Document 6wNVgq958YQELkgqJrEdZBv53

8a) OECD 407, Repeated dose Range Finding Study 132-006 Repeated Dose Range-Finding Toxiclty Sludy in Sprague-Dawley Rats Sludy Number: 132-006 A REPEATED DOSE RANGE-FINDINGTOXICITY STUDY OF T-7485 IN SPRAGUE-DAWLEY RATS STUDY NUMBER: 132-006 SPONSOR 3M Pharmaceuticals 3M Center, Building 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3320 DEC 0 9 2003 SPONSOR'S REPRESENTATIVE Ph.D., DABT, CIH Senior Laboratow Manager Telephone: FAX: TESTING FACILITY Primedica Redfield Mail: P.O. Box 308 Deliveries: 100 East Boone St. Redfield, AR 72132 Telephone: 501-397-2540 FAX: 501-397-2002 STUDY DATES Study Initiation: June 26, 2000 Animal Phase Initiation: June 27, 2000 Animal Phase Completion: July 7, 2000 Study Completion: October 11, 2000 Page 1 Repeated Dose Ranqe-Findinq Toxicity Study in Spraque-Dawley Rats QUALITY ASSURANCE STATEMENT Study Number: 132-006 Study Number: 132-006 DATEOF AUDIT~~NSPECT~ON TYPE OF AUDIT~~NSPECTION 06/26/00 0711 7/00 1011 1/00 Protocol Amendment #1 Final Report APPROVED BY: Val Gartner, B.A. Quality Assurance Auditor Primedica Redfield /,I@ 10 Date DATESREPORTETDO STUDY DIRECTORAND MANAGEMENT 06/26/00 07/17/00 1 011 1 /oo Page 2 Repeated Dose Range-Finding Toxictty Study in Sprague-Dawley Rats Study Number: 132-006 TITLE A REPEATED DOSE RANGE-FINDING TOXICITY STUDY OF T-7485 IN SPRAGUE-DAWLEY RATS APPROVED BY: Study Director Primedica Redfield Page 3 Repeated Dose Ranqe-Finding Toxicity Study in Spraque-Dawley Rats Studv Number: 132-006 TABLE OF CONTENTS Pase Cover Page....................................................................................................................................... i Quality Assurance Statement........................................................................................................... 2 Title and Signature Page................................... :.............................................................................. 3 Table of Contents ............................................................................................................................. 4 Study Abstract .................................................................................................................................. 5 Objective ........................................................................................................................................... 7 Materials and Methods ..................................................................................................................... 7 Archival Statement............................................................................................................................ 9 Results............................................................................................................................................ i o Discussion ...................................................................................................................................... 12 Conclusion ...................................................................................................................................... 12 Tables ............................................................................................................................................. 13 Table 1 - Summary of Clinical Observations............................................................................ 14 Table 2 - Summary of Body Weights ....................................................................................... 17 Table 3 - Summary of Body Weight Changes.......................................................................... 20 Table 4 - Summary of Absolute Feed Consumption................................................................ 23 Table 5 - Summary of Hematology Data.................................................................................. 26 Table 6 - Summary of Clinical Chemistry Data ........................................................................ 34 Table 7 - Summary of Organ Weights ..................................................................................... 42 Appendices ..................................................................................................................................... 56 Appendix 1 - Methods Used For Clinical Pathology Procedures............................................. 57 Appendix 2 - Pathology Rep0rt................................................................................................ 61 Appendix 3 - Individual Clinical Observations........................................................................ 123 Appendix 4 - Individual Body Weights ................................................................................... 126 Appendix 5 - Individual Feed Consumption ........................................................................... 129 Appendix 6 - Individual Hematology Data.............................................................................. 132 Appendix 7 - Individual Clinical Chemistry Data .................................................................... 162 Appendix 8 - Individual Organ Weights.................................................................................. 176 Appendix 9 - Formulated Test Material Stability Data ........................................................... 185 Appendix 10 - Certificate of Analysis ..................................................................................... 193 Appendix 11 - Protocol and Amendments ............................................................................. 195 Appendix 12 - Protocol Deviations......................................................................................... 207 Appendix 13 - Key Personnel................................................................................................. 209 Page 4 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 A REPEATED DOSE RANGE-FINDING TOXICITY STUDY OF T-7485 IN SPRAGUE-DAWLEY RATS STUDY ABSTRACT The objective of this study was to identify a maximum tolerated dose of T-7485 and identify dose levels for a 28-day repeated dose study. Healthy male and female rats were acclimated for seven days prior to Study Day 1 and were examined by the Staff Veterinarian prior to being released for use on the study. Twenty males and twenty females were randomly assigned to groups, and doses were administered via oral gavage for ten consecutive days. The study design was as follows: Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). During the ten-day repeated dose range-finding study of T-7485, there was no mortality recorded at any level of treatment for either male or female rats. Clinical observations were recorded once daily, and no adverse clinical observations were recorded at any level of treatment for either male or female rats. Body weights were recorded on Study Days 1, 7, 10, and 11. There were no significant effects of T-7485 treatment on male or female rat body weights. On Study Day 10, mean male body weights ranged between 275.8 and 286.2 grams, while female rats ranged between 193.2 and 206.2 grams. There were no significant effects of T-7485 treatment on male or female body weight changes. Between Study Days 1 and 10, the range of mean male body weight change was 62.4 to 75.0 grams, while in females the range was 27.8 to 32.4 grams. Feed consumption was measured on Study Days 1, 7, and 10. Relative to body weights and body weight changes, there was no adverse effect of T-7485 on absolute feed consumption for either male or female rats. On Study Days 1 through 10, the mean feed consumption ranged in males from 28.0 to 29.5 gramdday, while in female rats the mean feed consumption ranged from 20.8 to 12.9 gramdday. All hematological measurements in test material treated groups were comp?rablF to the vehicle control Tith the exception of elevated segmented neutrophils (1.2 f 0.3410 /mm vs. 0.5 f 0.25 103/mm vehicle control) in females that received 1000 mg/kg/day T-7485. At 1000 mg/kg/day of T-7485, female mean glucose levels were significantly increased when compared to the vehicle control (163 f 23.5 vs. 107 f 17.1 mg/dL, respectively). Both the hematologicaland clinical chemistry changes reported in female rats that received 1000 mg/kg/day T-7485 appear to be isolated changes that occurred in one gender and.had no clear association with other reported findings (including histopathology). Therefore, it is unclear if these marginal changes are related to the test material. Necropsy observations included: diverticulum of the small intestine, liver foci, small seminal vesicles, enlarged mandibular lymph nodes, lung foci, and kidney dilatation. Many of the gross findings correlated with microscopic findings. Three singular incidences of enlarged mandibular lymph nodes were noted in males that received vehicle and females that received 300 or 1000 mg/kg/day of T-7485. Enlarged mandibular lymph nodes correlated with either mild Page 5 Repeated Dose Range-FindingToxiclty Study in Sprague-Dawley Rats Study Number: 132-006 lymphoid hyperplasia (vehicle control males and high dose females, respectively) or mild plasmacytosis (females that received 300 mg/kg/day T-7485). In males that received 100 mg/kg/day of T-7485, kidney dilatation and diverticulum of the small intestine were correlated microscopically with unilateral moderate hydronephrosis and diverticulum, respectively. In males that received 1000 mg/kg/day of T-7485, lung foci was correlated microscopically with moderate focal hemorrhage. Finally, in females that received 1000 mg/kg/day T-7485, lung foci were correlated microscopically with mild chronic focal inflammation accompanied with mild focal crystalline material. Organ weights were recorded for the following organs prior to fixation: adrenal glands, brain, heart, kidneys, liver, ovaries, spleen, testes, and thymus. Paired organs were weighed together. Absolute and relative organ weights were similar to the corresponding vehicle control groups for both male and female rats with the exception of absolute and relative liver weights. Mean percent increases in absolute, % body weight, and % brain weight liver weight changes in males (female percentages are in parenthesis) that received 1000 mg/kg/day/day were 36% (22%), 34% ( I 8%), and 40% (22%), respectively. Histopathology was performed on the above listed tissues from the vehicle control and high-dose group animals and gross lesions from all animals. Non-neoplastic microscopic lesions included the following: necrosis of the heart; inflammation and hemorrhage of the esophagus; inflammation and edema of the stomach; ultimobronchialcyst of the thyroid; lymphoid hyperplasia and plasmacytosis of the mandibular lymph node; hemorrhage of the thymus; inflammation, hemorrhage, and crystalline material of the lung; and tubular degeneration, mineralization, and hydronephrosis of the kidneys. None of the microscopic lesions appear to be related to administration of the test material. The most common lesion was focal/multifocal necrosis of the cardiac muscle of the heart. This necrosis occurred in both the vehicle control animals and rats that received 1000 mg/kg/day T-7485. This lesion probably represents early cardiomyopathy, which is a common spontaneous lesion in Sprague-Dawley rats. In summary, clinical observations, body weights, body weight changes, feed consumption, gross findings, and histopathology failed to reveal any adverse effects associated with ten days of administration of sl000 mg/kg/day T-7485. Both the hematological and clinical chemistry changes reported in female rats that received 1000 mg/kg/day T-7485 appear to be isolated changes in one gender and had no clear association with other reported findings (including histopathology). It is therefore unclear if these marginal changes are related to the test material. Furthermore, in the absence of supportive clinical chemistry and histopathology, the marginal but significant differences in absolute and relative liver weights for both male and female rats that received 1000 mg/kg/day T-7485 were not considered biologically or toxicologically significant. In conclusion, these data suggest 11000 mg/kg/day T-7485 will be well-tolerated in rats that receive repeated doses of test material for 28 days. Page 6 Repealed Dose Range-FindingToxicity Study in Sprague-Dawley Rats Study Number: 132-006 OBJECTIVE The objective of this study was to identify a maximum tolerated dose of T-7485 and identify dose levels for a 28-day repeated dose study. MATERIALS AND METHODS TEST AND CONTROL MATERIALS: The test material and vehicle were identified as follows: Test Material: Lot Number: Date Received: Physical Description: Storage: Amount Received: T-7485 (Potassium Perfluoro Butane Sulfonate) 2 April 6, 2000 White Powder Room temperature, protected from light 500 grams Vehicle: Supplier: Lot Number: , Carboxymethylcellulose(medium viscosity) Sigma Chemical Co. 69H0028 The Sponsor assumed responsibility for characterization determinations of the test material. The test material was inventoried when received at Primedica Redfield, and a record of all test material usage was maintained. The Certificate of Analysis for the test material is in Appendix 10. Doses were prepared by Primedica Redfield on the day of use. The vehicle was prepared once to a concentration of 1% by mixing 10 grams of powdered medium viscosity carboxymethylcellulosewith deionized water using a Waring blender. Additional deionized water was added to yield 1000 mL of prepared vehicle. The vehicle was stored refrigerated when not in use. The test material was dissolved daily in the prepared vehicle to the required concentrations. The formulated test materials were stored refrigerated. Samples of the formulated test materials were collected and frozen at approximately -20C. These samples have not been shipped for analysis. From a single preparation, samples for stability analysis were collected immediately after the first preparation and 7, 14, 21, 28, and 32 days from the date of the first preparation and frozen at -20C. These samples were shipped to Serquest (Birmingham, Alabama) for analysis. TEST ANIMALS: Healthy male and female Crl: CD@(SD) IBS BR Stock out-bred albino rats were received from Charles River Laboratories, Inc., for use on the study. The animals were individually housed in screen-bottom stainless steel cages. Animal identification consisted of uniquely numbered ear tags and color-coded cage cards. On Study Day 1, the animals were approximately seven weeks old. The males weighed 196 to 223 grams, and the females weighed 156 to 183 grams. Teklad Certified Rodent Diet #8728 and filtered tap water were provided ad libitum. The feed and water were routinely analyzed for contaminants. There were no known contaminants in the feed or water that would be expected to affect the results of the study. The results of these analyses are on file at Primedica Redfield. Environmental controls were set to maintain temperatures of 18" to 26C (64" to 79F) with a relative humidity of 30% to 70%. These parameters were recorded at least once daily. A 12:12 hour 1ight:dark cycle was maintained in the animal room. GROUP DESIGNATION AND TREATMENT: The animals were acclimated for seven days prior to Study Day 1 and were examined by the Staff Veterinarian prior to being released for use on the Page 7 Repeated Dose Range-FindinqToxicity Study in Spraque-Dawley Rats Study Number: 132406 study. Twenty males and twenty females were randomly assigned to groups by a computer generated weight-ordered distribution such that individual body weights did not exceed G O % of the mean weight for each sex. The study design was as follows: Doses were administered via oral gavage for ten consecutive days. The first day of dosing was designated as Study Day 1. Individual dose volumes were calculated based on the most recently collected body weight for each animal. JUSTIFICATION FOR SPECIES SELECTION, ROUTE OF ADMINISTRATION, AND DOSE LEVEL: Rats are an animal model for acute toxicity studies of this type. In the assessment and evaluation of the toxic characteristics of a substance, determination of acute oral toxicity is the usual initial step. Dose levels were chosen by the Sponsor. CLINICAL OBSERVATIONS: Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). Clinical observations were recorded once daily. BODY WEIGHTS: Body weights were recorded on Study Days 1, 7, 10, and 11. The Study Day 11 body weights were only used for relative organ weight calculations. FEED CONSUMPTION: feed consumption was measured on Study Days 1, 7, and 10. HEMATOLOGY Blood samples for hematological analysis were obtained via cardiac puncture prior to necropsy. The animals were anesthetized with carbon dioxide for the collection. Feed was withheld Overnight prior to collection. The blood samples were evaluated for the following parameters: total and relative differential leukocyte counts, erythrocyte counts, hemoglobin concentration, hematocrit value, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelet counts, prothrombin time, and activated partial thromboplastin time. Specific methods for hematology analyses are located in Appendix 1. CLINICAL CHEMISTRY: Blood samples were obtained via cardiac puncture prior to necropsy. The animals were anesthetized with carbon dioxide for the collection. Feed was withheld overnight prior to collection. The samples were processed and evaluated for the following parameters: total protein, albumin, globulin, albumin/globulin ratio, glucose, cholesterol, triglycerides, total bilirubin, urea nitrogen, creatinine, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, gamma-glutamyltransferase, calcium, phosphorus, sodium, potassium, and chloride. Specific methods for clinical chemistry analyses are located in Appendix 1. NECROPSY AND ORGAN WEIGHTS: On Study Day 11, the animals were humanely euthanized via anesthesia with carbon dioxide followed by exsanguination. All animals were submitted for a complete necropsy examination. A complete necropsy was defined as examination of the external surface of the body, all orifices, and the cranial, thoracic, and abdominal cavities and their contents. Organ weights were recorded for the following organs prior to fixation: adrenal glands, brain, heart, kidneys, liver, ovaries, spleen, testes, and thymus. Paired organs were weighed together. Page 8 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats Study Number: 132-006 The following organs and tissues were examined in situ, dissected free, and fixed in 10% neutral buffered formalin: adrenal glands, aorta, bone marrow (sternum), brain (brain stem, cerebellum, cerebrum), cervix, epididymides, esophagus, eyes (with optic nerve), femur (with articular surface), gross lesions, Harderian gland, heart, large intestine (cecum, colon, rectum), small intestine (duodenum, ileum, jejunum), kidneys, lacrimal gland, liver, lungs (with bronchi), lymph nodes (mandibular, mesenteric), mammary gland, ovaries, oviducts, pancreas, pituitary gland, prostate gland, salivary gland (mandibular), sciatic nerve, seminal vesicles, skeletal muscle, skin (abdominal), spinal cord (cervical, thoracic, lumbar), spleen, stomach (forestomach, glandular), testes, thymus, thyroid/parathyroid, tongue, trachea, uterus, urinary bladder, Zymbal's gland, and vagina. HISTOPATHOLOGY: Fixed tissues were trimmed, embedded, sectioned, and stained with hematoxylin and eosin. The slides were delivered to Toxicology Pathology Associates for microscopic evaluation by Charles Frith, D.V.M., Ph.D. Histopathologywas performed on all tissues from the vehicle control and high-dose group animals and gross lesions from all animals. STATISTICS: Means and standard deviations were calculated for all quantitative data. Treated groups of the sam'e sex were compared to Group 1 at common time points using an ANOVA with a post hoc Dunnett's t Test (Dunnett, C.W., J. Amer. Statis. Assoc., 50:1096-1121, 1955). A twosided a of 0.05 or 0.01 was used to determine if statistically significant differences existed between the vehicle control and the treated groups. If no significant differences existed at the 0.05 level, then no comparison was necessary at the 0.01 level. ARCHIVAL STATEMENT All original data and the original final report will be retained at Primedica Redfield's archives, located at 100 East Boone Street, Redfield, Arkansas, for a period of five years after issuance of the final report. Wet tissues, slides, and blocks (as appropriate) will be retained for at least one year. After this time, the Sponsor will be contacted for disposition instructions. Page 9 Repeated Dose Range-Finding Toxicdy Study in Sprague-Dawley Rats Study Number: 132-006 RESULTS FORMULATED TEST MATERIAL STABILITY: From a single preparation, samples for stability analysis were collected immediately after the first preparation and 7, 14, 21, 28, and 32 days from the date of the first preparation and frozen at -20C. Data for the stability of the formulated test material is included in Appendix 9. With the exception of Study Day 1, stability data indicate the actual versus the theoretical concentrations for 4, 12, 40 mg/mL were within 10%. On Study Day 1, the 4 and 12 mg/mL actual values were 14% and 32% of their theoretical concentrations, respectively. It is unclear why these values are outside of 10% theoretical concentrations but these data support that at s40 mg/mL T-7485 does not decline but is stable for 32 days after the initial formulation. MORTALITY: The summary of mortality is in Table 1. The individual observations are in Appendix 3. Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). During the study, there were no mortality recorded at any level of treatment for either male or female rats. CLINICAL OBSERVATIONS: The summary of clinical observations is in Table 1. The individual observations are in Appendix 3. Clinical observations were recorded once daily, and no adverse clinical observations were recorded at any level of treatment for either male or female rats. BODY WEIGHTS AND BODY WEIGHT CHANGES: The summaries of body weights and body weight changes are in Tables 2 and 3, respectively.. The individual body weights are in Appendix 4. Body weights were recorded on Study Days 1, 7, 10, and 11. There were no significant effects of T-7485 treatment on male or female body weights. On Study Day 10, mean male body weights ranged between 275.8 and 286.2 grams, while female rats ranged between 193.2 and 206.2 grams. There were no significant effects of T-7485 treatment on male or female rat body weight changes. Between Study Days 1 and 10, the range of mean male body weight change was 62.4 to 75.0 grams, while in females the range was 27.8 to 32.4 grams. FEED CONSUMPTION: The summary of absolute feed consumption is in Table 4. The individual data are in Appendix 5. Feed consumption was measured on Study Days 1, 7, and 10. Relative to body weights and body weight changes, there was no adverse effect of T-7485 on absolute feed consumption for either male or female rats. On Study Days 1 through 10, the mean feed consumption ranged in males from 28.0 to 29.5 grams/day, while in female rats the mean feed consumption ranged from 20.8 to 12.9 gramdday. HEMATOLOGY: The summary of hematology data is in Table 5. The individual hematology data are in Appendix 6. Values in parenthesis indicate mean f one standard deviation. Blood samples for hematological analysis were obtained via cardiac puncture prior to necropsy. The animals were anesthetized with carbon dioxide for the collection. Feed was withheld overnight prior to collection. All hematological measurements in test material-treatedgroups were comparable to the vehicle control with the exception of elevated segmented neutrophils (1.2 f 0.341O h m 3vs. 0.5 f 0.25 103/mm3vehicle control) in females that received 1000 mg/kg/day T-7485. This elevation in segmented neutrophils appears to be an isolated Page 10 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 change that occurred only in one gender and had no clear association with other reported findings (including histopathology). It is therefore, unclear if this marginal change is related to the test material. CLINICAL CHEMISTRY: The summary of clinical chemistry data is in Table 6. The individual clinical chemistry data are in Appendix 7. Values in parenthesis indicate mean f one standard deviation. 0 At 100 mg/kg/day/day of T-7485, male mean albumin levels were significantly decreased when compared to the vehicle control (3.9 f 0.39 vs. 4.4 f 0.10 g/dL, respectively). 0 At 300 mg/kg/day of T-7485, male mean gamma-glutamyltransferase(GGT) was significantly elevated when compared to the vehicle control (4 f 2.4 vs. 0 f 0 U/L, respectively). 0 At 100 and 300 mg/kg/day T-7485, male mean chloride levels were significantly elevated when compared to the vehicle control (89 f 2.2, 90 f 1.5, and 87 f 1.1; respectively). 0 At 300 mg/kg/day T-7485, female mean alkaline phosphatase levels were significantly decreased when compared to the vehicle control (140 f 41.2 vs. 221 f 26.3, respectively). 0 At 1000 mg/kg/day T-7485, female mean glucose levels were significantly increased when compared to the vehicle control (163 f 23.5 vs. 107 f 17.1 mg/dL, respectively). In summary, with the exception of the elevated glucose level in females that received 1000 mg/kg/day, T-7485 had no dose-dependent effect on clinical chemistry parameters. The elevated glucose level was an isolated, gender specific change and it is unclear if this is related to the test material. , NECROPSY: The individual necropsy observations are included in the pathology report in Appendix 2. Necropsy observations included: diverticulum of the small intestine, liver foci, small seminal vesicles, enlarged mandibular lymph nodes, lung foci, and kidney dilatation. Many of the gross findings correlated with microscopic findings. Three singular incidences of enlarged mandibular lymph nodes were noted in males that received vehicle control and females that received 300 or 1000 mg/kg/day T-7485. Enlarged mandibular lymph nodes correlated with either mild lymphoid hyperplasia (vehicle control male and high dose female, respectively) or mild plasmacytosis (females that received 300 mg/kg/day T-7485). In males that received 100 mg/kg/day T-7485, kidney dilatation and diverticulum of the small intestine were correlated microscopically with unilateral moderate hydronephrosisand diverticulum, respectively. In males that received 1000 mg/kg/day T-7485, lung foci was correlated microscopicallywith moderate focal hemorrhage. Finally, in females that received 1000 mg/kg/day T-7485, lung foci were correlated microscopicallywith mild chronic focal inflammationaccompanied by mild focal crystalline material. ORGAN WEIGHTS: The summary of organ weights is in Table 7. The individual organ weights are in Appendix 8. Organ weights were recorded for the following organs prior to fixation: adrenal glands, brain, heart, kidneys, liver, ovaries, spleen, testes, and thymus. Paired organs were weighed together. Absolute and relative organ weights were similar to the correspondingvehicle control groups for both male and female rats with the exception of absolute and relative liver weights. Mean percent increases in absolute, % body weight, and % brain weight liver weights in males (female percentages are in parenthesis) that received 1000 mglkgldaylday were 36% (22%), 34% (18%), and 40% (22%), respectively. Page 11 Repeated Dose Ranae-Finding Toxicity Study in Spraque-Dawley Rats Study Number: 132-006 HISTOPATHOLOGY: The pathology report is in Appendix 2. Histopathology was performed on all tissues from the vehicle control and highdose group animals and gross lesions from all animals. Non-neoplastic microscopic lesions included the following: necrosis of the heart; inflammation and hemorrhage of the esophagus; inflammation and edema of the stomach; ultimobranchial cyst of the thyroid; lymphoid hyperplasia and plasmacytosis of the mandibular lymph nodes; hemorrhage of the thymus; inflammation, hemorrhage, and crystalline material of the lung; and tubular degeneration, mineralization, and hydronephrosis of the kidneys. None of the microscopic lesions appear to be related to administration of the test material. The most common lesion was focal/multifocal necrosis of the cardiac muscle of the heart. This necrosis occurred in both the vehicle control rats and rats that received 1000 mg/kg/day T-7485. This lesion probably represents early cardiomyopathy, which is a common spontaneous lesion in Sprague-Dawley rats. DISCUSSION As indicated, clinical observations, body weights, body weight changes, feed consumption, gross findings, and histopathology failed to reveal any adverse effects associated with ten days of administration of 11000 mg/kg/day T-7485. Both the hematological and clinical chemistry changes reported in female rats that received 1000 mg/kg/day T-7485 appear to be isolated changes in one gender and had no clear association with other reported findings (including histopathology). Therefore, it is unclear if these marginal changes are related to the test material. In the absence of supportive clinical chemistry and histopathology, the marginal but significant absolute and relative liver weights recorded in both male and female rats that received 1000 mg/kg/day T-7485 are not considered biologically or toxicologically significant. CONCLUSION In conclusion, these data suggest ~1000mg/kg/day T-7485 will be well-tolerated in rats that receive repeated doses of test materialfor 28 days. Page 12 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats TABLES Study Number: 132-006 Page 13 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 1 SUMMARY OF CLINICAL OBSERVATIONS Page 14 PROTOCOL 132-006: A REPEATED DOSE RANGE-FINDING TOXICITY STUOY OF 7-7485 I N SPRAWE-OAYLEY TABLE 1: C L I N I C A L OBSERVATIONS - " M Y - MALE RATS RATS Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 2 SUMMARY OF BODY WEIGHTS Page 17 E rr ttE tr . i : & Repeated Dose Range-FindingToxicrty Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 3 SUMMARY OF BODY WEIGHT CHANGES Page 20 SD 1 - 7 SD 7 - 1 0 MEANfS.0. . MEANfS.D +C7.4 f 5.1 e 3 . 0 f 3.3 +44.4 f 13.6 +18.0 f 13.0 +49.6 f 3.4 +25.4 f 3.6 +47.0 f 5.1 +26.2 f 2.9 S_D _1_- 1 _0 __II_EAN_fS.0.___+_71.2_f _5.0____+62_.4 f_26._3 ___+7_5.0 _f _6.4 ___+_73.2_f _6.3_______---.------------ SD = S T W Y DAY There uere ma significant differences noted. 9(P (D PROTOCOL 132-006: A REPEATED DOSE RANGE-FINDING TOXICITY S T W Y OF 1 - 7 4 0 5 I N SPRAGUE-DAULEY RATS T_ABL_E 3_: _BOOY_UE_IGH_T C_HANG_ES _- S_W _R Y _- F_EMA_LE R_ATS________________________--------.---------------- DOSE GRWP 1 2 3 4 (MG/KG/DAY) 0 (CONTROL) 100 300 1000 Repeated Dose Range-Finding Toxicty Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 4 SUMMARY OF ABSOLUTE FEED CONSUMPTION Page 23 h) P s 0 lD Iu UI Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 5 SUMMARY OF HEMATOLOGY DATA Page 26 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 KEY TO HEMATOLOGY TABLES ABBREVIATION TERM1NOLOGY UNITS WBC White Blood Cells (Leukocytes) 103/cu mrn RBC Red Blood Cells (Erythrocytes) 106/cu mm HGB Hemoglobin g/dL HCT Hematocrit (Packed Cell Volume) % MCV Mean Corpuscular Volume fL or cu micron MCH Mean Corpuscular Hemoglobin pg or picograms MCHC Mean Corpuscular Hemoglobin % Concentration PLT Platelets 103/curnrn NRBC Nucleated Red Blood Cell Count Ly rnphocyte Lymphocytes 103/cu rnrn Segmented Segmented Neutrophils 103/cu rnm Bands 103/cumrn Monocytes 103/curnrn Eosinophil Eosinophils 103/curnrn Basophils 103/curnrn Abnormal L Abnormal Lymphocytes 103/cu rnrn APT Activated Partial Thromboplastin Time seconds PT Prothrornbin time seconds Other Other Cells 103/curnrn Page 27 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats STUDY I D : REPEATED DOSE/RAT STUDY NO: 132-006 TABLE 5 Study Report for Hematology SUMMARY REPORT PERIOD: TERMINAL ANALYSIS OF VARIANCE FOLLOUED BY DUNNETT'S PROCEDURE Study Number: 132-006 SEX: MALE Group: I-M MEAN SD N Group: 2-M MEAN SD N Group: 3-I4 MEAN SD N Group: MEAN SD N 4-14 10.4 6.13 5 12.7 4.87 .5 8.4 2.68 5 12.4 2.87 5 6.47 0.994 5 6.23 0.953 5 6.29 0.870 5 6.30 0.292 5 15.2 2.21 5 14.7 1.69 5 14.3 1.62 5 14.9 0.51 5 40.7 7.08 5 62.8 2.71 5 39.2 5.37 5 63.1 1.a3 5 39.2 4.89 5 62.5 1.18 5 40.2 1 .a8 5 63.8 0.74 S 23.5 0.94 5 23.7 1.20 5 22.9 0.89 5 23.7 0.46 5 37.5 1211 13.3 1.14 93.2 0.37 5 5 5 37.6 1097 13.8 1.05 186.9 0.89 5 5 S 36.6 102s 13.5 0.76 174.3 0.63 5 5 5 37.2 0.51 5 1265 113.2 5 13.2 0.44 5 Page 28 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats TABLE 5 Study Report or Hematology Study Number: 132-006 STUDY I D : REPEATED DOSE/RAT STUDY NO: 132-006 ANALYSIS OF VARIANCE FOLLOUED BY DUNNETT'S PROCEDURE SEX: HALE Group: I-M HEAN SD N Group: 2-U UEAN SD N Group: 3-U UEAN so N Group: 4-M UEAN SD N 21 .o 2.89 5 19.8 0.95 .5 23.7 6.07 5 23.9 5.43 5 0 9.2 0.0 5.47 5 5 0 10.8 0.0 4.15 5 5 0 7.6 0.0 2.64 5 5 0 11.4 0.5 2.72 5 5 1.1 0.82 5 1.6 0.65 5 0.8 0.29 5 1.o 0.29 5 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 0.0 0.09 5 0.2 0.30 5 0.0 0.05 5 0.0 0.00 5 0.0 0.00 5 0.1 0.09 5 0.0 0.00 5 0.0 0.04 5 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 0.0 0.04 5 0.0 0.00 5 0.0 0.00 5 Page 29 Repealed Dose Range-Finding Toxicty Study in Sprague-Dawley Rats Study Number: 132-006 Group: 2-H MEAN SD N Group: 3-tl MEAN SD N Group: 4-H MEAN SD N 0.0 0.00 5 0.0 0.00 5 0.0 ' 0.00 5 Page 30 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats TABLE 5 Study Report f o r Hematology Study Number: 132-006 Group: I - F MEAN SD N Group: 2-F REAN SD N Group: 3-F MEAN SD N Group: 4-F MEAN SD N 8.0 3.34 5 6.81 0.485 5 15.6 0.32 5 7.5 4.20 ,5 6.M) 0.324 5 15.3 0.70 5 9.5 3.37 5 6.55 0.258 5 15.2 0.73 5 9.5 4.27 5 6.36 0.640 5 15.0 1.30 5 42.6 1.14 5 62.7 3.22 5 41.7 2.07 5 63.2 0.91 5 41 .I 1.94 5 62.8 1.98 5 40.5 3.86 5 63.7 1.93 5 22.9 1.23 5 23.2 0.47 5 23.2 0.59 5 23.6 0.78 5 36.5 0.38 5 1225 270.7 5 36.7 0.24 5 1091 125.2 5 36.9 0.44 5 1151 102.7 5 37.0 0.45 5 1284 236.1 5 13.2 0.62 5 13.4 0.26 5 13.4 0.16 5 13.4 0.32 5 Page 31 Repeated Dose Range-FindingToxiclty Study in Sprague-Dawley Rats Study Number: 132-006 Group: I-F MEAN SO N Group: 2-F HEAN SO N Group: 3-F HEAN SO N Group: 4-F MEAN so N 19.3 3.82 5 20.4 3.79 .5 23.7 3.94 5 17.3 2.04 5 0 7.4 0.5 0.0 0.0 0.0 0.0 0.0 0.4 3.24 0.2s 0.00 0.04 0.05 0.00 0.04 5 5 5 5 5 5 5 5 0 6.8 0.6 0.0 0.0 0.0 0.0 0.0 0.4 3.99 0.38 0.00 0.04 0.04 0.00 0.00 5 5 5 5 5 5 5 5 0 8.8 0.6 0.0 0.0 0.1 0.0 0.0 0.0 3.38 0.24 0.00 0.04 0.13 0.00 0.00 5 5 5 5 5 5 5 5 0 8.3 1.m 0.0 0.0 0.0 0.0 0.0 0.0 4.16 0.34 0.00 0.04 0.00 0.00 0.00 5 5 5 5 5 5 5 5 LABCAT HE4.41C Page 32 2O-JUL-2OW TABLE 5 Study Report f o r Hematology Study Number: 132-006 Group: I-F MEAN SD N Grwp: 2-F MEAN SO N Group: 3-F REAN SO N Group: 4-F REAN SO N 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 Page 33 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 6 SUMMARY OF CLINICAL CHEMISTRY DATA Page 34 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 KEY TO CLINICAL CHEMISTRY TABLES ABBREVlAT1ON TERM1NOLOGY ALP Alkaline Phosphatase AST Aspartate Aminotransferase ALT Alanine Aminotransferase CHOL Cholesterol GLU Glucose BUN Blood Urea Nitrogen CREAT Creatinine TRIG Triglycerides PHOS Phosphorus (inorganic) TP Total Protein ALB Albumin GLOB Globulin NG Albumin/Globulin Ratio CA Calcium CL Chloride NA Sodium K GGT Potassium Gamma-glutamyltransferase T-BIL Total Bilirubin UNITS U/L U/L U/L mgldL mg/dL mg/dL mg/dL mg/dL mg/dL g/d L g/d L g/d L mg/dL mEq/L or mmol/L mEq/L or mmol/L mEq/L or mrnol/L UIL mg/dL Page 35 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats TABLE 6 Study Number: 132-006 Group: I - M MEAN SD N Group: 2-M MEAN SD N Group: 3-M MEAN SD N Group: 4-M MEAN SD N 6.3 0.28 5 4.4 0.10 5 174 37.3 5 48 0.1 6.0 0.05 5 5 5.9 0.42 .5 3.m 0.39 5 152 26.0 5 47 0.1 7.5 0.04 5 5 6.1 0.27 5 4.2 0.08 5 198 24.7 5 51 11.9 5 0.1 0.07 5 6.2 0.23 5 4.5 0.19 5 208 55.1 5 41 10.9 5 0.1 0.04 5 16 0.3 1.1 0.05 5 5 16 0.2 1.9 0.04 5 5 15 0.2 1.6 0.05 5 5 17 0.3 4.4 0.07 5 5 48 173 8.0 85.9 5 5 42 189 6.6 109.2 5 5 53 187 5.1 66.2 5 5 57 167 7.2 80.2 5 5 LABCAT CC4.41 c Page 36 20- JUL-2ooO Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 6 Study Report for Clinical Chemistry ~~ SUMMARY REPORT P E R I O D : TERMINAL Group: I - M MEAN SD N Group: 2-B BEAN SO N Group: 3-M BEAN so N Group: 4-tl MEAN SD N 320 60.1 5 291 86.7 5 291 53.7 5 351 42.7 5 0 11.9 0.0 0.88 5 5 3 12.0 1.1 0.74 5 5 4* 11.9 2.4 0.86 5 5 3 12.0 2.6 0.64 5 5 14.4 1.68 5 15.0 2.80 5 15.1 2.19 5 14.7 2.36 5 27 16.7 5 22 7.4 5 24 5.8 5 32 15.6 5 149 8.0 1.5 1.26 5 5 150 7.9 1.5 1.55 5 5 149 8.1 1.3 1.28 5 5 149 7.3 1.8 1.01 5 5 07 1.9 1.1 0.19 5 5 8- 2.0 2.2 0.34 5 5 m 1.5 5 1.9 0.24 5 88 1.7 1.3 0.12 5 5 LABCAT CC4.41~ Page 37 20- JUL-2000 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 6 Study Report f o r Clinical Chemistry Group: 1-n MEAN SD N Group: 2-M MEAN SD N Group: 3-M MEAN SD N Group: 4-M MEAN SD N 2.3 0.23 5 2.0 0.40 5 2.2 0.27 5 2.6 0.23 5 Page 38 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats TABLE 6 Study Number: 132-006 STUDY I D : REPEATED DOSE/RAT STUDY NO: 132-006 ANALYSIS OF VARIANCE FOLLOUED BY DUNNETT'S PROCEDURE SEX: FEMALE TEST( s) : ____ UNITS: Group: I - F MEAN SO N TP ALE GLU CHOL T-BIL BUN CREAT ALT AST ....9./d.L........g./.d.L......w../d.L.......w./.d.L.......w./.d.L.......w./.d.L.......w./.d.L.........U./L........U../L.. 6.5 0.30 5 4.6 0.35 5 107 17.7 5 56 13.4 5 0.2 0.05 5 16 0.3 0.8 0.04 5 5 48 212 7.0 101.0 5 5 Group: 2-F MEAN SD N 6.3 0.23 5 4.5 0.26 5 112 17.1 5 64 0.2 6.1 0.04 5 5 16 0.3 49 270 1.6 0.05 13.0 91.4 5 5 5 5 Group: 3-F MEAN SD N 6.2 0.44 5 4.3 0.32 5 118 18.1 5 59 0.1 8.1 0.05 5 5 16 0.3 2.4 0.04 5 5 49 241 8.6 113.0 5 5 Group: 4-F MEAN SD N 6.2 0.17 5 4.5 0.17 5 163* 23.5 5 66 15.9 5 0. I 0.05 5 16 0.2 48 243 1.7 0.05 10.5 157.6 5 5 5 5 LABCAT CC4.41~ Page 39 20- JUL-20" Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats TABLE 6 Study Number: 132-006 Group: 2-F MEAN so N Group: 3-f MEAN SD N Group: 4-F MEAN SO N 191 2 47.8 1.3 5 5 14On 2 41.2 1.6 5 5 179 2 31.6 2.0 5 5 11.8 0.78 5 12.9 0.55 5 12.8 0.69 5 14.1 1.92 5 16.6 2.10 5 15.8 1.66 5 26 6.2 5 26 6.2 5 36 16.2 5 149 0.5 1.9 1.79 5 5 148 0.8 2.0 1.45 5 5 148 8.0 0.9 1.22 5 5 93 i.a 1.9 0.19 5 5 92 1.9 1.3 0.13 5 5 91 1.8 1.5 0.26 5 5 Page 40 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats TABLE 6 Study Number: 132-006 Group: 2-F MEAN SD N Group: 3-F MEAN SD N Croup: 4-F MEAN SD N 2.6 0.33 5 2.3 0.11 5 , 2.6 0.44 5 Page 41 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 7 SUMMARY OF ORGAN WEIGHTS Page 42 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 7 ORGAN WEIGHT SUMMARY (ABSOLUTE) Page 43 Repeated Dose Range-Finding Toxicdy Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 BOOY E I G H T (G)(ABSOLUTE) MEAN 250.000 so ii.7a98 N 5 ADRENAL GLANDS (G)(ABSOLUTE) MEAN 0.070 SD 0.0043 N 5 ' BRAIN (G)(AEMLUTE) MEAN SO N 1.%2 0.0991 5 HEART (G)(ABSOLUTE) MEAN SO N 1.158 0.0457 5 KIDNEYS (G)(ABSOLUTE) MEAN SO N 2.580 0.1713 5 LIVER (G)(ABSOLUTE) MEAN SD N 9.338 0.7433 5 SPLEEN (G)(ABSOLUTE) MEAN SO N 0.597 0.0896 5 246.200 26.281 2 5 0.084 0.0160 5 1.968 0.1001 5 1.156 0.1230 5 2.665 0.2169 5 10.012 1.0423 5 0.693 0.1664 5 254.800 9. a59o 5 0.0n 0.0165 5 1.998 0.0981 5 1 .on 0.1114 5 2.581 0.1544 5 10.573 0.6783 5 0.652 0.0874 5 254.200 a.8ia 5 0.076 0.0059 5 1 .a% 0.0449 5 1.134 0.0740 5 2.557 0.1218 5 12.R51.6508 5 0.650 0.0826 5 ___--_-..___-__-_..------.-.-----.-.------.-----.-----.-.---- ** - Significant difference Pq.01 REDFIELD LABORATORIES STUDY NUMBER 132-006 Page 44 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 ___________GRC_UP:___1-M___2-M___3-_M ___4-M_______________---- TESTES ( G I (ABSOLUTEI MEAN 2.818 2.838 2.671 2.917 SO 0.2088 0.1144 0.1927 0.1408 N 5 5 5 5 THYWS (G)(AESOLUTE) MEAN SD N 0.667 0.1450 5 0.592 0.1051 5 0.563 0.1421 5 0.620 0.0542 5 Page 45 Repeated Dose Range-Finding Toxictty Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 BWY N I G H T (G)(ABSOLUTE) MEAN 179.400 so 10.7842 N 5 ADRENAL GLANDS (G)(ABSOLUTE) MEAN 0.076 u) 0.0144 N 5 BRA1N (G) (ABSOLUTE) MEAN SO N 1.832 0.0937 5 HEART (G)(ABSOLUTE) MEAN SD N 0.810 0.0770 5 KIDNEYS (G)(ABU)LUTE) MEAN SO N 1.809 0.2449 5 LIVER (G)(ABSOLUTE) MEAN SD N 6.815 0.7907 5 OVARIES (Gl(ABSOLUTE) MEAN 0.089 SD 0.0204 N 5 173.200 8.3487 5 0.071 0.0154 5 1 A03 0.0598 5 0.810 0.0758 5 1.745 0.1428 5 6.570 0.3965 5 0.0n 0.0170 5 176.800 8.2280 5 0.070 0.0051 5 1 A70 0.0749 5 0.809 0.1050 5 1.797 O.OR8 5 7.161 1.0154 5 0.087 0.0157 5 186.000 6.4031 5 0.081 0.0098 5 1.842 0.0385 5 0.901 0.0943 5 1.982 0.2484 5 8.359. 0.8842 5 0.097 0. oom 5 Page 46 Repeated Dose Range-Finding Toxicty Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 ______.___GR_OUP_: -_1-F__-2-- F ----3-F----4-- F---------.--------------- SPLEEN (G)(ABSOLUTE) MEAN 0.444 0.422 0.464 0.507 SO 0.0559 0.0336 0.0495 0.0694 N 5 5 5 5 THYWS (G)(ABSOLUTE) MEAN SO N 0.606 0.0632 5 0.497 0.0945 5 0.569 0.0766 5 0.571 0.0849 5 Page 47 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 7 ORGAN WEIGHT SUMMARY (% BODY WEIGHT) Page 48 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 ADRENAL GLANDS ( X BODY EIGHT) MEAN 0.028 u) 0.0019 N 5 0.034 0.0067 5 BRAIN ( X BCOY MIGHT) MEAN 0.785 SO 0.0347 N 5 0.809 0.1160 5 HEART ( X BODY MIGHT) MEAN 0.464 SO 0.0158 N 5 0.471 0.0415 5 KIDNEYS ( X BOOY EIGHT) MEAN 1.032 u) 0.0295 N 5 1.089 0.1088 5 LIVER ( X BODY MIGHT) MEAN 3.731 SO 0.136a N 5 4.072 0.1828 5 SPLEEN ( X BODY E I G H T ) HUN 0.238 u) 0.0266 N 5 0.280 0.04% 5 TESTES ( X BWY E I G H T ) MEAN 1.129 SO 0.1045 N 5 1.165 0.1542 5 0.030 0.0061 5 0.784 0.0188 5 0.421 0.0352 5 1.013 0.0381 5 4.156 0.3373 5 0.256 0.0318 5 1.049 0.0670 5 0.030 0.0029 5 0.747 0.0362 5 0.446 0.0255 5 1.007 0.0556 5 5.0060.6324 5 0.255 0.0293 5 1.148 0.0466 5 ....................................................................................................................... * - Significant difference Pe.01 REOFIELD LABORATORIES S N D Y NUMBER 132-006 Page 49 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 Page 50 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 ADRENAL GLANDS ( X BWY UEIGHT) MEAN 0.042 SO 0.0056 N 5 0.041 0.0079 5 BRAIN ( X BOOY E I G H T ) MEAN 1.025 SO 0.0903 N 5 1.042 0.0353 5 ' HEART ( X BCQY WEIGHT) MEAN 0.451 SD 0.0206 N 5 0.468 0.0358 5 KIDNEYS ( X BODY WEIGHT) MEAN 1.006 Y) 0.0864 N 5 1 .om o.om 5 LIVER ( X BOOY WEIGHT) MEAN 3.709 SD 0.2092 N 5 3.793 0.1152 5 OVARIES ( X BODY E I G H T ) MAN 0.050 SD 0.0120 N 5 0.044 0. om2 5 SPLEEN ( X 8Q)Y WEIGHT) MEAN 0.247 SD 0.0259 N 5 0.245 0.0254 5 0.039 0.0027 5 1.059 0.0639 5 0.457 0.0437 5 1.017 0.0415 5 4.041 0.4551 5 0.049 0.0087 5 0.262 0.0217 5 0.043 0.0048 5 0.991 0.0354 5 0.484 0.0357 5 1.063 0.0978 5 4.4870.3388 ' 5 0.052 0.0034 5 O.2R 0.0353 5 _____.__._____._____-------------.-----------..------- tt - Significant difference Pq.01 REOFIELO LABORATORIES STWY NWBER 132-Ow Page 51 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 ________._G_RW_P: __1-F__.2--F----3--F ----4-F------------------.----.-- THYWS ( X B W Y MIGHT) MEAN 0.337 0.287 0.322 0.307 SO 0.0182 0.0508 0.0378 0.0437 N 5 5 5 5 Page 52 RepeatedDose Range-FindingToxicity Study in Sprague-Dawley Rats Study Number: 132-006 TABLE 7 ORGAN WEIGHT SUMMARY (% BRAIN WEIGHT) Page 53 Repealed Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 ADRENAL GLANDS ( X BRAIN WEIGHT) MEAN 3.588 SO 0.3627 N 5 4.290 0.9244 5 3.014 0.6R4 5 4.000 0.XZ 5 HEART ( X BRAIN M I G H T ) MEAN 59.130 SO 3.3619 N 5 50.926 7.9012 5 53.660 4.1340 5 59.790 3.8433 5 KIDNEYS ( X BRAIN E I G H T ) MEAN 131.- SO 0.7120 N 5 136.106 17.35n 5 129.250 6.2149 5 134.066 6.3555 5 LIVER ( X BRAIN M I G H T ) MEAN 475.020 SO 24.4282 N 5 510.796 70.5064 5 531.090 53.1680 5 670.M** 76.01 71 5 SPLEEN ( X BRAIN E I G H T ) MEAN 30.402 SO 3.9486 N 5 35.3% 9.4905 5 32.630 3.9612 5 34.336 4.0245 5 TESTES ( X BRAIN M I G H T ) MEAN 143.944 SO 13.7108 N 5 144.602 10.8629 5 133.760 0.4162 5 153.054 6.8824 5 THYWS ( X BRAIN UEICHT) MEAN 34.078 SO 7.6567 N 5 30.126 5.6885 5 28.054 6.0509 5 32.6% 2.6521 5 H - significant difference Ps.01 REDFIELD LABORATORIES STUDY NUMBER 132-006 Page 54 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats TABLE 7 Study Number: 132-006 .___-___-__G_ROU_P: __1-F-.---2-F-----3-F-----4-F---------------------.----. ADRENAL GLANDS ( X BRAIN E I G H T ) MEAN 4.176 3.m 3.744 4.388 SO 0.7894 0.8482 0.2554 0.5215 N 5 5 5 5 HEART ( X BRAIN E I G H T ) MEAN 44.2% so 4.4872 N 5 44.962 4.0631 5 43.382 6.6835 5 48.916 4.9685 5 KIDNEYS ( X BRAIN UEIGHT) MEAN 98.830 SO 12.6826 N 5 W.756 7.1336 5 96.280 7.3105 5 107.616 13.5825 5 LIVER ( X BRAIN UEIGHT) MEAN 373.052 SO 49.7104 N 5 364.316 17.6486 5 383.750 60.2532 5 453.8049 49.0385 5 OVARIES ( X BRAIN E I G H T ) MEAN 4.870 SO 1.02w N 5 4.280 0.9210 5 4.634 0.7124 5 5.254 0.3455 5 SPLEEN ( X BRAIN E I G H T ) MEAN 24.310 SO 3.2759 N 5 23.433 2.0511 5 24.852 2.8453 5 27.458 3.2939 5 THYMUS ( X BRAIN YEIGHT) MEAN 33.130 SO 3.8695 N 5 27.600 5.4349 5 30.374 3.1201 5 30.936 4.2188 5 ....................................................................................................................... - Significant difference P<.05 REOFIELD LABORATORIES STWY NUMBER 132-006 Page 55 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDICES Study Number: 132-006 Page 56 Repeated Dose Range-Finding Toxictty Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 1 METHODS USED FOR CLINICAL PATHOLOGY PROCEDURES Page 57 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 HEMATOLOGY The Baker System 9110 + Plus Automated Cell Counter is used for complete blood counts. White Blood Cell (WBC), Red Blood Cell (RBC) and Platelet counts - The instrument electronically counts WBC, RBC, and platelets by employing the impedance principle. This principle of electronic counting and sizing is based on the difference in conductivity between blood cells and the diluent in which they are suspended. Pulse editing is used to discriminate between red blood cells and platelets. Mean Corpuscular Volume (MCV) - Measurement of MCV is based on the RBC impedance pulse amplitude. RBC pulses are sized according to their voltage and an average is taken. The average is then compared to the MCV reference that is set during the Hematocrit calibration. Hemoglobin - The hemoglobin determination is accomplished by a cyanmethemoglobin procedure. The color intensity of this reaction is measured by a photometric cell. HCT = MCV X RBC 10 MCH = HGB X 10 RBC MCHC = HGB X 100 HCT Differentials and RBC morphology are done manually. Stain used - VOLU-SOL Wright's stain. Page 58 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 COAGULATION MCA 110 - a fully integrated microprocessor driven instrument designed to perform coagulation tests on plasma specimens. The MCA 110 operates on the principles of optical density measurement enhanced by electronic light amplification, differentiationand signal processing. Utilizing light generated by light diodes and solid state photo-detectors. Prothrombin Time Bio/Data Corporation Plastinex Thromboplastin Reagent No. 101158 Activated Partial Thromboplastin Time Bio/Data Corporation Cephalinex Activated PTT Reagent (Silica Activated) No. 101162 Page 59 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 CLINICAL CHEMISTRY The Boehringer Mannheim Hitachi - 717 Analyzer is a fully automated computerized chemistry analyzer. It is a multi-channel, discrete instrument capable of performing potentiometric and photometric assays of a wide range of analyses. The Hitachi - 717 utilizes a unique reagent and sampling system which allows the analysis of 750 tesffhour including Ion Selective Electrodes. Total Protein (Biuret) Colorimetric Determination BMC Albumin (Bromcresol Green) Colorimetric Determination BMC Glucose (Hexokinase) Enzymatic Determination BMC Cholesterol Enzymatic Determination BMC Blood Urea Nitrogen (RATE) Enzymatic Determination BMC Creatinine Kinetic Determination BMC Total Bilirubin Colorimetric Determination BMC Alanine Aminotransferase Kinetic Determination BMC Aspartate Aminotransferase Kinetic Determination BMC Alkaline Phosphatase Kinetic Determination BMC Creatine Kinase Kinetic Determination BMC Lactate Dehydrogenase Kinetic Determination BMC Gamma-Glutamy ltransferase Kinetic Determination BMC Triglyceride (GPO-Trinder) Enzymatic Determination BMC Calcium Colorimetric Determination BMC Phosphorus (Inorganic) Ultraviolet Determination BMC Direct Bilirubin Colorimetric Determination BMC Amy IaselEPS Enzymatic Colorimetric Test BMC Lipase Ultraviolet Determination BMC Sorbitol Dehydrogenase Enzymatic Determination Sigma Sodium - Ion Selective Electrode, Boehringer Mannheim Procedure Potassium - Ion Selective Electrode, Boehringer Mannheim Procedure Chloride - Ion Selective Electrode, Boehringer Mannheim Procedure BMC = Boehringer Mannheim Corporation Reagents Page 60 Study Number: 132-006 APPENDIX 2 PATHOLOGY REPORT Page 61 Repeated Dose Range-Finding Toxicty Study in Sprague-Dawley Rats Study Number: 132-006 TOXICOLOGY PATHOLOGYASSOCIATES . A REPEATED DOSE RANGE-FINDINGTOXICITY STUDY OF T-7485 IN SPRAGUE-DAWLEY RATS STUDY NUMBER: 132-006 SPONSOR TESTING FACILITY: EVALUATION FACILITY: 3M Pharmaceuticals 3M Center, Building 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3320 Tal: 651-733-1962 F a : 651-733-1773 Primedica Redfield P.O. Box 308 100 East Boone Street Redfield, Arkansas 72132 Tel: 501-397-2540 F a : 501-397-2002 Toxicology Pathology Associates 14201 Clarborne Ct. Little Rock, Arkansas 72211-5590 Tel: 501-224-5939 F a : 501-224-5939 E-mail: ISIS@Aristotle.net Page 62 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study 132-006 TABLE OF CONTENTS SUMMARY .......................................................................................................................... 3 INTRODUCTION................................................................................................................. 3 MATERIALS AND METHODS ........................................................................................... 3 RESULTS............................................................................................................................ 4 Gross Observations................................ "-..-................................................................................................ 4 Histopathological Observations............ ................................................................................................ 4 Non-neoplastic Microscopic Lesions ............................................................................................................................... 4 Correlationof Gross and MicroscopicFindings............................ "............................................... 5 _ DISCUSSION...................................................................................................................... 5 - -- Gross Observations ..._....l.-..._...... I........I....-....I_.- ..-....-.-..--.... ...l.......-..-ll-.I.. .......-.....-.- .-..5 Histopathologic Lesions............ "--.-.-- ....-..-................................................................ 5 CONCLUSION .................................................................................................................... 5 REPORTS ........................................................................................................................... 6 ABBREVIATIONS CODE TABLE...................................................................................... 8 2 Page 63 Repeafed Dose Range-FindingToxiclty Study in Sprague-Dawley Rats Study Number: 132-006 Study 132-006 Summary Treatment of male and female Sprague-Dawleyrats with a 14day repeated dose of T-7485 did not result in any gross or microscopic findings. Introduction The objective of this study was to identify a maximum tolerated dose of T-7485 and identify dose levels for a 28day repeated dose study. Materials and Methods Necropsy technicians at Primedica Redfield, Redfield, Arkansas, under the supervision of a pathologist, performed necropsies. The following tissues were routinely fixed in 10% neutral buffered formalin: adrenal glands, aorta, bone marrow (sternum). brain (brain stem), brain (cerebellum), brain (cerebrum), cervix, epididymides, esophagus, eyes with optic nerve. femur with articular surface, Harderian gland, heart, intestine, large (cecum), intestine, large (colon), intestine, large (rectum), intestine, small (duodenum), intestine, small (ileum), intestine, small (jejunum), kidneys, lacrimal gland, liver, lungs with bronchi, lymph nodes (mandibular), lymph nodes (mesenteric), mammary gland, ovaries, oviducts, pancreas, pituitary gland, prostate gland, salivary gland (mandibular), sciatic nerve, seminal vesicles. skeletal muscle. skin (abdominal), spinal cord (cervical), spinal cord (lumbar), spinal cord (thoracic), spleen, stomach (forestomach), stomach (glandular), testes, thymus, thyroid, parathyroid, tongue, trachea, urinary bladder, uterus, vagina, Zymbal's gland, and gross lesions. Fixed tissues were routinely trimmed, processed, embedded. sectioned, and stained with hematoxylin and eosin (H&E). The microscopic glass slides were delivered to Toxicology Pathology Associates, Little Rock, Arkansas, for microscopic evaluation. Tissues identified in the above paragraph from the vehicle control (Group 1) and the high dose (Group 4) animals, all gross lesions, and all animals found dead or sacrificed in extremis were processed for microscopic evaluation. Tissues were evaluated for the presence of nonneoplastic lesions. Non-neoplastic lesions were graded as to severity as trace (1). mild (2). moderate (3).and severe (4). 3 Page 64 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats Study 132406 The experimental design is outlined below: Study Number: 132-006 Results Gross Observations Gross observations were collected on all animals by necropsy technicians on a specially designed Individual Animal Necropsy Record (IANR) under the supervision of a pathologist, These findings were entered into the ADP' Computer Pathology system by the Study Pathologist for-all animals. It was necessary to enter the gross data into the ADP Pathology system in order to generate Gross Summary Tables and to correlate gross and microscopic findings. At the time of necropsy, the fate of the all animals was recorded on the IANR as scheduled sacrifice. Only a small number of gross observations were seen in the study, and none of them appeared to be related to administrationof the test article. Histopathological Observations Microscopic lesions were recorded on a Gateway Pentiurn computer utilizing the ADP automated pathology data collection system. For convenience of data collection and reporting, all data were collected utilizing the animal numbers provided by Primedica Redfield. Non-neoplastic Microscopic Lesions A small number of microscopic findings were seen, but none of them appeared to be related to administrationof the test article. `ISIS, Bellingham. Washington, markets ADP. 4 Page 65 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study 132-006 Correlation of Gross and Microscopic Findings The Gross and Microscopic Correlation Report correlates gross findings with microscopic findings by group. Discussion Gross Observations Only a small number of gross observations were seen in the study, and none of them ap- peared to be related to administration of the test article. Histopathologic Lesions A small number of microscopic findings were seen, but none of them appeared to be re- lated to administration of the test articles. The most common lesion was focaVmultifocal necrosis of the cardiac muscle of the heart. This necrosis occurred in both the vehicle control animals (Group 1) and the high dose (Group 4) animals. This lesion probably represents early cardiomyopathy, which is a common spontaneous lesion in Sprague-Dawley rats. Conclusion Treatment of male and female Sprague-Dawley rats with a 14-day repeated dose of T7485 did not result in any gross or microscopic findings. 5 Page 66 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study 132-006 Reports Study Number: 132-006 6 Page 67 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study 132-006 Reports The following reports are provided in the Reports Section. Study Groups and AssociatedAnimals - Lists the dose groups, dose levels, and identification - numbersof respectiveanimals in each group. . Gross Pathology Group Incidence Summary This report provides the number of organs examined and the incidenceof gross findings by group. Histopathology Non-neoplastic Morphology Incidence Summary - This report provides the number of organs examined and the incidenceof non-neoplasticfindings by group. Gross and Microscopic Correlation Report - This report correlates gross findings with micro- scopic findings by group. Individual Animal Necropsy Observation (Gross Pathology) - This report was generated by Artemis Computer Pathology System and includes all gross findings for a specific animal: IndividualAnimal Report - (Histopathology) This report includes all microscopic findings for a specific animal. 7 Page 68 ~~ Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats Study 132-006 Abbreviations Code Table NR = Not remarkable Correlate NP = Correlate Not Present Study Number: 132-006 8 Page 69 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawiey Rats Study Groups and Associated Animals Study Number: 132-006 Page 70 Study Number: 132-006 Study ID: 132-006 Compound: T-7485 Specier: Rac Pacholcgist: C.H. FriCh Dose Ranqe-Finding T o x ~ c i t yScudy in Sprague-Dawley PAC= SCudy Groups and The Aanociated M i m a l a Group: Group 1 Dose: 0 (rq/kq) Sex: Hale Number in Group: 5 6391 6393 6395 6397 6399 Group: Group 1 Dose: 0 (mglkg) Sex: Female Number in Group: 5 6392 6394 6396 6398 6400 Group: Group 2 Doae: 100 (mg/kg) Sex: Hale Number in Group: 5 6401 6403 6405 6407 6409 Group: Group 2 Dose: 100 (mg/kgl Sex: Female Number in Group: 5 6402 6404 6406 6408 6110 Group: Group 3 Dose: 300 (mg/kg) S e x : Hale Number m Group: 5 6411 6413 6415 6117 6419 Group: Group 3 Dose: 300 (mglkg) Sex: Female Number in Group: 5 6412 6114 6416 6418 6420 Group: Group 4 Dose: 1000 (mg/kg) Sex: Mala Number in Group: 5 6421 6123 6425 6427 6429 Group: Group 4 Dose: lo00 (mg/kgl Sex: Female Number i n Group: 5 6421 6424 6426 6420 6430 Page: 1 Date:07/25/00 Time: 11:04 ADP916 v3.0 Page 71 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Gross Pathology Group Incidence Summary Study Number: 132-006 Page 72 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Scudy ID: 132-006 Compound. T-1485 Spccres: Rae Patholcglsc: C.H. Frich Cardiovascular SyaCM Aorta Not Remarkable Hearc Not Remarkable Digeacive System ESOphaW8 NOC Remarkable. Incercine, large (cecum) NOC Remarkable Incesrine. large (colon) Nor Remarkable fncescine. large (rectum1 NOC Remarkable Incescine. small (duodenum) Noc Remarkable Intestine. small (ileum) NOC Remarkable lnccstine. small (jejunum) NoC Remarkable Remarkable Observation* Diverticulum OOse Range-Finding T o x x i t y Study in Sprague-Dawley Rats Gross Pathology Group Incidence Swnnary page: 1 Date:01/26/00 Time: 11:16 P9P401 V 3 . 0 Group case( W k g I sex Mimala On Scudy Animals Logged Group 1 0 H F 5 5 5 5 Group 7 100 5 5 5 5 Group 3 300 F 5 5 5 5 Group 4 1000 M J5 5 5 5 55555555 55555555 55555555 55555555 55555555 55555555 55555555 55555555 55555555 55555555 5555555'5 55555555 55555555 55555555 55555555 55555555 55555555 55455555 00100000 00100000 The cissue incidence rellccta the number ol cisaue. chat were processed: Remarkable. Noc Remarkable. missing. Aucolyzed or Notes Only. The morphology incidence retleccs Che number of animals in which che morphology occurred. Aucolvsia Only and NOCes Only are tabulaced i f no diagnoses are cendered. Page 73 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 study ID: 132-006 Compound: T-1485 Species: RaC Pathologist: C.H. frith D l g e s c l v e S p c m IContinuedJ Liver Not Remarkable Remarkable Observations FOCUS Pancreas NoC Renurkable Salivary Gland salivary gland (mandibular) Not Remarkable Stomach Stomach (forastomachl NoC Remarkable S t w c h (glandular) Not Remarkable sndocrine syatpn Adrenal gland- Not Remarkable Parathyroid Not Remarkable Picultaw gland Noc Remarkable Thyroid NoC Remarkable Dome Range-Fzndrng Toxicity Study rn Sprague-Davley Rats Gross Pathology Group Incidence SunvMry Page: 2 Datc;07/26/00 Time: 11:16 ADP401 V3.0 Group Dose( q / k g ) Sex .animals On Study Animals Logged Group 1 0 5 5 5 5 Group 2 100 - - - -- - -- - M f Group 3 300 MF Group I 1000 Mf 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 4 5 00 000010 0 0 0 0 0 0 1 0 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 s 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 s 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 55555555 The tismue incidence rellccta the number of tisaues thac were processed: Remarkable. Not Remarkable. Missing. Autolyzed o r Notea Only. The morphology incidence reflect. the number of animals in which the morphology occurred Auco1Ysis Onlr and Notea Only are tabulated if no diagnonea are tendered. Page 74 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: 132-006 Compound: T - 7 4 0 5 Species: RaC Pachologiac: C.H. FriCh Geni cal SyPCua Cervixlvagina Not Remarkable Epididymides NoC Remarkable nanmary gland NoC Remarkable ovaries NOC Remarkable WiduCCs Noc Remarkable ProsCaCe gland Noc Remarkable S C I I M ~ vesicles NoC Remarkable Remarkable Obacrvarions Small Terce8 Noc Remarkable uterua Not Remarkable Hematopoietic syscem Bone purrow Bone marrow (sternum) Not Rcwrkablt Dose Range-Finding Toxicicy Scudy in Spraguc-Davley RaCs Gross Pachology Group Incidence Swrvnary Page : 3 Date:07/26/00 Time: 11:16 ADP401 v3.0 Group Group 1 Dose( mg/kg I 0 SexlZ Animals On Scudy 5 5 Animals Logged 5 5 Group 2 100 M F 5 5 5 5 Group 3 300 F 5 5 5 5 Group 4 I000 H F 5 5 5 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 4 0 5 0 5 0 0 0 1 0 0 0 0 0 00100000 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 0 5 0 5 0 5 0 5 0 5 0 5 0 5 0 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 The ClSaUe lncidcnce r d l e c c s Che number o t CISaUes C h a C Were procasned: R e m a x M l e . HOC Rermrkable, nirsing. Aucolyred or mces only. The morphology incidence refleccm Che number o t animals in which Che morphology occurred. P.ucolvrir Onlx and Noces Only are tabulaced i f no diagnoses are tendered. Page 75 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: 132-006 Compound: T - 1 4 8 5 Species: Rae Patholcqist: C.H. Frith DOse Range-Finding T O X I C L ~S~tudy in Sprague-Dawley Rats Gross Pathology Group Incidence S m r y Page: 4 natC:oi/26/00 Time: 11:16 ADP401 V 3 . 0 Group mae( w l k g I Se Animala On Study Animals Logged Group 1 0 xKF 5 5 5 5 Group 2 100 M F 5 5 5 5 Group 3 300 AZ 5 5 5 5 Group 4 1000 KL 5 5 5 5 H-copAecic Syrcen lConcinucdl L p p h Node Mandibular lymph node Not Remarkable Remarkable Observatlona Enlarged Mesenteric lymph node Not Remarkable Spleen Not Remarkable Thymus Not Remarkable 5 5 5 5 5 5 5 5 4 5 5 5 5 4 5 4 10000101 1 0 0 0 0 1 0 1 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 Intequmentary System Skin ( a b d m l M 1 1 Not Remarkable 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 Wusculoakeletd Systea Bone ~ c m u r(including articular surface) Not Remarkable Skeletal muscle NOC Remarkable Tongue Noc Remarkable 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 The cissue Incidence reflects che number of tiaaues that were processed. Remarkable, Noc Remarkable. nimsing. Aucolyxed or Notea Only. The rorphology incidence reflects the number of animals in whlch the morphology occurred. AutOlYSis Onlr a d Notes Only are tabulaced if no diagnoses are tendered. Page 76 Repeated Dose Range-Finding Toxicdy Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: 137-006 compound: T-7185 speczes: Rat PathologisC: C.H. Frlth Nervous SysCMl Brain Brain (brain stem) NoC Remarkable Brain (cerebellum) Not Remarkable Brain (cerebrum) Not Remarkable Name Sciacic nerve NOC Remarkable Spinal cord s p m a l cord (cervical) NOC ReMrkdble Spinal cord (lumbar) NOC Remarkable Spinal cord (thoracic) NOC Remarkable Reapiraeory Systun Lungs (with bronchi) Not Remarkable Remarkable Obaervacions fOCU. Trachea Not Remarkable Special Senses Eyes Noc Remarkable Dose Range-Flndrng Toxicicy SCudy in Spraque-Dauley Racs Gross Pathology Group Incidence Sunvnary Page: 5 Datc:07/26/00 Time: 11:16 ADP401 V3.0 Croup Dose( W k g ) sex An~mlsOn Study Animals Logged Group 1 0 -M -f 5 5 5 5 Group 1 100 -M -f 5 5 5 5 Group 3 300 -M -F 5 5 5 5 Group 4 1000 -H -F 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 4 4 0 0 0 0 0 0 1 1 00000011 55555555 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 The cissue incidence reflects che nunbcr of Cmauel) chat were proceaned: Remarkable. Nor Remarkable. nimsing. Aucolyred or NoCes Only. The morphology incidence reflects che number of animal. in which the morphology occurred. AucolYsxII Only and NOcCa Onlr are tabulaced i f no diagnoses are Cendered. Page 77 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Scudy ID. 132-006 Compound: T-7485 Specles: Rat PaChologlsC: C.H. FrlCh special Senses lconcinuedl Harderian gland Noc Remarkable Lacrimal gland NOC Remarkable Opcic nerve N o t Rerarkable, Zymbal's gland Noc Remarkable urinary syscm Kidneys N o t Remarkable Remarkable Observations Dilatacion Urinary bladder NOC Remarkable DDse Range-Flnding Toxicicy scudy i n Spraguc-Dawley Rata Cross Pathology Croup Incidence S u m d r y page: 6 Date:01/26/00 Time: 11:16 ADP101 v 3 . 0 Group Group 1 mse( mg/kg I 0 sex& F Animals On Scudy 5 5 M i m a l s Lagged 5 5 Group 2 100 H F 5 5 5 5 Group 3 300 H F 5 5 5 5 Group I 1000 Az 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 4 5 5 5 5 5 00100000 00100000 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 The c1mmu1 incidence reflects the number 01 tir*ues that were processed: Remarkable. NOC ReMrkable, UiSsing. Aucolyzed Or Notes Only. The morphology incidence retlects Che number o t animals i n which che morphology occurred &ucolysls Only and Notes Only are Cabulated it M diagnoses are cendercd. Page 78 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Histopathology Non-neoplastic Morphology Incidence Summary Page 79 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Scudy ID: 132-006 Compound: T-7485 Species: RaC Pachologisc: C.H. frich DOse Range-Finding Toxiclcy Scudy in Sprague-Dawley Rats H1scopaChology Non-neoplascic Morphology Incidence Sumnary Page: 1 Date.07/26/00 Time: 11:07 ADP404 V3.0 Group mael wlkg 1 Sex mimals On Scudy M I ~ ~ragPged Group 1 Group 2 Group 3 -- 0100 300 - - - A & F Group 4 1000 MF 5 5 5 5 55 5 5 1 1 5 5 Cardzovascular system Hearc Nan-Neoplascic Observations Necrosis Digestive System Esophagus Non-Neoplascic Observations . Hemorrhage Inflamcion Inteacine. small ( ~ c i u n u m ) NOn-NeOplascIC Obsarvationa Drverciculum hver NOn-NeoplasciC Observaciona nononuclear Cell Infilcrace 55 0 4 1 0 41 0 55 0 00 0 00 0 oa 0 55 1 0 0 1 0 0 1 55 0 10 0 10 0 0 55 0 1 3 0 1 3 0 55 0 0 2 0 01 0 02 0 55 0 0 0 0 0 0 0 55 0 00 0 0 0 Scomach Sconuch (glandular) NOn-NeOplaaCic Obaervacions Edema Inflamcion Endocrine syacern Thyroid Non-Ncoplascic Observations Ultimobranchial cyst 55 a 00 0 00 0 0 0 0 55 0 0 0 0 00 0 a 5 5 0 02 0 01 0 01 0 55 0 1 1 0 I1 The czsaue incidence reLlecC8 che nurnber oL Ciasues ChaC were examined: Remarkable, Not Remarkable. or NoCca Only. me morphology incidence reflecca the nwnber of animals in whlch che morphology occurred. Page 80 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: 132-006 Compound: T-1405 Species:. Rat Pathologist: C . H . Frith Case Range-Finding Toxiclcy Study in Sprague-Dawley Rats Histopathology Non-neoplascic Uorphology Incidence Summary page: 2 Date:07/26/00 Time: 11:07 ADP404 V 3 . 0 H w C O p O i e c i C System Lymph Node Mandibular lymph node Non-Neoplastic Observations Lymphoid Hyperplasia PlasmacyCo-is Group Dose( w / k g ) sex Animals On Scudy Animals togged Group 1 Group 2 Group 3 Group 4 0 - - ---- - U F 100 - n 300 - F 1000 M F 5 5 5 5 5 5 5 5 3 1 5 5 4 4 0 30 0 10 0 2 0 0 1 4 5 1 0 5 0 02 1 0 3 Thymus Non-Neoplastic Observations Hemorrhage 55 0 10 0 10 0 0 55 0 01 0 01 Respiratory System Lungs (with bronchi) N O n - N e O p h ~ t i CObservations Czyrtallina material Hemorrhage Infl a m t i o n Non-Neoplastic Observations Inflanution 5 5 0 0 0 0 00 0 00 0 00 0 5 5 0 00 0 00 0 0 55 0 11 0 01 0 10 0 01 0 55 0 01 0 01 Urinary system Kidneys Non-Neoplaetic Observations Hydronephrosis uineraliration fubular osgeneration 5 '5 1 2 1 1 21 1 01 0 00 0 0 5 5 0 32 0 30 0 0 2 0 10 The tissue lncidance reflects the number o t tissues chat were examined: Remarkable, Not Remarkable. or Notes Only. The morphology incidence reflects the number of animals in which the morphology occurred. Page 81 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Gross and Microscopic Pathology Correlation Report Study Number: 132-006 Page 82 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: 132-006 Compound: T-7105 Specres: Rat Paehologist: C.H. FriCh Dose Range-Finding TOXxCiCy Study in Sprague-muley Rats Cross and Microscop~cPathology Correlation Report croup: C ~ O U D1 Dose: 0 (mq/kq) Sex: Males Animal ID: 6397 Gross Patholow Observations Mandibular l p p h node: Enlarged. Bilaceral. Correlated Microscopic PaCholoW Observations Lymphoid Hyperplasia. Mild. Page : 1 ~ace:07/26/00 Time: 11:20 ADP110 V 3 . 0 (Continuedl Page 83 ] Study Number: 132-006 Scudy ID: 132-006 Compound: T-7485 species: Rac Pacholcqiac: C.H. Frich Coee Range-Finding Toxicicy Study in Sprague-Dauley Racs Gross and Microscopic Pathology Correlation Reporc GrouD: GrOUR 2 Dose: 100 lns/kcr) Sex: Males Animal ID: 6401 Gross Pathology Obscrvacions Kidneya: Dilatatlon. RighC; Pelvia. 6407 Sunma1 vesicles: Small, Bilateral. 6409 Intestine. small IJejunwn): D ~ v e r t i c u l m i 15 x 5 EM. Correlated MicroacODiC Pathology Obserfacions Hydronephrosis. Moderace. unilateral. Corralace NP Diverciculwn. Page: 2 Dace:01/26/00 Time: 11;20 ADP110 v3.0 (Continuedl Page 84 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: 131-006 Compaund: T-7485 Species: Rat Pathologist: C.H. Frith Dose Range-Finding Toxicity study in Spraque-Oauley Rats Gross and Microscopic Pathology Correlation RepOrt Group: G r o w 3 Dose: 300 (mq/kg) Sex: Eemles Anlmal ID. 6112 Gross Pathology Observations Mandibular lymph node: Enlarged. Bilateral. Correlated Microscorric Pathology Observations Plasmacytosis, Mild. Page: 3 Date:07/26/00 T m e : 11:20 ADP110 V 3 . 0 (Continued) Page 85 Repealed Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: 132-006 Compound: T-7485 Specie.: Rat Pathologist: C.H. Frlth Duse Range-Finding TOXlCLty Study in Sprague-Dawley Rats Gross and Microscopic Pathology Correlation Report GTOUO: CrouD 4 DOSE: 1000 (ms/ks) Sex. Males h l M 1 ID: 6421 Gross P a t h o l o w Observations Correlated M~croscooic P a c h o l o w Observations L, "er' FOCUS; 2 x 2 HH Median Lobe White. Correlate NP 6427 Lungs ( w i t h bronchil: Focus: 20 x 20 HH Left LateralLobe Dark. Herrarrhage. Focal, Moderate. Page: 4 Date:01/26/00 Time: 11:Zo ADP110 v3.0 IContinuedI Page 86 Repeated Dose Range-Finding Toxicity Study in Sprague-Oawley Rats Study Number: 132-006 Study IO: 132-006 Compound: species: T-7485 Rat Pachalogisc: C.H. FrlCh Dose Range-Flndrng Toxicity Scudy in Sprague-Davley RaCa Gross and Microscopic Pathology Correlation Reporc Group: Group 4 Dose: 1000 (mq/kq) Sex: Females m i m i ID: 6422 Gross PaChologY Observations Correlated Microscooic P a c h o l w Observations Lungs (with bronchil: Focus; 4 x 3 IM Apical LObe Dark.- crystalline material, Focal, Mlld. I n f l a m t i o n . Focal, Chronic, Mild. 6424 Mandibular lymph node: Enlarged, Bilaceral. Lymphoid Hypcrplaoia. Mild. page: 5 Date:O7/26/00 Time: 11:20 ADP110 V3.0 (End OC Rsporc) Page 87 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Individual Animal Necropsy Observation (Gross Pathology) Page 88 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 REPORTS CODE TABLE TGL Trackable Gross Lesion REDFIELD LABORATORIES STUDY NUMBER 132-006 Page 89 I N O I V I W A L NECROPSY OBSERVATIONS 132-008 - A REPEATED DOSE RANGE-FINDINQ TOXICITY STUDY OF T - 7 4 8 5 I N SPRAGUE-DAWLEY RATS GROUP: 1 DOSE: D SEX: YALE ANIMAL W B E R YOM OF DEATH 6391 SCHEWLED SACRIFICE 6393 SCHEWLED SACRIFICE 6395 SCHEWLED SACRIFICE 8397 SCHEWLED SACRIFICE 6399 SCHEWLED SACRIFICE DEATH M Y (WEEK) OBSERVATION(S) 11 (2) NO V I S I B L E LESIONS 11 (2) NO V I S I B L E LESIONS 11 (2) NO V I S I B L E LESIONS 11 (2) 11 (2) LYUPH NODES-LUNDIBULAR; BILATERAL; ENLARGED (TGL) ,ANY REWINING PROTOCOL REQUIRED TISSUES, W I C H HAVE BEEN EXAMINED, NO V I S I B L E LESIONS HAVE NO V I S I B L E LESIONS I N D I V I W A L NECROPSY OBSERVATIONS 132-006 - A REPEATED DOSE RANGE-FINDING TOXICITY STUDY OF 1-7485 I N SPRAGUE-DAWLEY RATS GROUP: 1 DOSE: 0 SEX: FEUALE ANIYM NUYBER UODE OF DEATH 6392 SCHEWLEO SACRIFICE 6394 SCHEWLED SACRIFICE 6396 SCHEWLED SACRIFICE 8398 SCHEWLED SACRIFICE 6400 SCHEWLED SACRIFICE DEATH DAY (WEEK) OBSERVATION(S) 11 (2) NO VISIBLE LESIONS 11 (2) NO VISIBLE LESIONS 11 (2) NO VISIBLE LESIONS 11 (2) NO VISIBLE LESIONS 11 (2) NO VISIBLE LESIONS I N D I V I W A L NECROPSY OBSERVATIONS 132-006 -A REPEATED DOSE RANQE-FINDINQ TOXICITY STUDY OF T-7485 I N SPRAQUE-DAWLEY RATS QROUP: 2 DOSE: 30 SEX: W E ANIUAL - NUMBER WOE OF DEATH 6401 SCHEWLED SACRIFICE DEATH DAY (WEEK) OBSERVATION(S) 11 (2) KIDNEYS; RIGHT; DILATATION; PELVIS (TQL) ANY REWINING PROTOCOL REQUIRED TISSUES, WHICH HAVE BEEN EXAUINEO, HAVE NO VISIBLE LESIONS 6403 SCHEWLED SACRIFICE 11 (2) NO VISIBLE LESIONS 6405 SCHEDULED SACRIFICE 11 (2) No VISIBLE LESIONS 6407 SCHEDULED SACRIFICE 11 (2) SEMINAL VESICLES; BILATERAL; SMALL (TGL) tn-0 ANY REMAINING PROTOCOL REQUIRED TISSUES, WHICH HAVE BEEN EXAUINEO, HAVE NO VISIBLE LESIONS m (0 6409 SCHEWLED SACRIFICE 11 (2) SM. INTESTINE-JEJUNW; DIVERTICULUM (TQL): 15 X 5 UU ANY REMAINING PROTOCOL REWIRED TISSUES, WHICH HAVE BEEN EXAMINED, HAVE NO VISIBLE LESIONS 9(P ID INDIVIDUAL NECROPSY OBSERVATIONS 132-006 - A REPEATED W S E W O E - F I N D I N G TOXICITY STUDY OF T - 7 4 8 5 I N SPRAGUE-DAWLEY RATS ~ GROUP: 2 ~ ~~ DOSE: 30 ~ SEX: F E W ANIW - NWEER UODE OF DEATH 6402 SCHEDULED SACRIFICE DEATH M Y (WEEK) OBSERVATION(S) 11 ( 2 ) NO VISIBLE LESIONS 6404 SCHEDULED SACRIFICE 11 ( 2 ) NO VISIBLE LESIONS 6 4 0 6 SCHEOULED SACRIFICE 11 (2) NO VISIBLE LESIONS 6408 SCHEDULED SACRIFICE 11 ( 2 ) NO VISIBLE LESIONS 6410 SCHEDULED SACRIFICE 11 (2) NO VISIBLE LESIONS I N O I V I W A L NECROPSY OBSERVATIONS 132-006 -A REPEATED DOSE RANGE-FINDING TOXICITY STUDY OF T - 7 4 8 5 I N SPRAGUE-OAWCEY RATS GROUP: 3 DOSE: 1 0 0 SEX: W L E ANIMAL M E R WOE OF DEATH 6411 SCHEWLEO SACRIFICE 6413 SCHEWLED SACRIFICE 6415 SCHEWLEO SACRIFICE 6417 SCHEWLEO SACRIFICE 6 4 1 9 SCHEOULEO SACRIFICE DEATH OAY (WEEK) OBSERVATION(S) 11 (2) NO VISIBLE LESIONS 11 (2) NO VISIBLE LESIONS 11 (2) NO VISIBLE LESIONS 11 (2) NO VISIBLE LESIONS 11 (2) NO VISIBLE LESIONS P73 (D P 2 0 (D z 3 ln ? 3 3 a 3 ln 0 -I sE. ? a Y-. 3 cn sla c ? P i Y BIn 2 a Y z c 3:, 2 w Y 0 0 0 ) I N O I V I W A L NECROPSY OBSERVATIONS 132-008 - A REPEATED DOSE RANGE-FINDING TOXICITY STUDY OF 1 - 7 4 8 5 I N SPRAGUE-DAWLEY RATS OROUP: 3 DOSE: 100 SEX: FEUALE ANIUAL M E R UODE OF DEATH DEATH DAY (WEEK) OBSERVATION(S) 6412 SCHEWLED SACRIFICE 11 (2) LYUPH NODES-WDIBULAR; BILATERAL; ENLARGED (TGL) rVr, REUAINING PROTOCOL REWIRED TISSUES, WHICH HAVE BEEN EXAMINED, HAVE NO VISIBLE LESIONS 6414 SCHEWLED SACRIFICE 11 (2) NO V I S I B L E LESIONS 6416 SCHEWLED SACRIFICE 11 (2) NO VISIBLE LESIONS 6416 SCHEWLEO SACRIFICE 11 (2) NO V I S I B L E LESIONS 9 6420 SCHEWLEO SACRIFICE 11 (2) NO V I S I B L E LESIONS (D lD I N O I V I W A L NECROPSY OBSERVATIONS 132-006 - A REPEATED DOSE RANGE-FINDING TOXICITY STUDY OF T-7485 I N SPRAGUE-DAWLEY RATS GROUP: 4 DOSE: 300 SEX: MALE ANIWL NWBER W E OF DEATH DEATH M Y (WEEK) OBSERVATION(S) 6421 SCHEWLEO SACRIFICE 11 (2) LIVER; LlEOIAN LOBE; FOCUS; M I T E (TGL): 2 X 2 UU ANY REMAINING PROTOCOL REWIREO TISSUES, WHICH HAVE BEEN EXAUINEO, HAVE NO VISIBLE LESIONS 6423 SCHEWLEO SACRIFICE 11 (2) NO VISIBLE LESIONS 6425 SCHEWLED SACRIFICE 11 (2) NO VISIBLE LESIONS 6427 SCHEWLED SACRIFICE 11 (2) LUNGS WIBRONCHI; LEFT; FOCUS; DARK (TGL): 20 X 20 U 9 ANY RWAINING PROTOCOL REWIREO TISSUES, WHICH HAVE BEEN EXAUINEO, HAVE NO VISIBLE LESIONS (D (D 6429 SCHEWLED SACRIFICE 11 (2) NO VISIBLE LESIONS I N O I V I W A L NECROPSY OBSERVATIONS 132-006 - A REPEATED DOSE RANGE-FINDING TOXICITY STUDY OF T - 7 4 8 5 I N SPRAGUE-OAWLEY RATS GROUP: 4 DOSE: 300 SEX: FEUALE ANIUU NUUBER MODE OF DEATH DEATH MY (WEEK) OBSERVATION(S) 6422 SCHEWLED SACRIFICE 11 (2) LUNGS WIBRONCHI; APICAL LOBE; FOCUS; DARK (TGL): 4 X 3 UM ANY REUAINING PROTOCOL REWIRED TISSUES, WICH HAVE BEEN EXAMINED, HAVE NO VISIBLE LESIONS 6424 SCHEWLED SACRIFICE 11 (2) LYUPH NODES-WOIBULAR; BILATERAL; ENLARGE0 (TGL) ANY REUAINING PROTOCOL REQUIRED TISSUES, WHICH HAVE BEEN EXAMINED, HAVE NO VISIBLE LESIONS 6426 SCHEDULED SACRIFICE 11 (2) NO VISIBLE LESIONS 3 6428 SCHEWLED SACRIFICE 11 (2) NO VISIBLE LESIONS (P (D 6430 SCHEDULED SACRIFICE 11 (2) NO VISIBLE LESIONS Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats Individual Animal Report (Histopathology) Study Number: 132-006 Page 98 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified No Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 1 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 v3.0 - Sex Male Grour, Group 1 Dose ( mq/ks ) 0 Animal ID 6391 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followins histooatholow observations were noted: Heart. Necrosis, Multifocal, Mild. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, . Mandibular lymph node, Mesenteric lymph node, optic nerve, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular), Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal), Spinal cord (cervical),Spinal cord (lumbar),Spinal cord (thoracic), Spleen, Stomach (forestomach),Stomach (glandular),Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 99 Repeated Dose Range-Finding Toxicw Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID verified No Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 2 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-1485 ADP001 V3.0 Dose - Sex Male Grour, Group 1 1 ms/kq 0 Animal ID 6393 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 01/07/00 Terminal Sac The followins histoDatholow observations were noted: Heart. Necrosis, Multifocal, Trace. Esophagus, Normal. Liver. .Esophaguson Slide # 2 with lungs. Mononuclear Cell Infiltrate, Multifocal, Trace. Mandibular lymph node. Plasmacytosis, Mild. Thymus. Hemorrhage, Multifocal, Trace. Kidneys. Hydronephrosis, Trace, Unilateral. The followina tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum),Lacrimal gland, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, Optic nene, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular),Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal), Spinal cord (cervical), Spinal cord (lumbar), Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular),Testes, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 100 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 3 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 v3.0 - Sex Male Group Group 1 Dose ( ms/kq ) 0 Animal ID 6395 Age Days Date of in on Patholosist Removal Reason TeSt C.H. Frith 07/07/00 Terminal Sac The followinq histoDatholow observations were noted: Heart. Necrosis, Multifocal, Mild. Mandibular lymph node, Missinq. The followinq tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum), Brain (cerebrum), Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, Optic nene, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular),Sciatic nene, Seminal vesicles, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular), Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 101 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 4 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7405 ADP001 V3.0 - Sex Male Grour, Group 1 Dose ( mq/kq 0 Animal ID 6397 Age Days Patholosist Date of in Death Removal Reason on Test C.H. Frith 07/07/00 Terminal Sac The followinq histopatholow observations were noted: Mandibular lymph node. Lymphoid Hyperplasia, Mild, Correlate #l. Kidneys. Hydronephrosis, Trace, Unilateral. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum), Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, Optic nerve, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular),Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach). Stomach (glandular),Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 102 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 5 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:ZO T-7485 ADP001 V3.0 - Sex Male GrOUrJ Group 1 Dose ( ms/kq ) 0 Animal ID 6399 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followins histopatholow observations were noted: Heart. Necrosis, Multifocal, Trace. Pancreas, Missinq. Mandibular lymph n o d e . Plasmacytosis, Trace. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum), Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, Optic nerve, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular),Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal),Spinal cord (cenrical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular), Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 103 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain : Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 6 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 V3.0 Dose - Sex ( ms/ks ) Animal ID Female Group 1 0 6392 Age Days Patholoqist Date of in Oeath Removal Reason on Test C.H. Frith 07/07/00 Terminal Sac The followinq histopatholow observations were noted: Kidneys . Mineralization, Multifocal, Trace, Bilateral. The followinq tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Cervix/Vagina, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mandibular lymph node, Mesenteric lymph node, optic nerve, Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular),Sciatic nerve, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular),Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Uterus, Zymbal's gland. (End of Report) Page 104 Repeated Dose Range-FindingToxiclty Study in Sprague-Dawley Rats Study Number 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 7 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:ZO T-7485 ADP001 V3.0 - Sex Female Group 1 Dose ( mcr/ks ) 0 Animal ID 639 4 Age Days Patholocrist Date of in Death Removal Reason on Test C.H. Frith 01/07/00 Terminal Sac The followins histouatholosv observations were noted: No Remarkable Observations. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Cervix/Vagina, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mandibular lymph node, Mesenteric lymph node, Optic nerve, Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular),Sciatic nerve, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar), Spinal cord (thoracic), Spleen, Stomach (forestomach), Stomach (glandular), Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Uterus, Zymbal's gland. (End of Report) Page 105 Repeated Dose Range-Finding Toxicity Study in Sprague-Daw@ Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page: . 8 Rat Individual Animal Report Date:07/26/00 0 Histopathology Time: 11:ZO T-7485 AOPOOl v3.0 - Sex Female GTOUI) Group 1 Dose ( mq/kq ) 0 Animal ID 6396 Age Days Date of in on Patholoqist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followinq histouatholocw observations were noted: Mandibular lymph node, Missinq. The followinq tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum), Brain (cerebrum), Cervix/Vagina, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, Optic nerve, Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular),Sciatic nerve, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar), Spinal cord (thoracic), Spleen, Stomach (forestomach), Stomach (glandular),Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Uterus, Zymbal's gland. (End of Report) Page 106 ~ ~ ~~~ Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 9 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 v3.0 - Sex Female GTOUF, Group 1 Dose ( mq/kq ) 0 Animal ID 6398 Age Days Date of in on Patholoqist Death Removal Reason Tese C.H. Frith 07/07/00 Terminal Sac The followinq histopatholoqv observations were noted: No Remarkable Observations. The followinq tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Cervix/Vagina, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mandibular lymph node, Mesenteric lymph node, Optic nerve, Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular),Sciatic nerve, Skeletal muscle, Skin (abdominal),Spinal cord (cervical),Spinal cord (lumbar), Spinal cord (thoracic),Spleen, Stomach (forestomach),Stomach (glandular), Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Uterus, Zymbal's gland. (End of Report) Page 107 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body we0 Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 10 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 V3.0 - Sex Female Group Group 1 Dose ( ms/ks L 0 Animal ID 6400 Age Days Patholosist Date of in Death Removal Reason on Test C.H. Frith 07/07/00 Terminal Sac The followins histopatholow observations were noted: Heart. Necrosis, Multifocal, Trace. Kidneys. Hydronephrosis, Trace, Unilateral. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum), Cervix/Vagina, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mandibular lymph node, Mesenteric lymph node, Optic nerve. Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular), Sciatic nerve, Skeletal muscle, Skin (abdominal), Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular), Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Uterus, Zymbal's gland. (End of Report) Page 108 Repeated Dose Range-Finding Toxicdy Study in Sprague-Dawley Rats study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 11 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 V3.0 - Sex Male Grour, Group 2 Dose 1 mdks ) 100 Animal ID 6401 Age Days Patholosist Date of in Death Removal Reason on Test C.H. Frith 07/07/00 Terminal Sac The followins histoDatholosv observations were noted: Kidneys. Hydronephrosis, Moderate, Unilateral, Correlate #l. The followins protocol tissues were not examined: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mandibular lymph node, Mesenteric lymph node, Optic nerve, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular),Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar), Spinal cord (thoracic), Spleen, Stomach (forestomach), Stomach (glandular), Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 109 g Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 12 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:ZO T-7405 ADP001 v3.0 - Sex Male Group Group 2 Dose ( ms/ks L 100 Animal ID 6407 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followins histouatholow observations were noted: Seminal v e s i c l e s , Normal, Correlate NP. The followins tissues were marked as NR: Seminal vesicles. The followins urotocol tissues were not examined: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum), Brain (cerebrum), Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mandibular lymph node, Mesenteric lymph node, Optic nerve, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular), Sciatic nerve, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular),Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report 1 Page 110 Repealed Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified No Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 13 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 v3.0 - Sex Male Group 2 Dose ( mq/kq 1 100 Animal ID 6409 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followinq histooatholow observations were noted: I n t e s t i n e , small (jejunum). Diverticulum, Correlate #l. The followinq orotocol tissues were not examined: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (includingarticular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Manunary gland, Mandibular lymph node, Mesenteric lymph node, Optic nerve, Pancreas, Parathyroid, Pituitary glad, Prostate gland, Salivary gland (mandibular),Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal), Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular), Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 111 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats ~~ Study Number 132-006 Study ID: Species : Strain : Compound : ID Verified No Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 14 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: ll:20 T-1485 ADP001 v3.0 Dose _ Se. x ( mq/ks ) Animal ID Female Group 3 300 6412 Age Days Date of in on Patholoqist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followino histouatholow observations were noted: Mandibular l p p h node. Plasmacytosis, Mild, Correlate #1. The followinq protocol tissues were not examined: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum), Brain (cerebrum), Cervix/Vagina, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, Optic nerve, Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular),Sciatic nerve, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular), Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Uterus, Zymbal's gland. (End of Report) Page 112 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified No Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 15 Rat Individual Animal Report Date:01/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 V3.0 -Sex Male GTDtJ1) Group 4 Dose ( mq/ka 1 1000 Animal ID 6421 Age Days Date of in on Patholoqist Death Removal Reason Test C.H. Frith 01/01/00 Terminal Sac The followinq histouatholow observations were noted: Heart. Necrosis, Multifocal, Mild. Liver, Normal, Correlate NP. Sciatic nerve, Missinq. Kidneys . Hydronephrosis, Trace, Unilateral. The followinq tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain Stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mandibular lymph node, Mesenteric lymph node, Optic nerve, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular),Seminal vesicles, Skeletal muscle, Skin (abdominal), Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular),Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 113 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 16 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 V3.0 - Sex Male Group 4 Dose ( ms/ks ) 1000 Animal ID 6423 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followins histoDatholom observations were noted: No Remarkable Observations. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mandibular lymph node, Mesenteric lymph node, Optic nerve, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular), Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal), Spinal cord (cervical), Spinal cord (lumbar), Spinal cord (thoracic), Spleen, Stomach (forestomach), Stomach (glandular),Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 114 Repealed Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain : Compound : ID Verified No Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 17 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 V3.0 - Sex Grour, Dose ( mq/ks ) Animal ID Male Group 4 1000 6425 Age Days Date of in on Patholoqist Death Removal Reason TeSt C.H. Frith 07/07/00 Terminal Sac The followins histopatholoqv observations were noted: Heart. .Necrosis, Multifocal, Mild. Mandibular lymph node, Missinq. Kidneys. . Hydronephrosis, Trace, Unilateral. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum), Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine,.large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, optic nerve, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular),Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular), Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 115 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified No Body Wto Dose Range-Findhg Toxicity Study in Sprague-Dawley Rats 132-006 Page : 18 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 v3.0 - Sex Male GrouD Group 4 Dose 1 mq/ks 1 1000 Animal ID 6427 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followina histooatholow observations were noted: Heart. Necrosis, Multifocal, Mild. Lungs (with bronchi!. Hemorrhage, Focal, Moderate, Correlate #l. Kidneys. Hydronephrosis, Trace, Unilateral. Tubular Degeneration, Multifocal, Mild, Unilateral. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum). Intestine, small (ileum), Intestine, small (jejunum), Lacrimal gland, Liver, Mammary gland, Mandibular lymph node, Mesenteric lymph node! Optic nerve, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular),Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular),Testes, Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report) Page 116 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body w3J- Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 19 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 V3.0 -Sex Male Group 4 Dose ( ms/ks 1000 Animal ID 6429 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 01/07/00 Terminal Sac The followins histopatholow observations were noted: Heart. Necrosis. Multifocal, Trace. Thyroid . Ultimobranchial cyst, Focal, Trace, Unilateral. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum),Epididymides, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi.), Mammary gland, Mandibular lymph node, Mesenteric lymph node, optic nerve, Pancreas, Parathyroid, Pituitary gland, Prostate gland, Salivary gland (mandibular), Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin (abdominal), Spinal cord (cervical),Spinal cord (lumbar), Spinal cord (thoracic), Spleen, Stomach (forestomach),Stomach (glandular), Testes, Thymus, Tongue, Trachea, Urinary bladder, Zymbal's gland. (End of Report1 Page 117 Repeated Dose Range-Finding Toxictty Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 20 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 V3.0 -Sex Female GrOUp Group 4 Dose 1 ms/ks 1000 Animal ID 6422 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followins histoaatholosv observations were noted: Heart. Necrosis, Multifocal, Mild. Mandibular lymph node. Plasmaeytosis, Mild. Lungs fwith bronchi) . Crystalline material, Focal, Mild, Correlate #l. Inflammation, Focal, Chronic, Mild, Correlate #l. Kidneys. Mineralization, Multifocal, Trace, Bilateral. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum), Brain (brain Stem), Brain (cerebellum),Brain (cerebrum),Cervix/Vagina, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Lacrimal gland, Liver, Mammary gland, Mesenteric lymph node, optic nerve, Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular), Sciatic nerve, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Stomach (glandular),Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Uterus, Zymbal's gland. ... ~ . (End of Report) Page 118 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-FindingToxicity Study in Sprague-DawleyRats 132-006 Page : 21 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 v3.0 - sex Female GrOUp Group 4 Dose ( m/ks 1000 ) Animal ID 6424 Age Days Date of in on Patholoqist C.H. Frith Deaeh Removal Reason 07/07/00 Terminal Sac The followinq histouatholow observations were noted: stomach (glandular). Edema, Multifocal, Mild. Mandibular lymph node. Lymphoid Hyperplasia, Mild, Correlate #l. Thymus. Hemorrhage, Multifocal, Trace. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum), Brain (cerebrum), Cervix/Vayina, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mamary gland, Mesenteric lymph node, Optic nerve, Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular), Sciatic nerve, Skeletal muscle, Skin (abdominal), Spinal cord (cervical), Spinal cord (lumbar), Spinal cord (thoracic),Spleen, Stomach (forestomach), Thyroid, Tongue, Trachea, Urinary bladder, Uterus, Zymbal's gland. (End of Report) Page 119 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body Wto Dose Range-FindingToxicity Study in Sprague-DawleyRats 132-006 Page : 22 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 V3.0 -Sex Female Grour, Group 4 Dose ( ma/ks 1000 Animal ID 6426 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followins histopatholow observations were noted: Esophagus. Inflammation, Focal, Chronic, Mild; Muscle. Mandibular lymph node. Plasmacytosis, Mild. Trachea. Inflammation, Focal, Chronic, Mild. Xi dneys . Mineralization, Multifocal, Trace, Bilateral. The followins tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum), Brain (brain Stem), Brain (cerebellum), Brain (cerebrum), Cenix/Vagina, Eyes, Femur (including articular surface), Harderian gland, Heart, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, Optic nene, Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular),Sciatic nerve, Skeletal muscle, Skin (abdominal), Spinal cord (cervical), Spinal cord (lumbar), Spinal cord (thoracic), Spleen, Stomach (forestomach), Stomach (glandular), Thymus, Thyroid, Tongue, Urinary bladder, Uterus, Zymbal's gland. (End of Report) Page 120 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain: Compound : ID Verified NO Body wto Dose Range-FindingToxicity Study in Sprague-Dawley Rats 132-006 Page : 23 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 - Sex Female Group Group 4 Dose 1 mq/kq 1000 ADP001 V3.0 L Animal fD 6428 Age Days Date of in on Patholoqist Death Removal Reason Q=yg Test C.H. Frith 07/07/00 Terminal Sac The followinq histooatholocw observations were noted: Heart. Necrosis, Multifocal, Trace. Esophagus. Hemorrhage, Diffuse, Moderate. Inflammation, Diffuse, Subacute, Moderate. Parathyroid, Missinq. Thyroid . Ultimobranchial cyst, Focal, Trace, Bilateral. Mandibular lymph node. Plasmacytosis, Trace. The followinq tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum), Brain (brain stem), Brain (cerebellum), Brain (cerebrum),Cervix/Vagina, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, Optic nerve, Ovaries, Oviducts, Pancreas, Pituitary gland, Salivary gland (mandibular),Sciatic nerve, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar), Spinal cord (thoracic), Spleen, Stomach (forestomach), Stomach (glandular),Thymus, Tongue, Trachea,-Urinarybladder, Uterus, Zymbal's gland. (End of Report) Page 121 Repeated Dose Range-Finding Toxicdy Study in Sprague-Dawley Rats Study Number: 132-006 Study ID: Species : Strain : Compound : ID Verified NO Body Wto Dose Range-Finding Toxicity Study in Sprague-Dawley Rats 132-006 Page : 24 Rat Individual Animal Report Date:07/26/00 CD Histopathology Time: 11:20 T-7485 ADP001 v 3 . 0 Dose - Sex ( ms/ks ) Animal ID Female Group 4 1000 6430 Age Days Date of in on Patholosist Death Removal Reason Test C.H. Frith 07/07/00 Terminal Sac The followino histouatholow observations were noted: Heart. Necrosis, Multifocal, Mild. Stomach .(glandular). Inflammation, Focal, Acute, Mild. Mandibular lymph node. Lymphoid Hyperplasia, Mild. The followino tissues were marked as NR: Adrenal glands, Aorta, Bone marrow (sternum),Brain (brain stem), Brain (cerebellum),Brain (cerebrum), Cervix/Vagina, Esophagus, Eyes, Femur (including articular surface), Harderian gland, Intestine, large (cecum), Intestine, large (colon), Intestine, large (rectum), Intestine, small (duodenum), Intestine, small (ileum), Intestine, small (jejunum), Kidneys, Lacrimal gland, Liver, Lungs (with bronchi), Mammary gland, Mesenteric lymph node, Optic nerve, Ovaries, Oviducts, Pancreas, Parathyroid, Pituitary gland, Salivary gland (mandibular),Sciatic nerve, Skeletal muscle, Skin (abdominal),Spinal cord (cervical), Spinal cord (lumbar),Spinal cord (thoracic),Spleen, Stomach (forestomach), Thymus, Thyroid, Tongue, Trachea, Urinary bladder, Uterus, Zymbal's gland. (End of Report) Page 122 Repeated Dose Range-Finding Toxicty Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 3 INDIVIDUAL CLINICAL OBSERVATIONS Page 123 -0 fu u3 CD 3 N UI Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 4 INDIVIDUAL BODY WEIGHTS Page 126 9(mP _ _ _ _ _ _ _ ~~ ~ ~ ~ ~~~ ~ Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 5 INDIVIDUAL FEED CONSUMPTION Page 129 yam BTRee cores 0 rm 6. & Repeated Dose Range-Finding Toxictty Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 6 INDIVIDUAL HEMATOLOGY DATA Page 132 Repeated Dose Range-Finding ToxicQ Study in Sprague-Dawley Rats Study Number: 132-006 KEY TO HEMATOLOGY TABLES ABBR EVIATlON TERMINOLOGY UNITS WBC White Blood Cells (Leukocytes) 1o3/cUmm RBC Red Blood Cells (Erythrocytes) 106/cumm HGB Hemoglobin gld L HCT Hematocrit (Packed Cell Volume) % MCV Mean Corpuscular Volume fL or cu micron MCH Mean Corpuscular Hemoglobin pg or picograms MCHC Mean Corpuscular Hemoglobin Yo Concentration PLT Platelets 1o3/cUmm NRBC Nucleated Red Blood Cell Count Lymphocyte Lymphocytes 103/cumm Segmented Segmented Neutrophils 103/cumm Bands 103/cumm Monocytes 103/cumm Eosinophil Eosinophils 103/cumm Basophils 103/cu mm Abnormal L Abnormal Lymphocytes 103/cu mm APlT Activated Partial ThromboplastinTime seconds PT Prothrombin time seconds Other Other Cells 103/cumm Page 133 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 6 Study Report or Hematology INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL STUDY I D : REPEATED DOSE/RAT STUDY NO: 132-006 Animal I D uec THSN/CU HH RBC HILL/CU HH HGB GRAHS/DL HCT HCV HCH X CU MICRONS P I C 0 GRAMS _______-___-----I---_________I__________------------------------------------------------------------- GROUP: 1-M 6391 4.1 6.64 14.7 39.1 58.9 22.1 6393 4.1 6.13 14.7 38.2 62.3 24.0 6395 14.1 7.00 16.2 44.0 62.9 23.1 6397 17.8 7.60 18.2 50.5 66.5 23.9 6399 11.9 4.97 12.2 31.5 63.3 24.5 MEAN 10.4 ' 6.47 15.2 40.7 62.8 23.5 SD 6.13 0.994 2.21 7.08 2.71 0.94 N 5 5 5 5 5 5 SEX: HALE HCHC X PLT THSN/CU HM 37.6 38.5 36.8 36.0 38.7 37.5 1.14 5 1075 12n 1313 1215 1173 1211 93.2 5 GROUP: 2-M 6401 6403 6405 6407 6409 MEAN SD N 13.0 9.0 17.2 17.7 6.7 12.7 4.87 5 6.56 4.53 6.79 6.61 6.65 6.23 0.953 5 15.2 11.7 16.0 15.2 15.2 14.7 1.69 5 40.3 29.8 43.5 40.8 41.4 39.2 5.37 5 61.5 65.8 64.1 61.8 62.2 63.1 1.83 5 23.2 25.8 23.6 23.0 22.9 23.7 1 .M 5 37.7 1326 39.3 873 36.8 1251 37.3 1042 36.7 595 37.6 1097 1.05 186.9 5 5 _____________-_-___________l___________l----_-_-__-___I_ GROUP: 3-H 641 1 9.4 6.18 14.5 39.3 63.6 23.5 36.9 1313 661 3 8.5 4.83 11.5 30.8 63.8 23.8 37.3 896 641 5 5.3 7.01 15.1 42.8 61 . O 21.5 35.3 1054 64'17 12.3 6.75 15.4 42.0 62.2 22.8 36.7 890 641 9 6.7 6.68 15.2 41.3 61.9 22.8 36.8 971 MEAN 8.4 6.29 14.3 39.2 62.5 22.9 36.6 1025 SD 2.68 0.870 1.62 4.89 1.18 0.89 0.76 174.3 N 5 5 5 5 5 5 5 5 Page 134 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 GRWP: 4-M MI 6423 6425 6427 6429 MEAN so N 8.9 14.7 11.9 10.5 15.8 12.4 2.87 5 6.24 6.19 6.44 5.93 6.71 6.30 0.292 5 14.7 14.7 15.0 14.5 15.8 14.9 0.51 5 39.6 39.1 40.8 54.4 43.2 63.5 23.6 63.1 23.7 63.3 23.3 64.8 24.5 64.4 23.5 40.2 63.8 23.7 1.aa 0.74 0.46 5 5 5 37.1 37.6 36.8 37.8 36.6 37.2 0.51 5 1243 1084 1284 1341 1374 1265 113.2 5 Page 135 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 STUDY I D : REPEATED DOSE/RAT STUDY NO: 132-006 SEX: HALE GROUP: 1-M 6391 6393 6395 6397 6399 MEAN SD N 13.1 13.4 13.9 13.0 13.1 13.3 0.37 5 17.2 24.4 23.2 19.4 20.9 21 .o 2.89 5 0 3.6 0 3.4 0 12.5 0 15.5 0 11.2 0 9.2 0.0 5.47 5 5 0.5 0.6 1.6 2.3 0.5 1.1 0.82 5 0.0 0.0 0.0 0.0 0.0 0.0 0.00 5 0.0 0.0 0.0 0.0 0.2 0.0 0.09 5 0.0 0.0 0.0 0.0 0.0 0.0 0.00 5 GROUP: 2-tl 6401 6403 6405 6407 6409 MEAN SD N 12.8 13.5 13.9 15.2 13.5 13.8 0.89 5 20.6 19.3 18.5 20.8 19.6 19.8 0.95 5 0 10.8 2.0 0.0 0.1 0.0 0 8.0 1.0 0.0 0.0 0.0 0 15.3 . 1.7 0.0 0.0 0.2 0 14.5 2.5 0.0 0.7 0.0 0 5.6 1.0 . 0.0 0.0 0.1 0 10.8 1.6 0.0 0.2 0.1 0.0 4.15 0.65 0.00 0.30 0.09 5 5 5 5 5 5 _I________________-_________I_I_________--------------____-____- GROUP: 3-M 6411 13.3 20.4 6413 13.5 20.9 6415 6417 14.6 34.0 13.4 19.0 6419 12.9 24.2 0 8.8 0.6 0.0 0.0 0.0 0 7.7 0.9 0.0 0.0 0.0 0 4.2 1.o 0.0 0.1 0.0 0 11.2 1.1 0.0 0.0 0.0 0 6.2 0.4 0.0 0.1 0.0 MEAN SD N 13.5 0.63 5 23.7 6.07 5 0 7.6 0.8 0.0 0.0 0.0 0.0 2.64 0.29 0.00 0.05 0.00 5 5 5 5 5 5 Page 136 Repeated Dose Range-FindingToxicrty Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 STUDY ID: REPEATED DOSE/RAT SEX: MALE . .S.T.U.D.Y.N.O.:. .1.3.2.-0.0.6. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . AnimaL I D PT APTT NRBC Lymphocyte Segmented Bands Monocytes Eosinophi L seconds seconds COUNT THSN/CU M M THSN/CU M M THSN/CU M M THSN/CU M M THSN/CU M M GROUP: 4-H 6421 6423 6425 6427 , 6429 MEAN SD N 13.0 12.8 13.9 12.9 13.2 13.2 0.44 5 23.3 20.5 33.3 22.4 20.0 23.9 5.43 5 0 0.0 0 13.8 1 10.7 1 9.9 0 14.5 0 11.4 0.5 2.72 5 5 0.8 0.9 1.2 0.6 1.3 1.o 0.29 5 0.0 0.0 0.0 0.0 0.0 0.0 0.w 5 0.0 0.0 0.0 0.0 0.0 0.0 0.00 5 0.1 0.0 0.0 0.0 0.0 0.0 0.04 5 Page 137 Repeated Dose Range-Finding Toxicdy Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 Study Report for Hematology INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL STUDY ID: REPEATED DOSE/RAT STUDY NO: 1324% SEX: #ALE NEAN SD N 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 MEAN SD N 0.0 0.00 5 0.0 0.04 5 . 0.0 0.00 5 MEAN SD N 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 Page 138 APPENDIX 6 Study Report for Hematology Study Number: 132-006 HEAN SD N 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 Page 139 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats APPENDIX 6 Study Report for Hematology STUDY ID: REPEATED DOSE/RAT STUDY NO: 132-006 Study Number: 132-006 SEX: FEUALE MEAN 8.0 6.81 15.6 42.6 62.7 22.9 36.5 1225 SD 3.34 0.485 0.32 1.14 3.22 1.23 0.38 270.7 N 5 5 5 5 5 5 5 5 GROUP: 2-F 6402 6404 6406 6408 641 0 4.8 4.2 5.2 14.2 9.0 6.53 7.11 6.22 6.50 6.64 14.9 16.4 14.6 15.5 15.1 40.7 45.1 39.6 41.9 41.3 MEAN SD N 7.5 4.20 5 6.60 0.324 5 15.3 0.70 5 41 -7 2.07 5 ------------- ------ ___----__I- GROUP: 3-F 641 2 641 4 641 6 6418 6420 5.9 8.7 12.8 13.2 6.8 6.26 6.65 6.63 6.88 6.31 14.1 15.8 15.1 15.9 15.0 38.4 42.8 40.2 43.1 41.2 UEAN 9.5 6.55 15.2 41.1 SD 3.37 0.258 0.73 1.94 N 5 5 5 5 62.4 22.8 36.6 1199 63.5 23.1 36.4 1001 63.6 23.5 36.9 1245 64.4 23.8 37.0 959 62.2 22.7 36.6 1049 63.2 23.2 0.91 0.47 5 5 - - I _ - - - - 36.7 1091 0.24 125.2 5 5 ---- --- I- -- I ---- 61.3 64.3 60.6 62.6 65.3 62.8 1.98 5 22.5 23.8 22.8 23.1 23.0 23.2 0.59 5 36.7 36.9 37.6 36.9 36.4 36.9 0.44 5 1050 1317 1100 1172 1114 1151 102.7 5 Page 140 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 STUDY I D : REPEATED DOSEJRAT STUDY NO: 132-006 Animal I D UBC THSN/CU MM RBC HILL/CU HM GROUP: 4-F 6422 7.6 6424 3.0 6426 11.3 6428 13.5 6430 12.3 5.96 6.93 6.25 5.58 7.09 MEAN SO N 9.5 6.36 4.27 0.640 5 5 HGB GRAMS/DL 13.8 15.6 14.9 13.7 16.8 15.0 1.30 5 HCT ncv MCH X CU MICRONS PIC0 GRAMS 37.3 42.5 23.2 42.1 M.8 22.5 40.7 65.1 23.8 36.3 65.1 24.6 45.9 64.0 23.7 40.5 63.7 23.6 3.86 1.93 0.78 5 5 5 SEX: FEMALE MCHC X PLT THSN/CU MM 37.0 37.1 36.6 37.7 36.6 37.0 0.45 5 1363 1197 1547 1389 925 1284 236.1 5 Page 141 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 GROUP: 1-F 6392 6394 63% 6398 6400 MEAN SO N 13.5 12.3 13.2 14.0 13.2 13.2 0.62 5 23.6 18.2 22.9 17.2 14.7 19.3 3.82 5 0 11.3 0 2.7 0 8.8 0 8.3 1 6.0 0 7.4 0.4 3.24 5 5 0.5 0.3 0.9 0.3 0.4 0.5 0.25 5 0.0 0.0 0.0 0.0 0.0 0.0 0.00 5 0.0 0.1 0.0 0.0 0.0 0.0 0.04 5 0.0 0.0 0.0 0.1 0.1 0.0 0.05 5 ________-1___--_1--_1_____1___________1_--_----------__ GROUP: 2-F 6402 13.7 18.9 6404 13.5 25.8 6406 13.0 21.4 6408 13.3 20.5 6410 13.4 15.4 0 4.3 0.5 0.0 0.0 0.0 0 4.0 0.2 0.0 0.0 0.0 1 4.2 1.o 0.0 0.0 0.0 0 13.1 1.o 0.0 0.0 0.1 0 8.6 0.3 0.0 0.1 0.0 MEAN SO N 13.4 0.26 5 20.4 3.79 5 0 6.8 0.6 0.0 0.0 0.0 0.4 3.99 0.38 0.00 0.04 0.04 5 5 5 5 5 5 ________---_---___________-__l___l__l----------ll____l___ GROUP: 3-F 6412 6414 6416 6418 6420 13.4 13.1 13.4 13.5 13.5 21.2 20.8 21 .8 30.3 24.3 0 5.3 0.5 0.0 0.0 0.1 0 7.9 0.8 0.0 0.0 0.0 0 11.9 0.9 0.0 0.0 0.0 0 12.7 0.3 0.0 0.0 0.3 0 6.0 0.7 0.0 0.1 0.0 MEAN SD N 13.4 0.16 5 23.7 3.94 5 0 8.8 0.0 3.38 5 5 0.6 0.24 5 0.0 0.00 5 0.0 0.04 5 0.1 0.13 5 Page 142 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 hEAN SO N 13.4 17.3 0.32 2.04 5 5 0 8.3 1.2 0.0 0.0 0.0 0.0 4.16 0.34 0.00 0.04' 0.00 5 5 5 5 5 5 Page 143 S t u d y Report f o r Hematology INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL MEAN SD N 0.0 0.00 5 0.0 0.04 5 0.0 0.00 5 ................................................................... GROUP: 2-F 6402 0.0 0.0 0.0 6404 0.0 0.0 0.0 6406 0.0 0.0 0.0 6400 0.0 0.0 0.0 6410 0.0 0.0 0.0 MEAN SD N 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 MEAN SD N 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 Page 144 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 6 S t u d y Report f o r Hematology INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL MEAN SO N 0.0 0.00 5 0.0 0.00 5 0.0 0.00 5 Page 145 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6393 63% Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophils Basophi 1s Abnormal Lyllphocytes Other UEC Nucleated Red Cells Ly-cytes Segmented Neutrophi 1s Bands nonocytes Eosinophi 1s Basophi 1s A b n o r r a l Lymphocytes Other UBC 0 84 3.4 14 0.6 0 0.0 1 0.0 0 0.0 0 0.0 1 0.0 0 0.0 4.1 0 a9 12.5 11 1.6 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 14.1 Page 146 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6397 6399 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Bascphi 1s Abnormal Lymphocytes Other UBC Nucleated R e d Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophils 0asophi 1s Abnormal Lymphocytes Other WBC 0 a7 15.5 13 2.3 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 17.8 0 94 11.2 4 0.5 0 0.0 2 0.2 0 0.0 0 0.0 0 0.0 0 0.0 11.9 Page 147 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6403 6405 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands bonocytes Eosinophi 1s B a m i 1s Ahorma1 Lymphocytes Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Mcfmcytes Eosirophi1s Basophi 1s A b n o r u l Lymphocytes Other UBC 0 09 0.0 11 1.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 9.0 0 09 10 0 0 1 0, 0 0 15.3 1.7 0.0 0.0 0.2 0.0 0.0 0.0 17.2 Page 148 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6407 6409 Nucleated Red Cells Lymphocytes Segmented Neutrophi1s Bands Wonocytes Easinophi 1s Elasophi 1s Abnormal Lymphocytes Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophi1s Bands Monocytes Eosinophi 1s Basophi 1s Abnorual Lymphocytes Other UBC 0 a2 14.5 14 2.5 0 0.0 4 0.7 0 0.0 0 0.0 0 0.0 0 0.0 17.7 0 84 5.6 15 1 .o 0 0.0 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 6.7 Page 149 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6411 6413 6415 Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophils Easophi 1s Abnormal Lynphcytes Other UEC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophils Basophi 1s Abnormal Lymphocytes Other UEC Nucleated Red Cells Lymphocytes Segaented Neutrophi 1s Bands hocytes Eosinophi 1s Basophi 1s A b n o r m a l Lymphocytes Other UEC 0 94 8.8 6 0.6 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 9.4 0 90 7.7 10 0.9 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0, 8.5 0 80 4.2 l a 1.o 0 0.0 2 0.1 0 0.0 0 0.0 0 0.0 0 0.0 5.3 Page 150 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6417 641 9 -\ Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Uonocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other YBC Nucleated Red Cells Lylphocytes Segmented Neutrophi1s Bands Uonocytes Eosinophi 1s Barophi 1s Abnormal Lymphocytes Other YBC 0 91 11.2 9 1.1 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 12.3 0 93 6.2 6 0.4 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 0 0.0 6.7 Page 151 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6421 6423 6425 Nucleated R e d Cells Lylnphocytes Segmented Neutrophi1s Bands Mocytes Eosinophi 1s Basophi 1s Abnormal Lyaphocytes Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophi1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other UBC 0 90 8.0 9 0.8 0 0.0 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 8.9 0 94 13.8 6 0.9 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 14.7 1 90 10.7 10 1.2 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 11.9 Page 152 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6427 6429 Nucleated Red Celts Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other UBC Nucleated Red Celts Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s AbnOrML LynphoCytes Other UBC 1 94 9.9 6 0.6 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 10.5 0 92 14.5 a 1.3 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 15.8 Page 153 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6392 6394 63% Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Rands Monocytes Eosinophils Basophi 1s Abnormal Lyrphocytes Other UBC Nucleated Red Cells Lymphocytes SegHnted Neutrophi 1s Bands monocytes Eosinophi 1s Rasophi 1s A b n o m a 1 Lymphocytes Other UBC 0 9s 11.3 4 0.5 0 0.0 0 0.0 0 0.0 0 0.0 1 0.1 0 0.0 11.9 0 88 2.7 9 0.3 0 0.0 2 0.1 1 0.0 0 0.0 0 0.0 0 0.0 3.1 0 91 8.0 9 0.9 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 9.7 Page 154 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6398 6400 Nucleated R e d Cells Lymphocytes Segmented Neutrophils bnds Monocytes Eosinophils Basophi 1s Abnormal Lymphocytes Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other UBC 0 96 8.3 3 0.3 0 0.0 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 8.6 1 93 6.0 6 0.4 0 0.0 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 6.5 Page 155 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6402 644 6406 Nucleated Red Cells Lywhocytes Segmented Neutrophils Bands Monocytes Eosinophils Basophi Is Abnormal Lymphocytes Other UBC Nucleated Red Cells Lynphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basphi 1s Abnormal Lynphocytes Other UBC Nucleated R e d Cells Lynphocytes Segmented Neutrophils Bad, Monocytes Eosinophils B a s p h i 1s A b n o r m a l Lynphocytes Other UBC 0 a9 4.3 io 0.5 0 0.0 0 0.0 1 0.0 0 0.0 0 0.0 0 0.0 4.8 0 95 4.0 4 0.2 0 0.0 0 0.0 1 0.0 0 0.0 0 0.0 0 0.0 4.2 1 80 4.2 20 1.o 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 5.2 Page 156 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6410 Nucleated R e d Cells Lymphocytes Segmented Neutrophi 1s Bands Ronocyter Eosinophils Basophi 1s Abnormal Lymphocytes Other UBC 0 96 8.6 3 0.3 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 0 0.0 9.0 Page 157 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6412 6414 6416 Nucleated Red Cells Lyaphocytes Segmented Neutrophi1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands Monocytes Eorinophi 1s Basophi 1s Abnormal Lyrnphocyter Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophi 1s Bands nonc.cytts Eosinophi 1s B a r o p h i 1s Abnormal Lymphocytes Other UBC 0 90 5.3 9 0.5 0 0.0 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 5.9 0 91 7.9 9 0.8 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 E.? 0 93 11.9 7 0.9 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 12.8 Page 158 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6410 6420 Nucleated Red Cells Lymphocytes Segmented Neutrophi1s Bands Monocytes Eosinophils Basophi 1s Abnormal Lymphocytes Other uec Nucleated Red Cells Lymphocytes Segmented Neutrophi1s Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other uec 0 96 12.7 2 0.3 0 0.0 0 0.0 2 0.3 0 0.0 0 0.0 0 0.0 13.2 0 88 6.0 11 0.7 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 0 0.0 6.8 Page 159 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 6 Study Number: 132-006 6424 6426 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands Monocytes Eosinophi 1s Basophi 1s Abnormal Lymphocytes Other uec Nucleated Red Cells Lynphocytes Segmented Neutrophi 1s Bands Monocytes Eosinophi 1s Basophi 1s A b n o r m a l Lyaphocytcs Other UBC 0 66 2.0 33 1.0 0 0.0 0 0.0 1 0.0 0 0.0 0 0.0 0 0.0 3.0 0 a4 9.5 16 1.8 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 11.3 Page 160 Repeated Dose Range-Finding Toxictty Study in Sprague-Dawley Rats APPENDIX 6 Study Report or Hematology ~~ Study Number: 132-006 6428 6430 Nucleated Red Cells Lymphocytes Segmented Neutrophils Bands nonocytes Eosinophi 1s Barophi 1s Abnormal Lymphocytes Other UBC Nucleated Red Cells Lynphocytes Segmented Neutrophi 1s Bands nonocytes Eorinophi 1s Basophi Ls Abnormal Lyaphocytes Other UBC 0 91 12.3 8 1.1 0 0.0 1 0.1 0 0.0 0 0.0 0 0.0 0 0.0 13.5 0 92 11.3 a 1.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 12.3 Page 161 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 7 INDIVIDUAL CLINICAL CHEMISTRY DATA Page 162 Repeated Dose Range-Finding Toxictty Study in Sprague-Dawley Rats Study Number: 132-006 KEY TO CLINICAL CHEMISTRY TABLES AB BR EVlAT10 N TERM1NOLOGY ALP Alkaline Phosphatase AST Aspartate Aminotransferase ALT Alanine Aminotransferase CHOL Cholesterol GLU Glucose BUN Blood Urea Nitrogen C R EAT Creatinine TRIG Triglycerides PHOS Phosphorus (inorganic) TP Total Protein ALB Albumin GLOB Globulin NG Albumin/Globulin Ratio CA Calcium CL Chloride NA Sodium K Potassium GGT Gamma-glutamyltransferase T-BIL Total Bilirubin UNITS U/L U/L UIL mg/dL mg/dL mgldL mgldL mg/dL mgldL g/d L g/d L g/d L mg/dL mEq/L or mmol/L mEq/L or mmol/L mEq/L or mmol/L U/L mg/dL Page 163 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 7 Study Number: 132-006 STUDY I D : REPEATED DOSE/RAT STUDY NO: 132-006 SEX: BALE GROUP: I - R 6391 6393 6395 6397 6399 MEAN SD N 5.9 6.2 6.6 6.4 6.5 6.3 0.28 5 GROUP: 2-R 6401 6.4 6403 5.3 6405 6.1 6407 5.7 6409 5.8 BEAN SD N 5.9 0.42 5 ----- ---_------------ GROUP: 3-N 6411 6.0 6413 5.7 6415 6.3 6417 6.3 6419 6.3 MEAN SD N 6.1 0.27 5 4.3 4.3 4.5 4.4 4.5 4.4 0.10 5 4.2 3.7 4.2 3.3 4.1 3.9 0.39 5 4.2 4.1 4.1 4.3 4.2 4.2 0.08 5 124 45 0.1 14 0.2 45 147 52 0.1 16 0.3 36 im 39 0.2 15 0.3 50 199 50 0.1 16 0.3 49 21 2 54 0.2 17 0.2 58 174 48 0.1 16 0.3 48 37.3 6.0 0.05 1.1 0.05 8.0 5 5 5 5 5 5 184 39 135 42 119 50 154 44 169 5a 152 47 26.0 7.5 5 5 ------------ 181 53 182 31 241 49 192 60 1% 60 198 24.7 5 51 11.9 5 0.1 0.1 . 0.1 0.1 0.2 0.1 0.04 5 0.0 0.2 0.1 0.1 0.1 0.1 0.07 5 ia 0.2 34 13 0.2 39 16 0.3 43 17 0.2 52 16 0.2 43 16 0.2 42 1.9 0.04 6.6 5 5 5 ......................... 16 0.3 54 13 0.2 47 17 0.3 59 14 0.2 49 14 0.2 57 15 0.2 53 1.6 0.05 5.1 5 5 5 Page 164 Repeated Dose Range-Finding Toxicdy Study in Sprague-Dawley Rats APPENDIX 7 STUDY I D : REPEATED DOSE/RAT STUDY NO: 132-006 Study Number: 132-006 SEX: MALE MEAN SD N 6.2 4.5 208 41 0.1 17 0.3 57 0.23 0.19 55.1 10.9 0.04 4.4 0.07 7.2 5 5 5 5 5 5 5 5 Page 165 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 7 Study Number: 132-006 UEAN SO N 173 85.9 5 320 60.1 5 0 11.9 14.4 27 0.0 0.88 1.68 16.7 5 5 5 5 149 8.0 1.5 1.26 5 5 UEAN SD N 189 109.2 5 CROUP: 3-U 6411 6413 641 5 6417 6419 MEAN SD N 92 209 265 2'16 IS4 187 66.2 5 291 86.7 5 265 283 254 385 267 291 53.7 5 3 12.0 15.0 1.1 0.74 2.80 5 5 5 6 10.8 11.8 6 11.3 14.2 4 12.1 16.0 4 12.5 17.5 0 12.9 16.1 4 11.9 15.1 2.4 0.86 2.19 5 5 5 22 150 7.9 7.4 1.5 1.55 5 5 5 16 150 6.7 22 147 7.8 22 149 7.5 28 150 8.5 31 148 10.1 24 149 8.1 5.8 1.3 1.28 5 5 5 Page 166 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 7 Study Report o r Clinical Chemistry MEAN SD N 167 80.2 5 351 42.7 5 3 12.0 14.7 32 2.6 0.64 2.36 15.6 5 5 5 5 149 7.3 1.8 1.01 5 5 Page 167 Repealed Dose Range-FindingToxiclty Study in Sprague-Dawley Rats APPENDIX 7 Study Number: 132-006 MEAN 07 SD 1.1 N 5 GRWP: 2-M 6401 92 6403 09 6405 90 6407 w 6409 a6 MEAN 89 SO 2.2 N 5 1.9 0.19 5 2.2 1.6 1.9 2.4 1.7 2.0 0.34 5 2.3 0.23 5 1.9 2.3 2.2 1.4 2.4 2.0 0.40 5 MEAN SD N w 1.9 2.2 1.5 0.24 0.27 5 5 5 Page 168 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats APPENDIX 7 Study Number: 132-006 MEAN SD N a0 1.7 2.6 1.3 0.12 0.23 5 5 5 Page 169 Repeated Dose Range-FindingToxiclty Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 7 Study Report f o r Clinical Chemistry ............................................................................................................. INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL MEAN 6.5 4.6 107 56 0.2 16 0.3 48 SD 0.30 0.35 17.7 13.4 0.05 0.8 0.04 7.0 N 5 5 5 5 5 5 5 5 MEAN SD N 6.3 0.23 5 GROUP: 3-F 641 2 641 4 641 6 641 8 6420 MEAN SD N 6.1 6.5 5.5 6.5 6.5 6.2 0.44 5 4.5 0.26 5 4.3 4.6 3.8 4.5 4.5 4.3 0.32 5 112 17.1 5 106 147 109 123 1 03 118 18.1 5 64 0.2 6.1 0.04 5 5 71 0.2 63 0.1 53 0.1 58 0.2 51 0.1 59 0.1 8.1 0.05 5 5 16 0.3 49 1.6 0.05 13.0 5 5 5 19 0.3 41 16 0.3 44 18 0.3 46 13 0.3 63 15 0.2 49 16 0.3 49 2.4 0.04 8.6 5 5 5 Page 170 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 7 Study Number: 132-006 MEAN SD N 6.2 ' 0.17 5 4.5 0.17 5 163 23.5 5 66 15.9 5 0.1 0.05 5 16 0.2 48 1.7 0.0s 10.5 5 5 5 LABCAT CC4.41~ Page 171 20-JUL-2oOo Repeated Dose Range-Finding Toxicity Study in Sprague-Dawiey Rats APPENDIX 7 STUDY I D : REPEATED DOSEIRAT STUDY NO: 132-006 Study Number: 132-006 SEX: FEMALE MEAN SD N 212 101 .o 5 221 26.3 5 3 12.1 15.0 26 1.2 0.66 1.77 14.0 5 5 5 5 147 8.4 0.5 0.58 5 5 MEAN SO N 270 191 2 11.8 14.1 26 149 8.5 97.4 47.8 1.3 0.78 1.92 6.2 1.9 1.79 5 5 5 5 5 5 5 5 GROUP: 3-F 6412 252 164 6414 105 181 6416 185 149 641 8 41 2 74 6420 250 130 12.1 12.7 12.8 13.3 13.5 14.3 18.8 14.5 17.0 18.3 21 149 7.2 21 150 8.7 29 149 7.9 3s 147 9.3 22 145 11 .o MEAN 241 140 2 12.9 16.6 26 148 8.8 SD 113.0 41.2 1.6 0.55 2.10 6.2 2.0 1.45 N 5 5 5 5 5 5 5 5 Page 172 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 7 Study Number: 132-006 GROUP: 4-F 6422 6424 6426 6428 6430 MEAN SO N 146 298 a2 204 487 243 157.6 5 i9a 180 163 135 217 179 31.6 5 4 12.0 12.9 22 4 12.2 16.2 19 1 13.1 16.8 47 0 13.4 16.9 33 0 13.5 16.3 57 2 12.8 15.8 36 2.0 0.69 1.66 16.2 5 5 5 5 147 6.9 149 10.2 147 9.4 148 8.8 147 8.9 148 8.8 0.9 1.22 5 5 Page 173 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats APPENDIX 7 Study Number: 132-006 MEAN SD N 91 1.9 2.5 2.7 0.12 0.30 5 5 5 MEAN SD N 93 1.8 2.6 1.9 0.19 0.33 5 5 5 MEAN SD N 92 1.9 2.3 1.3 0.13 0.11 5 5 5 Page 174 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 7 Study Report or Clinical Chemistry GROUP: 4-F 6422 88 6424 92 6426 91 6428 91 6430 91 HEAN 91 SD 1.5 N 5 2.1 1.7 1.6 1.5 2.0 1.8 0.26 5 2.1 2.7 2.8 3.1 2.1 2.6 0.44 5 Page 175 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 8 INDIVIDUAL ORGAN WEIGHTS Page 176 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats APPENDIX 8 Study Number: 132-006 BWY W I G H T ( G ) ADRENAL GLANOS (C) % BCOY WEIGHT %BRAIN WIGHT BRAIN (GI X BCOY WEIGHT .HEART (G) % BODY UEIGHT % BRAIN WEIGHT KIDNEYS ( C ) % BODY WEIGHT X BRAIN WEIGHT LIVER (GI % BODY UEIGHT X BRAIN UEIGHT SPLEEN ( t i ) % BODY UEIGHT X BRAIN MIGHT TESTES (G) % BODY WEIGHT % BRAIN UEIGHT THYMJS ( G ) % BODY M I G H T % BRAIN UEIGHT 237.000 262.000 243.000 -245 000 263.000 0.066 0.028 3.393 0.075 0.029 3.874 0.074 0.030 4.011 0.070 0.029 3.564 0.066 0.025 3.115 1.945 0.821 1.936 0.739 1.845 0.159 1.964 0.802 2.119 0.806 1.079 0.455 55.476 1.188 0.453 61.364 1.159 0.477 62.818 1.185 0.484 60.336 1.180 0.449 55.687 2.445 1.032 125.707 2.835 1.082 146.436 2.448 1.007 132.683 2.4% 1.019 127.068 2.678 1.018 126.380 8.607 3.a2 442.519 9.m 3.716 502.893 8.870 3.650 480.759 9.041 3.690 460.336 10.4% 3.%9 492.591 0.550 0.232 28.278 0.708 0.270 36.570 0.480 0.198 26.016 0.590 0.241 30.041 0.659 0.251 31.100 2.672 1.127 137.378 2.563 0.978 132.386 3.088 1.271 167.371 2.824 1.153 143.788 2.941 1.118 138.792 0.458 0.193 23.548 0.781 0.298 40.341 0.643 0.265 34.851 0.826 0.337 42.057 0.627 0.236 29.589 Page 177 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats APPENDIX 8 Study Number: 132-006 B W Y WEIGHT (GI ADRENAL GLANOS (G) X B W Y UEIGHT X BRAIN UEIGHT BRAIN (G) X BOOY E I G H T .HEART ( G I X B W Y YEIGHT X BRAIN WIGHT KIDNEYS (GI X BODY UEIGHT X BRAIN UEIGHT LIVER (GI X BODY UEIGHT X BRAIN UEIGHT SPLEEN (G) X BODY UEIGHT X BRAIN UEIGHT TESTES (G) X BODY E I G H T X BRAIN UEIGHT THYWS (GI X BODY E I G H T X BRAIN WIGHT 231.000 269.000 274.000 211.000 246.000 0.075 0.032 3.904 0.077 0.029 4.096 0.109 0.040 5.758 0.091 0.043 4.461 0.069 0.028 3.275 1.921 0.832 1.880 0.699 1.893 0.691 2.040 0.967 2.107 0.857 0.981 0.425 51.067 1.258 0.468 66.915 1.288 0.470 68.040 1.135 0.538 55.637 1.116 0.454 52.966. 2.739 1.186 142.582 2.956 1.099 157.234 2.710 0.989 143.159 2.541 1.204 124.559 2.381 0.968 113.004 9.011 3.901 469.079 10.622 3.949 565.000 11.484 4.191 606.656 9.140 4.332 468.039 9.802 3.985 465.211 0.458 0.198 23.842 0.845 0.314 44.947 0.862 0.315 45.536 0.641 0.304 31.422 0.658 0.267 31.229 2.886 .1.249 150.234 2.816 1.047 149.787 2.887 1.054 152.509 2.950 1.398 144.608 2.652 1.078 125.866 0.566 0.236 28.423 0.493 0.18) 26.223 0.753 0.275 39.778 0.528 0.639 0.250 0.260 2 5 . a ~ ~ 30.327 Page 178 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 8 Study Number: 132-006 BWY EIGHT (GI 263.000 263.000 241 .OOO 259.000 248.000 ADRENAL GLANDS (G) X BODY WEIGHT X BRAIN E I G H T 0.091 0.035 4.367 0.096 0.037 4.580 0.074 0.031 3.842 0.064 0.025 3.182 0.058 0.023 3.100 BRAIN (G) X BODY E I G H T 2.084 0.792 2.096 0.797 1.926 0.799 2.011 0.776 1 A71 0.754 HEART (G) X B W Y WEIGHT X BRAIN M I G H T 1.012 0.385 48.560 1 .M 0.469 58.874 1.010 0.419 52.440 1.142 0.441 56.788 0.967 0.390 51.684 KIDNEYS (G) X BODY WEIGHT X BRAIN E I G H T 2.533 0.963 121.545 2.796 1.063 133.397 2.383 0.989 123.728 2.656 1.025 132.074 2.536 1.023 135.543 LIVER (G) X B W Y WEIGHT X BRAIN E I G H T 10.029 3.813 481 -238 9.944 3.781 474.428 10.308 4.277 535.203 11 A43 4.418 569.020 11.143 4.493 595.564 SPLEEN (G) 0.583 0.708 0.637 0.771 0.561 I. X BWY MIGHT 0.222 0.269 0.264 0.298 0.226 X BRAIN WEIGHT 27.975 3 3 . m 33.074 33.339 29.984 TESTES (G) X BODY M I G H T X BRAIN WEIGH1 2.663 1.013 127.783 2.675 1.017 127.624 2.619 1.087 135.981 2.967 1.146 147.539 2.430 0.980 129.871 T H W S (G) X BODY WEIGHT X BRAIN M I G H T 0.599 0.228 28.743 0.793 0.302 37.034 0.515 0.214 26.739 0.435 0.168 21.631 0.474 0.191 25.334 Page 179 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats APPENDIX 8 Study Number: 132-006 BWY E I G H T ( G I ADRENAL GLANDS (GI % B W Y UEIGHT % BRAIN WEIGHT BRAIN (GI % BODY E I G H T .HEART (G) % BWY MIGHT % BRAIN WEIGHT KIDNEYS (G) % BWY UEIGHT % BRAIN M I G H T LIVER (G) % BWY EIGHT % BRAIN WEIGHT SPLEEN ( G ) % B W Y WEIGHT % BRAIN E I G H T TESTES ( C ) % BODY S I G H T % BRAIN M I G H T THYWS (G) % BODY WEIGHT X BRAIN UEIGHT 242.000 260.000 251.000 253.000 265.000 0.084 0.035 4.428 0.072 0.028 3.863 0.070 0.028 3.714 0.073 0.029 3.702 0.080 0.030 4.296 1.897 0.784 1 .E64 0.717 1.885 0.751 1.972 0.779 1.862 0.703 1.112 0.460 58.619 1.148 0.442 61.588 1.017 0.405 53.952 1.1% 0.472 60.598 1.196 0.451 64.232 2.608 1 .on 137.480 2.545 0.979 136.534 2.350 0.936 124.660 2.641 1.044 133.925 2.639 0.996 141.729 10.846 4.482 571.745 11-523 4.432 618.187 13.150 5.239 697.613 15.106 5.971 766.024 13.000 4.906 698.174 0.6n 0.280 35.741 0.664 0.255 35 -622 0.548 0.218 29.OR 0.5% 0.236 30.223 0.764 0.288 41.031 2.767 1.143 145.862 3.021 1.162 162.071 2.782 1.108 147.586 3.083 1.219 156.339 2.931 1.106 157.411 0.573 0.237 30.206 0.550 0.212 29.506 0.660 0.263 35.013 0.665 0.263 33.722 0.652 0.246 35.016 Page 180 Repeated Dose Range-FindingToxiclty Study in Sprague-Dawley Rats APPENDIX 8 Study Number: 132-006 BOOY E I G H T ( G I ADRENAL GLANDS (G) X BCOY E I G H T X BRAIN MIGHT BRAIN (G) X 800'1 UEIGHT HEART (G) X BOOY M I G H T ' X BRAIN E I G H T KIDNEYS (G) X BOOY WEIGHT X BRAIN WIGHT LIVER (GI X BOOY M I G H T XBRAIN WIGHT OVARIES (G) X BOOY M I G H T X BRAIN WIGHT SPLEEN ( G I X BCOY WEIGHT X BRAIN E I G H T THYMJS (G) X BOOY M I G H T XBRAIN MIGHT 173.000 195.000 169.000 186.000 174.000 0.064 0.037 3.482 0.100 0.051 5.297 0.074 0.044 3.909 0.07a 0.042 4.673 0.066 0.038 3.524 1.838 1.062 1.888 0.968 1 .893 1.120 1.669 0.897 1 .E73 1.076 0.786 0.454 42.764 0.944 0.484 50.000 0.758 0.449 40.062 0.802 0.431 48.053 0.761 0.437 40.630 1.759 1.017 95.702 2.241 1.149 118.697 1.645 0.9TJ 86.899 1 .710 0.924 102.936 1.684 0.968 89.909 6.366 3.680 346.355 8.073 4.140 427.595 6.189 3.662 326.941 7.116 3.826 426.363 6.331 3.639 338.014 0.076 0.044 4.135 0.103 0.053 5.456 0.118 0.070 6.233 0.081 0.044 4.853 0.069 0.060 3.684 0.494 0.286 26. 8 T I 0.509 0.261 26.960 0.394 0.233 20.814 0.438 0.235 26.243 0.387 0.222 20.662 0.565 0.327 30.740 0.709 0.364 37.553 0.584 0.346 30.851 0.619 0.333 37.088 0.551 0.317 29.418 Page 181 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 8 Study Number: 132-006 BODY UEIGHT ( G ) ADRENAL GLANDS ( G ) X BCOY WEIGHT X BRAIN WEIGHT BRAIN (GI X BODY M I G H T HEART ( G ) X BCOY M I G H T ' X BRAIN WEIGHT KIDNEYS ( G ) X BODY H I G H 1 X BRAIN WEIGHT LIVER (G) X BODY WEIGHT X BRAIN M I G H T OVARIES (GI x Bmy UEIGHT X BRAIN UEIGHT SPLEEN ( G ) X BODY WEIGHT X BRAIN H I G H 1 THYWS (GI x Bmy MIGHT X BRAIN M I G H T 164.000 176.000 178.000 183.000 165.000 0.069 0.042 4.012 1 .no 1.049 0.060 0.034 3.343 1.795 1.020 0.064 0.036 3.393 1.886 1.060 0.098 0.054 5.379 1 .a22 0.996 0.064 0.039 3.569 1.793 1.087 0.826 0.504 48.023 0.739 0.420 41.170 0.849 0.477 45.016 0.908 0.496 49.835 0.729 0.442 40.658 1.648 1.005 95.814 1.567 0.890 87.290 1 .TI0 0.w4 93. M9 1.936 1 .OS8 106.257 1.803 1.093 100.558 6.377 3.888 370.756 6.389 3.630 355.933 6.869 3.859 M4.210 7.090 3.874 389.133 6.124 3.712 341.551 0.076 0.046 4.419 0.075 0.043 4.176 0.079 0.044 4.189 0.102 0.056 5.598 0.054 0.033 3.012 0.398 0.243 23.140 0.413 0.235 23.OoB 0.390 0.219 20.679 0.437 0.239 23.985 0.473 0.287 26.380 0.560 0.341 32.558 0.429 0.244 23.900 0.488 0.274 25.875 0.620 0.339 34.029 0.W 0.235 21.640 Page 182 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats APPENDIX 8 Study Number: 132-006 B W Y WEIGHT (G) ADRENAL GLANDS (G) % B W Y WEIGHT % BRAIN WEIGHT BRAIN (GI % BWY EIGHT HEART (G) X B W Y WEIGHT ' % BRAIN WEIGHT KIDNEYS (G) % B W Y WEIGHT % BRAIN E I G H T LIVER (G) % B W Y UEIGHT % BRAIN E I G H T OVARIES CG) % BWY EIGHT % BRAIN E I G H T SPLEEN ( G I % BWY EIGHT % B R A I N UEIGHT THYWS (G) % B W Y WEIGHT %BRAIN EIGHT 168.000 187.000 169.000 182.000 178.000 0.071 0.042 3.667 0.077 0.041 3.951 0.068 0.040 3.738 0.071 0.039 4.002 0.063 0.035 3.364 1.936 1.152 1.949 1.042 1 .819 1.076 1 .774 0.975 1 .B73 1 .OS2 0.700 0.417 36.157 0.837 0.448 42.945 0.709 0.420 38.9TI 0.950 0.522 53.551 0.848 0.476 45.275 1.723 1.026 88.998 1.803 0.W 92.509 1.790 1.059 98.406 1.914 1.052 107.892 1.753 0.985 93.593 6.362 3.787 328.616 7.367 3.940 377.989 5.870 3.473 322.705 8.102 4.452 456.706 8.105 4.553 632.728 0.087 0.052 4.494 0.102 0.055 5.233 0.081 0.048 4.453 0.064 0 035 3.606 0.101 0.057 5.392 0.440 0.262 22.727 0.479 0.256 24.577 0.391 0.231 21 A95 0.695 0.272 27.903 0.516 0.290 27.549 0.597 0.355 30.837 0.669 0.358 34.325 0.470 0.278 25.838 0.520 0.286 29.312 0.591 0.332 31.554 Page 183 Repeated Dose Range-Finding Toxicdy Study in Sprague-Dawley Rats APPENDIX 8 Study Number: 132-006 B W Y WEIGHT (GI ADRENAL GLANDS (GI X E W Y WEIGHT X BRAIN YEIGHT .HEART (GI X B W Y WEIGHT X ERAIN MIGHT KIDNEYS (GI X E W Y WEIGHT X BRAIN WEIGHT LIVER (G) X B W Y WEIGHT X BRAIN MIGHT OVARIES (G) X E W Y WEIGHT X BRAIN MIGHT SPLEEN (G) X B W Y MIGHT X BRAIN MIGHT T H Y W S (G) X E W Y WEIGHT X BRAIN WEIGHT 178.000 181 .ooo 191 .OOO 193.000 187.000 0.066 0.037 3.616 0.083 0.046 4.568 0.077 0.040 4.100 0.087 0.045 4.825 0.091 0.049 4.817 1A25 1.025 1.817 1.004 1 .878 0.983 1 .803 0.934 1 .a89 1.010 0.759 0.426 41.589 0.854 0.472 47.001 0.961 0.503 51.171 0.989 0.512 54.853 0.944 0.505 49.974 1.697 0.953 92.986 1 .SO5 0.997 99.340 2.204 1.154 117.359 2.270 1.176 125.901 1.936 1.035 102.488 7.776 4.369 426.082 7.265 4.014 399.835 8.820 4.618 469.649 9.532 4.939 528.674 8.402 4.493 444.786 0.099 0.056 5.425 0.085 0.047 4.6711 0.104 0.054 5.538 0.098 0.051 5.435 0.098 0.052 5.188 0.475 0.267 26.027 0.440 0.243 24.216 0.506 0.265 26.944 0.489 0.253 27.121 0.623 0.333 32.980 0.5% 0.335 32.65a 0.466 0.257 25.647 0.695 0.364 37.007 0.529 0.274 29.340 0.567 0.303 30.016 Page 184 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats Study Number: 132406 APPENDIX 9 FORMULATED TEST MATERIAL STABILITY DATA Page 185 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Southern Research Institute Project: A098.1 32 Day Stability Analysis Summary PerfluombutaneSulfonate (7% CMC) Study Number: 132-006 3.74 3.80 ttt 3.89 1 1.48 11.47 11.54 11.62 ND = not detected Rerullr rgrcscnl avetags of duplicatcr analysir +b One ol h e Jay 14 conbola (0 mghnL)was Result based on sinole analysis. Na sample raceived 40.38 38.86 39.63 39.31 wntaminated during preparetbn. Page 186 Repeated Dose Range-Finding ToxicQ Study in Sprague-Dawley Rats T". - wnicm 1.0 ... PnMld: 07/20/00 00:27 Results T.- julyZ000:drLx7/19 dow: p(br day 21 d m 07/20/00 W:W Study Number: 132-006 2.4( 2.oc 1.60 1.20 0.80 0.40 )O 2000.00 4000.00 8000.00 RBS yaF,ICI n-vc s al&atic Aru- -1+x y = 1.8300.6 ' x 0.999770 I +O.O0044 * x + 0.09467 0.00000 Concentrakm Ratlo 8000.00 10000.00 Page 187 Repeated Dose Range-Finding ToxicQ Study in Sprague-Dawley Rats TuhoQ~n". nniar 1.O Plin(.d: 07/21100 12:42 R m i U T-: ... MKinto.h:TuboQwn:T~Files: U r n 07L?lm 1241 1ed 2.4( 2.0( 1.6C 1.20 0.80 0.40 0.00 2000.00 4000.00 6000.00 Study Number: 132-006 Concentration Ratlo 8000.00 10000.00 Page 188 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Twboawn". vanion 1.0 ... Printrd: 07R11W 0 7 : s R e m T U : top ~Oi&C)Ay2GUO:da1~7L?O:7/20 dou 07/21/00 07:U 3.2 2.8' 2.4( 2.a 1.6C I .2a 0.80 0.40 DO 2000.00 4000.0 0 6000.00 ~ e 9f a h , h d near . 8 Qrudmtk - -Am. w l o 1+x y 1.!232S.d 0.999886 I x +o.o#y3 . X + 0.06783 0.00000 Study Number: 132-006 8000.00 10000.00~ Page 189 Repeated Dose Range-FindingToxictty Study in Sprague-Dawley Rats Study Number: 132-006 Pag. 1 of 1 w.o n 0 oormn I 3.50. 3.00- 2.50. 2.00. 1 .so- l .oo. 0.50 / 0.00 2000.00 4000.00 8000.00 Fit Qwdrcuk f l r PwMWmC W.igMiW - E C o m M o n b . m d . n t (r): Am8R.pls -1+x Y = 7..0220.-(1 0.999946 msQylr 0.00000 x - x +0.00(#4 . X + O.I3Lu)5 Concm ifion Ratio 8000.00 10000.00 Page 190 Repeated Dose Range-FindingToxiclty Study in Sprague-Dawley Rats Tu~tmCh~mva~n.ion 1.O Pnntrd 0 7 R W W 3 2 Redb TaObo: ...Fdd~julyzM)o:data:7/28dose: 7/27 dose 07/28/00 08:lS .e2 4.5c 1 4.00 $ 3.50 3.00 2.50 2.00. 1.so* 1 .oo- I 0.00 2000.00 4000.00 6000.00 Fit: Fit P-U. Weightin0: Eqwriar Coml.Uon caMd.nt (3: msqurr cwdmtk -AruR.lk 1+x y I 1.5830.6 0.999926 0.00000 * x * x +0.ooo(11~. + 0.oeOs Study Number: 132-006 Printed Pag. 1 o f 1 oomun Concentration Ratio 8000.00 1OOPO.00 Page 191 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats Study Number: 132-006 I 0.00 2000.00 4000.00 6000.00 8000.00 10000.00 Page 192 Repeated Dose Range-Finding Toxiclty Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 10 CERTIFICATE OF ANALYSIS Page 193 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Certificate Of Analysis C89SO3-K' (PFBS), Lot 2 March 10,2000 This sample was analyzed using ! and elemental analyses techniques. The results of thae tats show the sample to contain the following weight pcrccnt composition: .. C4F9SQK+ 0.13 % 99.82 % CF,(CFzh-SQ- 'K 97.86 Yo 1.96 % Page 194 Repeated Dose Ranqe-Findinq Toxicity Study in Spraque-Dawley Rats Study Number: 132-006 APPENDIX 11 PROTOCOL AND AMENDMENTS Page 195 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Primedica Redfield Protocol ProrocolNwnbrc 332- A REPEATED DOSE RANGEANDING TOXICITY STUDY OF T-7485 IN S P R A G U E - D A W RATS PROTOCOL NO.: 132-006 OBJECTIVE: To identify a maximum tolerateddose of T-7485 and identify dose levels for a 2 B d q repeated dose study. LOCATION OF STUDY AND CONDITIONS OF TESTING: It is the intention of 3M Pharmaceuticals,through the conduct of this study,to generate animal safety data that may be submittedto regulatory authorities. Primedica Redfield, 100 East Boone Street,Redfield, Arkansas, 72132, is accredited by AAALAC and licensed by the United States Department of Agricutture to conduct research in laboratory animals. All the conditionsof testing will conformto the A n i d Welfare Act (CFR 9) and its amendments. Primedica Redfieldwill follaw all requirements specified in this approved protocol as well as Primedica Redfield's Standard Operating Procedures. Changes in the protocol may be made by consultation with and approval from 3M Pharmaceuticals followed by mitten verification of the change. 3M Pharmaceuticals reserves the right to inspect facilities and procedures used for this study by means of announced or unannounced site visits. Primedica Redfield will nobfy 3M Pharmaceuticalspromptly by telephone prior to release of any data for review. SPONSOR: 3M Pharmaceuticals 3M Center, Building 220-2E-M P.O. Box 33220 St Paul, MN 55133-3320 SPONSOR'S REPRESENTATIVE: i TESTING FACILTTY: Primedica Redfield Mail: P.O. Box 308 Ddieries: 100 East BooneSt Redfield. AR 72132 Telephone: 501-397-2540 F M 501-397-2002 PERSONNEL: Study Director: John Seng, Ph.D. Principle Investigator: Karen Tranter. B.A, LATG Veterinarian: Allan Manus, D.V.M., M.Sc.,A C L W Clinical Pathology: Phyllis Powell, B.S.. M.T., ASCP Head Technician: Jennifer Dedman. B.S. PROPOSED STUDY DATES: Experimental Start Date: June 27,2000 Experimental Termination Date: July 7,2000 Draft Report Date: August 15,2000 P&8C 1 of 9 06/26/00 Page 196 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Primedica Redfield Protocol Protocol Number: 332- 1. REGUIA~ORCVOMWANCAEND QUALITAYSSURANCE This study will not be requiredto be conducted under GLP guidelines, however, the protocol, amendments, and the final report win be audited by the Quality Assurance Unit 2. lNSmuTlONAL ANIMALCARE AN0 USE COMMITTEE REWEW The protocolwill be reviewedand approved by PrimedicaRedfield's InstitutionalAnimal Care and Use Committee(IACUC)for compliance with regulations prior to study initiation. In the opinion of the Sponsor, indicatedby the signature on this protocol, the study does not unnecessarily duplicate any previouswork 3. MATawLs TESTMATERIAL T-7485 (Potassium perfluorobutane sulfonate, PFBS) lDENnFlCA 77ON The lot numberwill be listed in the raw data and final report PHYSICADLEsCRlpnoN white powder VEHICLE 1%carboxymethylcellulose(aqueous, mediumviscosityj bENnFlCATlON The lot number will be listed in the raw data and final report PHYSICADLESCRIPT~OwNhite powder STORAGE CorVofTloNS The bulk test material will be stored in its original container and will be stored at room temperature. The formulatedtest and controlmaterialwill be stored at 2" to 8OC. AUALMICALCHEMISTRY The Sponsor assumes responsibilii for characterization (identity, purity, and stability) ofthe bulk test material(includingunder test conditions). Informationon the compositionand methodof synthesis of the bulk test materialwill be held by the Sponsor. The doses, unless otherwisestated, will be calculated assumingthe test materialto be 100%pure. DOSE PREPARATIONAMD cOhK-snmu77ON Doses will be prepared by Primedica Redfield. The carboxymethylcellulosewill be pmpared at 1% in deionized water and may be prepared prior to Study Day 1. The test material container@)will be inventoriedwhen receivedat the testing laboratory, and a recordof all test materialuse will be maintained. The techniciandosing the animalswill: 1) Record the weight or volume of the test material container immediately before and after dosing; 2) Compute and record the amount used and compare it to the theoretical amount immediately after dosing; 3) Keepthe formulatedtest material use log in the animal study books at all times; 4) Assure the dose used is within f10% of theoretical. Page 2 of9 06/28/00 Page 197 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Primedica Redfield Protocol Protocol N u d e r : 1324x6 AlWLYTlCAL SAMPLES A sample of the formulated test material will be retained and frozen at approximately -20C for possibleanalysis. If no analysis is performed, the samplewill be discardeduponfinalization of the study report.or as authorized by the Sponsor. TESTMATERIALR~TENTION Unused test material may be returned to the Sponsor or designee at the termination of the study, or retained for use on future studies The Sponsorwill be notifiedin advanceof shipping, and a transmittalletterwill accompany the shipment. The material will be packed in a suitable container to maintain the conditions specified by the Sponsor during transit plus an adequatemargin of safety forany transit delays. SAFETYPRECAU~ONS General safety precautions as required by Primedica Redfield's policies and procedureswill be followed. The Sponsor's Representativewill be notifiedof any personnelexposures requiring a physician's examination or care. 4. ANIMALS Species: SbaidSource: Age at Initiation: Number and Sex: Identification: Rat CRCD*(SD) BR VAFWlus wt-bred albino rats, Charles River Laboratories, Inc. Six to eight weeks 20 male and 20 female (female nulliparous and non pregnant) Uniquely numberedear tag and cage card ANIMAL HUSBANDRY FEED WATER The animalswill be individually housed in stainless steel cages. The cages conform to standards set forth in the Guide for the Care and Use of LaboratowAnimals, NationalAcademy Press, Washington, D.C., 1996. Teklad Certified Rodent Diet #8728 will be provided adlibitum. This diet is routinely analyzed by the manufacturer for nutritionalcomponentsand environmentalcontaminants. Results of the manufacturer's analysesrn on file at Primedica Redfeld. Filtered tap water will be provided ad libihrm. Samplesof the water are analyzed for total dissolved solids, hardness, specified microbiological content, and for environmental contaminants. Results of these analyses are on file at Primedica Redfield. Page 198 Repeated Dose Range-Finding Toxicty Study in Sprague-Dawley Rats Study Number: 132-006 Primedica Redfield Protocol Protocol Nimber: 132- There are no known contaminants in the feed or water that would be expected to interfere with this study. ENVIRONMENT Environmentalcontrols are set to maintain a temperature of 18' to 26C (64' to 79OF) with a relative humidlty of 30% to 70%. These parameten are recorded at least once daily. A 1292 hour lightdark cyde is maintained. ACWMTION Animals will be acclimated for a minimum of seven days prior to the study start An adequate number of extra animals will be purchased so that no animal in obviously poor health is placed on test. At the request of the Study Director, excess animals may be used as replacementanimals. Animals will be examined by the Staff Veterinarian prior to release of that shipment of animals for use on the study. JUSTIFICATION FOR SPECIES SELECTIONAND NUMBEROFANIMLS Rats are an animal model for acute toxicity studies of this type. The number of animals assigned to this study represents the minimum required to meet the objective(s) of the study. 5. E ~ ~ ~ ? ~ M EMN~TOA DL O ~ O G Y STUDY DESIGN r I Group Group I Dosage I Dosage I Dosage I 1 Level Concentration Volu& , Number of Animals Number Designation (rng/kg) (WmL) (ml/kg) Males Females 1 Control 0 0 25 5 5 2 Mid-dOS8 10- 0_ 4 25 5 5 ~ 3 1 Midhighdose I 300 I 12 I 25 1 5 1 5 4 1 High-dose I 1000 I 40 25 1 5 1 5 ASSIGNMENT To GROUPS Animals will be randomly assignedto groups by a weightordered distributionsuch that individualbody weights will not exceed Y20% of meanweight for each sex FREQUENCYAND DURATION OF ADMINISTRATION Doses will be administered via oral gavage for ten consecutive days. The dose volume will be 25 rnVkg. The first day of dosing on the study will be Study Day 1. Individualdose volumes will be cakulated using the most recently collected body weights. JUSTIFlcATloNK I R ROUTE OF ADMINlSTRATlONAND DOSE LEVEL In the assessment and evaluation of the toxic characteristics of a substance, determination of acute oral toxicity is usually an initial step. Dose levels were chosen by the Sponsor. Page 4 af 9 06R6100 Page 199 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Primedica Redfield Protocol ~ ~~ ~ R O t o d Number: 132-006 ~~ -~ ANTEMORTEM OBSERVAllONS CLINICAL OSSERVATIONS Observations will be performed and recorded twice daily (a.m. and pm.) for moribundity and mortality. BODY WEIGHTS FEED CONSUMPllW Clinical observations will be performed, according to Primedica Redfield's SOPS, and recorded once daily and when a change is noted. Body weights will be recorded on Study Days 1,7, 10, and 11. Feed consumption will be measured on Study Days 1,7, and 10, and will be reported as an average daily consumption. CLINICAL cHEMlSTRY Blood samples will be obtained prior to necropsy while under anesthesia (CO,)via cardiac puncture. Feedwill be withheld overnight prior to collection. Blood samples collected (using EDTA for hematology and sodium citratefor coagulationindices) will be evaluatedfor the parameters specified below: Total and relativedifferentialleukocytecounts Erythrocyte counts Hemoglobin concentration 0 Hematocrit value Mean corpuscular volume Mean corpuscular hemoglobin Mean corpuscular hemoglobin concentration Platelet counts Prothrombin time 0 Activated partial thromboplastin time Blood samples will be obtained prior to neaopsy while under anesthesia (CO,) via cardiac puncture. Feed will be withheld overnight prior to collection. Blood samplescollectedfor clinicalchemistrywill be processed and evaluated for the parameters specified below: Total Protein Albumin Glut~~e Triglycerides UreaNitrogen Alanine Aminotransferase Alkaline Phosphatase Calaum Sodium Potassium Globulin AlbuminlGlobulin ratio Cholesterol Total Bilirubin Creatinine Aspartate Aminotransferase Gamma-glutamyltranskrase Phosphorus Chloride Page 5 of 9 06Rm Page 200 Repeated Dose Range-FindingToxicity Study in Sprague-DawleyRats Study Number: 132-006 Primedica Redfield Protocol RorocolNwnkc 1324% POSThU2REM OSSERVATlONS MORIBUNDA N I ~ L ~ALNDS ANIMLS FOUND DEAD Animals unlikely to survive until the next scheduled observationwill be weighed, euthanued, and necropsied. Animals found dead will be weighed and necropsied. In either case,bloodsamples will not be collected and a complete necropsywill be performedas detailed below. Euthanasiaof moribundanimals will be authorized by the Study Director or Staff Veterinarianwith the wncurrenm of the Study Director. EUTHANASIA All animalswill be humanely euthanized via anesthesiawith carbon dioxide to effect followed by exsanguination. Euthanasiawill be performedin accordancewith acceptedAmerican Veterinary Medical Association (AVMA) guidelines IJ. Amer. Vet. Med. Assoc., 202229-249, 19931. NECROPSY On Study Day 11, the animals will be sacrificed rotating sequentially through groups for each sex. All animals that de during the study,or are killed at study termination, will be subjectedto a necropsy examination. A complete necropsy is defined as examinationof the externalsurface of the body, all orifices, -and the cranial, thoracic, and abdominal cavities (and their contents). The following organs will be weighed prior to fixation: adrenal glands, brain, hearf kidneys. liver, ovaries, spleen, testes, and thymus. Paired organs will be weighed together. Organ weights will not be taken from animals that de or are euthanized prior to scheduled necropsy. Organs (or samples of organs) and tissues listed below will be examined in situ, dissected free, and fixed in 10% neutral buffered formalin or other suitable fixative for possible histopathologicalexamination. Observations notedat necropsywill be recordedon the IndividualAnimal Necropsy Record. .rn Adrenal glands Aorta rn Bone marrow sternum rn Brain brain stem cerebellum cerebrum Cervix Epididymides Esophagus Eyes with optic nerve rn Lymph nodes mandibular mesenteric Mammary gland Ovaries oviducts Pancreas pituitary gland Prostate gland Salivary gland mandibular rn Sciaticnave rn Seminal vesicles Page 6of9 06/26/00 Page 201 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 Primedica Redfield Protocol Proroc01 Number: 132- HISTOPATHOLOGY Femur with articular surface Gross lesions Harderian gland Heart Intestine, large cecum colon rectum Intestine, small duodenum ileum jejunum Kidneys Lacrimal gland LNW Lungs with bronchi Animal identificationwill be saved. Skeletal muscle Skin abdominal Spinal cord cervical lumbar thoracic Spleen stomach forestomach glandular Testes Thymus Thymidparathyroid Tongue Trachea Uterus Urinary bladder Zymbal's gland Vagina Histopathology will be performed on all tissues from the vehicle control and high dose group animals, all gross lesions. and all animals found dead or sacrificed inextremis. Additional histopathologywill be done at an additional cost to the Sponsor. Tissues will be trimmed, embedded, and sectioned. Slides will be stained with hematoxylin and eosin. Special stains may be used at the direction of the Study Pathologist, at the approval of the Sponsor, and at additional cost Slides will be read and interpreted by a qualified veterinary , pathologist Data will be summarized in tabular form and will be accompanied by a narrativeprepared by the Study Pathologist Means and standard deviations will be calculated for all quantitative data. Treated groups of the same sex will be compared to Group 1at common time points using an ANOVA with a post hoc Dunnett's t Test (Dunnett, C.W., J. Amer. Statis. Assoc., 50:1096-1121, 1955) oranother appropriatetest A two-sided a of 0.05/0.01 will be used to determine if statisticallysignificantdifferencesexist between the vehicle control and treated groups. If no significantdifferences exist at the 0.05 level, then no comparisonis necessary at the 0.01 level. Statisticalprogram names and compile dates are available in the Primedii Research Laboratories Computer Operations Department The SAS version is 6.12, from the SAS Institute, Inc., SAS Campus Drive, Cary, North Carolina, 27513. Page 7 of 9 06/26/00 Page 202 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats Study Number: 132-006 Primedica Redfield Protocol RO~ON&rC: O~132-006 7. REPORT A draft report will be sent to the Sponsor. Revisions to the initial draft report will be provided to the Sponsor by mail or fax transmission as printed copies of the corrected pages only. Additional revisionsor completecopies of the reviseddraft reports will be providedat additional expense to the Sponsor. Three copies of the approved final report, two bound and one unbound, will be submitted to the Sponsor approximately two weeks after approval of the draft report. The final reportwill include all elements requirWrecommendedby the regulations and guidelines. The report will include, but is not limited to, those items listed below: Descriptive text of the study objective Summary Test material identification (Certificate of Analysis, stabilily data, and analytical analysis if any are performed) Methods Results and conclusions 0 Body weights and body weight changes Feed consumption NecroW report Histopathology report Organ weights Serum chemistry and hematology 0 A copy of the protocol, all protocolamendments. and all protocol deviations that may affect the integrity of the study 8. MAINTENANCE OF R4w DATAAND RECORDS Original data or copies thereof, will be available at Primedica Redfield to facilitate auditing the study during its progressand beforeacceptance of the final report. When the final report is completed, all original paper data and the original final report will be retained in the archives of the laboratory for at least five years. Wet tissues, slides, and blocks (if generated) will be retained at Primedica Redfield for one year. After one year, the storage of the wet tissues, slides, and blocks will be negotiated with the Sponsor. After five years, the storage of the paper data will be negotiated with the Sponsor. The Sponsor will be notified prior to disposal of any original study data. 9. Q uASSU~RANCEREvrEw This is to certify that this protocol has been reviewed by Quality Assurance. P,m&vq& C Shevodya Noble1b.A Quality kurand;, Auditor Prirnedii Redfield I Dab Page 8 of9 06126/00 Page 203 Repeated Dose Range-Finding Toxicity Study in Sprague-Dawley Rats Primedica Redfield Protocol 10. APPROVALS Study Number: 132-006 Protocol Number: 1 3 2 a Senior Laboratory Manager 3M Corporate Toxicology Date John Sedg, Ph.D. Study Dhctor/Director of 'toxicology Primedica Redfield .. Date i Page 9 of9 Page 204 06R6100 Repeated Dose Range-FindingToxiclty Study in Sprague-Dawiey Rats Study Number: 132-006 Primedica Redfield Protocol Amendment Protocol Nwnbcr: 132co6 STUDY NUMBER: 132-006 AMENDMENT: 1 STUDY TITLE: A REPEATED DOSE RANGE-FINDINGTOXICIlY STUDY OF T-7485 IN SPRAGUE-DAWLEY RATS DATE ISSUED: July 17,2000 AMENDMENT # I ITEM I : Page 2, Section 1, Regulatory Compliance and Quality Assurance CHANGED FROM: This study will not be required to be conducted under GLP guidelines, however, the protocol, amendments, and the final report will be audited by the Quality Assurance Unit CHANGED TO: This study will not be required to be conducted under GLP guidelines, however, .. the protocol, amendments, and the final report will be reviewed by the Quality Assurance Unit REASON: Typographicalerror. non-GLP studies are not audited but are reviewed. ITEM 2: CHANGED FROM: CHANGED TO: REASON: Page 9, Approvals: John Seng. Ph.D. Date Study DirectorlDirectorof Toxicology Primedica Redfield \ John Seng, Ph.D. Study Director Prirnedica Redfield Date Typographicale m APPROVALS: Senior Laboratory Manager 3M Corporate Toxicology I ' %ate Study D i r k o r Prirnedica Redfield Page 205 Repeated Dose Range-Finding Toxicrty Study in Sprague-Dawley Rats Study Number: 132-006 Prirnedica Redfield I-Idoc01 Amendment Prolocoi M r : I32-c# STUDY NUMBER: 132-006 AMENDMENT: STUDY TITLE: 2 A REPEATED DOSE RANGE-FINDINGTOXICITY STUDY OF T-7485 IN SPRAGUE-DAWLEY RATS DATE ISSUED: September 19,2000 AMENDMENT # 2 ITEM 1: Page 2, Section 3, Materials ADD: STABILITY Samples will be taken from a single formulation of each concentration immediately after preparabonand 7,14,21,28. and 32 days after preparation. . . These samples will be stored at -20C until shipped to the following laboratory: Greg Gorman, Ph.D. Research Chemist 111 Southern Research Institution 2000 Ninth Avenue South Birmingham, AL 35205 Telephone: (205) 581-2725 FAX (205) 581-2726 The samples will be labeledas standard protocoland will include the appropriate theoretical concentrations of T-7485, the day post-preparation, and the date of sampling. A sample of 1% CMC will also be shipped as a control. REASON: Sponsor request for stability study prior to upcoming 26-day study. APPROVALS: Senior Laboratory Manager 3M Corporate Toxicology Date John deng, Ph.0 Study Director Primedica Redfield I Date 1 Page 206 Repeated Dose Range-Finding Toxicty Study in Sprague-Dawley Rats Study Number: 132-006 APPENDIX 12 PROTOCOL DEVIATIONS Page 207 Repeated Dose Range-Finding Toxtclty Study in Sprague-Dawley Rats Study Number: 132-006 PROTOCOL DEVIATIONS There were no protocol deviations that adversely affected the integrity of the study. Page 208 Repeated Dose Range-FindingToxicity Study in Sprague-Dawley Rats ~~~ ~~ Study Number: 132-006 APPENDIX 13 KEY PERSONNEL Page 209 Repeated Dose Ranae-Findinq Toxicity Study in Sprague-Dawley Rats Study Number: 132-006 KEY PERSONNEL Study Number: 132-006 Study Director................................................ Principle Investigator ..................................... Head Technician............................................ Veterinarian ................................................... Clinical Pathology.......................................... Quality Assurance.......................................... Report Coordination ...................................... Report Preparation ........................................ John Seng, Ph.D. Karen Tranter, B.A., LATG Jennifer Dedrnan, B.S. Allan Manus, D.V.M., M.Sc., ACLAM Phyllis Powell, B.S., M.T., ASCP Val Gartner, B.A. Amy Babb, B.S. Tammy Peckharn, Ad. Asst. Dana Rose, Ad. Asst. Page 210