorma 1 Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocytes Segmented Neutrophi 1s Bands Monocy tes Eosinophi 1s Basophi1s Abnormal Lymphocytes Other WBC 0 77 5.7 19 1.4 0 0.0 0 0.0 4 0.3 0 0.0 0 0.0 0 0.0 7.4 0 83 6.1 12 0.9 0 0.0 2 0.1 2 0.1 0 0.0 1 0.1 0 0.0 7.3 0 63 5.0 36 2.9 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 0 0.0 8.0 418-028:PAGE K-78 19044 19050 Nucleated R e d Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 69 7.3 29 3.1 0 0.0 I 0.1 1 0.1 0 0.0 0 0.0 0 0.0 10.6 0 74 6.1 22 1.8 0 0.0 3 0.2 1 0.1 0 0.0 0 0.0 0 0.0 8.2 LABCAT HE4.43 27- JUN-2002 418-028:PAGE K-79 19065 19068 Nucleated R e d Cells Lymphocytes Segmented Neutrophi 1s Bands Nonocytes Eosinophi 1s Basophi Ls Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi1s Abnormal Lymphocytes Other WBC 0 67 8.4 30 3.8 0 0.0 1 0.1 1 0.1 0 0.0 1 0.1 0 0.0 12.5 LABCAT HE4.43 27-JUN-2002 4 18-028:PAGE K-80 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19074 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 19075 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-81 19004 19009 19016 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 74 5.8 25 2.0 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 0 0.0 7.8 0 87 12.7 11 1.6 0 0.0 1 0.1 1 0.1 0 0.0 0 0.0 0 0.0 14.6 0 80 13.6 18 3.1 0 0.0 0 0.0 2 0.3 0 0.0 0 0.0 0 0.0 17.0 418-028:PAGEK-82 19026 19036 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Honocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 85 7.7 1s 1.4 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 9.1 0 84 15.1 12 2.2 0 0.0 2 0.4 2 0.4 0 0.0 0 0.0 0 0.0 18.0 418-028:PAGE K-83 19037 19043 19047 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocy tes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocy tes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 42 11.o 52 13.7 0 0.0 6 1.6 0 0.0 0 0.0 0 0.0 0 0.0 26.3 0 71 9.6 26 3.5 0 0.0 2 0.3 1 0.1 0 0.0 0 0.0 0 0.0 13.5 0 56 6.2 41 4.6 0 0.0 2 0.2 1 0.1 0 0.0 0 0.0 0 0.0 11.1 Study Report for Hematology 418-028:PAGE K-84 19048 Nucleated Red Cells 0 Lymphocytes 76 Segmented Neutrophi 1s 23 Bands 0 Monocytes 0 Eosinophi 1s 1 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19052 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19055 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-85 19067 19071 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi I s Basophi 1s Abnormal Lymphocytes Other WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGEK-86 19003 19007 19008 Nucleated Red Cells Lymphocytes Segmented Neutrophi1s Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 59 6.5 36 4.0 0 0.0 3 0.3 I 0.1 0 0.0 1 0.1 0 0.0 11 .I 0 72 5.7 25 2.0 0 0.0 2 0.2 0 0.0 0 0.0 1 0.1 0 0.0 7.9 0 72 5.0 27 1.9 0 0.0 0 0.0 0 0.0 0 0.0 1 0.1 0 0.0 7.0 418-028:PAGE K-87 1 901 3 1901 4 1901 5 Nucleated R e d Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi I s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi S Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 74 12.3 23 3.8 0 0.0 2 0.3 0 0.0 0 0.0 1 0.2 0 0.0 16.6 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 - -- 0 77 5.7 22 1.6 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 0 0.0 7.4 LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-88 Animal I D 19017 19024 19029 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC TERMINAL CNT ABS 0 76 9.8 20 2.6 0 0.0 1 0.1 3 0.4 0 0.0 0 0.0 0 0.0 12.9 0 70 7.4 25 2.6 0 0.0 2 0.2 2 0.2 0 0.0 1 0.1 0 0.0 10.5 0 82 10.3 17 2.1 0 0.0 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 12.5 418-028:PAGE K-89 19034 19038 19056 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated R e d Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 90 10.9 8 1.o 0 0.0 1 0.1 I 0.1 0 0.0 0 0.0 0 0.0 12.1 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- -- 0 80 10.2 17 2.2 0 0.0 3 0.4 0 0.0 0 0.0 0 0.0 0 0.0 12.7 LABCAT HE4.43 27- JUN-2002 Study Report for Hematology 418-028:PAGE K-90 19057 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19060 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19064 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC (--I - Data Unavailable LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-91 19002 19005 19035 Nucleated R e d Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi I s Abnormal Lymphocytes Other WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- -- 0 81 7.7 17 1.6 0 0.0 1 0.1 1 0.1 0 0.0 0 0.0 0 0.0 9.5 0 74 7. I 24 2.3 0 0.0 1 0.1 0 0.0 0 0.0 1 0.1 0 0.0 9.6 418-028:PAGE K-92 19040 19045 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 77 7.7 22 2.2 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 0 0.0 10.0 0 56 5.4 39 3.8 3 0.3 2 0.2 0 0.0 0 0.0 0 0.0 0 0.0 9.7 27- JUN-2002 418-028:PAGE K-93 19046 Nucleated R e d Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 1905 1 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnorma 1 Lymphocytes 0 Other 0 WBC 19054 Nucleated Red Cells 0 Lymphocytes 82 Segmented Neutrophils 13 Bands 0 Monocytes 3 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 2 Other 0 WBC LABCAT HE4.43 27- JUN-2002 418-028:PAGE K-94 19058 19062 19063 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 73 11.5 25 3.9 0 0.0 1 0.2 1 0.2 0 0.0 0 0.0 0 0.0 15.7 0 77 4.8 22 1.4 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 0 0.0 6.2 0 76 9.8 20 2.6 0 0.0 2 0.3 2 0.3 0 0.0 0 0.0 0 0.0 12.9 418-028:PAGE K-95 19066 Nucleated R e d Cells 0 Lymphocytes 84 Segmented Neutrophi1s 14 Bands 0 Monocytes 1 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 1 Other 0 WBC 19069 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19073 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27- JUN-2002 418-028:PAGE K-96 19001 19006 19011 Nucleated Red Cells Lymphocytes Segmented Neutraphils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red C e l l s Lymphocytes Segmented Neutrophi 1s Bands Monocyt es Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 67 10.1 28 4.2 0 0.0 3 0.5 2 0.3 0 0.0 0 0.0 0 0.0 15.1 0 61 4.6 36 2 .? 0 0.0 3 0.2 0 0.0 0 0.0 0 0.0 0 0.0 7.6 0 69 4.4 28 1.8 0 0.0 2 0.1 0 0.0 0 0.0 1 0.1 0 0.0 6.4 418-028:PAGE K-97 Animal I D 19020 19022 19025 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s BasophiI s Abnormal Lymphocytes Other WBC TERM I NAL 2 52 2.2 46 1.9 0 0.0 2 0.1 0 0.0 0 0.0 0 0.0 0 0.0 4.2 0 58 3.1 37 2.0 0 0.0 3 0.2 2 0.1 0 0.0 0 0.0 0 0.0 5.4 0 71 5.2 27 2.0 0 0.0 1 0.1 0 0.0 0 0.0 1 0.1 0 0.0 7.3 418-028:PAGE K-98 19028 19030 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other WBC 0 70 7.1 27 2.8 0 0.0 3 0.3 0 0.0 0 0.0 0 0.0 0 0.0 10.2 0 64 5.2 34 2.8 0 0.0 1 0.1 0 0.0 0 0.0 1 0.1 0 0.0 8.1 418-028PAGE K-99 19031 Nucleated R e d Cells 0 Lymphocytes 67 Segmented Neutrophi 1s 30 Bands 0 Monocytes 0 Eosi nophi 1s 3 Easophi Is 0 Abnormal Lymphocytes 0 Other 0 WBC 19032 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnorma 1 Lymphocytes 0 Other 0 WBC 19033 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Mono c y t e s 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-100 19049 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19059 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophi 1s 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC 19070 Nucleated Red Cells 0 Lymphocytes 0 Segmented Neutrophils 0 Bands 0 Monocytes 0 Eosinophi 1s 0 Basophi 1s 0 Abnormal Lymphocytes 0 Other 0 WBC LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-101 GROUP: 1-M 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 19143 19151 19157 19161 19167 MEAN SD N 5.9 6.5 5.8 6.0 6.2 6.5 6.5 6.1 6.3 7.0 6.3 0.36 10 3.9 4.4 4.0 4.1 4.3 4.3 4.2 4.3 4.3 4.7 4.3 0.22 10 164 165 119 134 140 169 128 141 140 153 145 16.8 10 46 0.1 62 0.2 61 0.2 44 0.1 62 0.1 56 0.3 59 0.1 51 0.1 56 0.2 -72- 0.2 57 0.2 8.3 0.07 10 10 16 0.3 263 19 0.4 164 16 0.3 330 14 0.3 105 15 0.3 177 15 0.4 260 18 0.3 385 15 0.3 250 15 0.3 722 15 -- 0.3 -- 221 16 0.3 288 1.5 0.04 172.6 10 10 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-102 GROUP: 2-M 19102 6.1 19106 6.2 19108 5.8 19110 6.1 19113 5.6 19120 6.5 19129 5.7 19136 6.2 I9138 6.3 -- 19139 6.1 19147 4. I 172 4.0 166 3.9 152 4.1 104 3.9 157 4.3 115 3.9 149 4.3 123 4.4 108 4-.2- 165 40 0.2 44 0.1 38 0.1 33 0.1 30 0.1 28 0.2 33 0.1 52 0.1 60 0.2 55 0.2 -- 16 0.3 ,169 17 0.3 151 15 0.3 235 16 0.3 307 16 0.3 133 17 0.4 350 17 0.3 292 17 0.3 134 15 0.4 478 -1-6 0.4 -- 213 -- MEAN SD N 6.1 0.28 10 4.1 0.19 10 141 26.0 10 41 17.1 I0 0.1 0.05 10 16 0.3 246 0.8 0.05 111.5 10 10 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-103 GROUP: 3-M 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 19155 19159 19172 19173 19174 MEAN SD N 6.2 H 6.1 5.8 6.3 6.3 6.6 6.2 5.7 6.7 6-.6- 6.3 0.33 10 4.0 H 4.3 4.1 4.3 4.3 4.4 4.4 4.1 4.4 4.4 -- 4.3 0.15 10 304 H 254 198 137 142 121 156 149 158 204 I 182 58.2 10 27 H 52 50 46 65 45 46 34 59 35 46 11.6 I0 0.3 H 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.06 10 18 H 0.4 H 129 H 16 0.3 118 15 0.3 132 12 0.3 286 16 0.3 250 18 0.4 289 17 0.2 329 17 0.3 480 17 0.3 200 16 0.3 -- -- 1-75- 16 0.3 239 I.8 0.06 112.9 10 10 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-104 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 SEX: MALE GROUP: 4-M 19100 6.0 19103 5.7 19104 5.9 19118 6.0 19133 6.3 19141 6.2 19142 6.6 19144 6.2 19148 6.5 19150 6.4 19156 19160 19162 MEAN SD N 6.2 0.28 10 4.0 4.0 4.1 4.2 4.4 4.3 4.4 4.1 4.4 4.4 -- 4.2 0.17 10 175 202 167 131 179 147 132 130 175 1-8-1 162 25 - 2 10 42 28 42 39 49 70 56 29 42 -30- 43 13.1 10 0.1 0.1 0.1 0.1 0.1 0.2 0.2 0.1 0.1 0-.2- 0.1 0.05 10 16 0.3 Ill 20 0.3 14s 16 0.3 196 20 0.4 187 17 0.3 146 16 0.3 182 17 0.3 270 18 0.3 242 16 0.3 267 18 -- 0.3 -- 218 -- _- 17 0.3 196 1.6 0.03 53.5 10 10 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-105 ______ ~~-~~ --__ Animal I D TP ALB G LU CHOL 1-BIL BUN CREAT CK __________g_/d-L----------g-/d-L---------W--/d--L--------w--/-d-L---------W--/d-L---------W--d--L---------W-/-d-L-----------U-/-L----------- GROUP: 5-M 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 19154 19158 19168 19169 19170 6.6 H 6.1 H 6.6 6.4 6.6 6.5 6.0 6.4 6.3 5-.8-- ---- 4.6 H 4.3 H 4.8 4.6 4.6 4.8 4.3 4.5 4.5 4-.2- -- ---- 173 H I77 H 226 165 156 181 110 134 195 132 -- ---- -- 38 H 0.2 H 37 H 0.3 H 25 0.1 29 0.1 47 0.2 35 0.2 24 0.2 27 0.2 38 0.2 3--1 0.2 -- -- -- -_ -- -- -- -- -- 26 H 0.4 H 211 H 23 H 0.3 H 269 H 20 0.4 94 20 0.3 156 22 0.3 379 21 0.4 472 18 0.3 304 19 0.3 239 19 0.3 175 19 -- 0.3 -- 1-2-8 -- -- -- -- -- -- -- -- -- -- -- -- MEAN SD N 6.3 0.28 10 4.5 0.20 10 165 33.7 10 33 0.2 7.2 0.06 10 10 21 0.3 , 243 2.4 0.05 117.4 10 10 10 ~-__ (--I - Data Unavailable LABCAT CC4.43 418-028:PAGE K-106 Study Report f o r Clinical Chemistry STUDY ID: ARGUS 418-028 STUDY NO: 060-069 SEX: FEMALE GROUP: I-F 1901 0 -- -- -- -- 19012 5.4 3.7 149 63 0.2 19019 6.3 4.3 139 92 0.1 19021 7.1 4.9 157 136 0.1 19023 6.1 4.3 156 85 0.2 19041 5.9 4.3 165 62 0.2 19042 6.3 4.3 15s 77 0.1 19044 6.6 4.4 144 93 0.3 19050 5.8 3.9 184 78 0.2 19053 5.5 3.7 108 43 0.1 19065 19068 5.7 3.5 163 -- -- -- 4-0- 0.1 -- 19072 MEAN SD N 6.1 0.52 10 4.1 0.42 10 152 19.8 10 77 27.8 10 0.2 0.07 10 -- -- -- 32 0.4 166 29 0.3 21 2 33 0.4 21 0 26 0.4 222 28 0.3 201 25 0.4 178 27 0.4 332 34 0.3 I 98 28 0.3 123 19 0.3 175 -- -- 28 0.4 202 4.4 0.05 54.1 10 10 10 LABCAT CC4.43 27- J UN-2002 418-028:PAGE K-I07 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 SEX: FEMALE GROUP: 2-F 19004 19009 19016 19018 19026 19036 19037 19043 19047 19048 19052 19055 I9067 19071 MEAN SD N 5.9 5.7 5.5 5.6 5.5 5.9 5.0 5.7 4.3 5.9 -- --- 5.5 0.50 I0 4.3 3.9 4.2 4.2 3.9 4.2 3.5 3.8 3.3 4.2 --- u -_ 4.0 0.34 10 178 166 173 IS2 170 203 IS3 149 135 1-4-2 162 20.0 +I0 56 62 83 77 85 84 54 92 57 -7-3 72 14.0 10 0.2 0.2 0.2 0.2 0.1 0.2 0.2 0.2 0.2 0.2 0.2 0.03 I0 20 0.3 144 26 0.4 311 26 0.3 237 27 0.4 151 32 0.3 208 22 0.4 228 22 0.3 369 21 0.4 269 16 0.3 392 29 0.4 186 -- -- -- -- 24 0.4 250 4.7 0.05 85.6 10 10 10 LABCAT CC4.43. 27-JUN-2002 418-028:PAGE K-108 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 SEX: FEMALE GROUP: 3- F 1 9003 5.6 3.9 179 54 0. I 36 0.3 199 1 9007 5.3 3.8 121 45 0.1 41 0.3 159 1 9008 5.7 4.0 171 56 0.1 20 0.3 206 1901 3 1901 4 5.8 4.1 146 -- -- -- 59 -- 0.-2- 24 -- 0.4 -- 160 -- 1901 5 5.6 3.7 155 61 0.2 28 0.3 212 1901 7 5.8 4.1 164 87 0.2 22 0.4 190 19024 6.7 4.5 140 96 0.2 33 0.4 21 5 19029 5.8 4.0 142 79 0.2 26 0.3 21 2 19034 19038 6.4 4.3 151 105 0.1 -- -- -- -- -- 23 0.4 169 -- -- -- 19056 19057 6 .I -- 4.-2- 179 63 -- 0. I -- 28 -- 0.3 -- I 97 _- MEAN SD N 5.9 0.41 10 4.1 0.24 10 155 18.7 I0 71 20.0 10 0.2 0.05 10 28 0.3 192 6.7 0.05 21.7 10 I0 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-109 Study Report f o r Clinical Chemistry STUDY ID: ARGUS 418-028 STUDY NO: 060-069 SEX: FEMALE GROUP: 4-F 19002 19005 19035 19039 19040 19045 19046 19051 19054 19058. 19062 19063 19066 19069 19073 MEAN SD N -- 6.1 5.7 5.5 5.8 5-.6- 6.3 5.6 6.0 5.2 5.9 5.8 0.32 I0 -- 4.3 4.0 4.0 3.9 3.7 -- -- 4.5 3.9 4.4 3.6 4.1 -- 4.0 0.29 10 -- 167 161 163 127 160 -- -- 129 150 127 145 132 -- 146 16.2 10 -- 70 47 71 63 91 -- - 41 64 49 62 63 -- 62 14.2 I0 -- 0.2 0.1 0.2 0.1 0-.2- -- 0.1 0.1 0.1 0.1 0.1 -- 0.1 0.05 10 -- -- -- 30 0.4 259 27 0.4 193 26 0.4 160 20 0.3 233 2-5- 0.5 398 -- -- -- -- 25 0.4 124 20 0.3 267 34 0.3 176 23 0.4 172 2-2- 0.2 -- 147 -- 25 0.4 213 4.4 0.08 80.4 10 10 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-110 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 SEX: FEMALE GROUP: 5-F 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 19032 19033 19049 19059 19070 MEAN SD N 5.5 5.7 5.8 5.6 5.8 5.8 5.5 5.9 6.3 6.3 -- 5.8 0.29 I0 4.0 4.0 4.3 3.5 4.1 3.7 3.8 4.0 4.5 4.4 4.0 0.31 10 165 119 154 53 132 133 152 159 180 157 I40 35.5 I0 55 58 48 33 60 49 71 70 58 108 61 19.8 10 0.1 0.1 0.1 0.2 0.1 0.1 0.1 0.2 0.1 0.2 0.1 0.05 10 23 19 29 73 25 28 27 23 26 25 -- 30 15.4 10 0.3 341 0.3 373 0.4 149 0.2 438 0.3 186 0.4 143 0.3 252 0.3 261 0.3 168 0.3 210 -- -- 0.3 0.06 10 252 101.6 I0 ___________ _ _ (--I - Data Unavailable LABCAT CC4.463 27-JUN-2002 418-028:PAGE E-111 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Animal I D ALT U/L GROUP: 1-M 19109 36 19115 38 19116 40 19119 33 19122 51 19125 49 19131 39 19132 42 19134 49 19135 43 AST U/L 78 77 93 83 86 113 116 80 146 85 ALP U/L 91 77 102 112 103 115 107 98 126 119 CA W/dL 10.6 11- 8 10.3 10.8 11.3 10.6 11.1 11 .I 10.4 11.4 PHOS mg/d L 12.0 11.6 9.7 7.9 9.4 8.? 7.9 9.8 8.5 9.2 TRIG mg/dL 23 48 59 43 28 65 93 45 36 76 NA mmol/L 143 146 145 747 146 145 146 146 147 148 SEX: MALE K mmol/L 10.3 10.5 6.6 4.6 5.9 5.5 4.9 6.2 5.7 5.5 MEAN SD N 42 96 105 10.9 9.5 52 6.0 22.4 14.3 0.48 1.40 21- 8 10 10 10 10 10 10 146 6.6 1.4 2.10 10 10 LABCAT CC4.43 27- JUN-2002 418-028:PAGE K-112 MEAN SD N 63 l59.9 10 121 92.6 10 111 36.9 10 10.7 0.44 10 9.4 1. I 1 10 47 17.2 10 146 6.1 1.3 0.89 10 10 (--I - Data Unavailable LABCAT CC4.43 27- JUN-2002 418-028:PAGE K-113 Study Report f o r Clinical Chemistry STUDY I D : ARGUS 418-028 STUDY NO: 060-069 SEX: MALE CROUP: 3-M 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 19155 19159 19172 60 H 35 35 45 141 42 101 55 44 4-2- 124 H 70 82 99 197 92 171 140 102 94 -- 101 H 107 97 92 108 88 80 124 108 96 -- 11.5 H 11.5 71.5 10.2 10.8 10.7 11.2 10.7 11- 3 11.3 -- 10.4 H 9.5 15.6 8.8 9.9 9.0 9.4 9.4 8.3 8-.8- 20 H 56 34 51 35 56 21 34 22 -35- 143 H 147 148 147 145 147 I46 148 146 148 -- 8.9 H 5.6 8.4 6.5 7.0 5.5 6.1 5.4 5.7 5.4 -- MEAN SO N 60 34.4 I0 117 40.9 10 100 12.4 I0 11 .I 0.44 10 9.9 2.09 10 36 13.8 10 147 6.5 1.6 1.27 10 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-114 Study Report for Clinical Chemistry STUDY I D : ARGUS 418-028 STUDY NO: 060-069 SEX: MALE GROUP: 4-M 19100 51 91 122 10.9 8.3 14 I46 5.5 19103 19104 37 83 50 107 76 11- 0 13.3 9a 11. s 8.6 19 147 6.6 3s I4a 5.6 19118 439 887 154 10.8 9.0 23 148 6.9 19133 54 106 152 11- 7 9.3 32 147 5.3 19141 46 91 93 10.7 10-7 113 145 6.4 19142 45 108 98 10.7 8.8 35 148 5.3 19144 48 119 124 11.2 9.8 30 147 5.5 1914a 138 277 112 11.2 9.2 28 147 6.1 19150 19156 19160 46 107 I23 11.3 9-.3- 3-2- 146 -- 6.5 -- -- MEAN SD N 95 124.1 10 198 248.7 10 115 25.1 10 11 .I 0.34 10 9.7 1.43 10 36 27.9 10 147 6.0 1.o 0.60 10 10 LABCAT CC4.43 27- JUN-2002 418-028:PAGE K-115 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 SEX: MALE GROUP: 5-M 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 19154 19158 38 H 49 H 33 52 49 50 40 54 40 42 -- 86 H 114 H 76 88 104 112 95 12s 87 87 115 H 153 H 204 159 182 83 169 137 140 99 11.7 H 11.0 H 11.9 11.3 11.8 11.6 10.9 11.3 12.0 11.8 -- 12.9 H 9.2 H 9.8 9.6 11 .I 8.4 9.0 10.8 10.2 11.3 -- 7H 18 H 2 17 21 17 23 16 16 31 -- 142 H 144 H 147 146 I47 148 148 143 147 148 10.7 H 6.6 H 5.5 6.8 7.1 6.0 5.5 7.9 7. I 5.8 -- MEAN SD N 45 97 144 71.5 10.2 17 146 6.9 7.0 15.6 37.5 0.38 1.33 8.0 2.2 1.55 10 10 10 10 10 10 10 10 418-028:PAGE K-116 Study Report for Clinical Chemistry STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Animal I D GROUP: I - F 19010 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 19068 19072 19074 ALT U/L -- 144 136 177 115 142 104 153 156 87 80 -- AST U/L -- 123 182 167 114 154 121 149 157 123 1-3-3 MEAN SD N 129 31.7 10 142 22.8 10 ALP UIL -- 217 77 345 89 171 82 100 202 92 73 145 88.7 10 CA w/dL -- 10.5 10.9 11.8 10.3 9.9 10.2 11.7 10.5 9.6 10.1 -- 10.6 0.72 10 PHOS mg/dL 6.8 11.4 8.3 8.6 9.2 8.7 8.3 10.4 7.6 8-.2- 8.8 1.33 10 TRIG %/dL -- 70 65 47 91 46 44 70 51 40 20 -- SEX: FEMALE NA mmol/L -- 143 142 159 140 140 138 149 140 140 143 -- K mmol/L -- 6.0 5.8 5.4 5.5 6.2 5.7 4.8 7.4 5.7 5-.6- 54 19.9 10 143 5.8 6.3 0.67 10 10 (--I - Data Unavailable LABCAT CC4.43 ~~~ ~ ~_ 27- JUN-2002 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 418-028:PAGE K1-117 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL SEX: FEMALE GROUP: 2- F 19004 97 133 80 11.3 8.4 19009 116 110 193 11.7 7.6 19016 150 141 242 11.6 7.8 19018 136 102 240 11.6 6.7 19026 145 142 183 10.7 9.6 19036 176 125 200 11.9 8.5 19037 115 181 48 10.9 5 -3 19043 150 139 252 11.5 8.3 19047 117 131 46 10.0 5.3 19048 132 I47 102 10.4 11.8 45 139 5.3 60 144 5.9 68 147 5.4 45 144 5.0 45 141 5.0 47 145 5.4 43 143 5.2 55 142 6.1 55 142 5.6 32 140 6.9 MEAN SD N 133 23.0 10 135 21.6 10 159 81.7 10 11.2 0.63 10 7.9 1.94 10 -- 50 10.2 10 143 5.6 2.4 0.58 10 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-118 Study Report for Clinical Chemistry STUDY I D : ARGUS 418-028 STUDY NO: 060-069 SEX: FEMALE GROUP: 3-F 19003 182 134 101 9.6 9.7 26 19007 90 113 60 9.3 9.2 17 19008 107 143 64 9.7 7.3 28 19013 19014 1-3-7 1-2-5 154 -- 10-.6- 5.6 -- 65 -- 19015 I78 137 97 9.3 14.3 116 19017 166 153 41 8 11.3 5.9 33 19024 121 139 60 11.3 11.5 46 19029 189 151 15% 11- 5 6.1 53 1199003384 1-2-4 14-0- 10--8 11-.o- 10-.5- -72- 19056 19057 1-3-0 133 -- Ill 10.7 -- 9-.9- 34 -- MEAN SD N 142 34.2 10 137 11.a 10 136 107.3 10 10.4 0.87 10 9.0 2.80 10 49 29.4 10 142 5.5 139 5.8 139 5.7 1-4-4 5-.4- 141 4.9 145 5.4 142 6.0 147 5.4 141 5.5 -- -- I42 5.9 -- 142 5.6 2.5 0.32 10 10 LABCAT CC4.43 27- JUN-2002 418-028:PAGE K-119 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Animal I D GROUP: 4-F 19002 19005 19035 19039 19040 19045 19046 1905 1 19054 19058 19062 19063 19066 19069 19073 MEAN SD N ALT U/L -- 134 115 174 134 157 -- 94 124 98 119 116 -- 127 24.7 10 AST U/L -- 127 128 125 132 149 -- 107 117 129 114 1-1-2 124 12.1 10 ALP "/L -- 108 107 228 90 432 -- 70 171 102 388 -8-8 178 131.O 10 CA mg/dL -- 10.6 10.0 11.o 9.7 11 .I -- 10.0 10.7 10.3 10.7 10.3 -- 10.4 0.46 10 PHOS mg/dL -- 9.6 9.2 4.4 10.8 8.6 --- 10.1 5.4 10.7 5.0 13.2 -- 8.7 2.88 10 TRIG W/dL -- 49 67 62 77 -5-5 -- 28 30 18 34 3-3- 4s 19.5 10 NA mmol/L -- I42 142 147 138 1-4-5 -- 139 142 144 143 1-4-2 142 2.6 10 SEX: FEMALE K mmol/L -- 6.2 5.5 5.8 7.0 5.9 6.5 5.3 7.1 5.1 6-.6-- 6.1 0.70 10 LABCAT CC4.43 27- JUN-2002 418-028:PAGE K-120 Study Report for Clinical Chemistry STUDY I D : ARGUS 418-028 STUDY NO: 060-069 SEX: FEMALE MEAN SD N 145 20.0 I0 156 40.7 10 128 44.7 I0 10.4 0.82 10 10.1 1.64 10 44 18.3 10 142 6.2 2.6 0.90 10 10 (--I - Data Unavailable LABCAT CC4.43 ~~ ~~~-~~~ 27-JUN-2002 418-028:PAGE K-121 MEAN SD N 98 2.0 2.1 2.7 0.19 0.18 10 10 10 LABCAT CC4.43 , 27- JUN-2002 4 18-028:PAGE K- 122 s t u d y Report f o r Clinical Chemistry GROUP: 2-M 19102 98 191 06 99 191 08 101 19130 101 19113 98 19120 96 1 91 29 99 191 36 99 19138 99 1 91 39 98 191 47 -- 19153 19164 191 65 191 71 MEAN 99 SD 1.5 N 10 2.0 2.2 1.9 2.0 1 .? 2.2 I .8 1.9 1.9 1.9 -- 2.0 0.16 10 2.1 1.8 2.1 2.1 2.3 2.0 2.2 2.3 2.3 2.2 -- 2.1 0.16 10 LABCAT CC4.43 27-JUN-2002 ZOOZ-NnP-LZ 01 01 01 9Z'O 92'0 E'Z 2'2 0'2 00 1 N as NW3W 3 l W W :x3s 418-028:PAGE K-124 Study Report for C l i n i c a l Chemistry STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Animal I D GROUP: 4-M 19100 19103 19104 19118 19133 19141 19142 19144 19148 19150 19156 I9160 19162 19163 CL mmol/L 104 104 700 103 98 99 96 99 98 99 -- ---- MEAN 100 SD 2.7 N I0 GLOB g/dL 2.0 1.7 1.8 1.8 1.9 1.9 2.2 2.1 2.1 2.0 -- ---- 2.0 0.16 10 A/G none 2.0 2.4 2.3 2.3 2.3 2.3 2.0 2.0 2.1 2-.2- 2.2 0.15 10 SEX: MALE LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-125 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Animal I D GROUP: 5-M 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 19154 19158 19168 19169 19170 MEAN SD N CL mmol/L 101 H 101 H 99 100 101 98 101 98 7 00 99 -- 100 1.2 10 GLOB 9/dL 2.0 H 1.8 H 1.8 1.8 2.0 1.7 1.7 1.9 1.8 1.6 -- 1.8 0.13 10 A/G none 2.3 H 2.4 H 2.7 2.6 2.3 2.8 2.5 2.4 2.5 2.6 2.5 0.17 10 SEX: MALE LABCAT CC4.43 27-JUN-2002 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 4 18-028:PAGE K- 126 SEX: FEMALE 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 19068 19072 MEAN SD N 96 I.7 2.2 92 2.0 2.2 100 2.2 2.2 91 1.8 2.4 93 1.6 2.7 92 2.0 2.2 96 2.2 2.0 99 1.9 2.1 99 1.8 2.1 -9-8 2.2 1.6 96 1.9 2.2 3.4 0.22 0.28 10 10 10 LABCAT CC4.43 27- J UN-2002 418-028:PAGE K-127 Study Report f o r Clinical Chemistry STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Animal I D GROUP: 2-F 19004 19009 19016 19018 19026 19036 19037 19043 19047 19048 19052 19055 19061 19067 19071 MEAN SD N CL mmol/L 89 93 98 97 94 95 96 91 93 95 -- 94 2.7 10 GLOB g/dL I.6 1.% 1.3 1.4 1.6 1.7 1.5 1.9 1.o 1.7 -- 1.6 0.26 10 A/G none 2.7 2.2 3.2 3.0 2.4 2.5 2.3 2.0 3.3 2.5 -- 2.6 0.43 10 SEX: FEMALE (--I - Data Unavailable LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-128 MEAN SD N 95 1.8 2.2 3.8 0.21 0.21 10 10 10 LABCAT CC4.43 27- JUN-2002 4 18-028:PAGE K-9129 Study Report f o r Clinical Chemistry STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal I D GROUP: 4-F 19002 19005 19035 19039 19040 19045 19046 19051 19054 19058 19062 19063 19066 19069 19073 MEAN SD N CL mmol/L -- 90 96 101 91 97 --- 94 97 98 97 93 95 3.4 10 GLOB g/dL -- I .8 1.7 1.5 1.9 1.9 --- I .8 1.7 1.6 1.6 1-.8- I .7 0.13 10 A/G none -- 2.4 2.4 2.7 2.1 I .9 -- -- 2.5 2.3 2.8 2.3 2.3 -- 2.4 0.26 10 SEX: FEMALE LABCAT CC4.43 27- JUN-2002 418-028:PAGE K-130 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Animal I D GROUP: 5-F 19001 19006 1901 1 19020 19022 19025 1 9027 19028 19030 19031 19032 19033 19049 19059 19070 MEAN SD N CL mmol/L 91 93 92 91 100 94 96 95 95 89 --- 94 3.1 10 GLOB g/dL I .5 1.7 1.5 2.1 1.7 2.1 1.7 1.9 1.8 1.9 -- 1.8 0.21 10 A/G none 2.7 2.4 2.9 1.7 2.4 1.8 2.2 2.1 2.5 2.3 -- 2.3 0.37 10 SEX: FEMALE (--I - D a t a Unavailable LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-131 Group: I - M MEAN SD N Group: 2-M MEAN SD N Group: 3-M MEAN SD N Group: 4-M MEAN SD N Group: 5-M MEAN SD N 6.3 0.36 10 6.1 0.28 I0 6.3 0.33 10 6.2 0.28 10 6.3 0.28 10 4.3 0.22 10 4.1 0.19 I0 4.3 0.15 10 4.2 0.17 10 4.5* 0.20 10 145 16.8 10 141 26.0 10 182 58.2 10 162 25.2 10 165 33.7 10 57 0.2 8.3 0.07 10 I0 41* 11 .I 10 0.1 0.05 10 46 11.6 10 0.1 0.06 10 43* 13.1 10 0.1 0.05 I0 33* 0.2 7.2 0.06 10 10 16 0.3 288 42 1.5 0.04 172.6 6.0 10 10 10 10 16 0.3 246 63 0.8 0.05 111.5 69.9 10 I0 10 I0 16 0.3 239 60 1.a 0.06 112.9 34.4 10 10 10 10 17 0.3 196 95 1.6 0.03 53.5 124.1 10 10 10 10 21lM 0.3 243 45 2.4 0.05 117.4 7.0 10 10 I0 10 _------------------------------------------------------------------------------------------------------------- *-Significant Difference from Control P < -05 w-Significant Difference from Control P < .01 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-132 study Report for Clinical Chemistry SUMMARY REPORT PERIOD: TERMINAL STUDY I D : ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: MALE TEST(S) : UNITS: Group: I - M MEAN SD N AST U/L 96 22.4 10 ALP CA PHOS TRIG NA K CL U/L w/dL w/dL mg/dL mmol/L mmol/L mmol/L 105 10.9 9.5 52 146 6.6 98 14.3 0.48 1.40 21.8 1.4 2.10 2.7 10 I0 10 10 10 10 I0 GLOB g/dL 2.0 0.19 10 Group: 2-M MEAN SD N 121 92.6 10 111 36.9 10 10.7 0.44 10 9.4 1.I1 10 47 17.2 10 146 6.1 1.3 0.89 10 10 99 2.0 1.5 0.16 10 10 Group: 3-M MEAN SD N 117 40.9 10 100 12.4 10 11 .I 0.44 10 9.9 2.09 70 36 13.8 10 147 6.5 1.6 1.27 10 10 100 2.0 2.3 0.24 10 10 Group: 4-M MEAN SD N 198 248.7 10 115 25.1 10 11 .I 0.34 10 9.7 1.43 10 36 27.9 10 147 6.0 1.o 0.60 10 10 100 2.0 2.7 0.16 10 10 Group: 5-M MEAN SD N 97 15.4 10 144H 37.5 10 11.w 0.38 10 10.2 1.33 10 17* 146 6.9 8.0 2.2 1.55 10 10 10 100 1.8 1.2 0.13 10 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-133 Group: 2-M MEAN SD N Group: 3-M MEAN SD N Group: 4-M MEAN SD N Group: 5-M MEAN SD N 2.1 0.16 10 2.2 0.26 10 2.2 0.15 10 2.5* 0.17 10 LABCAT CC4.43 27- JUN-2002 4 18-028:PAGE K-134 Study Report for Clinical Chemistry STUDY I D : ARGUS 418-028 SEX: MALE STUDY NO: 060-069 ____________________-A-N-A-L-Y-S-IS--O-F--V-A-R-IA-N-C-E---FO-L-L-O-W-E-D--B-Y--D-U-N-N-E-T-T-'S--P-R-O-C-E-D-U-R-E--------------------------------------- Total Protein __Group . Std DUNNETT' S DUNNETT'S RANGES Degree Sum o f Mean N Tot_al___M_e_an--__-D-e-v-.__-_It-'-------L-O----9-5-%----H-I-----L-O---9-9-%-----H-I --~---S-o-u-r-c-e-----Fd-m------S-q-u-a-r-e-s-----S-q-u-a-re------ 1-M 10 62.8 6.3 0.36 TREATMENTS 4 0.44 0.11 2-M 3-M 10 60.6 10 62.5 6.1 0.28 1.60 6.3 0.33 0.22 5.9 6.6 5.9 6.6 5.9 5.9 6.7 6.7 -----E-R-R-O-R----4-5--------4-.-2-6--------0-.-0-9-- 4-M 10 61.8 6.2 0.28 0.73 5.9 6.6 5.9 6.7 TOTAL 49 4.70 5-M I0 63.3 6.3 0.28 0.36 5.9 6.6 5.9 6.7 F Ratio = Coeff. Var. % = 6.35 F ' table values F.01 = 3.78 F.05 = 2.58 4.399 __Du_n_ne_tt_'s--I_T '--ta--b-l-e--v-a-lu-e-s------P-.-0-1--=-------3--.1-2------P-.-0-5--=--------2-.5-1------------------------------ Glucose _-_-----_------------------------------------------------------------------------------------------ L___________ I-M 10 1453 145 16.8 TREATMENTS 4 10942 2735 2-M 3-M 10 1411 141 26.0 0.27 10 1823 182 58.2 2.37 106 185 106 185 96 194 ERROR 45 55071 1224 96 194 ------------------------------------- 4-M 10 1619 162 25.2 1.06 106 185 96 194 TOTAL 49 66013 5-M 10 1649 165 33.7 1.25 106 185 96 194 F Ratio = Coeff. Var. % = 2.24 21.988 I F ' table values Dunnett's I T ' table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-135 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: MALE F Ratio = Coeff. Var. % = 6.76 I F ' table values F.01 = 3.78 F.05 = 2.58 23.859 Dunnett's ' T ' table values P.01 = 3.12 P.05 = 2.51 ___--_------------------------------------------------------------------------------------------- ___ T-o_t-_a-l--B--i--l--i-r--u--b--i-n---------------------------------------------------------------------------------- __ 1-M 10 1.6 0.2 0.07 TREATMENTS 4 0.031 0.008 2-M 3-M 10 1.4 0.1 0.05 0.76 10 1.2 0.1 0.06 1.53 0.1 0.2 0.1 0.2 0.1 0.1 0.2 0.2 -----E-R-R-O-R----4-5--------0-.1-5-4-------0-.-0-0-3- 4-M 10 1.3 0. I 0.05 1.15 0.1 0.2 0.1 0.2 TOTAL 49 0.185 5-M I0 1.9 0.2 0.06 1.15 0.1 0.2 0.1 0.2 F Ratio = 2.25 I F ' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 39.527 Dunnett's ' T I table values P.01 = 3.12 P.05 = 2.51 _________-__------------------------------------------------------------------------------------- .........B..l.o..o..d...U..r..e.a....N..i.t..r..o..g.e..n............................................................................ 1-M 10 158 16 1.5 TREATMENTS 4 162.32 40.58 2-M 10 162 16 0.8 0.53 14 18 13 18 ERROR 45 129.30 2.87 3-M 10 162 16 1.8 0.53 14 18 13 18 ---------- 4-M 10 174 17 1.6 2.11 14 18 13 18 TOTAL 49 291.62 5-M 10 207 21 2.4 6.46 14 18* 13 18- LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-136 Creatinine Group N Total Mean 1-M 10 3.2 0.3 2-M 10 3.3 0.3 3-M 10 3.1 0.3 4-M 10 3.1 0.3 5-M 10 3.3 0.3 Std. DUNNETT'S Dev . 't' 0.04 0.05 0.06 0.03 0.05 0.48 0.48 0.48 0.48 DUNNETT'S RANGES LO -95%- H I LO -99%-HI Degree Source Fdm Sum of Squares Mean Square 0.3 0.4 0.3 0.4 0.3 0.4 0.3 0.4 TREATMENTS 4 0.0040 0.0010 0.3 0.3 0.4 0.4 -----E-R-R-O-R----4-5-------0-.0-9--6-0------0-.0-0-2-1- 0.3 0.4 0.3 0.4 TOTAL 49 0.1000 F Ratio = Coeff. Var. % = 1.05 107.598 I F ' table values Dunnett's ' T I table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 LABCAT CC4.43 27-JUN-2002 4 18-028:PAGE K- 137 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: MALE Aspartate Aminotransferase Std. DUNNETT'S DUNNETT'S RANGES Degree Sum o f Mean __G__ro_up __N __T_o_ta_l ___Me_an-____De_v._----I-t-'-------L-O----9-5-%----H-I-----L-O----9-9%-----H-I-------S-o-u-r-c-e----F-d-m------S-q-u-a-r-e-s----S-q-u-a-r-e------ 1-M 10 957 96 22.4 TREATMENTS 4 69637 17409 2-M 10 1213 121 92.6 0.47 -40 231 -73 264 ERROR 45 655378 14564 3-M 10 1171 117 40.9 0.40 -40 231 -73 264 ------------------------------------ 4-M 10 1976 198 248.7 1.89 -40 231 -73 264 TOTAL 49 725015 5-M 10 974 97 15.6 0.03 -40 231 -73 264 F Ratio = 3.94 ' F ' table values F.01 = 3.78 F.05 = 2.58 __C_o_e_f_f.___V_a_r_.__% = 23.795 Dunnett's ' T ' t a b l e values P.01 = 3.12 P.05 = 2.51 _______------------------------------------------------------------------------------------------ _____C_a_l_c_i_u_m_________------------------------------------------------------------------------------------------ 1-M 10 109.4 10.9 0.48 TREATMENTS 4 3.45 0.86 2-M 3-M 10 107.3 10 110.7 10.7 11.1 0.44 0.44 1.12 0.69 10.5 10.5 11.4 11.4 10.4 10.4 11.5 11.5 ------ER-R-O--R---4-5---------7-.-9-7--------0-.-1-8 4-M 10 111.0 11.1 0.34 0.85 10.5 11.4 10.4 11.5 TOTAL 49 11.42 5-M 10 115.3 11.5 0.38 3.14 10.5 11.4* 10.4 11.5* F Ratio = Coeff. Var. % = 4.87 3.800 'F' table values Dunnett's ' T I table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 *-Significant Difference from Control P < -05 *-Significant Difference from Control P < -01 Error-within groups LABCAT CC4.43 Source-Source o f Variation Treatments-between groups ~~ ________ 27- J UN-2002 4 18-028:PAGE K- 138 STUDY I D : ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: MALE Phosphorus St d . DUNNETT' S DUNNETT'S RANGES Degree Sum o f Mean _-G-r-o-u-p------N-----T-o-t-a-l-----M--ea-n------- Dev.___'_t -' -------LO----9-5-%----H-I-----L-O----99-%-~-H-I-------S-o-u-r-c-e-~-F-d-m------S-q-u-a-r-e-s----S-q-u-a-_r-_e-_-_-_- I -M 10 94.7 9.5 1.40 TREATMENTS 4 4.80 1.20 2-M 3-M 10 93.7 9.4 1.11 0.15 10 99.1 9.9 2.09 0.65 7.8 11.2 7.8 11.2 7.4 7.4 11.6 11.6 ......E.R.R.O.R.....4.5........1.0.2...3.2.........2.27 4-M 10 97.1 9.7 1.43 0.36 7.8 11.2 7.4 11.6 TOTAL 49 107.12 5-M 10 102.3 10.2 1.33 1.13 7.8 11.2 7.4 11.6 F Ratio = Coeff. Var. % = 4.98 50.584 I F ' table values Dunnett's ' T I table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-139 Study Report for Clinical Chemistry Potassium Group N Total Mean 1-M 10 65.7 6.6 2-M 10 61.0 6.1 3-M 10 64.5 6.5 4-M 10 59.7 6.0 5-M 10 69.0 6.9 S t d . DUNNETT'S Dev. It' 2.10 0.89 1.27 0.60 1.55 0.76 0.19 0.97 0.53 DUNNETT'S RANGES LO -95%- H I LO -99%- HI Degree S o u r c e Fdm Sum of Squares Mean Square 5.0 8.1 5.0 8.1 5.0 8.1 5.0 8.1 TREATMENTS 4 5.56 1.39 4.6 4.6 8.5 8.5 -----E-R-R-O-R----4-5-_-----8-6-.-2-1--------1-.-9-2-- 4.6 8.5 4.6 8.5 TOTAL 49 91.77 LABCAT CC4.43 27- JUN-2002 418-028:PAGE K-140 I-M 10 21.2 2.1 0.18 TREATMENTS 4 1.02 0.25 2-M 3-M 10 21.4 2.1 0.16 0.24 10 21.9 2.2 0.26 0.84 1.9 2.3 1.9 2.3 1.9 1.9 2.4 2.4 -----E-R-R-O-R----4-5---------1-.5--7-------0--.0-3-- 4-M 10 21.9 2.2 0.15 0.84 1.9 2.3 1.9 2.4 TOTAL 49 2.59 5-M I0 25.1 2.5 0.17 4.67 1 . 9 2.3* 1.9 2.4* LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-141 Group: I - F MEAN SD N Group: 2-F MEAN SD N Group: 3-F MEAN SD N Group: 4-F MEAN SD N Group: 5-F MEAN SD N 6.1 0.52 10 5.5 0.50 10 5.9 0.41 10 5.8 0.32 10 5.8 0.29 10 4.1 0.42 10 4.0 0.34 10 4.1 0.24 10 4.0 0.29 10 4.0 0.31 10 152 19.8 10 162 20.0 10 155 18.7 10 146 16.2 10 140 35.5 10 77 27.8 10 72 14.0 10 71 20.0 10 62 14.2 10 61 19.8 10 0.2 0.07 10 0.2 0.03 10 0.2 0.05 10 0.1 0.05 10 0.1 0.05 10 28 4.4 10 24 4.7 10 28 6.7 10 25 4.4 10 30 15.4 10 0.4 0.05 10 202 54.1 10 0.4 0.05 10 250 85.6 10 0.3 0.05 10 192 21.7 10 0.4 0.08 10 213 80.4 10 0.3 0.06 10 252 101.6 10 129 31.7 10 133 23.0 I0 142 34.2 10 127 24.7 10 145 20.0 10 LABCAT CC4.43 ______I 27- J UN-2002 4 18-028:PAGE K- 142 study Report f o r Clinical Chemistry STUDY ID: ARGUS 418-028 STUDY NO: 060-069 TEST(s) : UNITS: Group: I-F MEAN SD N Group: 2-F MEAN SD N Group: 3-F MEAN SD N Group: 4-F MEAN SD N Group: 5-f MEAN SD N AST U/L 142 22.8 10 135 21.6 I0 137 11.a 10 124 12.1 10 156 40.7 10 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE ALP CA PHOS TR 1G NA K U/L mg/dL mg/dL mg/dL mmol/L mmol/L SEX: FEMALE CL mmol/L ~ ~~~ ~--- GLOB g/dL 145 88.7 10 10.6 0.72 10 8.8 1.33 10 54 19.9 10 143 5.8 6.3 0.67 10 10 96 1.9 3.4 0.22 10 10 159 81.7 10 11.2 0.63 10 7.9 1.94 10 50 10.2 10 143 5.6 2.4 0.58 10 10 94 1.6* 2.7 0.26 10 10 136 107.3 10 10.4 0.87 10 9.0 2.80 10 49 29.4 10 142 5.6 2.5 0.32 10 10 95 1.8 3.8 0.21 10 10 178 131.O 10 10.4 0.46 10 8.7 2.88 10 45 19.5 10 142 6. I 95 1.7 2.6 0.70 3.4 0.13 10 I0 10 10 128 44.7 10 10.4 0.82 10 10.1 1.64 10 44 18.3 10 142 6.2 2.6 0.90 10 10 94 * 1.a 3.1 0.21 I0 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-143 Group: 2-F MEAN SD N Group: 3-F MEAN SD N Group: 4-F MEAN SD N Group: 5-F MEAN SD N 2.6* 0.43 10 2.2 0.21 10 2.4 0.26 10 2.3 0.37 I0 LABCAT CC4.43 27-J UN-2002 4 18-028:PAGE K- 144 LABCAT CC4.43 27- JUN-2002 418-028:PAGE K-145 F Ratio = Coeff. Var. % = 1.18 28.929 ' F ' table values Dunnett's I T ' table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 I-F 10 1.6 0.2 0.07 TREATMENTS 4 0.025 0.006 2- F 10 I.9 0.2 0.03 1.30 0.1 0.2 0.1 0.2 ERROR 45 0.120 0.003 3- F 10 1- 5 0.2 0.05 0.43 0.1 0.2 0.1 0.2 ----------------------------------- 4- F 10 1.3 0.1 0.05 1.30 0.1 0.2 0.1 0.2 TOTAL 49 0.145 5- F 10 1.3 0.1 0.05 1.30 0.1 0.2 0.1 0.2 F Ratio = Coeff. Var. 2 = 0.79 30.672 I F ' table values Dunnett's ' T I table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 Error-within groups Source-Sour ce o f Varia t ion LABCAT CC4.43 Treatments-between groups ~~~ ~ _ _ _ _ _ _ 27-JUN-2002 418-028:PAGE IS-146 Study Report f o r Clinical Chemistry STUDY I D : ARGUS 418-028 STUDY NO: 060-069 Creatinine Group N Total Mean I-F 10 3.5 0.4 2- F 10 3.5 0.4 3- F 10 3.4 0.3 4- F 10 3.6 0.4 5- F 10 3.1 0.3 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: FEMALE Std. DUNNETT'S Dev . 't' 0.05 0.05 0.05 0.08 0.06 0.00 0.37 0.37 1.47 DUNNETT'S RANGES LO -95%- H I LO -9936- HI Degree Source Fdrn Sum o f Squares Mean Square 0.3 0.4 0.3 0.4 0.3 0.4 0.3 0.4 TREATMENTS 4 0.015 0- 004 0.3 0.3 0.4 0.4 -----E-R-R-O-R----4-5--------0-.-1-6-7-------0-.0-0-4-- 0.3 0.4 0.3 0.4 TOTAL 49 0.182 ~~ ~~----_____--- I-F 10 2017 202 54.1 TREATMENTS 4 30624 7656 23--FF 1100 21499159 215902 0251..67 01..4340 i1i1a8 228855 9988 330055 ------ER-R-O-R----4-5-------2-4-7-6-1-9--------5-5-0-3- 4- F 10 2129 21 3 80.4 0.34 118 285 98 305 TOTAL 49 278244 5-F 10 2521 252 101.6 1.52 118 285 98 305 F Ratio = 1.39 ' F ' table values F.01 = 3.78 F.05 = 2.58 Coeff. V_a_r_. __%__=____ 33.472 Dunnett's ' T ' table values P.01 = 3.12 P.05 = 2.51 ____------------------------------------------------------------------------------------- Alanine Aminotransferase __I_______----______---------------------------------------------------------------------- I-F 10 1294 129 31.7 TREATMENTS 4 2546 637 2- F 3- F 10 1334 133 23.0 0.33 10 1424 142 34.2 1.07 99 160 99 160 91 91 167 167 ------E-R-R-O-R---4-5--------3--3-3-9-6- 742 4- F 10 1265 127 24.7 0.24 99 160 91 167 TOTAL 49 35942 5-F 10 1447 145 20.0 1.26 99 160 91 167 F Ratio = Coeff. Var. % = 0.86 20.138 ' F ' table values Dunnett's ' T I table values F.01 = P.01 = 3.78 3.12 F.05 = P.05 = 2.58 2.51 LABCAT CC4.43 418-028:PAGE K-147 Study Report for Clinical Chemistry STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: FEMALE Aspartate Adnotransferase Group N Total Mean Std. DUNNETT' S Dev . It' DUNNETT'S RANGES LO -95%- HI LO -99X- HI Degree Source Fdm Sum o f Squares Mean Square I-F 10 1423 142 22.8 TREATMENTS 4 5 587 1397 2- F 3- F 10 1351 135 21.6 0.67 10 1368 137 11.8 0.51 115 169 115 169 109 109 176 176 ------ER-R-O--R---4-5---------2-6-3-5-9--------5-8-6 4- F 10 1240 124 12.1 1.69 115 169 109 176 TOTAL 49 31946 5- F 10 1564 156 40.7 1.30 115 169 109 176 F Ratio = 2.38 I F ' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. X = 17.422 Dunnett's ' T ' table values P.01 = 3.12 P.05 = 2.51 -_----------------------------------------------------------------------------------------------------------- Alkaline Phosphatase ~ ~-------------- I-F 10 1448 145 88.7 TREATMENTS 4 15762 3941 2- F 10 1586 159 81.7 0.32 38 252 12 277 ERROR 45 406995 9044 3-F 10 1363 136 107.3 0.20 38 252 12 277 -_------------------------------------ 4- F 10 1784 178 131.0 0.79 38 252 12 277 TOTAL 49 422757 5- F 10 1280 128 44.7 0.40 38 252 12 277 F Ratio = 1.94 I F ' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 6.760 Dunnett's I T ' table values P.01 = 3.12 P.05 = 2.51 Error-within groups Source-Source of Variation LABCAT CC4.43 Treatment s-bet ween groups ~~___ 27-JUN-2002 418-028:PAGE K-148 I-F 10 87.5 8.8 1.33 2- F 10 79.3 7.9 1.94 0.83 6.3 11.2 3- F 10 90.0 9.0 2.80 0.25 6.3 11.2 4- F 10 87.0 8.7 2.88 0.05 6.3 11.2 5- F I0 101.0 10.1 1.64 1.37 6.3 11.2 F Ratio = Coeff. Var. % = 1.26 24.803 I F ' table values Dunnett's ' T I table values F.01 = P.01 = 5.7 5.7 11.8 11.8 TREATMENTS 4 ERROR 45 24.53 219.09 --------------------_____I___________ 6.13 4.87 5.7 11.8 5.7 11.8 TOTAL 49 243.62 3.78 F.05 = 2.58 3.12 P.05 = 2.51 I-F 10 544 54 19.9 TREATMENTS 4 693 173 2- F 3- F 10 495 10 490 50 10.2 0.54 49 29.4 0.59 32 32 77 77 26 26 83 83 -----E-R-R-O-R----4-5-------1-8-7-1-1---------4-1-6-- 4- F 10 453 45 19.5 1-00 32 77 26 83 TOTAL 49 19404 5-F 10 437 44 18.3 1.17 32 77 26 83 ~ ~ I-F 2- F 3- F 4- F 5-F _ _ _ _ _ ___ _ _ _ _ _~_ ~ 10 1434 10 1427 I0 1422 10 1424 10 1421 ~ ~ 143 143 142 142 142 ~ 6.3 2.4 2.5 2.6 2.6 0.43 0.74 0.62 0.80 139' 147 139 147 139 147 139 147 F Ratio = Coeff. Var. % = 0.21 2.534 ' F ' table values Dunnett's ' T I table values F.01 = P.01 = 138 138 138 138 148 If& 148 148 ~ ~--_-_____ TREATMENTS 4 ERROR 45 10.92 587.40 2.73 13.05 ---------------I------- TOTAL 49 598.32 3.78 F.05 = 2.58 3.12 P.05 = 2.51 LABCAT CC4.43 418-028:PAGE K-149 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Potassium Group N Total Mean I-F 10 58.1 5.8 2- F 10 55.8 5.6 3- F 10 55.5 5.6 4- F 10 61.0 6.1 5- F 10 61.6 6.2 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: FEMALE S t d . DUNNETT'S Dev . It' 0.67 0.58 0.32 0.70 0.90 0.78 0.88 0.98 1.18 DUNNETT'S RANGES LO -95%- H I LO -wX- H I Degree Source Fdm Sum o f Squares Mean Square 5.1 6.6 5.1 6.6 5.1 6.6 5.1 6.6 TREATMENTS 4 3.23 0.81 4.9 4.9 6.7 6.7 ------E-RR-O-R-----45---------1-9-.6-7--------0-.-44 4.9 6.7 4.9 6.7 TOTAL 49 22.90 ~~-- I-F 10 19.4 1.9 0.22 TREATMENTS 4 0.82 0.20 2- F 3- F 10 15.5 1.6 0.26 4.12 10 18.2 1.8 0.21 1.27 1.7 2.2* 1.7 2.2 1.6 1.6 2.22.2 ERROR 45 2.02 --------------------l___________l_ 0.04 4-F 10 17.3 1.7 0.13 2.22 1.7 2.2 1.6 2.2 TOTAL 49 2.83 5- F 10 17.9 1.8 0.21 1.59 1.7 2.2 1.6 2.2 LABCAT CC4.43 27- JUN-2002. 418-028:PAGE K-150 Study Report for Clinical Chemistry STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWED BY DUNNETT'S PROCEDURE SEX: FEMALE Albumin/Globulin Ratio Group N Total Mean . S t d . DUNNETT'S Dev It' DUNNETT'S RANGES LO -95%- H I LO -WA-HI Degree S o u r c e Fdm Sum of Squares Mean Square I-F 10 21.7 2.2 0.28 TREATMENTS 4 1.16 0.29 2-F 3-F 10 26.1 2.6 0.43 3.07 10 22.3 2.2 0.27 0.42 1.8 2.5* 1.8 2.5 1.7 7.7 2.6 2.6 -----E-R-R-O-R----4-5---------4-.6-3--------0-.-1-0-- 4- F 10 23.7 2.4 0.26 1.39 1.8 2.5 1.7 2.6 TOTAL 49 5.80 5- F 10 23.0 2.3 0.37 0.91 1.8 2.5 I.? 2.6 F Ratio = Coeff. Var. % = 2.83 13.734 IF' table values D u n n e t t ' s I T ' table values F.01 = P.01 = 3.78 3.12 F.05 = P.05 = 2.58 2.51 LABCAT CC4.43 27- JUN-2002 APPENDIX L STATEMENT OF THE STUDY DIRECTOR 418-028:PAGE L-1 PROTOCOL 418-0281 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/ DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 STATEMENT OF THE STUDY DIRECTOR This final report accurately reflects the raw data obtained during the performance of the study. No deviations from the U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations; Final Rulea, the Japanese Ministry of Health and Welfare (MHW) Good Laboratory Practice Standardfor Safety Studies on Drugsb,the Organisation for Economic Co-operation and Development (OECD). The Revised OECD Principles of Good Laboratory Practices' and the Organisation for Economic Co-operation and Development (OECD), The OECD Guideline for Testing of Chemicalsdoccurred that affected the quality or integrity of the study. Associate Director of RW esearch Study Director Argus Research ................................ a. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. b. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance No. 21, March 26, 1997. c. Organisation for Economic Co-operation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186lFinall. d. Organisation for Economic Co-operation and Development (1996). OECD Guidelinefor Testing of Chemicals. Section 4, No. 422: Combined Repeated Dose Toxicity Study with the ReproductiodDevelopmental Toxicity Screening Test, adopted 22 March 1996. APPENDIX M QUALITY ASSURANCE STATEMENT 418-028:PAGE M-1 QUALITY ASSURANCE STATEMENT Argus Protocol: 418-028 Sponsor's Study Number: T-7706.1 Study Director: Raymond G. York, Ph.D., DABT The protocol, critical phases, raw data and draft final report were inspected by the Quality Assurance Unit (QAU), to assure conformance with: Organisation for Economic Co-operation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Final]. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. The undersigned indicate that the report is an accurate representation of the raw data. Data provided by the Sponsor or a subcontractor were not audited by the Argus Research Quality Assurance Unit. 418-028:PAGE M-2 The QAU inspection and report audit dates are listed below: Inspection Phase Inspection Date(s) Protocol 22 MAR 02 Test Substance Administration 04 APR 02 Test Substance Preparation 05 APR 02 Motor Activity 08 MAY 02 Blood Collection 09 MAY 02 Fetal Blood Collection 09 MAY 02 Fetal Liver Collection 09 MAY 02 Satellite Caesarean- Sectioning 09 MAY 02 Male Sacrifice 14 MAY 02 Male Blood Collection 14 MAY 02 Sperm Evaluation 14 MAY 02 Litter Observations 17 MAY 02 Functional Observational Battery 24 MAY 02 Blood Collection 30 MAY 02 Dam/Litter Sacrifice 30 MAY 02 In-Life Data 11-20 SEP 02 Necropsy Data 13-20SEP 02 Formulation Data 17 SEP 02 Report Tables 16-23 SEP 02 24 SEP 02 Report Text 19,22-23 SEP 02 25 SEP 02 Revised Report 06 NOV 02 Date(s) Findings Submitted to Study Director 22 MAR 02 05 APR 02 06 APR 02 16 MAY 02 10 MAY 02 13 MAY 02 13 MAY 02 13 MAY 02 14 MAY 02 14 MAY 02 14 MAY 02 17 MAY 02 24 MAY 02 04 JUN 02 04 JUN 02 23 SEP 02 20 SEP 02 17 SEP 02 23 SEP 02 24 SEP 02 23 SEP 02 25 SEP 02 06 NOV 02 Date(s) Findings Submitted to Management 22 MAR 02 05 APR 02 06 APR 02 16MAY 02 10 MAY 02 13 MAY 02 13 MAY 02 13 MAY 02 14 MAY 02 14 MAY 02 14 MAY 02 17 MAY 02 24 MAY 02 04 JUN 02 04 JUN 02 23 SEP 02 20 SEP 02 17 SEP 02 23 SEP 02 24 SEP 02 23 SEP 02 25 SEP 02 06 NOV 02 Matthew J. Vaneman, B.S. Date Manager of Regulatory Compliance