Document NG5pzmmp3b7NJN6pekonVe4rp

ARQ26 - 06Sh PFOS Pharmacokinetic - 3 000186 - CRNoordnhi.nng H59u41s5o53n70n7c7545 . aDoetnuzearnsen3M3a01dKoia1n$B3l33.8 onan Fax `Sponsor: 3M St. Paul, Minnesota FINAL REPORT CORNING Hazleton Study Title: Single-Dose Intravenous Pharmacokinetic Study of T-6684 in Rabbits Author: F. Bud W. McDonald Study Completion Date: December 31. 1997 Performing Laboratory: 33C0o1mKiinngsmHaaznleBtoounleIvnacrd Madison. Wisconsin 53704 Laboratory Project Identification: CHW 6329-199 Page 1 036 Corning Pharmaceutical Services 000187 QUALITY ASSURANCE STATEMENT CHW 6320-199 `aTchcisorredpaonrctehwaistbhetehnerFeovoidewaenddbDyrtuhgeAQdumailniitsytrAastsiuorna(nFceDAU)niGtooofdCoLambionragtHoarzylePtraocntiIcnec.,in Regulations, 21 CFR 58. The following inspections were conducted and findings reported to the Study Director and management. Inspection Dates Eom To 1272419 012897 021797 0327/97 1224/9 0128/97 0217/97 03/3197 Phas Protocol Review Necropsy Protocol Amendment Data/Report Review Date Reportedto SudyDimctr 1212419 0128/97 021797 0331/97 Date Reporied to Management 12724196 0128/97 0217/97 03/31/97 dan Quality Assurance Unit 1231.97 Date 2 000188 _-- cicawwe63m29-a19y9 STUDY IDENTIFICATION Test Material Single-Dose IOnftrTa-v6e6n8o4usiPnhRaarbmbaictoskinetic Study T6684 Sponsor Sponsor's Representative TMomxicology Service 3MMeCdeinctaelr,DeBlpdagr.tm2e2n0t-2-02 PS.tO.PaBulo,xM3N322505133-3220 RMoger G. Perkins, PhD, DABT ToMxeidcioclaolgyDeSpearrvitcmeent P3.MO.CBenotxer3,3B2l2d0g. 220-2E.02 6St1. 2Pa7u3l,3M32N2255133-3220 Study Director Study Location StSutduydyTiImneittiaabtlieon Date IEnx-pleifreimEenndtaDlat(eIn-life) Stat Date SEtxupdeyriCmoemnptlaeltTieornmiDnaatteion Date CF.orBnuidngW.HazMlceDtoonnaIlncd. MP.aOd.isBoonx, 7W5I4553707-7545 (608) 242-7901 3C3o0m1inKignHsamzalnetBoonuIlnec.vard Madison, WI 53704 JJaannuuaarryy 31,4,11999977 FFeebbrruuaarryy 1111,, 11999977 December 31, 1997 3 000189 _-- Acute Studies Steven M. Glaza Manager F. Bud W. McDonald Study Director Jeffrey B. Hicks In-lfe Supervisor Rose M. Bridge Administrative Supervisor Toxicology Support Kathy Myers Manager Calvin L. Horton Supervisor KEY PERSONNEL Quality Assurance Nancy M. Centanni Manager cCHHwW 6e325m-1y99 Laboratory Animal Medicine DDiopnlnoamaJt. eC,lAemCoLnsA,MDVM Supervisor Anatomical Pathology Deborah L. Pirkel LauriJe. Schuller Supervisors Necropsy 4 000190 CONTENTS CHW 6329-199 QUALITY ASSURANCE STATEMENT... --1 STUDY IDENTIFICATION... $i KEY PERSONNEL cosmos OBJECTIVE c.osssmsmsssssmmssmsmsmssess 8 REGULATORY COMPLIANCE cc TEST AND CONTROL MATERIALS... BQU 0 SGIY eLmY coon SATEPRCGRUIONS..cosmsnes 10 TESTSYSTEM costes se 10 ASRC D OFTH SHABO Mi AISN .....e c.cec s se ee]10 JustifforiSpecciaestSeilecotinon..........._....___._._ 11 PROCEDURES... DosePrepaandArdmainitstiratoionn... 12 RDOelaoBsoodSnSfCaomIrpRlVoeOuRFCtAoeUlloAOefcIAtNidoSnMsIS.Rhi.Ip.Sm.IencRtOc.N.c.....................c.o.o.m.s.s.........-ooo1r 3123 Fe ..... oor 13 SSIMSHCHLIOAFRBRPiIYlSeM EaS0 To EISN SUEs t S. o 13 LocationofRaw Data, Records, and Final Report... 14 14 5 000191 - chwemi (CHnICal OBSEIVANONS....rvve essere 14 A DISCUSSION c.crrssssssssssmmssmsmmmsmmsssssmmsmmssnssssssons TABLE 1 2 IInnddiiviidduuaall CBHoNdIyCaWlEISIBGhNSS r(8r) rvorrei1186 3 Individual Animal TissueWeightsandBile VOIUMES ............c...corn 20 APPEND csr] PROIOCON DEVIBONS ve. senses nm--rik PrOIOCOL ATENMENt NO. 1... A 6 000192 SUMMARY CHW6329-199 This study was done toassessthe levelofsystemic exposureofT-6684 when administered by a single intravenous injection to rabbits. `The study was conducted using four male and four female acclimated rabbitsofthe Hrai(NZW)SPF strainforeach treatment group as follows: Control/Test | Dose Level | NumberofAnimals' Group Material | (mghg)' | Male Female [Tem | o --} | 2[rem[ow [eT [r [we [e eo] | [ores[we[eT [2[mew[so [ J<7] DMSO - Dimethyl Sulfoxide a Administered at a dose volumeof 0.5 mL/kg. b Two animals/sex/dose level were sacrificed on Day 15. The remaining animals (two animals/sex/dose level) were sacrificed on Day 29. `The animals received a single intravenous injectionofthe control or test material at the indicated dose level into the marginal earveinofthe right ear. The test material was mixed with Dimethyl Sulfoxide (DMSO) to a specific concentration for each dose level in Groups 2-5. The respective control material and test mixtures were administered ata dose volumeof0.5 mL/kgofbody weight. Two animal/sex/dose level were sacrificed on Day 15 and the remaining animals (two animals/sex/dose level) were sacrificed on Day 29. Clinical observations were conducted predose and at approximately 0.5, 2.0, and 4.0 hours afer intravenous injection. Additional clinical observatiaonds twice a day 7 000193 CHW 6329-199 `mortality checks were conducted daily thereafter until the scheduled sacrifice interval (am. mortality check only on daysofscheduled sacrifice). Body weights were determined on Day -7forrandomization purposes, beforetestor control material administration (Day 1), andatthe scheduled sacrifice interval (Day 15orDay 29). A blood sample (approximately 4-mL) was collected fromeachanimal on the day before: controlortest material administration. Blood samples (approximately 4-mL)werealso collected from the animals at 4-, 8, 12-,24-,and 48-hours post-injection,andon Day 8, with the exceptionofone Group 1 maleatthe 8-hour collection interval from which only 1.7 mLofblood was collected. An approximate 4-mL blood sample wasalso collected on Days 15 and 22 from each animal scheduled forsacrificeon Day 29. In addition, at the timeofthe scheduled sacrifice (Day 15 or Day 29), approximately 20 to 40 mL of blood was obtained from each control animal and approximately 20mL of blood was obtained from each Group 2-5 animal. All samwerpecelntreifusged and separate `samples of serum and cellular fractions were obtained and sent frozen on dry ic to the Sponsor. On Day 15 or 29, the animals were anesthetized with sodium pentobarbital, bled via the posterior vena cava, and exsanguinated. An abbreviated gross necropsy examination was not done, however, tissueswerecollected. The whole liver, bile, and both kidneys from each animal were collected, weighed (volume only determined for bile), and sent frozen to the Sponsor. After being intravenously injected with T-6684 or the DMSO control, all animals appeared clinically normal and gained weight during the study, with the exception of one: Group 3 female (3.0 mg/kg) which exhibited an insignificant loss in weighta its termination interval (Day 15). OBJECTIVE `The objectiveofthis study was to assess the level of systemic exposure 10 the test `material, T-6684, when administered as asingle intravenous injection to rabbits. 8 000194 REGULATORY COMPLIANCE CHW 6320-199 `This study was conducted in accordance with the U.S. Food and Drug Administration's Good Laboratory Practice Regulations for Nonclinical Laboratory Studies, 21 CFR 58, with the exception that analysisofthe test mixtures for concentration, homogeneity/solubility, and stability was not conducted. All procedures usedinthis study wereincompliance with the Animal Welfare Act Regulations. In the opinion of the Sponsor and study director, the study didnotunnecessarily duplicate any previous work. TEST AND CONTROL MATERIALS Identification "The test material was identifiedas T-6684anddescribed as an off-white liquid. The control material was Dimethyl Sulfoxide, (Sigma Chemical Company, Lot No. 64H1007; Expiration March 19, 1997), and was described as a clear, colorless liquid. 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 stabilityofthe control material were considered to be adequate for the purposesofthis study. Storage and Retention `The control and test materials were stored at room temperature. Reserve samplesofthe test and control 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 CHW for 1 year. `The test material reservesamplewas sent to the Sponsor. Any unused test material will be retumed to the Sponsor. Any remaining control material may be used for other testing `and will not be discardedafter issuanceofthe final report. 9 000195 CHW 6325-199 Safety Precautions `The testandcontrol material handling procedures were according to CHW SOPs and policies. TEST SYSTEM Test Animal Adult albino rabbitsofthe Hra:(NZW)SPFstrainwere reccived from HRP, Inc., Kalamazoo, Michigan on December 11, 1996 and maintained at the CHW facility at 3301 Kinsman Boulevard, Madison, Wisconsin. Housing After receipt, the animals were acclimated foar periodofat least 7 days. During acclimation and throughout the study, the animals were individually housed in suspended stainless steel cages in temperature- and humidity- controlled quarters. The only exception to this was one Group | male which was found outside its cage at the time of the a.m. mortality check on Day 2ofthe study. Environmental controls for the animal room were set to maintain a temperature of 19 to 23C,a relative humidityof50% 20%, and a 12-hourlight/I -hour dark lighting cycle. The dark cycle was interrupted to conduct in-ife procedures. In cases where variations from these 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 waatd leibirtum 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 thewaterare periodically analyzed. There were no known contaminants in the feed or water at levels that would be expected to interfere with or affect the resultsofthe study. 10 000196 CHW6320-199 Selection of Test Animals "The animals were identified by animal nuanmdcobrresepondringeartag aad were placed into study groups basedon astratified body weight randomization program. The randomization body weights were determined on Day -7. Study Design Animals weighing from 2,372 to 2,800 g and approximately 17 weeksofage at initiation oftreatment were placed into the following study groups: Group Control/Test | Dose Level | NumberofAnimals" Material (mg/kg) Male Female Tem|mE |wT] [2 [re [ow [oT] [ [T r [ooww [e e +] | + [ore [oo[oT] a Administered at a dose volume of 0.5 mL/kg. b Two animalsisex/dose level were sacrificed on Day 15. The remaining animals (two animals/sex/dose level) weresacrificedon Day 29. Justification for Species Selection Historically, the New Zealand White albino rabbit has been the animal of choice because: ofthe large amountofbackground information on this species. n 000197 PROCEDURES CHW 6329-199 Dose Preparation and Administration `Thetestmaterialwasdiluted with DMSO to achievae specific concentration for each dose level in Groups 2.10 5. An individual doseofthe control material or respective test `mixture was calculated for each animal based on its body weight on the dayoftreatment (Day 1). The control material or respectivetest mixture was administered by intravenous injection into the marginal ear veinofthe rightear over aperiodof31 to 53 seconds. The prepared test mixtures were storedat100m temperature until administered. After administration, anyremainingtest mixtures were discarded. Reason for RouteofAdministration Intravenous injection is an acceptable route to assess systemic exposure. Observationsof Animals Clinical observations were conducted predose and at approximately 0.5, 2.0, and 4.0 hours after intravenous injection. Additional clinical observations and twice a day mortality checks were conducted daily thereafter until the scheduled sacrifice interval (a.m. mortality check only for animals sacrificed on Days 15 or 29). Body weights were determined on Day -7 for randomization purposes, before test or control material administration (Day 1), and at the scheduled sacrifice interval (Day 15 or Day 29). Blood Sample Collections/Shipment A blood sample (approximately 4-mL) was collected from a marginal ear vein (left car) ofeach animal on Day -4. Blood samples (approximately 4-mL cach) werethencollected from a marginal carveinofeach animal at 4-, 8-, 12-, 24-, and 48-hours post-injection, and on Day 8, with the exceptionofaGroup | male from which a 1.7-mL blood sample was collected at the 8-hour interval. Anapproximate 4-mL blood samplewasalso collected on Days 15 and 22 from amarginal ear vein (either ear) ofeach animal 12 000198 CHW 6329-199 scheduled for sacrifice on Day 29. In addition, at the timeofthe scheduled sacrifice (Day 15 or Day 29) and via the posterior vena cava, approximately 20 to 40 mLofblood `was obtained from each control animal and approximately 20mLof blood was obtained from each animalin Groups 2-5. Allsampleswere stoartreoodm temperature until centrifuged. After centrifugation, separate samplesofserum and cellular fractions were obtained. The serum and cellular fraction samples obtained pre-injection through Day 15 were stored in a freezersetto maintain a temperatureof-20C 10C until shipped 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)on Day 22. The serum and cellular fraction samples obtained on Days 22 and 29 were stored at CHW and shipped to the Sponsothre day after experimental (in-lfe) termination in the same manner as the samples obtained prior to Day 22. Pathology On Day 15, the first two animals/sex assigned to each dose level (based on the group assignment randomization) were anesthetized with sodium pentobarbital (via injection in the marginalearvein), bled via the posterior vena cava, and exsanguinated. An abbreviated gross necropsy examination was not done, however, tissues were collected. `The whole liver, bile, and both kidneys from each animal were collected, weighed (volume only determined for bile), and immediately placed in a freezersetto maintain a temperature of-20C 10C. After tissue/bile collection, the animals were discarded. `The remaining two animals/sex/dose level were anesthetized, bled, and exsanguinated on Day 29 in the same mannears the animals sacrificed on Day 15. Shipment of Bile and Tissues `The tissues (whole livers and kidneys) and bile collected on Days 15 and 29, along with documentationoftheir corresponding weights or volumes, were sent frozen (on dry icc) to the Sponsor (James D. Johnson, 3M ET. & S, Bldg. 2-3E-09, 935 Bush Avenue, St. Paul, MN, 55106) onDay21 and six days after in-life termination, respectively. The `Sponsor is responsible for the retention and dispositionofthe samples. CHW does not accept any responsibility fortheanalysisofthe samples collected in this study nor are these results presented in this report. 13 000199 Statistical Analyses No statistical analyses were required by the protocol. cw 6329-199 Location of Raw Data, Records, and Final Report The raw data, records, and an original signed copyofthe final report will be retained in the archivesofCHW in accordance with CHW SOP. RESULTS Body Weights Individual body weights are in Table 1. All animals gained weight during the study, with the exceptionof one Group 3 female (3.0 mg/kg) which exhibited an insignificant loss in weight (18 g) at its termination interval (Day 15). Clinical Observations Individual clinical sigas are in Table 2. All animals appeared normal throughout the study. Pathology Individual animal tissue weights and bile volumes are in Table 3. The animals were not examined grossly, although tissues were saved. DISCUSSION `The levelof systemic exposureofT-6684 was evaluated in male and female albino rabbits when administeraesd a single intravenous injection at levelsof 1.0, 3.0, 10.0, and 30.0 mg/kg. Al animals appeared clinically normal and gained weightduringthe study, with the exceptionof one Group 3 female (3.0 mg/kg) which exhibited an insignificant Toss in weight at its termination interval (Day 15). 1" 000200 we SIGNATURE FVutAW SDWolidnton) Sty Director cue Studies Bae I es 15 000201 cwenim Table IndiBvodiyWdeiughtas l W sec Member FDaya n)m Toy o ToT To Group 1 (Control-) DMSO (0.0 mg/kg) vale RFeeisiss Z26o0n0 Foisze Yes i26w8 Si S2754090 " :. 200 Feisas im Zor : Son Female FoFiesess Z22t5 Folsaa Suis S23omus 22amm Zs " 20:.91 Feises Sear pd Si Group2-T-(6LO6m8i4) aie FFeeiisszsr pr25e0t Feiszs sm 2Zs Tes 2Zea:em -. 3029 Foiszo Sito 55 : 27s Female FFeellsiesn Zasamm Feises Zs01 2bp0] Zio S20a91 i .: 2550 Foisis im i : Sos Grou3p T-6684 G0 mee) ae FoFiisssi 228550 Fels Tim 235050s am 2B020y0 si.: s Feiss Zen pH : 2m Female FoFiesissis 2parm 2par 2i66n0 :. FFeaissss: Zpei Iwr % pZr a AenimalrsaoinDayd15 16 000202 . ch esas-1s9 Table 1 Continued) IndiBvodiyWdeiughtasl) TSeT Neumeber mDaiyn)e oalD Ow emma ow Group4 T-6(1600m8g/k4g) Male FFolesl3ss 2205199 Fo1s36 241 22587060 229058 2087 3 .. 2818 Fo1537 23m 2 : 28m Female FFoeisssse 2525m5 Fo1356 249 2246816 204 2235073 : .. 2m Foss Zant 2s : S002 Group 5-7-(630.06m8e)4 Mie For1e5s3i9 2Z.a6 25250609 26204 Foss 26546 2610 : .. 2060 Foss 2594 2609 : 2360 Fete FFoeiiss 22603m 22463 226m0 .. Fe6i1s5s60 225m8 2626150 :: 2390294 + ATnimealswacrificed onDay 15. 1" 000203 ---------------------------------- A-- A Table2 Individus) Clinical Sigas TT Amma Sex Number Observation 05 Rew2d0 wl 40bm 3% i 515 WH Group 1 (Control) - DMSO (0.0 mg/kg) Male F6ISZ2 F612 AAppppeeaarreeddNNoommaall 4 4 4 FF6IIs2s5 AAppppeeaarrecddNNoommaall 4v44 4J vvoosor. v vvs7 Female F6IS@2 FOIS AAppppeeawrreeddNNoommaall 4/ 4/ /J FFSiISs4tS AAppppeeaarrccddNNoommaall v 4 44 / vvooss v svr Group 2.-T-6684 (1.0mg/kg) Male F656 F121 AAppppeeawreeddNNoommaall v v7/vJ 661152298 AAppppeeaarrccddNNoommaall 44 44 vv o v. vvo or Female FF61Is4d7s AAppppeeaarrccddNNoommaall 44 47 J 6F611558489 AAppppeeaarrccddNNoommaall 4v74 vvo os vvooorr Group 3-7-(63.06mk8s4) Male F6I1SI0 F613 AAppppeeaarrccddNNoommaall 4 44 4 FIs AppewedNomal FOI AppewedNomal v/ 7/ J4 o vors v v rv Female F6ISSO AppearcdNomal 4 4 / FF6oIsSSs2 AAppppeeaarrecddNNoommaall 44 44 / FOISS) AppeedNomal 4 Jf vo vv o v voor + AEnaicmhaalnismaaclrsipfpiecaoerndedDnaoyrmISa.l prio ts control or est mater sdmmsEaon + Condition existed. 18 000204 `T2a(Cobntilnueed) IndividualClinical Signs TT Sex Ama Number Observation 3w52o0w 40 Group 4- T6684 (100 mpke) Male FFO6IISSIMS AAppppeeawreeddNNoommaall FF66IS1I3S AAppppeeaarrccddNNoommaall 44 4/ // 4 4 /J 4/ Female F6ISs4 F6ISSS AAppppeeawreeddNNoommaall FF6OISSSS6 AAppppeeaarrccddNNoommaall 4 J /J 44/J 4/ Group 5.T-6684 (30.0mye) Mile FF66IS3I98 ApApppeeaarrceddNNoommaall FF661140 AAppppeeaarrceddNNoommaall 4v74 v / // v/ Female F6ISSS F6ISSO AppearedNomal AppewcdNomal FFoIISSESlO AAppppeeaarrecddNNoommaall 44 v v/ /v v v/v 3"AnEaicmhaalnsiamcarlifaipcpeedaornedDnaoyr1m5alprior tetmaterial smmEaon. 7 Conditionexisted CHW 6325-199 23b 51ses 16% v vo s. . v v vv vvooss. v sv v v vo o. vPE os vos vvoo vv 19 000205 eID 0, Be vy En POR OBMOBM OMEpGi B om 5} mms mun nam one Polommt nam mum pee POG BMOCIE OMEi1a B MB oD om BUM om HEME comm Hm bom o FER OBER NEES DoE Em oa iE : APPENDIX Protocol Deviations Protocol TP6797 Protocol Amendment No. 1 CHW 6325-199 2 000207 Protocol Deviations CHW 6329-199 Protocol Page 5, 7. Experimental Design, A. Animals, (7) Husbandry, (2) Housing. Individually, in suspended stainless steel cages Page 8, 7. Experimental Design, D. Observationof Animals, (3) Blood Sample Collections, (5) Method of Collection/NumberofAnimals, first paragraph, first sentence. Blood samples (approximately 4 mL)... and from the marginal ear vein (lef car) at 4-hours postdose through Day 8. Page 9, 7. Experimental Design, E. Termination (3) Sample Collection. `The whole liver, bile, and both kidneys (collected as one sample) from each animal wil be collected, weighed (volume only determined for bile), and immediately placed into a freezer set to maintain a temperature of 20C 10C. Actual Procedure OneGroup 1 male was found outside itscageat the timeofthe a.m. check onDay2. Blood samples were collected at the following intervals (postdose) from the right marginal ear veinofthe following animals (animal `number/grouplsex): 12- and 24-hour: F61524 (IM), F61557 (4F). 48-hour and Day 8: F61524 (1M), F61549 (2F), F61555 (4F), F61557 @P. Only one kidney for Animal No. F61531 (Group 3M) was weighed at the time of tissue collection. The other kidney was weighed the next day. `These study. deviations are not considered to have had an adverse effect on the outcomeofthe 2 000208 CHW 6329-199 Sponsor: St. Pau,mMMisoesots CORNING Hazleton PROTOCOL TP797 Study Tike: Single-Dose IOnfrTav6e6n8o4usiPnhRaarbmbaictoskinetic Study Date: January 3, 1997 Performing Laberatory: 33C0o1mKiinngsimasnzcBaosnlenvc.ard Madison, Wisconsin $1704 Laboratory Project emtificstion: cw 629.199 2 000209 CHW 6329-199 STUDY IDENTIFICATION Single-Dose IOnftTra6v6en8o4uisnPRhaabmiactoskineic Stdy anwaoseimrm CHW No. Test Material Sponsor `Sponsor's Representative Study Director Study Locaion PrExoppeorsiemdeSnttualdSy tTirmeDattbele DrEaxpfetrRiempeonrtatlDTaetmeination Dae: 25199 Tess TMomxicology Service 3MMedCeinatlerD,eBpladr,me2n2t0.2602 SPtO.PaBulo,xM3N3225051333220 RTMoger . Perkins, PAD, DABT ToMxeidcioclaolgDyeSpearrvimceent PIOM.CBeontxr3,3B2l2d0g 20.26.02 St 7Pau3l, M3N3$5133.3220 CFo.mBiundgWH.azMlceDtoonnanlcd. PMaOd.isBoonx, 7W5i4553707-7545 (608) 262.7901 C33o0m1inKginHsamzalnetBoonuolecvard Madison, WI 53704 WWeekeooffJeFaenburkaurayry1310,,19199797 WeofMearckh24, 1997 El 00021.0 CHW 6329-199 1 S`tuSdyDiosneIngtralveneous Pharmacokinetic Study in Rabbits cw 2o91e9 ees 2 Purpose s"iTnogalsesienstsrtahveelneovuesilnojfecstyisotnemtiorcaebxbpiotssure whenthefstmaterial sadministeredass 3. R"eThgiuslsatutdoyrwyiClolmbpelcioannduccetedin accordance withthefollowingGoodLaboratory tPersatctmiacteerReigaumlatiionxsSftotarncdoaunrcdesrn/tGrueaitdieoslni,nessolwubiitlhitthy,eehxocmeopgteinoeintthya,tanandasltyasbiisloitfytwhiell ot be conducted. [[) CoansaNd onseguulaicedSttudy [[X]] 2410CCFFRR51860((FEDPAA)-FIFRA) [) 40CFR 792 (EPATSCA) [[1][) 5C98N1o3h0Sa(nFNinoa.l)3(8O5E0C(Dl)spanese MAFF) (1) Notification Nos. 313 and870 (Japanese MOHW) A`Rlelguplraotcioendsu.resInitnhetoipsipnriootoncoofltharee SinpocnosmoprlaianndcsetuwdityhditrheectAonri,mtahlesWteuldfyadroeeAscntot unnecessarily duplicate any previous work. 4 Quality Assurance "`TAhsesuprraontcoecoUln,isttiundaycccoonrdduacntc,eanwditthhethfeinWailsrceopnosrtinwiflalcibleitayoudfitCeodmbiyngthHeaQzulaeltiotny Inc. (CHW) Standard Operating Procedures (SOPs)and polices. 5. Test Material A MTdoenetisfication B. (PThoysbiecadlocDuemscernitpetdioinn therawdata) C.. PTuhreitSypoannsdoSruabsisluimteys responsiblity for purity and stability determinations including undertestconditions). Sampolfetesst materialivhicle mixture(s) 2 000211 CHW 6329-199 cHweszTsriesyn Pes tfaokrecnonbceefnotrreataidomni,nihsotmroagtieonneuintlyessoslrueiquletsyt,eadnodtshtearbwiliisteyabnyatlShypsoesnwsiollrn.tTbheese samples(faken)willbe senttothe Sponsorafer experimentaltermination. D. RSotoormagteemperature E RReesseerrvveesSaammppllees) of cachbatehot of estmaterial willb takenforthisstudy. mTahientteasitnmatteermipalerreasterevoe fsa2m0pleC(s)10ilCabensdttohreendraettuCrnHeWd1i0ntahefSrpezoenrssoerta1t0er completionofthe niephaseof the study. F. RAentyenutniuosned tes material willbe retumed othe Sponsor ate completionofall related nie testing. G.. SAasferteyquPirreedcabuytiCoHnsW SOPs and polices 6. Control Material A `WDeinmteitfhyclatSiuolnfoxide (DMSO); Lot mmber, source, and expiration dae 1 be: recorded inthe raw dataan included inthefinal port. B. P(ThoybseicdaocDuemsecrniipetdioinntherawdata) C.. "PTuoribteydaoncudmSeanbtiedltbyy CHW (information from the supplier) D. SRtooormagteemperature: 26 000212 CHW 6320-199 wens es E. RReesseerrvveesSaampmlpesioffesac)h boteofvconitrool material willbe taken orthis andy. "m Theconatroltmi eamtpeerrin salresoefrve2s0amCpl4e1s0)Cw.illbestoredat CHWin freezersetto F. AnReyternetmiaoinningcontrolmaterialmaybeusedfooes esting ndwill notbe discarded ahr issuaonfcthee ial report. G.. SraefqeutiyrePdrbecyaCutHiWonSsOPs and policies 7. Experimental Design A Animals SRpaebcbiiets @ HSatr(inNsZSWouIrScPeFHRP, Inc. A`Agdeulatt tiation @ 2We5i0gh3t5aktsIniiation ) N20emmableesr2anndd2S0exfemales 1Inedniivifdcosniuomnbered ea tag. Hosbandry, @) HInoduisviidnugally, in suspended sanless sel cages 27 000213 CHW 6329-199 cwaesrm (4 FAomoeadsuredamountofLaboratoryRabbitDietHF #5326(PMI Fefeordnsut,rIitnicon.alTchoemfpooondeinstrsouatnidneelnyviarnoanlmyeznetdablyctohnetammainnuafnatcst.urer Water `Aadnlablyizneadfmofrrtoomtaalndsissoolmvedassotlyisdits,ecmsp.ecSifiead mimcorfoptbhieollwotgieecraalsre ccholnotreinnta,tseedlheycdtreodccalrebmoennst.,beavymetals,organophosphates, and @ "TChoenrteaamriennaonktnsowncontaminants i thefoodoFwaterthatwould interfere with this study. ( `EEnavviirroonmaemnetnatlcontrolsortheanimalroomwillbesttomaintain aantdem3pe1r2a-thuoruer olfi1g9hC1t01232d-Ca,rhkacoryeulcalrtei.veThhuemiddariktycoyfclSe0m%ay2b0e%, interrupted due 10 in-life procedures. AAlcecalsitm7atdiaoyns, () SBealseecdtioonnohefalTtehstanAdnibmoadlys.weight according0CHW SOPs. An adequate `ponoruhemaolfbtheexitsrrpalaacneidmaolnstwesit.l bTehepuarncihmaaslesdw0iltlhabte polacaendimianloisntoubdyv.iously. agryosuopsfussuidngy insittriaattiiofni.ed body weight randomizaion program within nine. (9) JHistuoriscaltly,iftfoherNiSepecwcZiaeeasltSaenlidecotWihnointealbino rabbit hasbeen the animal of chaice becauseofthe large amount ofbackground information on this: species. 28 000214 J avi03n = Tmeonrn IfoCm:| ]c Cono3tr5oslTe[st||oDoSswteLyeve[L l|| NaopuemTofBbaAniesm]aarle re | [ r R p= re m =e sT, n+a ] PR 0 pBeEeErpR,smiirte iet a EEten A ze PR i A a d e a i ,1 ee 000215 CHW 6329-199 aweman art asttardooosmetvempoeratluofreu0.unmLl isdekmi.niTshteerperde.pared testmitureswillbe stored D. ObservadonofAnimals: (0) BCelfiosrieeatleOsbtsoecrovnattrioolnmsaterialadennistsion, tapproximately0.5, 2.0, anCldini4c.a0 hsiogunrss,paosttwiincejdeic(ltDoa(ayn.m1.) faonrdcplimni)caflorimgonr,adliatlyyutnthilrheeafoer s0c1hDedauylsed1s5aocrri2fi9cweililntbeervaclbs(eDrevye1d5anoryDaanyc2e9f)o.rmTohreaalniitmyaolnstshaocrsieficed resstpuedcytdiivreecdtaoyr.s. Obsemarybveexatentdediwheondnicsedbythe @ BFoordyranWdeoimgihztastion, before est control mateial injection (Day 1. at dtehaetshcahnedduslaecdrisfaiccreisfi(cwehietnesvuravi(vDaalyex1c5eeordDsa|yd2a9y)),or a unscheduled 3) Blood Sample Coletions PFrreeiqnueecntciyon (anime from up tofourdays before test material osdumrisnspiocsaiinoencttoioDna,yo1n),Datyaspp,ro1x5i,m2a2t,ealnyd4a Bt,he1s2c-h,e2d4u-lend d 48sacrifice interval (Day 15 or Day 29) (8) MBleotohdosdamopflCeas(lspeprcoixiomant/eyN4oumfrLA)msibwimleallrbse collected fromthe morarcgoinntraollcmaratveeriinal(iitnhjeectciaonr,)oanfdalfraonmitmhalsmaorngitnhealdeoayrbveefionre(ctefs `aand)ditaito&na-loubrlosopdosstadmopsleetsharpopurgohxiDmaayte8.lyOnmDLa)yswil1l5b3endco2l2l.cted sfcihoemdutlheedmafrogrisnaaelicnacsevoeniD(y1c29a.rifpossbi) ofthe animals bFcroobmstihenpdosreorimorecvehnaancaivmaa, sparcorixfiimceadtienl3ym2o0rimbLuondfbcloondoidtiwoinll iffopmoscsaicblhceo.ntarpoplroaxniimmaatlelsyac2r0if0ic4ed0omn LDoafysbl1o5oodr w2i9l(bheebund maximo volume posible will be obained). and spprosimately 30 000216 CHW 6320-199 HwaTeoenms 2S0acmriLfiocefdbolnooDdaywisl15boerob2t9a.inedfromeachGroup2 animal c"eTnhtesfaumgpelde,swainldltbhee ssetpoarreadteatsoeroumm t5emdpecreatlulraefnradcttihonesnstoredin anafdeccezleuslaertfroacmtailoontsabiin nateemdpterraotuugrehoDfay2015Cwi1l0lbCe.seTthefrsoezreunm otnerdmriynactieont. ThheeSpsoenrsuomaronndceloltlwrowfeaecktisopnrsioobrtainiendiaffecDay 1a5fwerilnlbiefsetnetmfirnoazdeonno.ndTyheicSpoontshoerSipsorresspoornwsiibtlheifnorontehwe.eek. retention and disposiotfitohne samples. "Theserumandcelular fractionsamples willbeshippedto: JSaMmeEsTD.&JoShnson 9Bl2d5gB.u2s3h6A0ve9nue Spal, MN $5106 JShaimpemsen.toJfothhenssaoamnplheiss. sirte willb notified regarding the E Termination (1) AUnnsycahneidmualleddyiSnagcrdiufriicensgatnhedsDteuadtyohssacrificed in a moribund condition `wSilblnabremsaulbijeesctweidltlobaenreschobrrdeedv.iseAdrriomsas ninecrmoopsryibeuxnadmicnoantdiiotnioanndwilallbe: avneeisnt)hebtliezdedviwaitthhevseondaicoamvpae,noabnadrebxisaalng(uviniatiendj.ectTiiosnsiunetsh,3smdaersgeinrailbeedar dinyisnegctdiuorni7n.g2th.s3t)uSadmyploer sCaoclrilfciicoedn,inwialmlobreicboulnldecctoenddiftoiomn.anAyftneirna necropsy, the animals will be discarded. @) OScnhDedauyle1d.Stahceriffiisctetswoanimals signed o ach dos level (based oSondtihuemgrpeonutpobaasrsbiigtnalme(nitarainnjdeocmtiizoantiinotnh)ewmialrgbienaanlecsatrhevteiizne)d, wblietdh via the vena cava, and exsanguinated. Theremainingtwo ammalssexidose 3 000217 CHW 6325-199 canTsre.tmo lmeavregliwnialllcbaeavneeisnt)h,etbiezdedvwiaitthhsevodeinucmapveso,taoabdarcbxisaalng(uviinnateecdtioonnDainyth2e9. sAuneasbbr[e1svidaetsecdrgirboesdsinescercotpisony 7ex.a3m)inSaatimopnlweiClolloletctbieond]onwei,lhobweever, collected. @ S"TahmepwlheoCloelllevcet,iboinl,andbothkidneys (colectedssanesale) fom. ebailceh),aannidmailmwmieldliabteecloylplleaccteedd,wnetioghefrde(evzoelsuemt1eo0nmlayndeitaermitneemdpfeorrature Of-20C 410C.Afrsamplecollection heanimalswillbediscarded. `TShpeonssaomrpwlietshiinveorn,ebwileee,knafdiKicdoalelyesc)twioin.lbTehesesnafrmopzelaneonsndtehyeiric 0the Sceocmteisopno7n.dDi/n(g3Xwb)e.iTghhetoSsvpoonlsuomresiwsirleslpboenssihbilpepfeodrttothheepeetrntsioonnasntded in dispositionofthe samples. F. SNtoatsiusttiisctailcaAlnaalynseas laryeresqueiresd. 8 RAefpionratreport including those tems sted below will be submited. D`Deessccrriippttiioonnoofftthhee eesstt asnydsecronel materials PDraotcsedoufreexsperimental ntistion snd termination GDreosscsippaitohnoloofagynyfitnodxiincgsefifeacptpslicable) GInrdoisvsidpuaatlhsolnoigmyltespoote(iwaepipglhitcaabnldebnilde rveoqluuemsetsed by te study director) 32 - 0002185 CHW 6329-199 caweoeseitmn 9. OLriogicoaaldtaofiaR,oaornwcoDpaiies,tRheesreeorfv,eiSlalmpbeleasv)ai,aRbelceosrtdCs,HsWatdoFfaicniallitRaetepsoirttingthe srteupdoyrdtusrcionmgpliesteprdo,grceosntsraonlmdabteefroiraelarcecseeprtvaenscaempolfetsh)e,ilall orreipgoireta.lpWapehdtahteea, incal ipnecrliuoddiongftohnoesyeeaermfoslloiwstiendgbseilgonwinwgiolfltbeereftianianlerdepionrtt.heOarncehyievaersaofirCsHiWgfnorionafg thteufrinaflcrweipolrltb,aelchoafrtgheeda.fTorheemeSnptoinosnoermdmaayteerleicatlw0ihalvbeetsheet taottehrelSsporentsaoirnend idna theee.CHtWheASrpcohnisvoers cfohroaonseasdtdoithiaovnealCpeHrWioddiosfpoismeoeafnthdeCmaHteWrwiiallsl,cahdairsgpeosasltofreage ill be charged PDroosteopcroelpaarndatpiroontroeccoolradsmendments IBnoidtyrweecoirgdhss DObosseervsadtmioinnsision `SShaimpppliengcarlelccotridosn records PSattuhdoylcoogmyeRsepcoonrddesnce Final epor (onginal signed copy) "wTihtehtfohlelsotwuidnyg sdautpsp.orting records will berine at CHW but will not be archived AWnaitemralanraelcyesiispuriesccolridmsaion records ARevfifmiagelrartooormantdemfpreercaaterurteemnpderahtuumriedirteycorredcsords Instrument calibration snd mainensnce records 3 000219 caw ens PROTOCOL APPROVAL avaonsennn ( SponsoprsReeprtFidDv.eOiRT s n EFiane4d) refine) ASctyeDSiurits Coning Hagen nc Lind, LZ QCoumainygAHisaveisnnlUnn.it st4 elle Bat[397 1-377 Bue 3 0002220 -_ ccwheemnws COVANCE Coeserm ee AMENNo.D170MTHEEPRONTOCTOL FrorocoL Tre? SingleDoeofTveno6usiPnoRaubeiersSey cw ens Sponsor Tatag Fuctity aTNnoxiiccoleogDyeSpearrviment SM Comer, Bg 3202802 . Co5M3me0idnKSgiinHoo1niee$n3v70orc.d SPCPlaxM3N3223031333220 Sponsor's Represent Study Distr _T Ro--ger. aki, PD, DABT e FB. McDonald Ths amendment mohedloien orsionofhe prot Eftectv Jauar3y, 1997 I ecIPonsamcgeoclneyes..eldriEixeenpemhrnaitmbeoetlernetcwlaoilthryDhoDeresSim iOgnr,,B0.a D1poveesAldsdwtutmioemydi.bs1)ewtTmessnatyGrirSoroeupnds, To 35 000221 CHW 6325-199 AmcadmentNo. 1 CHW 63"2T9e1er9997 Page2 Effectivedanaary 14, 1997 2 bPaaocgdceoymswm,eoi7d.ghaEttxrepbaenregiemeweionttfhahlgeDiefsorilalso,wAiin.gAlunnidevmraalllien,e(d)cohWauniggteh:itnaheasnttyonm.odTioy he 21035k8 PROTOCOL AMENDMENT APPROVAL. Sponsotr RaepgreseaaiDv,e TM oo 2 Fe99 FF5ouao.dMVeDeonlld ma@ Sty Director Ace Sudies ComingHazonIn. iAbhinsLin, QCuoimlitnygAzsianocne IUn.ni oeIZFEBT oeges1917 36 000222