Document NExgZBMgOYDB1M8jr9BmknBZQ

AR x x r o n i Draft Report 5-Daily Dose Dermal Absorption/Toxicity Study of T-6684 in Rabbits PREPA3RMED FOR: COVANCE6S3T29U-D20Y0 NUMBER: IJSuSlyUE11D, A19T9E7: 005501 Sponsor: 3M St. Paul, Minnesota DRAFT FINAL REPORT Study Title: 5-Daily Dose DofeTrm-6a6l8A4bisnorRpatbiobnit/sToxicity Study Author: F. Bud W. McDonald Study Completion Date: July 11,1997 Testing Facility: MC3a3od0vi1asnoKcnei,nWLsmaibsaocnoranBtsooirunilees5v3Ian7rcd0.4 Testing Facility Project Identification: Covance 6329-200 Page 1 of 52 05S02 QUALITY ASSURANCE STATEMENT (To be provided in the final report) Covance 6329-200 2 005803 STUDY IDENTIFICATION 5-Daily Dose DofeTrm-6a6l8A4bisnoRrpatbiobnit/sToxicity Study Covance 6329-200 Test Material Sponsor Sponsor's Representative Study Director Testing Facility StEIESSunttxxd-uuppylddieefyyTrreiiiICmmmEnoneieetmndntiaattpDaabtilllloaee(TtntIeienoDr-nmlaiDftieena)atetSitoanrtDDaatete T-6684 T3P3SMM.toMO.xP.ieCacBdeouionllc,oxtaegMl3ryD,3NBS2e2pled5r0agv5r.it1cm32e32esn-03t-222E0-02 R3Moger G. Perkins, PhD, DABT TP3M.oMOx.ieCcBdoeionlcoxtaeglr3yD,3BS2ep2eldr0agvr.itcm2e2esn0t-2E-02 (S6t.1P2a)7u3l,3M-3N22255133-3220 PMFC60..oOa8Bvd..a2uiBsn4doco2nWex.7,L.97Wa0M5b14Ioc5D5ra3ot7on0rai7led-s7I5n4c5. C3M3oa0vd1ainsKocinen,sLWmabaIon5rBa3to7o0uri4leesvaInrdc. (JM(ADDupaanartyetieel25f1f,i,in18n1a9,a91l9l99r7r7e9ep7poorrttiissssiiggnneedd)) 3 OC5S04 Acute Studies FS.tuBduydDWir.ecMtocrDonald JIne-fflirfeeySBu.pHerivcikssor RAodsmeiMnis.tBrartiidvgeeSupervisor Toxicology Support KMaatnhaygMeryers Calvin L. Horton Supervisor Quality Assurance MShaenrarygeRr. W. Petsel KEY PERSONNEL Covance 6329-200 Laboratory Animal Medicine DSCuiinppdleoyrmvJia.stoCer,arAyC, DLAVMM Anatomical Pathology TAhnoamtoamsiEca.lPPaalmtheorl,oPghisDt Deborah L. Pirkel/ JNSauecpckerorSvpeisrsyfoorrst 4 005305 CONTENTS Covance 6329-200 QUALITY ASSURANCE STATEMENT...........................................................................2 STUDY IDENTIFICATION.................................................................................................3 KEY PERSONNEL............................... 4 SUMMARY....................... 7 OBJECTIVE.......................................................................................................................... 9 REGULATORY COMPLIANCE.........................................................................................9 TEPISSSdutaToefrnreiAtattyyigNfeaiPcDnaradentCcidSoaOtRnuaN.teb.i.oTt.ie.lnR.in.st.Oty..i..o...L....n........M.............A............T............E........R............I....A............L............S............................................................................................................................................................................................................................................................................................................111. 99000 TEAJTHSSSuetenoTuslsiteudtmiScsyfAitiaYcinDlnoagSniDtem.Ti.sooi.iEe.ang.ftl.Mn...Tf...o....e.....r.....s.......t...S.........A.p.........e.....n.....c.....i.....im.....e.........s.a..........lS.....s..........e...........l.......e............c.........t.........i......o.............n.....................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................i.......................................11111110111100 PRPLDROBPSSOtarholeobaeoCticassthppoasieeEsmoadotrtiArDvlnioeaoScandntUafgatimotiolmyoRoronA.ifnp.ERnf.Rsonl.iBSe.osfa.oa.u.itlECw..flry.t.ea..exAo.s.tD..paeli.o.nl.o.onsa.e.fi..n.dstc.m..Aau....t....T,ir....da.o..e.R.i...lm.n.s..s.A.e..s..s...i.c....u/.n.r...oS.....ee..i...r..h.s..as..d...t...i.....r..s.p........a..,......m.....t....a.....i........oen..............nnd..............t..........F....................i...........n................a..............l..................R........................e...............p...................o.................r...........t.........................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................11111111113332422444 5 005806 Covance 6329-200 REBDCSoleUidrnmLyicaTWallSIeO.r.ir.bg.i.ts.ha.e.tt.rsi.vo....a.n...t...i...o......n......s.................................................................................................................................................................................................................................................................................................................................................................................................................................11115555 Pathology..................................... 15 DISCUSSION.......................................................................................................................16 SIGNATURE........................................................................................................................17 REFERENCE........... ...........................................................................................................17 TA4231BLIIIIEnnnnGGddddrriiiioovvvvuuiiiiddddppuuuu2aaaa1llll-(DaDCCTnloee-dirrn6nmmMt6irc8aaoae4lllla)IInS(rr-3rriBiiD.gtt0aaonitmtmdsiiooy.eg.nn.t/W.h.kSS.y.gecc.l./ioo.dSg.rr.haue.e.ytssl.s.f).o--..(....x.g.......i..)..d..........e......,........(.....0................0...............m.................g........../......k............g........../.....d..........a..........y..........)............................................................................................................................................................................................................................................122221811009 APIPnEdiNviDdIuXalAA.n..i.m...a..l..P..a..t.h..o..l..o..g..y...D...a..t.a.....................................................................................................................................................................2232 APAPPPrrmEoottNeoonDccdooImllXDTenBPetv6.N.i7.a.o9.t..i8.o..1.n....T.......o...........T......h......e.........P......r......o......t..o....c....o....l...................................................................................................................................................................................................................................................33351576 6 &05807 SUMMARY Covance 6329-200 This study was done to assess the systemic absoiption/toxicity and relative skin irritancy of T-6684 when applied to the skin of rabbits for five consecutive days. The study was conducted using three male and three female acclimated rabbits of the Hra:(NZW)SPF strain for each treatment group. Group 1 (Control) 2 TeCstoMntartoelr/ial SDTuim-lf6oe6xt8hi4dyel Dose Level (mg/kg/day) 0.0a 3.0a Number of Animals Males Females 33 33 a Administered at a dose volume of 0.1 mL/kg of body weight. The back of each rabbit was clipped free of hair and a single dose of the respective control or test material was administered to the skin of the rabbits for 5 consecutive days. The treatment sites remained intact. The area of application was covered with a gauze patch secured with paper tape around all edges and overwrapped with Saran Wrap and Elastoplast tape to provide an occlusive dressing for each approximate 23-hour exposure period. Clinical observations were conducted predose on Day 1 only and at approximately 1,2.5, and 4 hours after each control or test material administration on Days 1through 5. Additional clinical observations were conducted daily thereafter through Day 29. Mortality checks were conducted twice daily on Days 2 through 29 (a.m. mortality check only on Day 29). Body weights were determined on Day -5 for randomization purposes, before the first control or test material administration (Day 1), and at the scheduled sacrifice interval (Day 29). The initial dermal irritation reading was made before the first control or test material administration (recorded as the Day 1reading). Subsequent readings of dermal irritation were made after each patch removal on Days 2 through 6 and on Day 13. A blood sample (approximately 4 mL) was collected from each animal on 7 005808 Covance 6329-200 Days -3, 8,15, and 23. In addition, at the time ofnecropsy (Day 29), approximately 20 to 40 mL of blood was obtained from each control animal and approximately 20 mL of blood was obtained from each Group 2 animal. All samples were centrifuged and separate samples of serum and cellular fractions were obtained and sent frozen on dry ice to the Sponsor. On Day 29, the animals were anesthetized with sodium pentobarbital, bled via the posterior vena cava, exsanguinated and necropsied. The whole liver, bile, an approximate 1-cm x 1-cm section of the dermal application site, and both kidneys (collected as one sample) were collected from all animals, weighed (volume only determined for bile), and sent frozen to the Sponsor. Application of T-6684 did not result in any test material-related changes in body weight gain or macroscopic findings at necropsy. All animals appeared normal throughout the study with the exception of one Group 1 animal and two Group 2 animals which had decreased feed consumption for one to three days within the first nine days of the study. Slight dermal irritation was observed in one control animal while slight to moderate dermal irritation was observed in the other five control animals treated with dimethyl sulfoxide (DMSO). In the animals treated with T-6684, slight dermal irritation was observed in one animal while slight to moderate dermal irritation was observed in the other five animals. No macroscopic lesions were seen in any of the animals at necropsy. 8 005809 OBJECTIVE Covance 6329-200 The objective of this study was to assess the systemic absorption/toxicity and relative skin irritancy of test materials when applied to the skin ofrabbits for five consecutive days. REGULATORY COMPLIANCE This study was conducted in accordance with the United States Food and Drug Administration Good Laboratory Practice Regulations for Nonclinical Laboratory Studies, 21 CFR 58, with the exception that analysis of the test material mixtures for concentration, homogeneity/solubility, and stability was not conducted. All procedures used in this study were in compliance with the Animal Welfare Act Regulations. In the opinion ofthe Sponsor and study director, the study did not unnecessarily duplicate any previous work. All procedural times presented in this report fall within the acceptable ranges as specified in the Covance Laboratories Inc. (Covance) Standard Operating Procedures (SOPs). TEST AND CONTROL MATERIALS Identification The materials were identified and described as follows: Identification Physical Description T-6684 (test) off-white liquid DMSO (control) Clear colorless liquid Dimethyl sulfoxide was manufactured by Mallinckrodt Chemical. 9 005810 Covance 6329-200 Purity and Stability The Sponsor assumes responsibility for test material purity and stability determinations (including under test conditions). Analysis of the test material mixtures for concentration, homogeneity/solubility, and stability was not conducted. The purity and stability of the control material were considered to be adequate for the purposes of this study. Storage and Retention The control and test materials were stored at room temperature. A reserve sample of each the control and test materials were taken and stored in a freezer set to maintain a temperature of -20C 10C. The control material reserve sample will be retained at Covance for one year. The test material reserve sample will be sent to the Sponsor. Any unused test material will be returned to the Sponsor. Any remaining control material may be used for other testing and will not be discarded after issuance ofthe final report. Safety Precautions The control and test material handling procedures were according to Covance SOPs and policies. TEST SYSTEM Test Animal Adult albino rabbits of the Hra:(NZW)SPF strain were received from Covance Research Products Inc., Kalamazoo, Michigan on April 16,1997. Housing After receipt, the animals were acclimated for a period of at least 7 days. During acclimation and throughout the study, the animals were individually housed in suspended stainless steel cages. Environmental controls for the animal room were set to maintain a oossii 10 Covance 6329-200 temperature of 16 to 22C, a relative humidity of 50% 20%, and a 12-hour light/12-hour dark lighting cycle. In cases where variations from the required temperature and humidity conditions existed, they were documented and considered to have had no adverse effect on the study outcome. Animal Diet The animals were provided access to water ad libitum and a measured amount of Laboratory Rabbit Diet HF #5326, PMI Feeds, Inc. The feed is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. Samples of the water are periodically analyzed. There were no known contaminants in the feed or water at levels that could be expected to interfere with or affect the results of the study. Selection of Test Animals The animals were identified by animal number and corresponding ear tag and were placed into study groups using a stratified body weight randomization program. The randomization body weights were obtained on Day -5. Study Design Animals weighing from 2,067 to 2,500 g and approximately 15 weeks of age at initiation of treatment were placed into the following study groups: Group 1 (Control) 2 TeCstoMntarotelr/ial SDTuiml-f6oe6xt8hi4dyel Dose Level Dose Volume Number of Animals (mg/kg/day) (mL/kg/day) Males Females 0.0 0.1 33 3.0 0.1 33 Justification for Species Selection Historically, the New Zealand White albino rabbit has been the animal of choice because of the large amount of background information on this species. 11 005312 PROCEDURES Covance 6329-200 Preparation of Exposure Area On the day before the first control or test material application, the back and, if necessary (to obtain unblemished skin), the flanks of each rabbit was clipped free of hair with an electric clipper. The clipped area made up approximately 20% of the total body surface area. The test sites (intact skin) were inspected for interfering lesions, irritation, or defects that would preclude the use of any of the animals. The animals were clipped as needed throughout the study (Days 3, 6,13 and 29). Dose Administration All animals received five consecutive days of administration ofthe respective control or test material. The first day of treatment was designated as Day 1. Group 1. An individual dose was calculated and measured based on each animal's body weight on the first day of treatment. The control material (dimethyl sulfoxide) was applied to the test site at a dose volume of 0.1 mL/kg in a thin and uniform layer. Group 2. An individual dose of the test material mixture (T-6684 and dimethyl sulfoxide) at a concentration of 30 mg/mL was calculated and measured based on each animal's body weight on the first day of treatment. The test material mixture was applied to the test site at a dose volume of 0.1 mL/kg of body weight in a thin and uniform layer. Each area of application in Groups 1and 2 was covered with a 4-ply 5-cm x 5-cm gauze patch. Each gauze patch was secured with paper tape around all edges and overwrapped with Saran Wrap and Elastoplast tape to provide an occlusive dressing. Collars were used to restrain the animals during each approximate 23-hour exposure period. One Group 2 male was found without its collar on at the a.m. mortality check on Day 3. The collar was placed back on the animal at this time (see protocol deviations page). Approximately 23 hours after each control or test material application, the restraining collars and patches were removed and any residual material removed from the application sites using tap water and disposable paper towels. 12 O05S13 Reason for Route ofAdministration The dermal route is a potential route of exposure in humans. Covance 6329-200 Observations of Animals Clinical observations were conducted predose on Day 1 only and at approximately 1,2.5, and 4 hours after each control or test material application on Days 1through 5. Additional clinical observations were,conducted daily thereafter through Day 29. Mortality checks were conducted twice daily on Days 2 through 29 (a.m. mortality check only on Day 29). Body weights were determined on Day -5 for randomization purposes, before the first control or test material administration (Day 1), and at the scheduled sacrifice interval (Day 29). The initial dermal irritation reading was made before the first control or test material application according to the Draize1technique (recorded as the Day 1reading). Subsequent readings of dermal irritation were made within 30 to 37 minutes after each patch removal (Days 2 through 6) and on Day 13. Blood Sample Collections/Shipment A blood sample (approximately 4 mL) was collected from a marginal ear vein of each animal on Days -3, 8,15, and 23. In addition, at the time of necropsy (Day 29), approximately 20 to 40 mL of blood was obtained from each control animal and approximately 20 mL of blood was obtained from each Group 2 and 3 animal via the posterior vena cava. All samples were stored at room temperature until centrifuged. After centrifugation, separate samples of serum and cellular fractions were obtained and stored in a freezer set to maintain a temperature of -20C 10C until shipped to the Sponsor. The serum and cellular fraction samples obtained on Days -3,8, and 15 were shipped frozen (on dry ice) to the Sponsor (James D. Johnson, 3M E.T. & S, Bldg. 2-3E09, 935 Bush Avenue, St. Paul, MN, 55106) on Day 16. The serum and cellular fraction samples obtained on Days 23 and 29 were shipped to the Sponsor the day after the experimental (in-life) date of termination in the same manner as the samples obtained prior to Day 23. The Sponsor is responsible for the retention and disposition of the 13 0GSS14 Covance 6329-200 samples. Covance does not accept any responsibility for the analysis of the samples collected in this study nor are these results presented in this report. Pathology At termination of the in-life experimental phase (Day 29), animals were anesthetized with sodium pentobarbital, bled via the posterior vena cava, exsanguinated, and necropsied in random order. The sites of control or test material application were washed with warm tap water before the necropsy procedure. All animals were subjected to an abbreviated gross necropsy examination and any abnormalities were recorded. The whole liver, bile, an approximate 1-cm x 1-cm section ofthe dermal application site, and both kidneys (collected as one sample) were collected from all animals, weighed (volume only determined for bile), and placed in a freezer set to maintain a temperature of -20C 10. After necropsy, the animals were discarded. Shipment of Bile and Tissues One week after in-life experimental termination, the tissues (whole livers, dermal application sites, and kidneys) and bile were sent frozen (on dry ice) to the Sponsor (James D. Johnson, 3M E.T. & S, Bldg. 2-3E-09, 935 Bush Avenue, St. Paul, MN, 55106), along with documentation of their corresponding weights or volumes. The Sponsor is responsible for the retention and disposition ofthe samples. Covance does not accept any responsibility for the analysis of the samples collected in this study nor are these results presented in this report. Statistical Analyses No statistical analyses were required by the protocol. Location of Raw Data, Records, and Final Report The raw data, records, and an original signed copy of the final report will be retained in the archives of Covance in accordance with Covance SOP. 14 005S15 RESULTS Covance 6329-200 Body Weights Individual and mean body weights are in Table 1. All animals, with the exception of one control animal, exhibited a weight loss from randomization to initiation (in-life) which can be attributed to the acclimation of the animals to restraining collars conducted for 3 days (approximately 21-23 hours/day) before the initial control and test material administration. All animals gained weight from Day 1to Day 29. Clinical Observations Individual clinical signs are in Table 2. All animals appeared normal throughout the study with the exception of one Group 1 animal and two Group 2 animals which had decreased feed consumption for one to three days within the first nine days of the study. Dermal Irritation Individual dermal irritation scores are in Tables 3 and 4. Slight erythema, edema and Assuring reactions were observed in one control animal while slight to moderate erythema and edema and slight desquamation reactions were observed in the other five control animals treated with dimethyl sulfoxide. Ofthese five control animals, two animals also exhibited slight coriaceousness reactions. In the animals treated with T-6684, one exhibited slight erythema, edema and Assuring reactions, one exhibited slight erythema and slight to moderate edema reactions, one exhibited slight to moderate erythema and slight edema and Assuring reactions, one exhibited slight to moderate erythema and slight Assuring reactions, one exhibited slight to moderate erythema and edema and slight desquamation and coriaceousness reactions, and one exhibited slight to moderate erythema and edema and slight desquamation, coriaceousness, and Assuring reactions. 15 005816 DISCUSSION Covance 6329-200 The acute systemic absorption/toxicity and relative skin irritancy of T-6684 was evaluated in male and female albino rabbits when administered dermally for five consecutive days. Application of this material did not result in any test material-related changes in body weight gain or macroscopic findings at necropsy. All animals appeared normal throughout the study with the exception of one Group 1 animal and two Group 2 animals which had decreased feed consumption for one to three days within the first nine days of the study. Slight dermal irritation was observed in one control animal while slight to moderate dermal irritation was observed in the other five control animals treated with dimethyl sulfoxide. In the animals treated with T-6684, slight dermal irritation was observed in one animal while slight to moderate dermal irritation was observed in the other five animals. 16 005817 SIGNATURE Covance 6329-200 F. Bud W. McDonald Study Director Acute Studies Date REFERENCE 1. Draize, J. H., "Acute Dermal Toxicity (Single Exposure)," In: Appraisal o fthe Safety ofChemicals in Foods, Drugs and Cosmetics - Dermal Toxicity, Association of Food and Drug Officials ofthe U.S., pp. 54-56 (1959). 17 00S81S Table 1 Individual and Mean Body Weights (g) Covance 6329-200 Males Animal Random Number -ization Day 1 29 Females NAunmimbaelr Random -ization 1 Day 29 Group 1 (Control) - Dimethyl Sulfoxide (0.0 mg/kg/day) FFF666333000111132 222,,,344134570 222,,,523087807 222,,,775602869 FFF666333000111897 222,,,342682304 222,,,322374374 222,,,776851359 Mean 2,397 2,388 2,668 Mean 2,356 2,285 2,719 Group 2 - T-6684 (3.0 mg/kg/day) FFF666333000111564 2,404 22,,448457 22,,440424 2,390 222,,,877025524 FFF666333000222120 222,,,243963528 2,067 22,,342219 22,,580064 2,735 Mean 2,445 2,412 2,760 Mean 2,365 2,272 2,682 00SS19 18 Table 2 Individual Clinical Signs Covance 6329-200 Animal Number Sex Observation Day 1 Day 2 Day 3 Day 4 Day 5 Hour Hour Hour Hour Hour Day(s) I 2.5 4 1 2.5 4 1 2.5 4 1 2.5 4 1 2.5 4 6 7 8 9 10-15 16-29 Group 1 (Control) - Dimethyl Sulfoxide (0.0 mg/kg/day) F63011 M Appeared normal / / / / / / / / / / - - - / F63012 M ADcpeocpnreesauarsmeedpdtnifooornomdal / / / / / / / / / / / - F63013 M Appeared normal / / / / / / / / / - F63017 F Appeared normal / / / / / / / / / / / F63018 F Appeared normal / / / / / / / / / / / F63019 F Appeared normal / / / / / / / / / / / / / / Group 2 - T-6684 (3.0 mg/kg/day) F63014 M Appeared normal / / / / / // / / / F63015 M Appeared normal / / / / / - - - / / / / / F63016 M ADcpeocpnreesauarsmeedpdtnifooornomdal / / / / / /// / / // / / / F63020 F Appeared normal / / / / / - * - / / / / / - F63021 F ADcpeocpnreseauarsmeedpdtnifooornomdal / -// // --/ -/ / / F63022 F Appeared normal / / / / / / / / / // / - / / CCoonnddiittiioonn nexoitsetevdid. ent. 19 005820 Covance 6329-200 Table 3 Individual Dermal Irritation Scores Group I (Control) - Dimethyl Sulfoxide (0.0 mg/kg/day) Dermal Reaction ADEECrdoetyoesrtinqmhaiuceaaaemmoauastnioenss Fissuring AEEDCrdoteyoesrtniqmhaiuecaaamemoauastinoenss Fissuring Erythema AEDFCidotesoesrsniqmuaiurcaaiaenmoguastnioenss StuMdayleDsay 1 2 3 4 5 6 13 Animal No. F63011 000000 000000 000000 000011 220000 220000 220001 Animal No. F63012 0 00000 00001 0 000001 00001 0 00001 0 000011 00011 1 Animal No. F63013 000000 000001 000001 000011 200001 220000 000111 Females 1 2 Study Day 345 6 13 Animal No. F63017 000000 000000 000001 200001 220001 220000 000111 Animal No. F63018 00000 0 00011 0 00011 0 00011 0 200001 . 220001 20o011- Animal No. F63019 000000 000011 000011 000011 200001 220000 220001 20 005821 Table 4 InGdirvoiudpua2l-DTe-r6m68al4I(r3r.i0tamtiog/nkgS/cdoarye)s - Covance 6329-200 Dermal Reaction ADEECFirdoetsyoesrstniqmuhaiurecaaiaemnmogauastinoenss EADFECidroetsyoesrstniqmuhaiurceaaiaemnmogauastinoenss AEEDFCirdotesyoesrstnimquhaiurecaaiamenmogauastnioenss 1 2 S3tuMday4leDsay5___6____13_ Animal No. F63014 000000 0 00000 11 00 00 000011 2001 00 21 00 00 001 000 Animal No. F63015 000000 000001 000011 000011 000011 000011 000111 Animal No. F63016 0 00000 01 0000 001 000 000001 011 000 21 0000 11 0001 __1____2__ S3Ftuedmy4aDleasy5 6 .13 Animal No. F63020 0 00000 0000 00 0001 00 2 01 000 220 000 220 011 0111 00 Animal No. F63021 000000 000011 200001 200001 202000 200201 200011 Animal No. F63022 0 00000 00000 0 00011 0 11 0000 2001 00 22000 0 000111 21 005822 APPENDIX A Individual Animal Pathology Data Covance 6329-200 22 005823 Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 1 DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 DATE AND TIME OF NECROPSY: 06/02/97 8:59 PROSECTOR: DAVID SCHUETTE POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER TERMINAL BODY WEIGHT: NOT ENTERED RECORDER: KEVIN BILLINGS WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 1.2216 15.3938 67.8896 0.5000 ------- -...... WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * * * GROSS PATHOLOGY OBSERVATIONS * * * SEVERITY, KEYWORD (S) OR PHRASE FREE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) .................... -NO MACROSCOPIC LESIONS u>.NJ, --------------------------------A-B-B-R--N-EC"PERF-O--R-M-.------ 005824 I Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 2 ANIMAL NUMBER: F63012 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 8:35 PROSECTOR: DAVID SCHUETTE RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 1.1614 16.1562 61.6643 0.5000 ------..... . ....... ....... -------...... .... . -...... WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * * * GROSS PATHOLOGY OBSERVATIONS ** * SEVERITY, KEYWORD (S) OR PHRASE FREE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) .................... -NO MACROSCOPIC LESIONS A.rO,, ...........................................-.A..B..B.R...N..E..C...P..E.R..F.O..R..M........... 00582S Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 3 ANIMAL NUMBER: F63013 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 9:05 PROSECTOR: JILL PAUS RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 0.7738 15.1059 73.3509 0.6000 WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * * * GROSS PATHOLOGY OBSERVATIONS * * * SEVERITY, KEYWORD (S) OR PHRASE FREE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) .................... -NO MACROSCOPIC LESIONS .N , -ABBR NEC PERFORM. U ------------------ ------------------------------ 005826 Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 4 ANIMAL NUMBER: F63014 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 8:17 PROSECTOR: JILL PAUS RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 0.4833 12.5487 68.0339 0.9000 -...... ..... . WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * * * GROSS PATHOLOGY OBSERVATIONS * ** SEVERITY, KEYWORD (S) OR PHRASE FREE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) -NO MACROSCOPIC LESIONS K> -ABBR NEC PERFORM. On 005827 Com ing Hazleton Inc. Madison, Wisconsin USA Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 5 ANIMAL NUMBER: F6301S SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 8:25 PROSECTOR: DAVID SCHUETTE RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY <KD) LIVER (LI) BILE (VOLUME) (BI) 1.0044 15.9025 74.7248 1.9000 ....... .... -- - WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * ** GROSS PATHOLOGY OBSERVATIONS * * * SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) .................... -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. 005S28 Coming Hazleton Inc. Madison, Wisconsin USA Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 6 ANIMAL NUMBER: F63016 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 8:56 PROSECTOR: JILL PAUS RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT ( V ) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 0.6073 14.9890 61.8841 0.6000 WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * * * GROSS PATHOLOGY OBSERVATIONS * * * SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) .................... -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. 005829 Coming Hazleton Inc. Madison, Wisconsin USA Appendix A . Individual Animal Pathology Data Covance G329-200 PAGE: 7 ANIMAL NUMBER: FG3017 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: S TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 8:47 PROSECTOR: DAVID SCHUETTE RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 0.9965 14.5047 55.4966 1.4000 ..... . -...... WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * * * GROSS PATHOLOGY OBSERVATIONS * * * SEVERITY, KEYWORD (S) OR PHRASE FREE-TEXT COMMENTS AND NOTES to GENERAL COMMENT (GC) VO -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. 005830 Corning Hazleton Inc. Madison, Wisconsin USA Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 8 ANIMAL NUMBER: F63018 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 8:14 PROSECTOR: DAVID SCHUETTE RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 0.8229 13.4273 63.0821 1.6000 WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME *** G R O S S P A T H O L O G Y O B S E R V A T I O N S * * * SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES OU) GENERAL COMMENT (GC) -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. Corning Hazleton Inc. Madison, Wisconsin USA Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 9 ANIMAL NUMBER: F63019 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 8:23 PROSECTOR: JILL PAUS RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 0.6150 14.0486 51.5804 0.6000 .... ... ------...... ------- ...... ------....... ------- WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME *** G R O S S P A T H O L O G Y O B S E R V A T I O N S * * * SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) .......... ........ -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. 0SS32 Com ing Hazleton Inc. Madison, Wisconsin USA Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 10 ANIMAL NUMBER: P63020 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: S TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 8:49 PROSECTOR: JILL PAUS RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT ( V ) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 0.6370 13.4440 52.3928 0.8000 ---- -........................ WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * * * GROSS PATHOLOGY OBSERVATIONS * * * SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES U> GENERAL COMMENT (GC) -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. PC8S00 Com ing Hazleton Inc. Madison, Wisconsin USA Appendix A Individual Animal Pathology Data Covance 6329-200 PAGE: 11 ANIMAL NUMBER: F63021 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 8:37 PROSECTOR: JILL PAUS RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 0.8344 17.0195 53.0880 1.5000 .... --- ------- WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * * * GROSS PATHOLOGY OBSERVATIONS *** SEVERITY, KEYWORD (S) OR PHRASE FREE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) .................... -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. 00S834 Appendix A Individual Animal Pathology Data Covance 329-200 PAGE: 12 ANIMAL NUMBER: F63022 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT ENTERED DATE AND TIME OF NECROPSY: 06/02/97 9:07 PROSECTOR: DAVID SCHUETTE RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ORGAN NAME ABSOLUTE ORGAN WEIGHT (GRAMS) ORGAN WEIGHT RELATIVE TO BODY WEIGHT (%) ORGAN TO BRAIN WEIGHT RATIO ORGAN STATUS DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 1.0944 14.2647 60.3910 1.6000 WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN WEIGHT TAKEN ORGAN NAME * * * GROSS PATH OLOGY OBSERVATIONS * * * SEVERITY, KEYWORD (S) OR PHRASE FREE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. - scssoo APPENDIX B AmendmPPernorottotNoccooo.llD1TTePvo6ia7Tt9hio8enProtocol Covance 6329-200 35 S 8 3 6 Protocol Deviation Covance 6329-200 Protocol PrABaa.bdgDbmeioti7ssne,wis7Ait.lrldEabmtxieopinnceoi,rsilNtmlariarenetnitdohtadnSlu,eDr(in3net)gseiDngecoanecs,:eh The approximate 23-hour application period. Tsthuidsyd. eviation is not considered to have Actual Procedure tmrAwfieomnaomusirenmat.pdainallTawiletchydNietehdcoinpho.baeupFatctl6ccakih3kctso0eaon1cpno6opDntleh(lataaGehyrrreeao3odna.nuiptpmoaTpt2ahlhitlecahMavaectteaoiatlolh.elmn)aisr.switaes an adverse effect on the outcome of the 36 005837 Covance 6329-200 COVAfTC^ THE DEVELOPMENT SERVICES COMPANY Covane LaboratoriesIne. P.O. B o x 7545 Madison, W isconsin 53707*7545 Packages: 3301 Kinsm an Boulevard Madison, W isconsin 53704 Tel: 60B/241-4471 Fax: 608/241*7227 Sponsor: St. Paul,3MMinnesota PROTOCOL TP6798 Study Title: 5-Daily Dose oDfeTrm-6a6l8A4bisnoRrpatbiobnit/sToxicity Study Date: April 18,1997 Testing Facility: MC3a3od0vi1asnoKcnei,nWLsmaibsaocnoranBtsooirnuilees5v3Ian7rc0d.4 Testing Facility Project Identification: Covance 6329-200 THE AMERICAS EUROPE ASIA/PACIFIC AFRICA 37 005838 Covance 6329-200 Covance 63T2PP9a6-g72e90802 STUDY IDENTIFICATION 5-Daily Dose DofeTrm-6a6l8A4bisnorRpatibobnit/sToxicity Study Covance No. 6329-200 Test Material T-6684 Sponsor T33PSMMMt.o.OxPe.iCdcaBeiouconllao,xtlegM3rDy,3NeBS2pe2lad5r0rv5gtim1.c32ee32sn-0t3-222E0-02 Sponsor's Representative T(PS33R6MMMt.ooO.1xg2Pe.iCe)dcaBreiou7conGll3a,oxt.3legM-rP3Dy3,3Ne2eBS2r2pk2el2adi5r0nrvg5tsi.1m,c32ePe32snh-0tD3-22,2ED0-A02BT Study Director 6CMFP0..oOa8Bvd..a2uiBsn4doc2onWe.x7,L9.7Wa0M5b1I4oc5Dr5a3ot7on0rai7led-s7I5n4c5. Testing Facility C3M3oa0vd1aisnKocnien,LsWmabIaon5raB3t7oo0uri4leesvaInrdc. PEEDroxxrpappofeetsrreRiimmdeeepSnnotuttraatdllyDTSTatetairemrmteiDntaaabtteiloen Date WWWeeeeeekkk ooofffJJMuunlayye 1254,,11,1999999777 38 005839 Covance 6329-200 Covance 63T2PP9a6-g72e90803 1. S5-tDudayily Dose Dermal Absorption/Toxicity Study in Rabbits 2. mTPouartaepsrosiaeslesswthheensaypstpelmieidctaobtshoerpstkioinnoafnrdabtobxitisciftoyrafnivderecloantisveecustkivineidrraiytasncy of a test 3. TtnPReorhseatitgcsbmtueislcataucetteodoRrnyiredaywgluCucmilltloaeixbtdmite:oupncrlesoi/asnSndftcoauenrcdcteoadrndcisne/nGatcruacitodireodlnian,nescoselwuwbiitithlhittythh,eeheofxomclleoopgwteiionnnegitthGy,aotaoandndaLslatyabsboisirlaiottyfortwhyeill [[[X]]] 42C10onCCdFFuRRct51a86s0(aF(NEDPoAnA)r-eFgIuFlRatAed) Study [[[[ ]]]j N4C509o(8tNCi1fFoi)c3hRa0St7ai(o9Fnn2iNnN(aoElo.)Ps3(.AO83-5E1T03CS(DCaJna)Apd)a8n7e0se(JMapAaFnFes)e MOHW) AuRnellngepucrleaostciseoadnrusil.ryesIdnuintphlteihcioasptepinraoinotoyncpoorlfeatvhrieeouiSnspwcoonomsrokpr.liaanndcestwudityhdtihreecAtonri,mthael WstuedlfyardeoAesctnot 4. AOTQhspuseeaurplrairattoiynntcogAecPsoUsrluno, ircstateunidndcuyeareccscoo(nSrddOuacPntcs,)eaanwndidtthhpeoClofiicvniaeansl.creepLoarbtowrialtlobrieeasuIdnict.ed(CboyvathneceQ)uSatlaintydard 5. Test Material A. TId-6en68ti4fication B. OPhffy-swichaitleDlieqsucirdiption 39 0 0 5 8 4 0 Covance 6329-200 Covance 63T2PP9a6-g72e90084 C. TtP(faiohunkrecrecilnSuto.ydpniocanenngnsdtourrnSatadtiasoesbrnui,tlmeihsteoytsmcrooengsdpeinotienoisntiybs/)is.loitlSyuabfmoilrpitplyeu,sraiontyfdtaesnstadtbmsitlaaittbyeirlaiiantyla/vldyeeshteeiscrmlweiimnllaintxiootutnrbsee(s) D. SRtooormagteemperature E. RReesseerrvveesSaammpplele(ss) ofeach batch/Iot oftest material will be taken for this study. Tcmohameinpttelaesittnimoanatteoemfritpahleerraeintsu-erlrievfeeopfsah-2ma0spelCeo(fst)h1we0islCtlubdueyn.stitlorreedtuarnteCdotvoanthceeSinpoanfsroerezaeftresret to F. ARneytenuntiuosned test material will be returned to the Sponsor. G. ASasfreetyquPirreedcabuytiCoonvsance SOPs and policies 6. Control Material A. DIdiemnetithfiyclaStiuolnfoxide (DMSO) B. CPhleyasri,ccaollDorelesscsrilpiqtiuoind C. TPourbietydaoncudmSetnatbeidlitbyy Covance (information from the supplier) D. SRtooormagteemCpoenrdaittuiroens 40 005841 Covance 6329-200 Covance 63T2PP9a6-g72e90580 E. RReesseerrvveesSaammpplele(ss) of each batch/lot ofcontrol material will be taken. Ttohme caoinnttarionl amtaetmerpiaelrarteusreervoefs-2am0pCle(s1)0wiCll.be stored at Covance in a freezer set F. dARinesctyeanrredtmieodanianfitnegr icsosunatrnoclemoaftethriealfimnaalyrebpeourts.ed for other testing and will not be G. ASasfreetqyuPirreedcabuytiCoonvsance SOPs and policies 7. Experimental Design A. Animals (1) SRpaebcbiiets (2) SHtrraa:i(nN/ZSWou)rScPeF/Covance Research Products Inc. (3) AAgdeulatt Initiation (4) W2.0eitgoh3t.0atkIgnitiation (5) 6Nmumalbeseranadnd6Sfeemxales (6) IInddeinvtiidfiucaaltniounmbered ear tag (7) Husbandry (a) HInoduivsiidnugally, in suspended stainless steel cages 41 005842 Covance 6329-200 Covance 63T2PP9a6-g72e90806 (b) AFfFoeormeonddeusa,tsrIuintriceo.d)n.aalTmchooeumfnpotooondfeiLnstarsboaounrtaidntoeernlyyviRarnoaanblmbyizetenDdtaibelytcHothnFeta#mm5a3inn2au6nfa(tsPc.tMurIer (c) ApeWnedvrailitoirebodrinitcumamlelnyftraaonlmacloaynnzteaadmutfioonmransapttsie.ccisfyiestdemm.icrSoaomrgpalensisomfsthaendwater are (d) TiCnhoteenrrfteaeramereiwnnaiotnhktstnhoiswsntucdoyn.taminants in the food or water that would (c) aaiEEnnnnttdeevvmriairrrupoo1penn2rtmea-mhdteuoendrunteuartelolicftgooh1nit9tn/r1-oC2lli-sftheofoop2urr3rotdhcCeaedr,akuanrciermyesc.laalleti.rvoeTohmheumwdaiildrlkitbyceyoscfelet5t0mo%amyab2ine0t%ain, (f) AAtccleliamsta7tidoanys (8) SanppBdolreaaoelscaegqeencruddaitamimootiennantwlnohoiuientfsmahtToulibtnbedhevsynratiiongonAurdfeonsebdluixymoaptdyrspaaysoulsaooswnrifneihsmgiteguaaahdlltsstyhtawrciaincistliioltpfiribladeatediicnopebgnduo.trdoocynhCawtoseevesdtia.gnshTcotehrtheaSnaOatdnPoismm. aiAzlsantwioinll be (9) JscHpuhiesoscttiioicfeerisicc.baaetlciloyaun, sthfeoeorNfStehpweecZliaeersgaleSaenalmdecoWtuiohnnitteofablbaicnkogrraobubnidt hinafsobrmeeantitohneoannitmhiasl of 42 00S843 Covance 6329-200 Covance 63T2PP9a6-g27e09087 B. Dose Administration (1) Test Groups Group TeCstoMntarotelr/ial 1(Control) 2 DMSO T-6684 Dose Level (m g/kg/day) 0.0* 3.0* Number ofAnimals M aks Females 33 33 * To be administered at a dose volume ofO.l mL/kg ofbody weight (2) TPbraTOirarahhrrnebcieeeatbkptahitawtatraineeorniwialadnmdltita,,lmicalyooiolebfrsnnbnendsewtoetecfisicoffltliieeulrtEpesectbpsxetstesaeph(ardtcieoyphnlspfiait(rupnartteopocirwetetxeioaodoisAbmlkufattirlhaaesndetias)aenntiplrweyrauewenindl2cdlbiel0tulhbdc%edeomatenhinointtrsfrhoeshotpeluheleegdeucmchtsttsroeaeoikutcdtoeiatnrcfflti)olaha,birlpnetohpiyadsneeptytourpef.fdslrliaufytTcehnr.arehfkitaesniacognosenhifl,maaeertvsheaaiealecosd.hn. s, (3) awtteapo2rApbaoDrahoexnpac3evrdikloesoedptcptle-dpoceshpmhlorlaaehnuee2siwaonsncDesc.s)ruAutioariieitemvwMmgarvirtnAdmdveiopneaoiaaleSmDllpfappillnldlurnsOetscepaepirembdsonryeilsaawdeianiahnsiettcwttnsices1atcuahGlertoihdsilotrrbnoeitaierfneamhraDegloacose0tcfph.nuaouipSac.ohwo1pnnrpyeladTrpelndtiisedtriaemivlthe1aelaleyrut1ese.ndyihLlabtnoyoa.rfedTe/WaidmiankffmvcthT.fbsodgaaroieaeeibrhra.dtTvnc2msecpidecmetiutThro*ososwritraeilnthaeenseanawrdilseesdtapdlnyimretlsiatrdevoGlwcitwembpliorrolunpE.repainbilatmtoiattullih.esbTthiiauvremoetaeascpreahrpdtntdsaoieeeoa4.2edatrltpptidlo-olaaieo/Taatpleyrvfesanolrterlhstessh0eynaweattthm.deoa*1n5otimifpfhcdslidt.iflxmea0aalreaueastmbadp-traLutpaecpeiemmrernrnm/pdpoekiatigiilaalrousmxgnipieylxcene.tipwspaauswdadop5ltrltrTiicr'iei.ofleetsao0hlalohcvdtlltn-brwletbiiacbteioodueehvpm(iedadnelndGeeptlsoyddmgratorgsabooiieonewralleferxsuueusuneteiattphazdtmfeeinesoegodwdaadrhf1ttiatletl 43 005844 Covance 6329-200 Covance 63T2PP9a6-g27e09088 (4) TRheeasdoenrmfoalr rRoouuteteisoafpAodtemnitniailstrroauttieonofexposure in humans. (5) AtrowRhefepaemtmtpeeporsroaovttovxecandaihnmledDuosascatfiaeoynTnlngy2det9rws2cot3opalMtlrmheliaooraarruotsteertrwosrraiiaiaatnlhlfllteaaebpprtepipesrrlsaeoiuccmpeharotictiaveootesneltdleswocuaritnblicldosotbnatnehnptercwrooerla,ecsmseihifddeanuudtereaecrwl.ieamisltshaaatprweypra.ilraicmTlahwttieaiolpsnli,btees C. Observation of Animals (1) dwCBaDDnaehaladiifyylenoyns4i2rce(91dhaa-ioc.l5(rmouaeOn.frc.mosbttarers.aondceflmdtlrbeiovopnyrra.riemcttttaehaiao.lscel)itnhtssyfmsiotaguoradndntmmseyl,ryoiiddnaroiaitlrsnaiealtlyrciDdattymottahiryosi.entnr2aiesr9oatt)firf.natcegtoOironobfntnors,oreDalrctvalaiaaynnptdii2pcotarnaeolsnsxsdtmiimmgcanoaaysntt,eebtlriaeyninauedl1ixno,tt2wgen.ni5tcdo,eed (2) eBiS(wRrAatreeiucittfathhdtoaadiyrtncaiiennohD3timnghm0aeyeroatenoftia1.tnDd36i1i.t0eni)a.rgImlmnAwcdianodiilvunldlitItiderrbtoisureoilaatnmloafatdrtaleiedtordereneemsartamacmnlhdaailprtrreaeiirrtctricaoaihttlriadoratienedodmmnreaoircsnveoiatarshlddter(isanDDtwgiaoasiylynwls,b12itleh-lr6eemb)aeddaaeiminnrndmgatadoaienlned for (3) (FBDooardyrya1nW)d,oeomingihDztaastyio2n9,,boerfoartedtehaethin(iwtihalentessutrovrivcaolnetrxoclemedasteIridalaya)pplication (4) Blood Sample Collections (a) FPatedrrreemmdqioiunnseiaesntti(rcoaaynntiyo(Dtnimatoye2Df9rao)ym1u),potnoDfoauyrsd8a,y1s5b,e2f2o,reanthdeaitneixtipaelrdimoseental 44 00SS4S Covance 6329-200 Covance 63T2PP9a6-g72e90809 (b) mBDMlaaoeyrtogh2dion2sda.almoefapCrlevosel(ilneapcotpifroaonlxl/iNamnuaimmtelbayles4raomtftLhAeneaiinmcthea)rlvswaillslpbreedcoolsleecthterdoufgrohm a bvbF(Dfrileorofaoolympooumbodm2ets9tawesah.iicnebpihlelolpedscbo)so,efsinrtbaooetplrrbmepoitorwlaeoriaianxvlncelieimhdmbneaaafarntolceoiablsmmyvataaca2eilr,n0aisafecaiptdhcocp)erG,4rdiofa0rioxnocmniduemdpLDaapi2atonpeyaflraynob2mix9l2moi0om(oatrdmhlaibetswLeuamlnicyolrdlaif2xfbcbi0iecolmoenmodoudbLmditotainwoionfnielld tdTastcffherreeihoarfsernmrepcuzetSteormibeiipnnosfzlioaouneaottnoigsnrniosodedsonoddnebscr,rt.oateyoaatmfloTinnlitnucphdhemeleleeadtetaohtrssSoiteanfhtpwmrwttrosahaioeonpcliepulntslwieaSgobo-srhpere2na.eoi0stDssekntooassrCsbyeroepetsrrardp1uiwi5o1onmani0retwtsdhtaroiiCoinlbanlod.lifenbmtoTec-enfrelhotieslefDelreemuwntaslhttepyeaeeerrefrur1kmrrfo5ameraiztatnwufeecatanrnteinetiltriodlioooaninbncnnnse.ed-dlsalsriltTntfyeuohedhnlreiaeetncrde in to The serum and cellular fraction samples will be shipped to: J93BSa3Mtl.m5dPgeEBa.s.uu2TDls-.,3h.&MEJAo-NS0vh9enn5suo5en106 JsahmipemseDnt. Joofhthnesosnamorphleiss.alternate will be notified regarding the D. Pathology (1) aAwUbinnnlylsocbrahmeneisamduluibatlijleeeddscytwSeindailgcltrobdifueairncrieenacsgboabtrhnrdeedevdsiD.tauteAedadynthigomsrroasslasscnirniefciacreomdposirynibeauxnmadmocriinobanutdinoidtniocanonnwddiialtllilobne 45 005846 Covance 6329-200 Covance 63TP2aP9g6-e72901800 pa(acmAvapnolefalopleiltclnresleaiei)trbccv,hdnutaaebeeintitldnilicedoaztrdfonbeocrdlovpoasemisin)watfy,deri,teaaithtatelhnihlenozeasdaenvopnra,ebdpinsnwomireiuaotmetahmxcitlgaoisakmlhpvisdmdeeaaywdnn,taieetanii(onlynvgl1btsdoa-bdacl(iereuuncmb.xrmodiaistlixnaealstegncle1ocagm(t-ntrvhucedlpiemidyaenesdraadista.sntueeetjtcudodeetryn.crieoemtoiTnoosrihnfnaoseme-afi2dncwpt0hrflthieeohfCo)iredclwbeeemirdillm1laielvi0r)nageb,lrieCaan,n.abdlileear (2) ecrabO(eSlev-sxoxic1cnlciamsehoa0omam)Dnerp,nidCedniuagasneujyet.nsueadelcadict2er.Antmi-tid9opaigoTfopn,tletnSeahnlet.nehdcareoe)ieTe,cnnfrwdrwahaaetitnhthecnfhiieilnderoieldcaomtledosnbempruaeieasalmblarynismcrvjf,daegorwetaloecilrhsnllietm,eleeawzalbcdlepaintbirlelenptueelodasilmmibea,r(caantewbvnaaleteletlnoesisariaeoitnsbgmhvcwn))bhera,iaaorsteilbeiinllipdnzatvldsbeeetbi(ia,edadlevaieatdlnodevwl)nl,idiidaausianatcbmghtbntanrerhoearomosaedtnosprohepdsmvdpndekoeinrla.urnimoyealdmaixctdntuioriceemeporratyespedevasnserwotaymetr(,foic(l-ib1ovln2-alibe0clareedmbcCifttxeoadlr (3) - TtaTawsmlhiantshiieacdsttteinehrhsriddneuisfcniieaesisocmrnocperSahnrospSaeseesleamsdeitcnpwthsuitpdooeiel(loneneldesdindkaevoSimaes7nafhtar.gfhpt,Ccithpslebrw.eeir(mi(fs4leisicec)ifeafo,g.ernma(lohdblptwemept)pscr.lilmetloitilsTchora.baehnlvbee.soalsceplSSeuh)ppnamewlomtdiceniuflpasrslltoloeiwebzodrseeinilsncslsasoebrocilneterlnetesif,spcdwihatcoreiniyentpdd.hspiicakTeabetndilhdeudteotnnofiessotncyahtrhhmsetthe)ehpSedecplpuoeesrolalsreelsnmtedaoescnennotdetridon E. NStoatsitsattiicsatilcAalnaanlaylsyesses are required. 46 005847 Covance 6329-200 Covance 63TP2aP9g6-e27901801 8. ARefpinoarltreport including those items listed belowwill be submitted. PDTTDDDGGInraaareeredobbootsssiceuuscccsvesssrrrllidaaiiidoppppputtuiiaaftttrooaiiitteeooolhhnnxsnnnooapoollnoooeooffirfffmggmmiattmyyhhaneoeelrefayreitntnnttaipteetsdolaobsssiixltrtotunytiidasegcn(ynydsiwiesdatfwftitereafcaeieemtogiqcbionhgtuysntthers/stsdoaseatelnexnbrdmdmdyatabnaebstydlreeimlxrietdirhaiarovneilnstsoaaedSlttuitiloduoemondvnesyeeslDleivreecltor) 9. tptrtsdOfiLhhhneitienooreeestcruiaprcligasCfruoloaiintgddnrntsuoeieiaaavdfonoplellfayngoefefdrnroeedeotcwate.hpnuetfiiaooreswlIRAi,rslfnyctieoab,rltgoelchrweiaamtheibclertclioespDmvShfoploepcapeasfislhrroettltgoleaaosnhfiedgr,sowestgd,trrhRoeeei.acerandsdeforngscTo.cenbhrohaoasteeornreidflmogod,oddSlnsswweibe,pitmniniseoRolwtgalfintnooetiobseaolsnrloerlehefeirbrpaatadahvemlvevcemerearrcaieeieofSayClstipadtaeanoeebtirmaarnovlvlleineeafaepcrcdnltealtessiectpmait(ewoonoCmsefrd)ihtotpl,htia.lavhslaenveapbeOnd(neaoesdfrncsC)tisceehen,eFhoaeantoyivilolvntmelfareaatfntoeoasharlpcrtceioaeetoRgihfrlfrmiwteeiitneCta.paaiarSloltsotWllepesvrripcroatgeahihannatunepaaslcdinsrieoneing,rtrgteieafhdnaodoeaangrftidana,a ASISSFPDnOBRDairthno-nuomilaaobsoadptietnodfsslpypoeeedcylprimreoconerarvwolepmgeicdaccrpoaeomtrraioraniizelelgotrlirdacsednhanp(ptocsoititpsaorsisotrrtoditninhoorngsdaotnidoetrlniaoerncoaetcogclancoelysoraidrrgmdesncseeondrddcmsoepnyt)s 0S848 47 Covance 6329-200 Covance 63TP2aP9g6-e72901802 aTrhcehifvoeldlowwiitnhgtshuepsptourdtyindgartae.cords will be retained at Covance but will not be AARIWnennsafiittrmmreiugraamelalrenrraneoatcotloyermcsiaapilsnttie/bdramercfacpcrtoelieiorermdnzasetauartnriteodenmamnrpedaecirnhoaturtedumnsraiednriceteycorrerecdcosorrddss 00SS49 48 Covance 6329-200 PROTOCOL APPROVAL Covance 63TP2aP9g6-e72901803 3RSMpoognesrowr'.s PReerpkriensse, nPthaDtiv, eDABT ACFS.tocuBvudatuyendcDSeWtiurL.edaciMebtosocrDraotonraileds Inc. CRQoeupvaraleintsyceenAtLassatiubvroearnactoeriUesniItnc. 2 Date Date Date 49 O S 8 S o Covance 6329-200 Attachment 1 Scoring Scale for Acute Dermal Reactions Covance 63TP2aP9g6-e72901804 Erythem2301---a- NMSSeloiovgndeheerterate Edema2031---- NMSSeloiovgndeheertera((btreaari(esrleaydispemedrocarepeppttrhiobaxlneim1toamtwemley)ll1dmefmin)ed by definite raising) Atonia2031---- NSMMloiaognrdhkeeterda(stel(ing(oshlteoliwamstpriecatiiutrymrn)etnotnoofremlaaslt)icity) Desqua0231m----aNMSMtilooiaognnrdhkeeterd(ast(leipg(rshoctnasolceuasnlcianengdd)ffllaakkiensg) with denuded areas) Coriace0231o----uNsMMSnloaioegnrsdkheseetrda(dt(eetco(rlueegaahtsheaenirnydptbelrixiattbtulierlei)t)y) Fissurin2301g---- NMMSloiaognrdkheeetrda(dt(eecf(ricanrciatkecskcwsraiitnchkdsbelirenmeedispi)nidge)rmis) 50 OSSl Covance 6329-200 C O V T H tfc!?"' THE DEVELOPMENT SERVICES COMPANY AMENDMENT NO. 1TO THE PROTOCOL Covance Lab o rato ries Inc. P.O.Box 7545 Madison, W isconsin 53707*7545 Packages: 3301 Kinsm an Boulevard Madison, W isconsin 53704 Tel: 603/241*4471 Fax: 608/241*7227 PROTOCOL TP6798 5-Daily Dose DofeTrm-6a6l8A4bisnorRpatbiobnit/sToxicity Study Covance 6329-200 Sponsor Testing Facility T3SP3MMMt.oO.xPe.iCdacBeuioconllao,xtleMg3rDy,3NeBS2p2el5adr05rvgt1i.mc32e3e2sn-03t-222E0-02 3CM3oa0vd1aisnKocnien,LsWmabIaonr5aB3t7oo0uri4leesvaInrdc. Sponsor's Representative Study Director Roger G. Perkins, PhD, DABT F. Bud W. McDonald This amendment modifies the following portions ofthe protocol: Effective May 20,1997 TcporhonetdooucbcostlettrhoveamnDcoaevyeo2tf2hMebelDemeaodysr2iao2lnbDaleaheyodclsiodtioanycD,idameyso2dw3if.iyththDeayfo2ll2owofinthgetwstoudsye.ctiToonsavoofitdhehaving to 1. cCPhaoaglnleegc8et:,io7.nsE,x(pa)erFimreeqnuteanlcDye. sMigno,dCify. OthbissesrevcatitoionnwoitfhAthneimfoalllso,w(4in)gBulonodderSlinamedple taPedrremmdiionnsiaesttir(oaanntiyo(Dtnimatoye D2f9rao)ym1u),potno Dfoauyrsd8a,y1s5b,e2f1o,reanthdeaitneixtipael rdimoseental THE AMERICAS EUROPE ASIA/PACIFIC AFRICA 51 0 S 8 S 2 Covance 6329-200 Amendment No. 1 Covance 63T2PP9a6-g27e09082 2. pCPaaorglalegerc9at,pi7oh.nisEn,x(thpbi)esrMismeecetthniootnadlwoDiftehCsitoghlnele,fcCotli.looOnw/bNisneugrmvuabntdeireornolifnoAefdnAicnmhiamanlsag.les:M, (4o)diBfylotohde Sfiarsmt ple Bmlaorogdinsaalmeaprlevse(inapopfroalxlimanaimtelayls4amt tLheeaincthe)rvwaillslpbreedcoolsleecthterdoufrgohmDaay 21. PROTOCOL AMENDMENT APPROVAL F. Bud W. McDonald^ Study Director Acute Studies Covance Laboratories Inc. Representative Quality Assurance Unit Covance Laboratories Inc. (j DSate-2J-Q 1 DatSe-M -R 7 52 005853