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/ | R ja & o 5 3 2 . M il, Il III H I h lW lf c O T lM W if f iiW M g a a iM a a M l FinalaWniiMBI' j Mj Biological Phase Report Metabolism of N-Ethyl Perfluorooctane Sulfonemido Ethanol (Net-Fose; T-6316) in Cvnomolgus Monkeys Following Administration of a Single Capsule Dose PREPARE' !') I OR: 3M Chcnii-.Lil.s CO VAN CE S ! Cl >V NUMBER: 6329-226 C O V A itfC ^ THE DEVELOPMENT SERVICES COMPANY 04143 Sponsor: 3M Chemicals St. Paul, Minnesota Study Title: Metabolism ofN-Ethyl Perfluorooctane Sulfonamido Ethanol (Net-Fose; T-6316) in Cynomolgus Monkeys Following Administration of a Single Capsule Dose Data Requirement: None Authors: Frederic W. Thalacker, PhD Deborah K. Lee Study Completed on: June 2, 1999 Performing Laboratory: Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Project Identification: Covance 6329-226 Page 1 of 77 004144 2 Covance 6329-226 STATEMENT OF NO DATA CONFIDENTIALITY CLAIM Metabolism ofN-Ethyl Perfluorooctane Sulfonamido Ethanol (Net-Fose; T-6316) in Cynomolgus Monkeys Following Administration of a Single Capsule Dose No claim of confidentiality is made for any information contained in this biological phase on the basis of its falling within the scope of FIFRA 10(d)(A), (B), or (C). These data are the property of 3M Chemicals and as such are confidential for all purposes other than compliance with FIFRA Section 10. Submission of these data in compliance with FIFRA does not constitute a waiver of any right to confidentiality that may exist under any other statute or in any other country. Company: 3M Chemicals Company Representative: Andrew Seacat, PhD Title: Study Director Department: Signature Date 004145 3 Covance 6329-226 COMPLIANCE STATEMENT Metabolism ofN-Ethyl Perfluorooctane Sulfonamido Ethanol (Net-Fose; T-6316) in Cynomolgus Monkeys Following Administration of a Single Capsule Dose All aspects of this biological phase were conducted in accordance with the Environmental Protection Agency Pesticide Programs Good Laboratory Practice Standards (40 CFR 160). Andrew Seacat, PhD Study Director 3M Chemicals Study Submitted by Sponsor Date Date Date 004146 4 Covance 6329-226 QUALITY ASSURANCE STATEMENT This report has been reviewed by the Quality Assurance Unit of Covance Laboratories Inc., in accordance with the Environmental Protection Agency (EPA) Good Laboratory Practice Standards, 40 CFR 160. The following inspections were conducted and findings reported to the study director and study director management. Written status reports of inspections and findings are issued to Covance management according to standard operating procedures. Inspection Dates From To Phase Date Reported to Study Director and Study Director Management 04/22/98 05/13/98 06/02/98 10/09/98 11/06/98 11/06/98 11/06/98 05/19/99 06/01/99 04/22/98 05/13/98 06/02/98 10/09/98 11/16/98 11/16/98 11/16/98 05/19/99 06/01/99 Protocol Review Test Article Administration Protocol Amendment Review Protocol Amendment Review Data Review Report Review Data Review Report Review Report Review 05/04/98 05/13/98 06/03/98 10/09/98 04/06/99 04/06/99 04/06/99 06/02/99 06/02/99 Representative, .ali: Assurance Unit 004147 Metabolie Chemistry Frederic W. Thalacker, PhD Biological Phase Investigator Deborah K. Lee Study Coordinator Philip J. Teitelbaum, PhD Associate Director LeAnn Eastman Supervisor Radiochemical Analysis Kuntal Sinha Study Chemist 5 Covance 6329-226 KEY PERSONNEL Toxicology Operations Peter Kong Manager Cara Himrich Supervisor Metabolism In-Life Quality Assurance Unit Nancy M. Centanni Manager Laboratory Animal Medicine Donna J. Clemons, DVM, MS Diplomate, ACLAM Supervisor 004148 6 SIGNATURES Covance 6329-226 Frederic W. Thalacker, PhD Biological Phase Investigator Metabolic Chemistry Covance Laboratories Inc. Philip eitelbaum, PhD Associate Director Metabolic Chemistry Covance Laboratories Inc. Andrew Seacat, PhD Study Director 3M Chemicals 004149 7 Covance 6329-226 TABLE OF CONTENTS STATEMENT OF NO DATA CONFIDENTIALITY CLAIM COMPLIANCE STATEMENT QUALITY ASSURANCE STATEMENT KEY PERSONNEL SIGNATURES ABSTRACT TITLE OBJECTIVE REGULATORY COMPLIANCE QUALITY ASSURANCE EXPERIMENTAL DESIGN AND PROCEDURES Study Design Justification for Dosing Route and Dose level Test Substance Test Animals and Housing Animal Selection Justification Surgical Procedure Bile Replacement Dose Preparation and Analysis Dosing Procedure Antemortem Observations Physical Examinations Body Weights Clinical Pathology Clinical Pathology Tests Antemortem Sample Collection Termination Sample Identification, Retention, and Disposition Blood, Plasma, Serum, and Cellular Fraction Preparation and Storage Sample Transfer Disposition of Raw Data, Records, Samples, and the Final Report RESULTS AND DISCUSSION Quarantine and Acclimation Page 2 3 4 5 6 9 10 10 10 10 10 10 11 11 12 12 12 12 13 13 13 13 14 14 14 14 15 16 17 17 17 18 18 18 004150 8 Covance 6329-226 Clinical Pathology Body Weights and Doses Administered Antemortem Observations CONCLUSIONS TABLES Table 1. Individual Body Weights of Cynomolgus Monkeys Reported in Kilograms Table 2. Individual Body Weights of and Dose Weights Administered in One Capsule to Cynomolgus Monkeys (50 mg/kg) Table 3.1% Dextrose in Lactated Ringers Solution and Bile Salt Replacement Solution Infusion Times APPENDIX A Protocol CMS 22672A1, Protocol Amendment No. 1, and Protocol Deviations APPENDIX B Summary of Antemortem Observations, Clinical Pathology Codes and Abbreviations, and Clinical Pathology Data Tables 19 19 19 20 21 21 22 23 24 24 42 42 004151 9 Covance 6329-226 ABSTRACT Tissue, plasma, serum, bile, feces, and urine specimens were provided to obtain information on the metabolism of N-ethyl perfluorooctane sulfonamido ethanol (Net-Fose; T-6316) in cynomolgus monkeys following administration of a single oral dose. Sixteen cynomolgus male and female monkeys from Covance Research Products were dosed according to the following study design: Group Number and Sex of Animals 1 6 Male 6 Female Status Intact 2 2 Male Bile Duct 2 Female Cannulated Dose 50 mg/kg 50 mg/kg Collections Postdose Tissues from 2/sex on Days 2, 28, and 90. Urine and feces from 0-12 and 12-24 hours, then daily through 28 days, and weekly through 90 days. Blood predose, and at 1, 2, 4, 6, 12, 24, 36, and 48 hours, 7 days, and weekly through 90 days. Tissues at 28 days. Bile from 0-12 and 12-24 hours, and daily through 28 days. Blood predose, and at 1, 2, 4, 6, 12, 24, 36 and 48 hours,7 days, and weekly through 28 days. Separate blood samples were collected for preparation of plasma and serum. Plasma was prepared by centrifugation of the blood. Serum was prepared by allowing the blood to clot and centrifuging it to separate the cellular debris. All specimens were shipped to the Sponsor for analysis and the results of these analyses will be reported separately by the Sponsor. 004152 10 Covance 6329-226 TITLE Metabolism ofN-Ethyl Perfluorooctane Sulfonamido Ethanol (Net-Fose; T-6316) in Cynomolgus Monkeys Following Administration of a Single Capsule Dose OBJECTIVE The purpose of this study was to administer a perfluorooctane test substance to male and female cynomolgus monkeys and collect samples for determination of absorption, distribution, metabolism, and excretion. REGULATORY COMPLIANCE The biological phase of this study was conducted in accordance with the Environmental Protection Agency (EPA), Good Laboratory Practice Standards, 40 CFR 160 and by Covance Laboratories Inc., 3301 Kinsman Boulevard, Madison, Wisconsin, in accordance with Covance Protocol CMS 22672A1 dated April 21, 1998, and Covance Standard Operating Procedures (SOPs). The protocol, protocol amendment, and protocol deviations are in Appendix A. All procedures in this study are in compliance with the Animal Welfare Act Regulations, 9 CFR 3, dated October 30, 1989, and as modified on March 18, 1991. In the opinion of the Sponsor and the study director, the study did not unnecessarily duplicate any previous work. QUALITY ASSURANCE The protocol, study conduct, data, and biological phase final report were audited by the Covance Quality Assurance Unit (QAU) in accordance with Covance SOPs. All study activities performed at 3M will be the responsibility of the 3M Quality Assurance Department. EXPERIMENTAL DESIGN AND PROCEDURES Study Design Animals were assigned to two groups, one was the bile duct cannulated group and the other was intact. The group designation, number of animals, specimen collection, and target dose level were as follows: 004153 11 Covance 6329-226 Number and Sex Group of Animals Status Dose Frequency, Route, and Level Collections Postdose 6 Male 6 Female Intact One Capsule Oral 50 mg/kg Tissues from 2/sex on Days 2, 28, and 90. Urine and feces from 0-12 and 12-24 hours, then daily through 28 days, and weekly through 90 days. Blood predose, and at 1,2, 4, 6, 12, 24, 36, and 48 hours, 7 days, and weekly through 90 days. 2 Male Bile Duct 2 Female Cannulated One Capsule Oral 50 mg/kg Tissues at 28 days. Bile from 0-12 and 12-24 hours, and daily through 28 days. Blood predose, and at 1, 2, 4, 6, 12,24,36 and 48 hours,7 days, and weekly through 28 days. Justification for Dosing Route and Dose level The oral route has been used for evaluating systemic toxicity of the compound in cynomolgus monkeys. The 50 mg/kg dose was chosen because it provides the maximum non-toxic single dose amount necessary to recover measurable concentrations of the test substance in minor tissues. Test Substance The test substance, T-6316, was received in the Department of Metabolic Chemistry on May 8,1998 from the Covance Department of Toxicology. Stability and retention of a sample as specified in 40 CFR 160.105 of the test substance is the responsibility of the Sponsor. Information on synthesis methods, composition, or other characteristics that define the test substance are on file with the Sponsor. The following information was obtained from the label on the test substance container: Substance No.: Container No.: Test Substance: Lot Nos.: Date Combined: Purity/Concentration: Expiration Date: Storage Requirements: Physical State: 1084A3 65 T-6316 (Narrow Range N-Ethyl Perfluorooctanesulfonamido Ethanol, NEtFOSE) 30035, 30037, and 30039 07-31-1997 98.1% On file with the Sponsor Room temperature Amber waxy solid, slight odor Test Animals and Housing 12 Covance 6329-226 The in-life portion of the study was conducted May 13, 1998, through August 11, 1998. Eleven male and eleven female cynomolgus monkeys (nonhuman primates) from Covance Research Products were received on March 24, 1998. The animals were young adult to adult and weighed approximately 3 to 4 kg at arrival. Upon arrival, each animal was assigned a temporary identification number. Before being placed on test, each animal was randomly assigned a permanent identification number and identified with a neck tag attached to the collar. All animals were housed in individual, suspended, stainless steel, wire-mesh cages during acclimation and housed in individual metabolism cages designed for the separation and collection of bile, urine, and feces while on test. The primates were fed Certified Primate Diet #8726C (Harlan Teklad) ad libitum, except when fasted for surgery and prior to test substance administration through approximately 4 hours postdose. Diets were supplemented with appropriate fruits and cereals. Water was provided fresh daily and ad libitum. Environmental controls for the animal room were set to maintain 18 to 29C, 50% 20% relative humidity, and a 12-hour light/12-hour dark cycle. The 12-hour dark cycle was interrupted, as necessary, to accommodate study procedures. Animal Selection Clinical chemistry and hematology test results and physical examination results were used to select test animals. In addition, animals were selected for the surgical procedure based on the best apparent adjustment to the jacket and tether system. Before dose administration, a laboratory animal veterinarian evaluated the health of the animals and approved their use on the study. Justification Studies using live animals are essential for characterizing the disposition of chemicals in animals. No acceptable non-live animal models were available. The cynomolgus monkey has been used as the non-rodent species to evaluate toxicity. This study will aid in the evaluation of test results obtained in toxicology studies conducted in monkeys and will also provide a basis for the extrapolation of safety data from animals to humans. The number of animals was the minimum number necessary to provide scientifically valid results. Surgical Procedure The surgical procedure was conducted in accordance with Covance Standard Operating Procedures. The animals were fasted overnight before surgery and sedated with 004155 13 Covance 6329-226 ketamine. An appropriate antibiotic was administered. Surgical anesthesia was maintained using oxygen and isoflurane. Lactated Ringer's solution was administered through a saphenous catheter throughout the surgical procedure or as determined by a staff veterinarian. A sterile surgical scrub was performed. The common bile duct and duodenum was cannulated and the gallbladder was removed and discarded. Analgesics were administered as recommended by the staff veterinarian. Additional antibiotics and/or fluids were administered as recommended by the staff veterinarian. The animals had at least 10 days to recover from the surgical procedure prior to dose administration. Bile Replacement Beginning the day after surgery through 3 days post surgery, a solution of 1% dextrose in Lactated Ringer's solution was administered via the duodenal cannula for approximately 8 hours per day at approximately 20 mL/hour to prevent dehydration. Beginning on the fourth post surgical day until sacrifice, a bile salts solution was infused via the duodenal cannula for approximately 8 hours per day at approximately 23 mL/kg/day. The bile salts solution was prepared by adding 18.0 g of cholic acid to 1 L of 0.9% sodium chloride solution. Sodium bicarbonate (1.3 g) was added to each 1 L of solution. The solution was mixed and the pH adjusted to 7.4 to 7.8 with 0.1N HC1 or sodium hydroxide, as appropriate. Infusion intervals and volumes administered were recorded (See Table 3). Dose Preparation and Analysis The capsules (gelatin, No. 2, 0.37 mL) for the dose preparation were received from Torpac, Inc. on April 9,1998. The test compound was homogenized. A No. 2 capsule was tared on an analytical balance and an appropriate amount of test substance, based on animal weight and dose level, was added using a spatula. The capsules were sealed and placed in separate 20-mL scintillation vials labeled with the animal number. The capsules were stored under ambient conditions. Dosing Procedure The animals were fasted overnight prior to dose administration through approximately 4 hours postdose. The capsule was given to the animals by a device specific for capsule administration. Antemortem Observations Mortality and moribundity checks were done twice daily (a.m. and p.m.). Cageside observation for general health and appearance were done once daily. Signs of poor health or abnormal behavior were recorded as they were observed. C04156 Physical Examinations 14 Covance 6329-226 Physical examination were performed twice during acclimation, once pre-surgery, and once post surgery before the initiation of treatment. Animals were anesthetized with ketamine hydrochloride according to Covance Standard Operating Procedures for the examination. Jackets were checked according to Covance Standard Operating Procedures. The animals were anesthetized as above for the examination; however, a lower dose of ketamine was administered for the limited examination. Body Weights Body weights were taken weekly during acclimation, within 5 days of dosing (to be used for dosing calculations), weekly after dosing, and on the day of sacrifice. Clinical Pathology The animals were not fasted. Blood was collected via the femoral vein before dose administration as follows: approximately 1 week before surgery from all animals and approximately 4 and 8 days post surgery from cannulated animals. When health problems developed, blood was collected for clinical pathology tests to assess health status as deemed necessary by the staff veterinarian. Clinical Pathology Tests The following are the tests performed by the Clinical Pathology Department: Hematology Red blood cell count Hemoglobin Hematocrit Mean corpuscular volume Mean corpuscular hemoglobin Mean corpuscular hemoglobin concentration Platelet count White blood cell count Differential blood cell count Blood cell morphology Prothrombin time Activated partial thromboplastin time 004157 15 Clinical Chemistry Glucose Urea nitrogen Creatinine Total protein Albumin Globulin Total bilirubin Serum bile acids Cholesterol Aspartate aminotransferase Alanine aminotransferase Alkaline phosphatase Creatine kinase Gamma glutamyltransferase Calcium Inorganic phosphorous Sodium Potassium Chloride Covance 6329-226 Antemortem Sample Collection No Teflon was used for this study. Blood (Groups 1 and 2). Approximately 2 mL of blood was collected from Group 1 animals predose and approximately 2 mL was collected at 1, 2,4, 6,12, 24, 36, and 48 hours, 7 days, and weekly through 90 days postdose. Approximately 2 mL of blood was collected from Group 2 animals predose and approximately 2 mL was collected at 1, 2, 4, 6, 12, 24, 36, and 48 hours, and on 7, 14, 21, and 28 days postdose. The blood was collected via a femoral vein into heparinized tubes and placed immediately on wet ice. Effective June 3,1998 (Study Day 21) and at the remaining time points, an additional 2-mL blood sample was collected into a glass tube without heparin and set aside at room temperature to clot. Bile (Group 2). Bile was collected from the Group 2 animals via the implanted cannula into plastic containers surrounded by dry ice predose (for at least 12 hours), and at 0-12, 12-24, and continuing at 24-hour intervals through 28 days postdose. Urine (Group 1). Urine was collected in plastic containers surrounded by dry ice at 0-12, 12-24, and continued at 24-hour intervals through 28 days, then weekly until 90 days postdose. 16 Covance 6329-226 Feces (Group 1). Feces were collected at 0-12, 12-24, and continued for 24-hour intervals through 28 days, then weekly until 90 days postdose. Specimens were transferred into plastic containers at the time of collection. Termination Scheduled Sacrifice. At the time of sacrifice, animals were anesthetized with sodium pentobarbital. Cerebrospinal fluid (CSF) was collected once the animal was fully anesthetized. CSF was placed on wet ice immediately after collection and collection times were recorded. Blood was collected via cardiac puncture into glass tubes with heparin (at least 20 mL) and into glass tubes without heparin (at least 20 mL), except Animal No. 10541 that had a short sample obtained via vena cava. Following blood collection, the animals were sacrificed via exsanguination under sodium pentobarbital. Residual urine from the bladder was added to the last urine sample. Stomach, large and small intestinal contents were collected from the two per sex Group 1 animals that were sacrificed at Day 2 only. All tissues collected were excised, rinsed with water, blotted dry, weighed, and placed in a plastic container surrounded by wet ice. After the tissues were removed, the residual carcasses were discarded. The following tissues were collected from all animals at sacrifice, except the gallbladder that was collected from Group 1 only: Adrenal glands Aorta Bone (femur) Bone marrow (from femur) Brain Diaphragm Epididymis Esophagus Eyes Fat Gallbladder Heart Kidneys Large intestine Liver Lungs Lymph nodes (cervical) Lymph nodes (mesenteric) Muscle (thigh) Ovaries Pancreas Prostate Salivary glands Seminal vesicles Skin (dorsal, shaved) Small intestine Spinal cord Spleen Stomach Testes Thymus Thyroid/Parathyroid Tongue Trachea Urinary bladder Uterus 004159 17 Sample Identification, Retention, and Disposition Covance 6329-226 Specimens were identified with the study number, sex, animal number, group, matrix, specimen number, and collection interval. Blood, Plasma, Serum, and Cellular Fraction Preparation and Storage Blood in glass tubes with heparin was stored at approximately 5C prior to analysis. At the individual time points, the blood was gently inverted several times to homogenize, then approximately 700 pL was taken and placed into a labeled vial for each individual time point. At sacrifice, approximately half of the blood was homogenized and placed into a labeled vial. The subsamples were placed in a freezer at a temperature of approximately -20C. The remaining blood was centrifuged at 2,400 rpm (1,300 x g) for approximately 10 minutes at approximately 5C. The resulting plasma and cellular fraction was transferred to separately labeled vials and placed in freezer at a temperature of approximately -20C until they were shipped. The additional 2-mL blood collection in glass tubes without heparin for the collection of serum was allowed to clot at room temperature for approximately 30 to 60 minutes. The sample was then centrifuged at 2,400 rpm (1,300 x g) for approximately 10 minutes at approximately 5C. The serum was removed and placed into a labeled vial. The cellular debris was discarded and the serum was stored in a freezer at a temperature of approximately -20C to await shipment. Sample Transfer All specimens were shipped on dry ice to the Sponsor on June 12,1998, and August 18, 1998: Dr. Kris Hansen Environmental Technology and Safety Services 935 Bush Avenue Building 2-E3-09 St Paul, Minnesota 55133-3331 Phone: (612)778-6018 Fax: (612)788-6176 The Sponsor will be responsible for further analysis, reporting, and archiving upon completion of the study. 004160 18 Disposition of Raw Data, Records, Samples, and the Final Report Covance 6329-226 The following data records to be maintained and transferred to the archives of Covance will include, but will not be limited to: protocol and amendments; study correspondence; test material receipt; final report. The following supporting records to be retained at Covance but not archived with the study data will include, but will not be limited to: feed analysis records; water analysis records; animal room temperature and humidity records; refrigerator/freezer temperature records; instrument calibration and maintenance records; United States Department of Agriculture animal records; stock records. The following data records to be transferred to the archives of 3M upon completion of the report will include, but will not be limited to: In-life records: Animal receipt Acclimation Animal room maintenance Antemortem observations Body weights Physical examination records Clinical pathology Clinical pathology slides Dose administration Sample collection Bile salts replacement solution infusion times and amounts All correspondence, documentation, records, protocol, and final report generated as a result of this study will be archived in the storage facilities of Covance for a period of one year following the signing of the final report. One year after signing the final report, all of the aforementioned materials will be sent to the Sponsor and a return fee will be charged. The Sponsor may elect to have the materials retained in the Covance Archives for an additional period of time and Covance will charge a storage fee. If the Sponsor chooses to have Covance dispose of the materials, a disposal fee will be charged. RESULTS AND DISCUSSION Quarantine and Acclimation All animals under quarantine and acclimation beginning March 24,1998, except Animal 105428, appeared clinically healthy and were released on April 24, 1998. Animal 105428 was observed to have diarrhea and was monitored until normal. At least one physical examination, one fecal examination, and three tuberculin tests were performed before 004161 19 Covance 6329-226 release from quarantine. Weekly body weights were recorded and animals were acclimated to the jacket and tether system required for bile duct cannulation. Clinical Pathology Results of presurgical clinical pathology tests indicated no obvious group or individual health abnormalities. Although several males in Group 1 had notably high leukocyte counts, these were attributed to excitement associated with blood collection procedures. A few females in Group 1 had mildly high values for alanine aminotransferase activity consistent with Hepatitis A infection, a subclinical condition commonly observed in cynomolgus monkeys that does not adversely effect liver function or animal health. Results of the clinical pathology tests following surgery demonstrated no critical complications secondary to bile duct cannulation. Four days post surgery, increased activities for aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma glutamyltransferase, and creatine kinase in Group 2 animals were consistent with the surgical procedure. The total bilirubin was notably increased for only one of the males (Animal No. 105436); and one of the females (Animal No. 105441) exhibited evidence of blood loss from the surgical procedure. Eight days post surgery, the high enzyme activities generally exhibited significant reductions indicative of continued recovery from the surgical procedure, and total bilirubin was no longer elevated for Animal No. 105436. Persistent, mildly to moderately elevated activities for alkaline phosphatase (males and females) and gamma glutamyltransferase (males only) at 8 days post surgery were characteristic of mildly increased biliary pressure secondary to bile duct cannulation, but normal values for total bilirubin and serum bile acids indicated hepatic function was likely not significantly compromised. Body Weights and Doses Administered Individual body weights and doses administered to Groups 1 and 2 are presented in Tables 1 and 2. Actual doses administered ranged from 100.2 % to 104.4% of the nominal dose of 50 mg/kg. Antemortem Observations The animals appeared normal during dose administration and appeared healthy and exhibited no overt signs of toxicity throughout the study. The noted observations are listed in Appendix B. None of these observations appeared to be test substance related. On May 22,1998, no bile flow was noted for Animals 105437 and 105441. The swivel was flushed for Animal 105437 and the bile started to flow again. The technician found that the swivel for Animal 105441was hooked up incorrectly on the previous day, so it was connected correctly and the pump restarted. Approximately 2 hours later, there was no bile flow, so the system was rechecked and bile began to flow. On May 26,1998 and 004162 20 Covance 6329-226 May 29,1998, blood was drawn from Animal 105441 for clinical chemistry analysis to monitor her health status. The results on May 29,1998, were indicative of cholestasis, but the results were improved from the results on May 26,1998. The Clinical Pathologist also indicated that the hepatic dysfunction appeared minimal and concluded very little hepatocellular degeneration or necrosis occurred. On June 4,1998, Animal 105449 did not have a screen above the pan to separate the urine from the feces. The feces was scraped from the pan and collected; however, some cross-contamination may have occurred. Bile flow slowed and was noted on Study Day 17,23, and 24 then stopped on Day 25 from Animal 105446. However, the animal exhibited no signs of compromised health. Upon sacrifice it was noted that a portion of the bile duct had re-grown, evidently providing an alternate path of bile flow. CONCLUSIONS Samples were provided to obtain information on the pharmacokinetics, excretion and tissue distribution of N-ethyl perfluorooctane sulfonamido ethanol (Net-Fose; T-6316) after a single casule dose in cynomolgus monkeys. Bile-duct cannulated as well as intact animals were used in the study. All samples were shipped to the Sponsor for analysis and results of these analyses will be reported separately by the Sponsor. No test substance effects were noted on the monkeys. 004163 Table 1. 21 Covance 6329-226 TABLES Individual Body Weights of Cynomolgus Monkeys Reported in Kilograms Animal Number 1 Day 3 8 9 16 22 23 31 35 GrouD 1 Male 105428 4.4 4.2 105430 3.9 3.9 105431 3.8 3.8 3.8 3.9 3.9 105432 4.2 4.2 4.2 4.2 4.2 105433 4.1 4.0 4.0 4.4 4.3 105435 4.0 3.9 4.1 4.3 4.3 4.4 4.4 4.3 4.3 Group 1 Female 105439 3.1 3.1 105440 3.1 3.1 105442 3.0 2.9 2.8 3.0 2.9 105444 3.0 2.9 2.9 2.9 2.8 105447 3.2 3.1 3.2 3.3 3.3 105449 3.3 3.3 3.4 3.3 3.2 3.5 3.6 3.3 3.3 105436 3.7 3.5 106437 3.7 3.7 Group 2 Male 3.5 3.6 3.6 3.8 3.8 3.6 105441 3.0 3.0 105446 4.2 4.1 Grout) 2 Female 2.8 3.2 3.8 4.2 3.2 4.1 Day 42 49 56 63 70 77 84 91 Grout) 1 Male 105433 4.5 4.6 4.6 4.7 4.8 4.8 4.9 4.9 105435 4.1 4.2 4.2 4.2 4.2 4.2 4.3 4.3 Group 1 Female 105447 3.5 3.5 3.5 3.5 3.6 3.6 3.7 3.6 105449 3.4 3.4 3.4 3.4 3.5 3.5 3.7 3.7 004164 Table 2. 22 Covance 6329-226 Individual Body Weights of and Dose Weights Administered in One Capsule to Cynomolgus Monkeys (50 mg/kg) Animal Number 105428 105430 105431 105432 105433 105435 105436 105437 105439 105440 105441 105442 105444 105446 105447 105449 Group 1 1 1 1 1 1 2 2 1 1 2 1 1 2 1 1 Animal Weight (kg) 4.4 3.9 3.8 4.2 4.1 4.0 3.7 3.7 3.1 3.1 3.0 3.0 3.0 4.2 3.2 3.3 Test Substance Administered (g) (mg/kg) % of Nominal 0.2207 50.2 100.3 0.1978 50.7 101.4 0.1930 50.8 101.6 0.2138 50.9 101.8 0.2082 50.8 101.6 0.2033 50.8 101.7 0.1884 50.9 101.8 0.1897 51.3 102.5 0.1573 50.7 101.5 0.1556 50.2 100.4 0.1522 50.7 101.5 0.1508 50.3 100.5 0.1566 52.2 104.4 0.2108 50.2 100.4 0.1616 50.5 101.0 0.1653 50.1 100.2 004165 23 Covance 6329-226 Table 3. 1% Dextrose in Lactated Ringers Solution and Bile Salt Replacement Solution Infusion Times Day3 Post Surgery 105436 AM Start PM Finish Animal Identification 105437 105441 Time AM PM AM PM Start Finish Start Finish 105446 AM Start PM Finish 1 7:07 14:55 5:59 13:58 6:00 14:01 6:26 15:03 2 5:58 13:59 6:26 14:32 6:26 14:32 6:15 14:15 3 6:26 14:31 6:15 14:00 6:15 14:17 6:22 14:01 4 6:15 14:18 6:22 14:01 6:22 14:01 7:35 15:42 5 6:22 14:01 7:35 15:42 7:35 15:42 6:38 14:36 6 7:35 15:42 6:38 14:36 6:38 14:36 6:37 14:40 7 6:38 14:40 6:37 14:40 6:37 14:40 6:23 14:24 8 6:37 14:40 6:23 14:24 6:23 14:24 7:00 14:57 9 6:23 14:28 7:00 14:57 7:00 14:57 6:24 14:12 10 7:00 15:03 6:24 14:12 6:24 14:12 6:23 14:15 11 6:24 14:12 6:23 14:15 6:23 14:15 6:42 14:44 12 6:23 14:15 6:42 14:47 6:42 14:49 6:55 15:38 13 6:42 14:44 6:55 15:38 6:55 15:38 5:21 13:23 14 6:55 15:38 5:21 13:21 5:21 13:23 6:51 14:58 15 5:21 13:22 6:51 14:58 6:51 14:58 6:16 14:25 16 6:51 14:58 6:16 14:24 6:16 14:25 5:46 13:56 17 6:16 14:24 5:46 13:56 5:46 13:56 6:48 14:51 18 5:46 13:56 6:48 14:51 6:48 14:51 6:39 14:41 19 6:48 14:51 6:39 14:41 6:39 14:40 6:38 14:46 20 6:39 14:41 6:38 14:46 6:38 14:46 6:36 14:36 21 6:38 14:46 6:36 14:38 6:36 14:37 6:53 15:01 22 6:36 14:36 6:53 15:01 6:53 15:01 6:17 14:14 23 6:53 15:01 6:17 14:18 6:17 14:20 5:50 13:47 24 6:17 14:18 5:50 13:47 5:50 13:47 5:43 13:48 25 5:50 13:47 5:43 13:48 5:43 13:48 5:46 13:44 26 5:43 13:48 5:46 13:47 5:46 13:47 6:49 14:53 27 5:46 13:48 6:49 14:53 6:49 14:53 6:39 14:41 28 6:49 14:53 6:39 14:41 6:39 14:41 6:55 14:57 29 6:39 14:41 6:58 14:57 6:55 14:57 6:36 14:42 30 6:55 14:57 6:36 14:35 6:36 14:35 6:39 14:31 31 6:36 14:35 6:39 14:31 6:39 14:31 7:02 14:59 32 6:39 14:31 7:02 15:00 7:02 14:59 6:45 14:46 33 7:02 15:00 6:45 14:46 6:45 14:46 6:41 14:39 34 6:45 14:46 6:41 14:40 6:41 14:39 6:41 14:44 35 6:41 14:40 6:41 14:44 6:41 14:44 6:45 14:05 36 6:41 14:44 6:45 14:05 6:45 14:05 7:01 14:58 37 6:45 14:05 7:01 14:58 7:01 14:58 NAb NA 38 7:01 14:58 5:41 13:40 5:41 13:40 NA NA 39 5:41 13:40 6:36 14:47 6:36 14:47 NA NA 40 6:36 14:47 6:38 14:48 6:38 14:45 NA NA 41 6:38 14:45 6:51 14:35 6:51 14:35 NA NA 42 6:51 14:35 a On Days 1 through 3, the animals were infused with 1% dextrose in Lactated Ringer's solution at approximately 20 mL/kg/hour for approximately 8 hours, and on the remaining days the animals were infused with bile salts at approximately 23 mL/kg/hour for approximately 8 hours. b Not applicable. Since no bile was recovered, no bile salts were infused. 24 Covance 6329-226 APPENDIX A Protocol CMS 22672A1, Protocol Amendment No. 1, and Protocol Deviations 004167 25 Covance 6329-226 PROTOCOL Sponsor 3M Chemicals St. Paul, Minnesota Protocol CMS 22672A1 Study Title Metabolism ofN-Ethyl Perfluorooctane Sulfonamido Ethanol (Net-Fose; T-6316) in Cynomolgus Monkeys Following Administration of a Single Capsule Dose Testing Guideline Environmental Protection Agency, Good Laboratory Practice Standards, 40 CFR 160 Date April 21, 1998 Performing Laboratory Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Project Identification Covance 6329-226 Page 1 of 13 004168 26 Covance 6329-226 STUDY IDENTIFICATION Metabolism of N-Ethyl Perfluorooctane Sulfonamido Ethanol (Net-Fose; T-6316) in Cynomolgus Monkeys Following Administration of a Single Capsule Dose Test Substance N-Ethyl Perfluorooctane Sulfonamido Ethanol (Net-Fose; T-6316) Sponsor 3M Chemicals 3M Center Building 220-2E-02 St. Paul, MN 55144-1000 Study Director Andrew Seacat, PhD 3M Chemicals 3M Center Building 220-2E-02 St. Paul, MN 55144-1000 Phone: (612) 575-3161 Fax: (612)733-1773 Principal Analytical Investigator Kris Hansen Environmental Technology and Safety Services 935 Bush Avenue Building 2-E3-09 St. Paul, Minnesota 55133-3331 Phone: (612) 778-6018 Fax: (612)788-6176 Biological Phase Investigator Frederic W. Thalacker, PhD Covance Laboratories Inc. P.O. Box 7545 Madison, Wisconsin 53707 Phone: (608) 242-2712 ext. 7370 Fax: (608)241-7412 Biological Phase Location Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Proposed Study Timetable In-Life Experimental Start Date In-Life End Date May 13,1998 August 11,1998 004169 27 Covance 6329-226 OBJECTIVE The purpose of this study is administer a Perfluorooctane test substance to male and female cynomolgus monkeys and collect samples for determination of absorption, distribution, metabolism, and excretion. REGULATORY COMPLIANCE Environmental Protection Agency, Good Laboratory Practice Standards, 40 CFR 160 All procedures in this protocol are in compliance with the Animal Welfare Act Regulations, 9 CFR 3, dated October 30, 1989, and as modified on March 18, 1991. In the opinion of the Sponsor and the study director, the study does not unnecessarily duplicate any previous work. The protocol, study conduct, data, and biological phase final report will be audited by the Covance Laboratories Inc. Quality Assurance Unit in accordance with Covance Standard Operating Procedures. All study activities performed at 3M will be the responsibility of the 3M Quality Assurance Department. TEST SUBSTANCE Source The test substance will be provided by the Sponsor. The Sponsor will be responsible for analysis of the test substance. Storage Conditions The test substance will be stored at approximately -20C until dosing. Any test substance remaining will be stored at approximately -20C. Disposition Unused test substance will be returned to the Sponsor or a recipient designated by the Sponsor, or discarded at the Sponsor's request. The Sponsor will assume responsibility for retention of a sample of the test material as specified in 40 CFR 160.195 if the study is longer than 4 weeks in duration. C04170 28 Covance 6329-226 Safety Precautions Safety precautions will be taken as required by Covance Policies and Procedures, in consideration of the Sponsor's Material Safety Data Sheet, or other relevant safety information provided by the Sponsor. EXPERIMENTAL DESIGN Animals Species/Strain. Nonhuman primate/cynomolgus Source. Covance Research Products (Alice, Texas, or Denver, Pennsylvania). The source of the animals will be documented in the raw data. Weight at Arrival. 3 kg or greater Age at Arrival. Young adult/adult Number and Sex. Eight males and eight females on test, six male and six female intact and two male and two female bile duct cannulated. One male and one female extra intact and two male and two female extra cannulated will be available as possible replacement animals. Identification. Individually numbered collar tags Housing For the acclimation period, the animals will be in individual, stainless steel cages. For the test period, the animals will be in individual metabolism cages designed for the separation and collection of bile, urine, and feces. Food Certified Primate Diet #8726C (Harlan Teklad) ad libitum, except when fasted for surgery. Food will also be withdrawn overnight prior to, and returned approximately 4 hours after dose administration. Diets will be supplemented with appropriate fruits and cereals. Water Ad libitum, provided fresh daily 004171 Contaminants 29 Covance 6329-226 There are no known contaminants in the food or water that would interfere with this study. Environment Environmental controls for the animal room will be set to maintain 18 to 29C, 50% 20% relative humidity, and a 12-hour light/12-hour dark cycle. The 12-hour dark cycle may be interrupted to accommodate study procedures. Quarantine and Acclimation Animals will be under quarantine for at least 30 days. At least one quality control physical, one fecal examination, and three tuberculin tests will be done before release from quarantine. Weekly body weights will be taken. Animals will be acclimated to the jacket/tether system required for bile duct cannulation. Animal Selection Clinical chemistry and hematology test results and physical examination results will be used to select test animals. Animals will be selected for the surgical procedure based on the best apparent adjustment to the jacket and tether system. In the case that all or most animals adjust well, animals will be selected in numerical order based on the individual collar tag numbers previously assigned. Before dose administration, a laboratory animal veterinarian will evaluate the health of the animals and approve their use on the study. Justification Studies using live animals are essential for characterizing the disposition of chemicals in animals. No acceptable non-live animal models are available. The cynomolgus monkey has been used as the non-rodent species to evaluate toxicity. This study will aid in the evaluation of test results obtained in toxicology studies conducted in monkeys and will also provide a basis for the extrapolation of safety data from animals to humans. The number of animals is the minimum number necessary to provide scientifically valid results. Surgical Procedure The surgical procedure will be conducted in accordance with Covance Standard Operating Procedures. The animals will be fasted overnight before surgery and sedated with ketamine. An appropriate antibiotic will be administered. Surgical anesthesia will be maintained using oxygen and isoflurane. Lactated Ringer's solution will be administered through a saphenous catheter throughout the surgical procedure or as 004172 30 Covance 6329-226 determined by a staff veterinarian. A sterile surgical scrub will be performed. The common bile duct and duodenum will be cannulated and the gallbladder removed and discarded. Analgesics will be administered as recommended by the staff veterinarian. Additional antibiotics and/or fluids may be administered as recommended by the staff veterinarian. The animals will have at least 10 days to recover from the surgical procedure prior to dose administration. Bile Replacement Beginning the day after surgery through 3 days post surgery, a solution of 1% dextrose in Lactated Ringer's solution will be administered via the duodenal cannula for approximately 8 hours per day (approximately 20 mL/hour) to prevent dehydration. Beginning on the fourth post surgical day until sacrifice, an appropriate amount of mixed bile salts solution will be infused via the duodenal cannula for approximately 8 hours per day. The volume of bile salts solution to be administered will be approximately 23 mL/kg/day. Bile salts solution will be prepared by adding 18.0 g of cholic acid to 1 L of 0.9% sodium chloride solution. Sodium bicarbonate (1.3 g) will be added to each 1 L of solution. The solution will be mixed and the pH adjusted to 7.4 to 7.8 with 0. IN HC1 or sodium hydroxide, as appropriate. Infusion intervals and volumes administered will be recorded. Group Designations and Dose Level Number Group and Sex Status Dose Frequency, Route, and Level Collections Postdose 1 6 Male 6 Female Intact One Capsule 50 mg/kg Tissues from 2/sex on Days 2, 28, and 90. Urine and feces from 0-12 and 12-24 hours, then daily through 28 days, and weekly through 90 days. Blood predose, and at 1 , 2 , 4 , 6 , 12, 24, 36, and 48 hours, 7 days, and weekly through 90 days. 2 2 Male Bile Duct One Capsule Tissues at 28 days. 2 Female Cannulated 50 mg/kg Bile from 0-12 and 12-24 hours, and daily through 28 days. Blood predose, and at 1, 2, 4, 6, 12, 24, 36 and 48 hours,7 days, and weekly through 28 days. Justification for Dosing Route/Dose Level. The oral route has been used for evaluating systemic toxicity of the compound in cynomolgus monkeys. The 50 mg/kg dose was chosen because it provides the maximum non-toxic single dose amount necessary to recover measurable concentrations in minor tissues. 004173 31 Covance 6329-226 Dosing Procedure Predose and Postdose Feeding. Animals will be fasted overnight prior to dose administration through approximately 4 hours postdose. Dose Administration. The capsule will be administered by a device specific for capsule administration. Observation of Animals Antemortem Observations. Mortality and moribundity checks will be done twice daily (a.m. and p.m.). Cageside observation for general health and appearance will be done once daily. Signs of poor health or abnormal behavior will be recorded as they are observed. Physical Examinations. Twice during acclimation, once pre-surgery, and once post surgery before the initiation of treatment. Animals will be anesthetized with ketamine hydrochloride according to Covance Standard Operating Procedures for the examination. Jackets will be checked according to Covance Standard Operating Procedures and will be replaced as necessary. The animals will be anesthetized as above for the examination; however, a lower dose of ketamine may be administered for the limited examination. Body Weights. Weekly during acclimation, on the day of dosing, weekly thereafter, and on the day of sacrifice. Additional body weights may be taken at the discretion of the Biological Phase Investigator or a staff veterinarian. Clinical Pathology. The animals will not be fasted. Blood will be collected via the femoral vein before dose administration as follows: approximately 1 week before surgery from all animals and approximately 4 and 8 days post surgery from cannulated animals. If health problems develop, blood will be collected for clinical pathology tests to assess health status as deemed necessary by the staff veterinarian. Any blood samples taken after modification of the ports for collection of bile will be collected before the administration of the bile salts replacement solution for the day. 004174 32 Covance 6329-226 Tests Clinical Chemistry Glucose Urea nitrogen Creatinine Total protein Albumin Globulin Total bilirubin Serum bile acids Cholesterol Aspartate aminotransferase Alanine aminotransferase Alkaline phosphatase Creatine kinase Gamma glutamyltransferase Calcium Inorganic phosphorous Sodium Potassium Chloride Hematolosv Red blood cell count Hemoglobin Hematocrit Mean corpuscular volume Mean corpuscular hemoglobin Mean corpuscular hemoglobin concentration Platelet count White blood cell count Differential blood cell count Blood cell morphology Prothrombin time Activated partial thromboplastin time Antemortem Sample Collection NO TEFLON IS TO BE USED FOR THIS STUDY. Blood (Groups 1 and 2). Approximately 2 mL of blood will be collected from Group 1 animals predose and approximately 2 mL will be collected at 1, 2,4, 6,12, 24, 36, and 48 hours, 7 days, and weekly through 90 days postdose. Approximately 2 mL of blood will be collected from Group 2 animals predose and approximately 2 mL will be collected at 1, 2, 4, 6,12, 24, 36, and 48 hours, and on 7, 14,21, and 28 days postdose. The blood will be collected via a femoral vein into heparinized tubes and placed immediately on wet ice. The animals may be re-hydrated with lactated Ringer's solution given subcutaneously or via the duodenal cannula at the discretion of the Biological Phase Investigator or Laboratory Animal Veterinarian following blood collections, if the total blood volumes collected exceed the recommendations in Covance Standard Operating Procedures. Bile (Group 2). Bile will be collected from the Group 2 animals via the implanted cannula into plastic containers surrounded by dry ice predose (for at least 12 hours), and at 0-12, 12-24, and continuing at 24-hour intervals through 28 days postdose. 004175 33 Covance 6329-226 Urine (Group 1). Urine will be collected in plastic containers surrounded by dry ice at 0-12, 12-24, and continuing at 24-hour intervals through 28 days, then weekly until 90 days postdose. Feces (Group 1). Feces will be collected at 0-12,12-24, and continuing at 24 hour intervals through 28 days, then weekly until 90 days postdose. Samples will be transferred into plastic containers at the time of collection. Termination Unscheduled Sacrifices and Deaths. The carcasses will be retained and may undergo postmortem examination for any gross changes. Further examination may be performed upon Sponsor's request. Scheduled Sacrifice. At the time of sacrifice, animals will be anesthetized with sodium pentobarbital. Cerebrospinal fluid (CSF) will be collected once the animal is fully anesthetized. CSF will be placed on wet ice immediately after collection and collection times will be recorded. Blood (at least 20 mL) will be collected via cardiac puncture into heparinized tubes. Following blood collection, the animals will be sacrificed via exsanguination under sodium pentobarbital. Residual urine from the bladder will be added to the last urine sample. Stomach, large and small intestinal contents will be collected from the two per sex Group 1 animals that are sacrificed at Day 2 only. All tissues collected will be excised, rinsed with water, blotted dry, weighed, and placed in a plastic container surrounded by ice. The following tissues will be collected from all animals at sacrifice, except the gallbladder will be collected from Group 1 only: 004176 Adrenal glands (~0.5 g) Aorta (~0.5 g) Bone (femur) Bone Marrow (from femur) Brain Diaphragm Epididymis Esophagus Eyes Fat Gallbladder (~0.5 g) Heart Kidneys Large Intestine Liver Lungs Lymph nodes (cervical) Lymph nodes (mesenteric) 34 Covance 6329-226 Muscle (thigh) Ovaries (-0.5 g) Pancreas Prostate Salivary Glands Seminal vesicles Skin (dorsal, shaved) Small Intestine Spinal Cord Spleen Stomach Testes Thymus Thyroid/Parathyroid (-0.75 g) Tongue Trachea Urinary Bladder Uterus Sample Identification, Retention and Disposition Specimens will be identified with the study number, sex, animal number, group, matrix, specimen number, and collection interval. Sample Preparation and Storage All specimens will be stored at approximately -20C until shipment. Blood samples will be stored at approximately 5C until centrifugation. A subsample, approximately 1 mL at the individual time points and approximately half of the sacrifice blood, will be removed and placed in a separate labeled vial. The remaining blood will be centrifuged to separate plasma. The blood, cellular fraction of the blood (RBC), and plasma will be frozen immediately and stored at approximately -20C. 004177 Sample Transfer 35 Covance 6329-226 All specimens will be shipped on dry ice to the Sponsor. The specimens will be shipped to the following person and address: Dr. Kris Hansen Environmental Technology and Safety Services 935 Bush Avenue Building 2-E3-09 St Paul, Minnesota 55133-3331 Phone: (612)778-6018 Fax: (612)788-6176 The Sponsor will be responsible for further analysis, reporting, and archiving upon completion of the study. REPORT A biological phase draft report including, but not limited to, those items listed below will be submitted to the Sponsor for review and comment. Then, a biological phase final report will be provided. Experimental Design and Methods As defined by the protocol, protocol amendments, and any protocol deviations. Data and Results Animal receipt and acclimation Antemortem observations Body weights Physical examinations Clinical pathology Dose administration Sample collection Bile salts replacement solution infusion times and amounts 004178 36 Covance 6329-226 Maintenance of Raw Data, Records, and Specimens The following data records to be maintained and transferred to the archives of Covance will include, but will not be limited to: Protocol and amendments Study correspondence Test material receipt Final report The following supporting records to be retained at Covance but not archived with the study data will include, but will not be limited to: Feed analysis records Water analysis records Animal room temperature and humidity records Refrigerator/freezer temperature records Instrument calibration and maintenance records United States Department of Agriculture animal records Stock records The following data records to be transferred to the archives of 3M will include, but will not be limited to: In-life records: Animal receipt Acclimation Animal room maintenance Antemortem observations Body weights Physical examination records Clinical pathology Clinical pathology slides Dose administration Sample collection Bile salts replacement solution infusion times and amounts All correspondence, documentation, records, protocol, and final report generated as a result of this study will be archived in the storage facilities of Covance for a period of one year following the signing of the final report. One year after signing the final report, all of the aforementioned materials will be sent to the Sponsor and a return fee will be charged. The Sponsor may elect to have the materials retained in the Covance Archives for an additional period of time and Covance will charge a storage fee. If the Sponsor chooses to have Covance dispose of the materials, a disposal fee will be charged. 0043.79 April 21,1998 37 PROTOCOL APPROVAL Covance 6329-226 Covance 6329-226 Page 13 o f 13 d u .' fit- Andrew Seacat, PhD Study Director 3M Chemicals Frederic W. Thalacker, PhD Biological Phase Investigator Metabolic Chemistry Covance Laboratories Inc. Associate Director Metabolic Chemistry Covance Laboratories Inc. //->/n Date Date Apri &!, IfK Date(J C04150 38 Covance 6329-226 PROTOCOL AMENDMENT NO. 1 Covance 6329-226 Metabolism ofN-Ethyl Perfluorooctane Sulfonamido Ethanol (Net-Fose; T-6316) in Cynomolgus Monkeys Following Administration of a Single Capsule Dose Sponsor 3M Chemicals 3M Center Building 220-2E-02 St. Paul, MN 55144-1000 Study Director Andrew Seacat, PhD 3M Center Building 220-2E-02 St. Paul, MN 55144-1000 Contractor Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Biological Phase Investigator Fred Thalacker, PhD Covance Laboratories Inc. P.O. Box 7545 Madison, Wisconsin 53707 This amendment modifies the following portions of the protocol: Effective May 11,1998 1. Page 7. EXPERIMENTAL DESIGN, Observation of Animals. Body Weights. To accurately reflect when the body weights will be taken, replace the first sentence with the following: Weekly during acclimation, on the day of randomization (to be used for dosing calculations), weekly after dosing, and on the day of sacrifice. 004181 39 Covance 6329-226 2. Page 9. EXPERIMENTAL DESIGN. Termination. Scheduled Sacrifice. To clarify the disposition of the animal carcass, please add a sentence following the first sentence of the third paragraph: After the tissues have been removed, the residual carcass will be discarded. 3- Page 10, EXPERIMENTAL DESIGN. Sample Preparation and Storage Since a subsample of less than 1 mL of whole blood may need to be removed, please replace the second sentence of paragraph two with the following: A subsample at the individual time points and approximately half of the sacrifice blood will be removed and placed in a separately labeled vial. Effective May 13,1998 4. Page 3. TEST SUBSTANCE, Storage Conditions Upon actual receipt of the test substance, it was noted that the storage was to be room temperature. Therefore, replace the first and second sentence with the following: The test substance and any remaining test substance will be stored at room temperature. Effective June 2,1998 To indicate that an additional 2 mL blood sample will be removed for serum collection for the remaining time points, the following changes to the protocol are necessary. 5. Page 8. EXPERIMENTAL DESIGN, Antemortem Sample Collection. Blood (Groups 1 and 21. Add the following sentence after the third line: Effective June 3,1998 (Study Day 21) and at the remaining time points, an additional 2 mL blood sample will be collected into a glass tube without heparin and set aside at room temperature to allow clotting. 6. Page 10, EXPERIMENTAL DESIGN, Sample Preparation and Storage Add a new third paragraph as follows: Remove the serum and place it into a separately labeled vial. Discard the cellular debris and store the serum at approximately -20C for shipment. 004182 40 Covance 6329-226 To indicate that additional blood will be removed for serum collection at sacrifice, the following changes to the protocol are necessary. 7. Page 9. EXPERIMENTAL DESIGN. Antemortem Sample Collection. Scheduled Sacrifice. Replace the fourth line with the following: Blood will be collected via cardiac puncture into glass tubes with heparin (at least 20 mL) and into glass tubes without heparin (at least 20 mL). PROTOCOL AMENDMENT APPROVAL Study Director 3M Chemicals Date Biological Phase Investigator Metabolic Chemistry Covance Laboratories Inc. Associate Director Metabolic Chemistry Covance Laboratories Inc. 0 0 4 1 .8 3 41 Covance 6329-226 PROTOCOL DEVIATIONS Protocol Actual Environmental controls for the animal room will be set to maintain a relative humidity of 50% 20% April 25,1998, ranged from 80.1% to 86.2% between 2:27 and 8:00 May 28,1998, ranged from 73.4% to 73.9% between 19:08 and 20:30 June 11,1998, ranged from 71.5% to73.6% between 7:30 and 10:00 and ranged from 75.3% to 82.6% between 17:19 and 21:00 June 20,1998, ranged from 70.2% to 72.8% between 13:19 and 15:00 June 27,1998, ranged from 71.3% to 78.1% between 6:38 and 9:00 At sacrifice, blood will be collected via cardiac puncture into glass tubes with heparin (at least 20 mL) and into glass tubes without heparin (at least 20 mL). A short sample was collected via the vena cava for serum and plasma. Body Weights. Weekly during The body weights were not taken on the acclimation, on the day of randomization day of randomization, because no (to be used for dosing calculations), randomization was needed. The animals weekly after dosing, and on the day of were selected by test results and sacrifice. adjustment to the jacket and tether system. The weights were taken within 5 days of dosing to be used for dosing calculations. These deviations would not be expected to have had an effect on the outcome of the study. 004184 42 Covance 6329-226 APPENDIX B Summary of Antemortem Observations, Clinical Pathology Codes and Abbreviations, and Clinical Pathology Data Tables 004185 43 Covance 6329-226 Table Bl. Animal ID Summary of Antemortem Observations Sacrifice Day and Sex Observation Noted Study Day Group 1 (Intact) 105428 2 Day, Male Urine appears to contain water 1 105430 2 Day, Male No observations noted 105431 28 Day, Male Urine discolored, brown 21 105432 28 Day, Male Urine appears to contain water Few feces 10 2 105433 91 Day, Male No observations noted 105435 91 Day, Male Urine appears to contain water Urine appears to contain water Urine discolored, red Low food consumption Low food consumption Low food consumption Low food consumption Low food consumption Low food consumption 19 64 66 72 73 74 75 76 77 105439 2 Day, Female No observations noted 105440 2 Day, Female Few feces 2 105442 28 Day, Female Urine appears to contain water Urine appears to contain water Second container used for urine Low food consumption Low food consumption 17 19 17 11 12 105444 28 Day, Female Urine appears to contain water Urine appears discolored, red Urine appears discolored, red Few feces Low food consumption Low food consumption 19 9 10 2 11 12 0Q41S6 44 Covance 6329-226 Table Bl. Continued Animal ID 105447 Sacrifice Day and Sex 91 Day, Female Observation Noted Urine contained feces Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Second container used for urine Second container used for urine Second container used for urine Second container used for urine Second container used for urine Urine appears discolored, red Discharge, appeared to be menstruating No feces Few feces Low food consumption Study Day 22 10 12 13 15 16 17 18 19 23 43 64 78 15 16 17 64 78 10 10 2 50 73 105449 91 Day, Female Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Urine appears to contain water Second container used for urine Second container used for urine Second container used for urine Few feces 10 43 50 57 64 71 78 57 64 78 2 C041S7 45 Covance 6329-226 Table Bl. Continued Animal ID Sacrifice Day and Sex Observation Noted Group 2 (Bile Duct Cannulated) 105436 Bile collected 28 Day Male Discharge, vomitus containing food Low food consumption 105437 Bile collected 2 Day Male Small amount of bile Liquid feces Liquid feces Non-formed feces Non-formed feces Non-formed feces Low food consumption Low food consumption Low food consumption No feces 105441 Bile collected 28 Day Female Small amount of bile Non-formed feces Non-formed feces Non-formed feces Low food consumption Low food consumption No feces 105446 Bile collected 2 Day Female Small amount of bile Small amount of bile Small amount of bile No bile present No bile present No bile present No bile present No bile present Non-formed feces Non-formed feces Non-formed feces Non-formed feces Study Day 8 6 10 12 13 6 7 23 10 11 12 10 10 6 22 23 10 11 10 17 23 24 25 26 27 28 29 1 2 5 7 004188 NS QS/QNS NR FS SC SH H SL L SI I u RE EE SE PC PD PI PL PA CO HB PLASMO NO AGG FR 46 Covance 6329-226 Codes for Clinical Pathology GENERAL CODES No sample Quantity not sufficient No repeat (sample volume not sufficient for repeat analysis) Fibrin strands Sample clotted Slightly hemolyzed Hemolyzed Slightly lipemic Lipemic Slightly icteric Icteric Unscheduled/moribund bleed Recording error (recorded incorrect data, e.g., wrong number, spelling error, incorrect date) Entry error (incorrect keyboard entry) Sampling error Platelets clumped Platelets decreased Platelets increased Platelets large Platelets appear adequate Color interferes with test Heinz bodies observed Plasmodium No aggregation Fractious CODES FOR BLOOD CELL MORPHOLOGY The following scale was used to measure the degree of anisocytosis (ANISO), poikilocytosis (POIK), polychromasia (POLY), hypochromasia (HYPO), or basophilic stippling (BASTIP) or the presence of Howell-Jolly bodies (HJBODY), toxic neutrophils (TOXNEUT), or atypical lymphocytes (ATYPLYM): Scale Deeree Presence - Normal for the species 1 Slight 2 Moderate 3 Marked 4 Not applicable Not present Rare Few Moderate Many 004180 47 Covance 6329-226 Abbreviations and Units for Clinical Hematology Test Red blood cell count Hemoglobin Hematocrit Mean corpuscular volume Mean corpuscular hemoglobin Mean corpuscular hemoglobin concentration Platelet count Prothrombin time Activated partial thromboplastin time White blood cell count Differential blood cell count Nucleated red blood cell count Segmented neutrophil count Band neutrophil count Lymphocyte count Monocyte count Eosinophil count Basophil count Anisocytosis Polychromasia Poikilocytosis Hypochromasia Toxic neutrophils Abbreviation (Units) RBC (E6/UL or X106/yuL) HGB (G/DL) HCT (%) MCV (FL) MCH (PG) MCHC (%) PLT (E3/ULorX103/AiL)) PT (SEC) PTT (SEC) WBC (E3/UL or XKP/uL) NRBC (/100 WBC) N-SEG (E3/UL or X103///L) and % N-BAND (E3/UL or XIOV/^L) and % LYMPH (E3/UL or XIOVL) and % MONO (E3/UL or X107mL) and % EOSIN (E3/UL or X103//iL) and % BASO (E3/UL or XIOV^L) and % ANISO (-,1,2,3) POLY (-,1,2,3) POIK (-,1,2,3) HYPO (-,1,2,3) TOXNEUT (-,1,2,3,4) Abbreviations and Units for Clinical Chemistry Test Glucose Urea nitrogen Creatinine Total protein Albumin Globulin Total bilirubin Cholesterol Triglycerides Aspartate aminotransferase Alanine aminotransferase Alkaline phosphatase Gamma glutamyltransferase Creatine kinase Calcium Inorganic phosphorus Sodium Potassium Chloride Serum bile acids Abbreviation (Units! GLU (MG/DL) UN (MG/DL) CREAT (MG/DL) T PRO (G/DL) ALB (G/DL) GLOB (G/DL) T BILI (MG/DL) CHOL (MG/DL) TRIG (MG/DL) AST/SGOT (IU/L) ALT/SGPT (IU/L) ALK PHOS (IU/L) GGT (IU/L) CK (IU/L) CA (MG/DL) I PHOS (MG/DL) NA (MMOL/L) K (MMOL/L) CL (MMOL/L) SBA (UMOL/L or MG/DL) 004190 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Males Presurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SOLFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL CHOL MG/DL SBA umol/L Group: 1 105428 105430 105431 105432 105433 105435 MEAN S.D. N Group: 2 105436 105437 MEAN S.D. N Extra 105429 105434 105438 MEAN S.D. N Dose Level: 50 134 16 77 17 80 20 124 20 99 17 118 13 105 23.7 6 17 2.6 6 Dose Level: 50 100 17 106 17 103 4.2 2 17 .0 2 124 105 103 111 11.6 3 19 16 18 18 1.5 3 Dosage Unit: mg/kg 1.1 8.8 1.0 8.0 1.0 8.4 1.1 9.1 1.0 8.6 1.1 8.8 1.0 .05 6 8.6 .38 6 Dosage Unit: mg/kg 1.3 8.4 1.0 8.4 1.2 .21 2 8.4 .00 2 4.6 4.4 4.6 4.8 4.6 4.7 4.6 .13 6 4.8 4.6 4.7 .14 2 1.3 1.2 1.3 1.3 .06 3 8.4 9.3 7.8 8.5 .75 3 4.8 4.7 4.7 4.7 .06 3 4.2 3.6 3.8 4.3 4.0 4.1 4.0 .26 6 3.6 3.8 3.7 .14 2 3.6 4.6 3.1 3.8 .76 3 .2 85 .2 95 .2 150 .3 128 .4 94 .3 137 13 10 8 7 10 12 .3 115 10 .08 26.9 2.3 666 .3 138 .1 146 14 11 .2 142 12 .14 5.7 2.1 222 .2 172 .2 133 .4 113 11 17 8 .3 139 12 .12 30.0 4.6 333 Covance 6329-226 004191 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Males Presurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SOLFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L CK IU/L CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group: 1 105428 105430 105431 105432 105433 105435 MEAN S.D. N Group : 2 105436 105437 MEAN S.D. N Extra 105429 105434 105438 MEAN S.D. N Dose Level:: 50 32 58 62 28 56 33 56 116 39 78 35 24 47 12.8 6 56 35.8 6 Dose Level:: 50 39 38 35 25 37 2.8 2 32 9.2 2 50 48 34 44 8.7 3 34 120 28 61 51.5 3 Dosage Unit: mg/kg 413 76 421 94 547 111 604 189 452 43 566 157 500 81.8 6 112 53.5 6 Dosage Unit: mg/kg 370 53 295 82 332 53.0 2 68 20.5 2 361 571 761 316 125 367 417 220.5 6 340 348 344 5.7 2 607 297 416 440 156.4 3 138 122 111 124 13.6 3 1055 1019 255 776 451.8 3 9.9 9.5 9.0 10.2 9.4 10.6 9.8 .58 6 10.9 10.2 10.6 .49 2 9.6 11.1 10.4 10.4 .75 3 6.2 3.6 6.2 7.9 5.8 6.5 6.0 1.40 6 6.8 4.4 5.6 1.70 2 6.4 5.9 5.5 5.9 .45 3 155 147 146 155 154 152 152 4.0 6 4.3 4.6 4.9 5.7 4.2 4.8 4.8 .54 6 160 152 156 5.7 2 5.1 4.3 4.7 .57 2 151 159 156 155 4.0 3 5.2 4.8 5.2 5.1 .23 3 101 100 97 107 106 99 102 4 6 105 99 102 4 2 103 103 109 105 3 3 Covance 6329-226 Z6T-V00 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Females Presurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGDS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL CHOL MG/DL SBA umol/L Group : 1 105439 105440 105442 105444 105447 105449 MEAN S.D. N Group: 2 105441 105446 MEAN S.D. N Extra 105443 105445 105448 MEAN S.D. N Dose Level : 50 112 17 116 18 128 19 111 18 78 22 114 26 110 16.8 6 20 3.4 6 cn o Dose Level 98 16 91 16 94 4.9 2 16 .0 2 88 94 114 99 13.6 3 16 14 12 14 2.0 3 Dosage Unit: mg/kg 1.1 8.4 1.2 9.7 .9 8.5 1.0 9.0 1.0 8.8 1.0 9.4 1.0 .10 6 9.0 .51 6 Dosage Unit: mg/kg 1.2 9.3 .8 8.3 1.0 .28 2 8.8 .71 2 4.6 4.7 4.4 4.6 3.6 4.5 4.4 .40 6 4.7 4.1 4.4 .42 2 .9 .9 1.1 1.0 .12 3 7.7 9.2 8.4 8.4 .75 3 4.0 4.3 4.4 4.2 .21 3 3.8 5.0 4.1 4.4 5.2 4.9 4.6 .55 6 4.6 4.2 4.4 .28 2 3.7 4.9 4.0 4.2 .62 3 .2 122 .4 182 .3 123 .3 148 .2 126 .3 141 26 49 9 9 12 13 .3 140 20 .08 23.0 15.7 666 .2 181 .3 201 11 7 .2 191 .07 14.1 22 9 2.8 2 .4 202 .2 155 .6 136 10 20 5 .4 164 12 .20 34.0 7.6 333 Covance 6329-226 G4193 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Females Presurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L CK IU/L CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group: 1 105439 105440 105442 105444 105447 105449 MEAN S.D. N Group : 2 105441 105446 MEAN S.D. N Extra 105443 105445 105448 MEAN S.D. N Dose Level:: 50 57 134 95 199 27 25 38 94 55 233 42 216 52 23.7 6 150 80.8 6 Dose Level :: 50 46 34 34 37 40 8.5 2 36 2.1 2 31 28 44 34 8.5 3 82 95 118 98 18.2 3 Dosage Unit: mg/kg 363 98 218 233 90 160 215 44 104 192 87 256 284 52 202 407 69 239 282 86.2 6 73 21.9 6 196 56.1 6 Dosage Unit: mg/kg 296 78 1061 184 92 134 240 79.2 2 85 9.9 2 598 655.5 2 160 308 195 221 77.3 3 76 64 108 83 22.7 3 147 205 652 335 276.3 3 10.2 10.7 9.2 10.1 9.1 10.0 9.9 .62 6 10.5 9.4 10.0 .78 2 9.1 11.1 9.5 9.9 1.06 3 6.3 6.2 4.7 6.0 4.3 4.6 5.4 .91 6 6.2 3.9 5.0 1.63 2 3.1 4.5 4.2 3.9 .74 3 155 157 153 156 149 158 155 3.3 6 4.8 4.2 5.1 4.3 4.0 4.3 4.4 .41 6 16 151 156 7.1 2 5.0 5.1 5.0 .07 2 153 159 152 155 3.8 3 3.7 4.5 4.5 4.2 .46 3 105 104 105 105 107 107 106 1.2 6 107 108 108 .7 2 106 109 106 107 1.7 3 Covance 6329-226 004194 S6T>00 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Males - 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB iG/DL GLOB G/DL T BILI MG/DL CHOL MG/DL SBA umol/L Group: 2 105436 105437 MEAN S.D. N Extra 105434 Dose Level : 50 75 27 99 18 87 17.0 2 22 6..4 2 84 15 Dosage Unit: mg/kg 1.2 7.8 .9 8.1 1.0 .21 2 8.0 .21 2 3.7 4.2 4.0 .35 2 1.1 8.3 3.9 4.1 3.9 4.0 .14 2 4.4 2.4 .5 1.4 1.34 2 2.1 181 81 131 70.,7 2 33 83 58 35.4 2 148 245 Covance 6329-226 c/> to Covance 6329-226 0041.96 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Males ~ 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L CK IU/L CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group : 2 105436 105437 MEAN S.D. N Extra 105434 Dose Level: 50 175 316 194 131 184 13.4 2 224 130.8 2 397 364 Dosage Unit: mg/kg 1231 453 267 1652 228 2328 842 550.1 2 248 27.6 2 1990 478.0 2 1278 237 3267 9.6 10.0 9.8 .28 2 9.8 5.5 4.0 4.8 1.06 2 5.1 153 149 151 2.8 2 4.9 4.5 4.7 .28 2 150 5.3 109 111 110 1.4 2 110 Lto Covance 6329-226 004197 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Females ~ 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL CHOL MG/DL SBA umol/L Group: 2 105441 105446 MEAN S.D. N Extra 105443 Dose Level : 50 91 15 108 25 100 12.0 2 20 7. 1 2 90 14 Dosage Unit: mg/kg 1.1 6.9 .9 8.5 1.0 .14 2 7.7 1.13 2 3.5 3.8 3.6 .21 2 .8 7.3 3.6 3.4 4.7 4.0 .92 2 3.7 .4 103 .4 153 .4 128 .00 35. 4 22 .9 139 6 2 4 2.8 2 1 -H . IPoO ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Females - 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGDS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L CK IU/L CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group: 2 105441 105446 MEAN S.D. N Extra 105443 Dose Level: 50 248 232 123 123 186 88. 4 2 178 77. 1 2 153 150 Dosage Unit: mg/kg 258 48 1802 246 67 2832 252 8.5 2 58 13.4 2 2317 728. 3 2 145 63 2669 9.6 10.2 9.9 .42 2 9.2 3.9 4.2 4.0 .21 2 3.7 156 159 158 2..1 2 4.6 4.9 4.8 .21 2 155 4.6 112 118 115 4.2 2 112 Covance 6329-226 tuyni Covance 6329-226 004199 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Males ~ 8 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGDS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL CHOL MG/DL SBA umol/L Group: 2 105436 105437 MEAN S.D. N Extra 105434 Dose Level : 50 85 24 98 18 92 9.2 2 21 4.,2 2 Dosage Unit: mg/kg 1.2 7.8 1.0 7.6 1.1 .14 2 7.7 .14 2 3.9 4.1 4.0 .14 2 110 14 1.1 8.5 3.9 3.9 3.5 3.7 .28 2 4.6 .6 .3 .4 .21 2 3.8 121 76 98 31.8 2 4 5 4 2 240 932 LCT/l\ Covance 6329-226 004200 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Males ~ 8 Days Postsurgery METABOLISM OF N-ETHYL PERFLOOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE AST/SGOT ALT/SGPT .ALK PHOS IU/L IU/L IU/L GGT IU/L CK IU/L CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group: 2 105436 105437 MEAN S.D. N Extra 105434 Dose Level: 50 36 141 27 63 32 6. 4 2 102 55.2 2 345 444 Dosage Unit: mg/kg 1141 690 281 260 916 318.9 2 270 14.8 2 468 214 341 179.6 2 4815 364 533 9.5 10.3 9.9 .57 2 10.2 5.5 3.0 4.2 1.77 2 4.2 149 146 148 2.1 2 4.7 4.4 4.6 .21 2 153 4.5 107 103 105 2.8 2 109 Vi Covance 6329-226 004201 ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Females - 8 Days Postsurgery METABOLISM OF N-ETHYL PERFL00R00CTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGOS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL CHOL MG/DL SBA umol/L Group : 2 105441 105446 MEAN S.D. N Extra 105443 Dose Level : 50 114 10 115 19 114 .7 2 14 6..4 2 74 6 Dosage Unit: mg/kg 1.2 7.4 .8 8.2 1.0 .28 2 7.8 .57 2 3.8 3.8 3.8 .00 2 .7 7.1 3.6 3.6 4.4 4.0 .57 2 3.5 .2 .4 .3 .14 2 1.6 63 152 108 62..9 2 5 5 5 2 135 382 U00h ANIMAL NUMBER Summary and Individual Clinical Chemistry Data Females - 8 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SOLFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L CK IU/L CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group: 2 105441 105446 MEAN S.D. N Extra 105443 Dose Level: 50 38 101 35 58 36 2. 1 2 80 30. 4 2 396 374 Dosage Unit: mg/kg 377 52 88 229 66 1048 303 104.7 2 59 9.9 2 568 678..8 2 841 262 617 10.7 10.1 10.4 .42 2 9.1 3.5 3.7 3.6 .14 2 3.0 156 144 150 8.5 2 5.8 5.7 5.8 .07 2 148 4.1 106 107 106 .7 2 109 sO Covance 6329-226 C 0 0 ANIMAL NUMBER Summary and Individual Clinical Hematology Data Males Presurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGOS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE RBC HGB HCT MCV MCH MCHC PLT PT XIOV/iL G/DL % FL PG i XlOV^iL SEC Group : 1 105428 105430 105431 105432 105433 105435 MEAN S.D. N Group: 2 105436 105437 MEAN S.D. N Extra 105429 105434 105438 MEAN S.D. N Dose Level: 50 6.55 5.97 5.35 5.90 6.14 6.41 12.8 11.6 9.9 12.4 11.1 12.7 6.05 .426 6 11.8 1.12 6 Dose Level: 50 6.80 6.21 12.9 11.5 6.50 .417 2 12.2 .99 2 7.85 6.29 6.05 6.73 .977 3 15.1 12.6 11.5 13.1 1.84 3 Dosage Unit: mg/kg 43.2 37.9 32.6 40.9 38.9 43.4 65.9 63.5 60.9 69.3 63.3 67.8 19.6 19.4 18.5 21.1 18.0 19.8 39.5 4.03 6 65.1 3.13 6 19.4 1.08 6 Dosage Unit: mg/kg 44.9 38.5 66.0 62.1 19.0 18.5 41.7 4.53 2 64.0 2.76 2 18.8 .35 2 48.7 42.6 38.7 43.3 5.04 3 62.0 67.7 63.9 64.5 2.90 3 19.3 20.1 19.0 19.5 .57 3 29.7 30.6 30.4 30.4 28.5 29.2 29.8 .83 6 28.8 29.8 29.3 .71 2 31.0 29.7 29.8 30.2 .72 3 754 352 529 375 371 653 506 169.0 6 9.6 8.7 9.3 8.9 8.6 9.7 9.1 .47 6 547 383 465 116.0 2 9.3 9.3 9.3 .00 2 491 545 390 475 78.7 3 9.3 9.4 9.6 9.4 .15 3 PTT SEC 15.9 13.9 18.6 13.4 15.3 15.9 15.5 1.84 6 12.9 14.3 13.6 .99 2 15.1 12.6 13.3 13.7 1.29 3 Covance 6329-226 o c 0 ANIMAL NUMBER Summary and Individual Clinical Hematology Data Males Presurgery METABOLISM OF N-ETHYL PERFLOOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGDS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE WBC XIOViiL N-SEG LYMPH X10V//L XlOV/iL MONO EOSIN XIOV^L XIOVaL BASO N-SEG% XIOViiL LYMPH% MONO* EOSIN* BASO* Group: 1 Dose Level : 50 Dosage Unit: mg/kg 105428 35.6 16.4 14.2 2.2 2.8 .1 46 40 6 8 0 105430 29.9 17.2 9.9 2.0 .8 .1 57 33 7 3 0 105431 12.7 5.5 5.6 1.2 .3 .0 43 44 10 2 0 105432 26.8 13.0 11.6 1.8 .2 .0 49 43 7 1 0 105433 27.0 14.6 10.2 1.8 .5 .1 54 38 7 2 0 105435 25.3 14.3 8.9 .9 1.1 .0 57 35 4 4 0 MEAN S.D. N 26.2 13.5 10.1 1.6 1.0 .0 51 39 7 3 0 7.56 4.20 2.86 .50 .96 .05 5.9 4.4 1.9 2.5 6 6 66 6 666 66 6 Group: 2 Dose Level : 50 Dosage Unit: mg/kg 105436 15.3 10.2 4.1 .9 .1 .0 67 27 6 1 0 105437 17.6 10.0 5.6 1.6 .5 .0 57 32 9 3 0 MEAN S.D. N 16.4 10.1 4.8 1.2 .3 .0 62 30 8 2 0 1.63 .14 1.06 .49 .28 .00 7.1 3.5 2.1 1.4 2 2222222222 Extra 105429 105434 105438 MEAN S.D. N 19.1 10.5 6.7 1.2 .7 .0 55 35 7 3 0 16.5 7.2 8.1 .8 .4 .0 44 49 5 2 0 16.7 9.8 4.7 1.8 .5 .0 58 28 11 3 0 17.4 9.2 6.5 1.3 .5 .0 52 37 8 3 0 1.45 1.74 1.71 .50 .15 .00 7.4 10.7 3.1 .6 33333333333 Covance 6329-226 004204 Individual Clinical Hematology Data Males Presurgery METABOLISM OF N-ETHYL PERFLOOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 1 105428 105430 a 105431 105432 105433 105435 Group: 2 105436 105437 Extra 105429 105434 105438 Dose Level: 50 - Dose Level: 50 - - Dosage Unit: mg/kg -- Dosage Unit: mg/kg -- - a Plasmodium was observed. Covance 6329-226 004205 Covance 6329-226 004206 ANIMAL NUMBER Summary and Individual Clinical Hematology Data Females Presurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGDS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE RBC HGB HCT MCV MCH MCHC PLT PT X107/ G/DL % FL PG % XIOVm L SEC Group : 1 105439 105440 105442 105444 105447 105449 MEAN S.D. N Group : 2 105441 105446 MEAN S.D. N Extra 105443 105445 105448 MEAN S.D. N Dose Level : 50 7.08 6.97 5.29 6.52 6.07 7.01 12.9 11.9 11.0 11.5 11.7 13.1 6.49 .702 6 12.0 .82 6 Dose Level: 50 7.64 5.58 14.1 10.4 6.61 1.457 2 12.2 2.62 2 6.37 6.29 5.99 6.22 .200 3 12.2 11.3 11.5 11.7 .47 3 Dosage Unit: mg/kg 44.3 41.9 35.4 39.7 39.2 45.8 62.5 60.2 66.8 60.9 64.7 65.3 18.3 17.1 20.8 17.6 19.2 18.7 41.1 3.77 6 63.4 2.61 6 18.6 1.31 6 Dosage Unit: mg/kg 50.8 33.6 66.6 60.2 18.5 18.7 42.2 12.16 2 63.4 4.53 2 18.6 .14 2 37.8 38.0 38.9 38.2 .59 3 59.3 60.5 64.9 61.6 2.95 3 19.2 17.9 19.2 18.8 .75 3 29.3 28.4 31.2 29.0 29.7 28.6 29.4 1.01 6 27.8 31.1 29.4 2.33 2 32.3 29.6 29.6 30.5 1.56 3 556 421 531 291 324 444 428 106.7 6 9.5 10.2 9.6 9.1 9.4 9.5 9.6 .36 6 449 446 448 2.1 2 10.0 9.6 9.8 .28 2 392 402 402 399 5.8 3 9.9 9.3 9.7 9.6 .31 3 PTT SEC 14.7 18.1 14.3 14.0 15.9 15.5 15.4 1.50 6 17.3 13.2 15.2 2.90 2 14.2 13.2 16.0 14.5 1.42 3 ANIMAL NUMBER WBC XIOV^L Summary and Individual Clinical Hematology Data Females Presurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE N-SEG LYMPH X10V/^L XIOV^L MONO EOSIN XlOV^L XlOV^iL BASO N -SEG% XIOViiL LYMPH% MONO% EOSIN% BASO% Group: i Dose Level : 50 Dosage Unit: mg/kg 105439 14.7 6.4 6.4 1.5 .4 .0 43 43 10 3 0 105440 9.7 2.4 6.5 .5 .2 .0 25 67 5 2 0 105442 21.0 6.4 13.4 .6 .6 .0 30 64 3 3 0 105444 12.2 5.0 5.4 1.6 .2 .0 41 44 13 2 0 105447 10.8 4.3 5.4 .6 .5 .0 40 50 6 5 0 105449 11.4 5.0 5.7 .6 .1 .0 44 50 51 0 MEAN S.D. N 13.3 4.9 7.1 .9 .3 .0 37 53 7 3 0 4.13 1.49 3.11 .51 .20 .00 7.8 10.2 3.7 1.4 6 6 666666 666 Group: 2 Dose Level;: 50 Dosage Unit: mg/kg 105441 105446 15.2 7.6 5.5 1.2 .9 .0 50 36 12.3 5.7 5.7 .6 .3 .0 46 46 86 52 0 0 MEAN S.D. N 13.8 6.6 5.6 .9 .6 .0 48 41 6 4 0 2.05 1.34 .14 .42 .42 .00 2.8 7.1 2.1 2.8 22222222222 Extra 105443 105445 105448 14.3 4.3 8.2 1.1 .6 .1 30 58 8 4 0 12.5 5.4 5.5 1.5 .2 .0 43 44 12 1 0 10.8 3.0 5.5 1.9 .3 .0 28 51 18 2 0 MEAN S.D. N 12.5 4.2 6.4 1.5 .4 .0 34 51 13 2 0 1.75 1.20 1.56 .40 .21 .06 8.1 7.0 5.0 1.5 33333333333 Covance 6329-226 o n o si Individual Clinical Hematology Data Females Presurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 1 Dose Level: 50 105439 - - - 105440 - - - 105442 - - - 105444 - 105447 - - - 105449 - Group: 2 Dose Level: 50 105441 105446 Extra 105443 105445 105448 Dosage Unit: mg/kg ---- -- Dosage Unit: mg/kg oo-n\ Covance 6329-226 o 0 1 Si o nI 602*00 ANIMAL NUMBER Summary and Individual Clinical Hematology Data Males - 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE RBC HGB HCT MCV MCH MCHC PLT PT X I O V aiL G/DL % FL PG % X I O V aiL SEC Group: 2 105436 105437 MEAN S.D. N Extra 105434 Dose Level: 50 6.46 6.62 11.9 11.9 6.54 .113 2 11.9 .00 2 5.81 11.2 Dosage Unit: mg/kg 39.7 39.8 61.4 60.1 18.4 17.9 39.8 .07 2 60.8 .92 2 18.2 .35 2 37.7 64.9 19.2 30.0 29.9 30.0 .07 2 29.6 679 519 599 113.1 2 9.0 8.9 9.0 .07 2 498 9.7 PTT SEC 15.8 14.1 15.0 1.20 2 15.3 Covance 6329-226 0CT\\ OTZkGQ ANIMAL NUMBER Group: 2 105436 105437 MEAN S.D. N Extra 105434 Summary and Individual Clinical Hematology Data Males ~ 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGOS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE WBC X10V//L N-SEG LYMPH XIOVaL XIOV^L MONO EOSIN XIOVaiL XIOVaL BASO N-SEG% XlOV/iL LYMPH% MONO% EOSIN% BASO% Dose Level : 50 Dosage Unit: mg/kg 14.7 10.2 3.6 .8 .1 .0 70 24 5 10 9.4 5.2 3.1 .9 .3 .0 55 32 9 3 0 12.0 7.7 3.4 .8 .2 .0 62 28 7 2 0 3.75 3.54 .35 .07 .14 .00 10.6 5.7 2.8 1.4 .0 22222222222 10.3 3.6 5.7 .7 .2 .1 35 55 7 2 1 Covance 6329-226 0-J\ Covance 6329-226 004211 Individual Clinical Hematology Data Males ~ 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 2 105436 105437 Extra 105434 Dose Level: 50 ___ ___ ___ Dosage Unit: mg/kg __ __ __ Ooos ANIMAL NUMBER Summary and Individual Clinical Hematology Data Females ~ 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE RBC HGB HCT MCV MCH MCHC PLT PT X I O V aL G/DL % FL PG % XIOV/iL SEC Group: 2 105441 105446 MEAN S.D. N Extra 105443 Dose Level: 50 3.77 5.76 7.3 10.7 4.76 1.407 2 9.0 2.40 2 6.37 11.3 Dosage Unit: mg/kg 25.8 35.6 68.6 61.7 19.5 18.5 30.7 6.93 2 65.2 4.88 2 19.0 .71 2 37.3 58.5 17.7 28.4 30.0 29.2 1.13 2 30.2 458 598 528 99.0 2 9.3 8.7 9.0 .42 2 522 9.1 PTT SEC 16.2 12.6 14.4 2.55 2 13.5 On NO Covance 6329-226 o c p ANIMAL NUMBER Summary and Individual Clinical Hematology Data Females ~ 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE WBC X I O V aiL N-SEG LYMPH XlOViiL XIOVaL MONO EOSIN X10V/AL XIOV/aL BASO N--SEG% XIOV/aL LYMPH% MONO'S EOSIN% BASO% Group : 2 Dose Level : 50 Dosage Unit: mg/kg 105441 12.4 5.3 5.4 . 1.4 .2 .2 43 44 11 2 1 105446 15.3 7.7 5.6 1.7 .2 .1 50 36 11 1 1 MEAN S.D. N 13.8 6.5 5.5 1.6 .2 .2 46 40 11 2 1 2.05 1.70 .14 .21 .00 .07 4.9 5.7 .0 .7 .0 22222222222 Extra 105443 10.4 3.8 5.5 .8 .2 .0 37 53 8 2 0 -4 Covance 6329-226 c o 0 P , Co Individual Clinical Hematology Data Females ~ 4 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 2 Dose Level: 50 105441 105446 1 _ 1 _ _ Extra 105443 _ _ _ Dosage Unit: mg/kg __ __ Covance 6329-226 * T Z Tr09 ANIMAL NUMBER Summary and Individual Clinical Hematology Data Males - 8 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE RBC HGB HCT MCV MCH MCHC PLT PT X I O V aiL G/DL % FL PG % XlOV/iL SEC Group: 2 105436 105437 MEAN S.D. N Extra 105434 Dose Level: 50 6.10 6.28 11.5 11.5 6.19 .127 2 11.5 .00 2 5.91 11.5 Dosage Unit: mg/kg 38.0 37.5 62.2 59.7 18.9 18.3 37.8 .35 2 60.9 1.77 2 18.6 .42 2 38.4 65.0 19.5 30.4 30.6 30.5 .14 2 30.0 651 424 538 160.5 2 9.1 8.7 8.9 .28 2 516 9.6 PTT SEC 15.2 14.0 14.6 .85 2 14.9 Covance 6329-226 o c N u\ Covance 6329-226 004216 ANIMAL NUMBER Group : 2 105436 105437 MEAN S.D. N Extra 105434 Summary and Individual Clinical Hematology Data Males ~ 8 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE WBC XIOV/JL N-SEG LYMPH XIOVllL XIOV^L MONO EOSIN XIOVk L XIOV/iL BASO N--SEG% XIOV/iL LYMPH% MONO% EOSIN% BASO Dose Level : 50 Dosage Unit: mg/kg 13.5 7.9 4.4 1.0 .2 .1 58 33 7 1 0 11.6 5.9 4.2 .9 .6 .0 51 36 8 5 0 12.6 6.9 4.3 1.0 .4 .0 54 34 8 3 0 1.34 1.41 .14 .07 .28 .07 4.9 2.1 ' 2.8 .0 2 2 2 222222 22 7.6 2.5 4.4 .6 .1 .0 33 58 8 1 0 LO Covance 6329-226 004217 Individual Clinical Hematology Data Males ~ 8 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SOLFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 2 105436 105437 Extra 105434 Dose Level: 50 ___ ___ Dosage Unit: mg/kg - __ __ ' U Covance 6329-226 004218 ANIMAL NUMBER Summary and Individual Clinical Hematology Data Females - 8 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGOS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE RBC HGB HCT MCV MCH MCHC PLT PT XIOV^L G/DL % FL PG % XlOVpiL SEC Group: 2 105441 105446 MEAN S.D. N Extra 105443 Dose Level: 50 4.80 5.91 9.4 10.8 5.36 .785 2 10.1 .99 2 5.76 10.7 Dosage Unit: mg/kg 33.9 35.8 70.7 60.5 19.5 18.3 34.9 1.34 2 65.6 7.21 2 18.9 .85 2 34.4 59.7 18.6 27.6 30.2 28.9 1.84 2 31.2 773 652 712 85.6 2 9.0 8.9 9.0 .07 2 497 10.0 PTT SEC 15.3 13.8 14.5 1.06 2 16.2 Covance 6329-226 .004219 ANIMAL NUMBER Group: 2 105441 105446 MEAN S.D. N Extra 105443 Summary and Individual Clinical Hematology Data Females ~ 8 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SDLFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE CAPSULE DOSE WBC XlOV/iL N-SEG LYMPH XlOV/iL XIOVaL MONO EOSIN XIOV^L XIOV^L BASO N--SEG% XIOVm L LYMPH% :MONO% EOSIN% BASO% Dose Level : 50 Dosage Unit: mg/kg 14.6 5.8 6.7 1.8 .2 .1 40 46 12 2 1 16.5 6.5 7.6 1.3 1.0 .1 39 46 8 6 1 15.6 6.2 7.2 1.6 .6 .1 40 46 10 4 1 1.34 .49 .64 .35 .57 .00 .7 .0 2.8 2.8 22222222222 9.9 3.3 5.5 .9 .2 .0 33 55 9 2 0 orzfroo Individual Clinical Hematology Data Females ~ 8 Days Postsurgery METABOLISM OF N-ETHYL PERFLUOROOCTANE SULFONAMIDO ETHANOL (NET-FOSE; T-6316) IN CYNOMOLGUS MONKEYS FOLLOWING ADMINISTRATION OF A SINGLE -CAPSULE DOSE ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 2 105441 105446 Extra 105443 Dose Level: 50 ___ ___ ___ Dosage Unit: mg/kg __ __ __ Covance 6329-226 -u -J C o v an ce is an in d e p e n d e n t, p u b lic ly h e ld c o m p a n y op eratin g in over IS countries and over 30 offices w orldw ide, w ith headquarters in P rinceton, N ew Jersey, U SA . 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