Document 7MDeMjeLm80wMMyVE70MJNK5R

AR 226 _ 065 PFOS Pharmacokinetic - y 000223 COVANCE > ome mai hag Le, Sponsor: 3M St. Paul. Minnesota FINAL REPORT Study Title: Daily Dose DoreTrm6a6l5A4bsnorRpaibobnisToxicity Study Author: . BudW. McDonald Study Completion Date: December 31, 1997 Testing Facility: C5o3v0a1nKcienLsambaornatBooruileesvaInrcd. Madison, Wisconsin 53704 Testing Facility Project Identification: Covance 6329-200 Page 1 of 52 tesa i sn Hv0z24 QUALITY ASSURANCE STATEMENT Covance 6329-30 "This Inc. rinepaocrctohradsanbceeenwirtehvitheeweFdoboyd tahnedQDuraluigtyAdAmsisnuirsatnracteioUnni(toFfDAC)ovGaonocde Laboratories Laboratory rPerpaoctritceed RteogtuhleatSitoundsy, D2i1reCcFtoRr 5a8n.dTmhaenafgoellmoewnitn.g iWnrsiptetcetniosntsatwuesrreecpoonrdtuocfitnesdpaencdtifoinnsdianngds findings are issued to Covance management according to standard operating procedures Inspection Dates Fom To 04/15/97 04/15/97 05/05/97 0505/97 0521/97 0521/97 0714/97 0715/97 Phase Protocol Review Dose Administration Protocol Amendment Daw/Report Review Date Study Reported Director and to Management 04/15/97 05/05/97 0321197 0715197 Mpuresednfa,tivep , Qua ality Assurance Unit 2 Da1e2737 000225 -_ CcCoovavnmcee6s32p9-a20m0 STUDY IDENTIFICATION 5-Daily Dose Dermal Absorption/Toxicity Study of T-6684 in Rabbits Test Material Sponsor Sponsors Representative: Study Director Testing Facility Study Timetable Study Initiation Date Experimental (In-life) Start Date n-life End Date Experimental Termination Date Study Completion Date T6684 3M Toxicology Services Medical Department 3M Center. Bldg. 220-2E-02 P.O. Box 33220 St.Paul, MN 55133-3220 Roger G. Perkins. PhD, DABT 3M Toxicology Services Medical Department 3M Center. Bldg. 220-2E-02 P.0. Box 33220 St. Paul. MN 55133-3220 (612)733-3222 F. Bud W. McDonald Covance Laboratories Inc. P.O. Box 7545 Madison, WI 53707-7545 608.242.7901 Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, WI 53704 April 18, 1997 May June 25,. 11999977 June 2, 1997 December 31. 1997 3 000226 _-- Acute Studies Steven M. Glaza Manager F. Bud W. McDonald Study Director Jeffrey B. Hicks In-life Supervisor Rose M. Bridge Administrative Supervisor Toxicology Support Kathy Myers Manager Calvin L. Horton Supervisor Quality Assurance Nancy M. Centanni Manager KEY PERSONNEL CoComvamncee s63m29a-20m0 Laboratory Animal Medicine DonnaJ. Clemons, DVM Diplomate, ACLAM Supervisor Anatomical Pathology Thomas E. Palmer, PhD Anatomical Pathologist Deborah L. Pirkel Laurie Schuller Supervisors Necropsy . 000227 CONTENTS Covance 6329-200 QUALITY ASSURANCE STATEMENT ....cccvcc smmm------_ STUDY IDENTIFICATION w-- ws 3 KEY PERSONNEL............. as-- smn SUMMARY. so---- s---- wren] OBJECTIVE s-------- sons end) REGULATORY COMPLIANCE ....c.cocr ss 9 TEST AND CONTROL MATERIALS... Identification. ........... c---- Purity and Stability snss-- Storage andRetention. s------------ Safety Precautions. as------ srl] -- wasnt A ssl TEST SYSTEM... ---------------------- 10 TeSt ANIMAL... ---- 10 LL m-- 10 ARTA)DIE. ---- n SeleofcTestt Ainimoalsn....... s---- 1 Study DeSIgN.....rre ---- n Justification for Species Selection. c-- n PROCEDURES woven mmm PrepaofrExapotsuireon Afea.............. " ST DOSEAAMUAISUBON crn 12 Reason for Route ofAAMIRISIQON..........rcrcrcror 13 ObservationsofAnimals ------------------ 13 B100d Sample COECHONS/SHIPMENL......rveverer ener ShiOpfBimleaendTnIStSUES... el 1 Location ofRaw Data. RecoarnddFisna,l REPOR.......orurernrenrre 1 S 000228 Connee 129200 [i -- meme RESULTS cc... Body Weights .............. Clinical Observations.d................. Dermal Irritation .................. Lt ataA Rat eens 15 sss | essere sores sss | missense |. DISCUSSION.....ccocrmrrrs rss nesses SA 16 SIGNATURE................ ---- weeerrranas cseeren - 17 REFERENCE. errs eseeresnas = -- snrmisssssessnssnnnnnsene: | 1 TABLE 1 Individual and Mean Body Weights (g). - ER 23 IInGnddriiovvuiidpduuaa|ll(DCCloeinrntimcraaollI-)SriiDgaintmsieot.n.h..y.S.l.c..oSreuseslsrsrfs(sr0ao.e 0xmgkig- iddaey).. SE ceseeeeesssae -- 18 en 19 GIrnoudpu2 DTe-r6m6a8l4I(3T.H0tmoukngSiCdOarye)s roo r 20 APIPndEiNviDdIuaXlA Animal Patv hology Dae ta... ne 2212 Amendment No. | To The Protocol... APPENDIX B...... rrssmenesssssn ress Protocol Deviation... sins - sssrerens 35 Protocol TP6798.......................s . -- -s ss sn 36. 31 i ` 000229 | _-- coCmovmanccee6m32s9-a20m0 SUMMARY This study Of T-6684 was done to asses when applied to the the systemic absorption/toxicity and relative skinofrabbits for five consecutive days. skin iritancy The study was conducted Hra:(NZW)SPF strain for using each three male and treatment group. three female acclimated rabbitsof the ~ ~_ -- GCrrouopp 1 (Control) 2 _--_-- Comrol Dose Level Number ofAnimals TTeesttMaterriiaall __((mmyykkgg/daayy))~--MMaalleess Females _ Dimethyl Sulfoxide 00* 3 3 T6684 30 3 3 a Administered at a dose volume of 0.1 mL/kg ofbody weight `The backofeach rabbit was clipped free of hair and a single doseof the respective Tcohnetrtorleaotrmteenstt smiattesriraelmawiansedadimnitnacits.terTehdetao retaheofskaipnploifctahteionrawbbaistscofvoreredcownistehcautigvaeuzdeays. patch secured with paper tape around all edges and overwrapped with Saran Wrap and Elastoplast period. tape to provide an occlusive dressing for each approximate 23-hour exposure aClnidni4cahlouorbssearfvtaetrieoancshwceornetrcolonodructetsetdmpatreerdioasleaodnmiDniasytr1atoinolnyoanndDaatysap|prtohxriomuagthe5l.y 1. 2.5, 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 boenfloyroenthDeafyirs2t9)c.ontBroodlyorweteisgthmtastwereirael daedtmeirnimsitnreadtioonn D(aDyay-51)f,oranrdanadtotmhiezastcihoendupluerdposes, csaocnrtirfoilceorintteesrtvmaalt(erDiaayl 2a9d)m.inTishteraitniiotanl(dreercmoradlediraristatthieonDraeyad1inrgeawdiansg)m.adSeubbseefqoureentthe first orenaDdianygs13o.fdeArbmlaolodiistaamtpiloen (waeprperomxaidmeataefltyer4eamcLh)pwatacshcroellmeocvtaeld ofnroDmaeyasch2 atnhirmouaglho6n and 7 000230 --_-- cCoovavnmcee6e32m9-a20m0 4D0aymsL-3o,f8b.l1o5o,dawnads23o.btaIinnaedddiftrioonm.eaatcthhecotnitmroeol fanniemcarlopasnyd(aDpapyro2x9i)m.aatpeplryo2x0immatLeloyf20to sbelpoaordatweassaombptlaeisnoefdsferroummeaacnhd Gcerloluulpar2farnaicmtailo.nsAwlelresaombptlaeisnewdearnedcesnenttriffruogzeednaonnddry ice 0 the Sponsor. On Day 29, the animals were anesthetized with sodium pentobarbital, abplpedrovxiiamtahteepo1s-tcemrixorIv-eenmasceacvtai,onexofstahngeuidneartmeadl aanppdlinceactrioposniseidt.e. aTnhdebwohtohlkeildinveery,sbile, an (collected as one sample) were collected from all animals. determined for bile), and sent frozen to the Sponsor. weighed (volume only aApipnliocratmiaocnroofscTo-p6i6c8f4inddiidngnsotatrenseuclrtopisnya.nyAtlelstanmiatmearlisala-prpeelaatreedd cnhoarnmgaels tihnrboougdhyouwteitghhet study with the decreased feed exception of one consumption for Group one to 1 animal and two three days within Group 2 th fist animals which had ine days of the study. Slight dermal irritation was observed in one control animal while slight to moderate dermal imitation was observed in the other five control animals treated with dimethyl osbuslfeorxvieddei(nDoMnSeOa)n.imaInl twhheialneismlailgshtttroeamtoeddewriattheTd-e6r6m8a4l. isrlriigthattidoenrwmaasl oibristeartvieodn winatshe 2 other five animals. No macroscopic lesions were seen in anyofthe animals at necropsy. . 000231 -_-- coComvamneces6m32s9a.2m0 OBJECTIVE 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 analysisofthe 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 opinionof the 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 Tdentification PhysicalDescription T-6684 (tes) off-white liquid DMSO (control) Clear colorless liquid Dimethyl sulfoxide was manufactured by Mallinckrodt Chemical. 5 . 000232 -_ CCoovmanmcee 6s32s9-a20m0 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 purposesofths study. Storage and Retention The control and test materials were stored at room temperature. A reserve sampleof each the control and test materials were taken and stored in a freezer set to maintaian temperature of -20C =10C. The control material reserve sample will be retained at Covance for one year. The test material reserve sample was 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 ot be discarded aftr issuance of the final report. Safety Precautions The control and test material handling procedures were according to Covance SOPs and policies TEST SYSTEM LH] 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 Aer 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. Environmental controls for the animal room were set to maintain a i temperature of 16 to 22C. a relative humidityof 50% =20%, anad 12-hour light/12-hour 10 000233 Covance 6320-200 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 adlibitum 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 would be expected to interfere with or affect the results ofthe study. Selectionof 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 i Animals weighing from 2.067 to 2,500g and approximately 15 weeks of age at initiation oftreatment were placed into the following study groups: Control/ Dose Level Group __ Test Material _ (mg/kg/day) 1 (Control) Dimethyl 00 Sulfoxide 2 T6684 30 Dose Volume _(mi/kg/day) 01 ol _NumobfAenrimals Males Females 3 3 5 3 Justification for Species Selection Historically, the New Zealand White albino rabbit has been the animal of choice because ofthe large amount of background information on this species. n 000234 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 flanksof each rabbit was clipped freeofhair 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, imitation. or defects that would preclude the useofany of the animals. The animals were clipped as needed throughout the study (Days 3.6. 13 and 29) Dose Administration Al animals received five consecutive daysofadministration of the respective control or test material. The first day of treatment was designated as Day | 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 concentrationof 30 mg/mL was calculated and measured based on ach animal's body weight on the first day of treatment. The test material mixture was appliedtothe tes site at a dose volume of0.1 mL/kgofbody weight in a thin and uniform layer. Each areaofapplication in Groups | and 2 was covered with a4-ply 5-cm x S-cm gauze patch. Each gauze patch was secured with paper tape around all edges and overwrapped with Saran Wrap and Elastoplast tape to provide anocclusivedressing. Collars were used to restrain the animals during each approximate 23-hour exposure period. One Group 2 male was found without ts 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. " 000235 : E Reason for RouteofAdministration The dermal route i a potential route of exposure in humans. Covance 6329200 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 1 through. 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 Draize' technique (recorded as the Day 1 reading). Subsequent readingsofdermal 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 car veinof each animal on Days -3. 8, 15. and 23. In addition.a the timeofnecropsy (Day 29). approximately 20 to 40 mLofblood was obtained from each control animal and approximately 20 mL of blood was obtained from each Group 2 animal via the posterior vena cava. All samples were stored at room temperature until centrifuged. Afier centrifugation. separate samplesofserum and cellular fractions were obtained and stored ina freezer set to maintain a temperature of -20C =10C until shippedto 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 ET. & S. Bldg. 2-3E-09. 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 dispositionofthe samples. Covance . v 000236 ----------------------E-- Covance 6329200 does not accept any responsibility for the analysisofthe samples collected in this study norare 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 sitesofcontrol or test material application were washed with warm tap water before the necropsy procedure. All animals were subjected to an abbreviated ross necropsy examination and any abnormalities were recorded. The whole liver. bile. an approximate 1-cm x 1-cm sectionofthe 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 freezerset 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 ET. & S. Bldg. 2-3E-09, 935 Bush Avenue. St. Paul. MN. 55106). along with documentationoftheir corresponding weights or volumes. The Sponsor is responsible for the retention and disposition ofthe samples. Covance does not accept any responsibility for the analysisof 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. " 000237 RESULTS Counce 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 ofthe 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 | to Day 29. Clinical Observations Individual clinical signs are in Table 2. All animals appeared normal throughout the study with the exceptionof one Group | animal and two Group 2 animals which had decreased feed consumption for one to three days within the first nine days ofthe study. Dermal Irritation Individual dermal irritation scores are in Tables 3 and 4. Slight erythema, edema and fissuring erythema raenadcteiodnesmwaearnedosblsiegrhvteddesinquoanmeatcoinotnrorleaacntiimoanls wwehrileeosblsiegrhvtetdo imnodtheeraotteher five control anima treated with dimethyl sulfoxide. Ofthese ive control animals. two animals also exhibited slight coriaceousness reactions. In the animals treated with T-6684, one exhibited slight erythema, edema and fissuring reactions. one exhibited slight erythema and slight to moderate edema reactions, one exhibited slight to moderate: erythema erythema and and slight slight edema isuri and ng r fissuring reactions, one eactions, one exhibited l iegxhhtibtiotemdodselirgahttetoermyotdehreamtae:a n d edema and slight desquamation and coriaceousness reactions. and one exhibited slight ( moderate erythema and edemaand light desquamation. corlaceousness. and fisuring reactions. * 000238 Covance 6329200 R Pathology Individual animal pathology data (issue weights, bile volumes, and gross pathology observations) are presented in Appendix A. No macroscopic lesions were observed in the animals at necropsy. DISCUSSION "The acute systemic absorption/toxicity and relative skin irritancyof T-6684 was evaluated in male and female albino rabbits when administered dermal for five i consecutive days. Application ofthis material did not result in any test material-related changes in body weight gain or macroscopic findings at necropsy. Al animals appeared i normal throughout the study with the exceptionofone Group | animal and two Group 2 H animals which had decreased feed consumption for one to three days within the first nine 4 days of the study. Slight dermal iitation was observed inone control animal while ' slight to moderate dermal irritation was observed in the other ive control animals treated with dimethyl sulfoxide. In the animals treated with T-6684. light dermal iritation was observed in one animal while slight to moderate dermal irritation was observed in the other five animals. 16 00023 9 SIGNATURE Counce 6329:200 F B7udBW.uMdcDo1na)l.dmal) Study Director | Acute Studies Dae 12/31/97 i REFERENCE : I. Draize. J. H.. "Acute Dermal Toxicity (Single Exposure)." In: Appraisalof the Safety ofChemicals in Foods, Drugs and Cosmetics - Dermal Tosicit. Association of Food i and Drug Officialsofthe UsS.. pp. 54-56 (1959). ! i i i i] 000240 Tabl1e Individual and Mean Body Weights ) -- Animal Number Males Random _-ization Day 1 29 Animal Number Females Random _-ization Day 7 29 Group 1 (Control) - Dimethyl Sulfoxide (0.0 mg/kg/day) FF0O1I2 22433105 22520707 2275696 FR0O1T8 2234800 222377 2277588 Fes 2447 2388 2708 FOI 2263 2244 2619 Men 2397 2388 2668 Mem 23% 2285 2719 Group 2-T-6684 (3.0 mekg/day) FFGO0IOS 22448054 22440424 2277524 FFeEwaOl 22246308 2204697 22850046 Fewls 2447 2390 2805 Few 239 2320 27 Mean 2445 2412 2760 Men 2365 20m 268 i ; . i{ } | ! s 000271 f---- -- oe -- e-- e h-- ee -- ow a ne Ll iii SO A RE EE re } game A - 9 000242 Covance 6320-200 Tabl3e Individual Dermal Irritation Scores Group 1 (Control-) Dimethyl Sulfoxide (0.0 mg/kg/day) DermalReaction vam Study Day 1 3 3 4 5 6 13 Females Study Day 1 2 5 4 5 613 Animal No. F63011 Animal No. F63017 Erythema EAtdoenmiaa Desquamation Coriaceousness Fissuring 0001222 00000001020202 0000001 0000000 0000000 001 2221 0001 221 0000000 00000 01 0 0 0 0 1 00 0000000 Animal No. F63012 Animal No. F63018 z i Erythema Edema 0 1 tn 0000 0 I 1 01 11122 01112 21 3i DeAstoqnuiaamation 00000000000000 0000000 0000001 Coriaceousness 0 0 0 0 0 0 0 0 0 0 0 0 1 0 Fissuring 00000 01 0000000 i Animal No. F63013 Animal No. F63019 Erythema Edema or 12 00 01 221 01 11222 01 11122 3 Atonia Desquamation 00000000000001 00000000000001 2 i Coriaceousness Fissuring 0000000 0000000 0000000 0000000 | - 2 000243 -_ cCoovauncmee63m29a.2m00 Tabled InGdrivoiudpua2l-DTe6rm6a8l4I(r3r.i0tamtgi/okngS/cdoarye)s - DT eDmeamlaRiReeaacction 7 T235SuM 45 dyDia 5y661s 13-- 4 TT3ST5 u3dT44e5 bma5 ysr6T Animal No. F63014 Animal No. F63020 EdEreymthaema Atonia 0000 11 1112120 0000000 000010122222 , 0000000 3 CDoersiqaucaemoautsinoenss~~ 00. 00 00 00 00 00 00 Fissuring 0000000 00 00 00 00 00 o| oi 0000010 i Animal No. F3015 Animal No. F63021 {: EErdyetmhaema Atonia O 0011 1 LT 0000000 0 0112121222 2 0000000 ii CDoersiqaucacmoautsinoenss~~ 00 .00 00 00 00 00 00 Fissuring 0000001 00 00 00 00 00 o| oi 0000000 : Animal No. F63016 Animal No. F63022 EdEreymthaema Atonia 0O 001 0 12 1 1 0000000 0000 11 1112221 0000000 CDoersiaqcueaomusanoenss~~ 0000 00 00 00 00 0I Fissuring 0000000 00 00 00 00 00 00 0o 000000 o-- e ------------------------ ee au 0200244 APPENDIX A Individual Animal Pathology Data i i N 000245 | 2 ou 23H R3 222 833 -- | es AR 22ES 83@ Eh 4 30S AN 223 8%= Yig . 3g BE nivias v58g3 8aa O-- I ----re---- matte 282 8a8 Lg888 &4 282 8&2 i ] EE a = RAL aT A EE : HE ile an ii g388g a4 HEE APPENDIX B Conse 6325:200 Protocol Deviation AmendmePnrtotNooc.ol1 TTPo67T9h8e Protocol i 000258 _-- Protocol Deviation cCoovvanmcee6e32n9a20m0 Protocol Actual Procedure PB.agDeos7e,7A.dmEixnpiesrtirmaetnitoanl, D(e3)siDgons,e Aradbmbiintisswtirlaltbieonc,olNlairnetdhdSuernitnegnccaec:h The approximate 23-hour application period. AfonuinmdalwiNtoh.ouFt6i3t0s1c6oll(aGrroonupat2tMhealae.)m.was mwoarstaplliatcyedchbeacckkoonn Dtahey a3.nimTahleactotlhliasr time. The patch appeared to have remained in place on the application site | st`Tuhdiys.deviation is not considered to have had an adverse effect on the outcome of the ii | | " 000259 COVANCETM Covance 6329200 Samii. Sponsor | an | St.Paul.Minnesota | ] sRorocCoL T7678 | Study Tite: | Ii 5-Daily Dose Tsim Dermal Absorption/Toxicity Study i| } Dac: i i ! April 18.1997 ! i Testing Facility: CSo3r0a)nKceoLnaabonratBoorieees nd Nadion, Wisconsin $3708 |i TestingFacility Prec dentition: | Cone 6529200 1 | . 000260 - coComnnesee29n2m00 5i . ----Tn i STUDY IDENTIFICATION 5-DayDose TDemaatsArnioonnToiiy Sty Counce aman Tear Tan --. TwNmoeniioaSpennices S20hCaoamnnesiitsysi 252 220 ! Soars tpi Ror. Fin, 30,08 TNNMoeiCsolsnoaesyBSeaoynip0n2502 Sn forrn aynesstsizz Sut Dirir r C7o0rehs L7r 5i8ars. pNrieernewst 57027505 Tein Fy CSfoorereaLtnoreBroend. PECxpoeormiSauen!sTSTeiamamieanotilaoen Die VWeeckkootnteys5,117971 Or eon Wekothy 1.1997 " 000261 - cCoovamnceme63n29.00 [--iods iPli| SiDoeDl ermal Avorn ToitStyin Ri 2 Purpose HE i Tmatoermialewtheenyapiplis 0tehde se kin onfrdarbobvitispyfoar fdivielcionvseencutive sire days of tes 3 RPTergiiussletaRyregwCalaormaepnlsiamSncmeiadsin ctartdeiwwiithelclepoiosn sGoosd yLtsrioroy tomrmbans fo Soeemon so. hoe et ot x1))Cocrnmesiromness Suc {1 40CFR160 (EPA-FIFRA) 11[[]]1 40 CFR792 (EPA-TSCA) 5NC5(a8Ni1o)en3as0(iiFnoinnvaNlo)o3(5O3E31CDmp) s$7s3 VpAoTME)OH) ARespimricnssd.usntinosrironcoeweerSin cpoomapneswcithdheoAnniel WtselosActt. 4 ToAqeuiasrynoeacsUrantnyewciontd,sawnidttheConrneLpraivlobn eCsoteyQtuyet ering re SOF ne kpe 5 Terran A aldnensiteaion 5. PhOfyfsaiceDteaseo i| 000262 Counce 329.200 3 Cometioion i!} C.TphueriSpnoodrSub.iays sponyfo putsndsabilydetemintions || : {(haercyuoinncennets. homoangea)slSaaym.saodSeaba Sose iOntObe) b. sSuroe pa E ReRveesreessuammpele)of sch cotof st teil illbe ken or is dy. ji Teheitnessmaetepr esreeofs3p0l21i0lClaietsmtedCo0n0e Sinpforre6es |i! Commie oFoe speofey. | F. ARetenutsioend et mt wil eum the Spr. | G. SatfereeredcabmyiConosveSOPs splice i 6. Conte Saterit i A DiemietcaiSoanft OM50) 8. PhpysiicsDtesorinio)n : C.Topbureidnoamebditebyy Covance (normanfrom te spp) 5. RStoormagceaCpoentins 000263 ! 40 Covance 6329-200 2 Comme sitsa00 i acs ii E ReRseseerrvveeSsaampmlepslofeesac)h beh ofcontol mers wil be taken. | ! t"oThmeacionnttarionl matteemrpiearlarteusreervoefs2sm0plCfs=)10wC.ibeslorled st Covance ina feeze set. F. ARentyenrteimoanining control material maybe used for thr testing and will no be discardedafer sseanceofthe fin report, ! G.. S`aAstereyuPirreecdaubtyiCoonvsance SOPs snd policies 7. Experimental Design A Animals SRpaebcbiites @ Hesawi(nNsZoWuIrSePeFCovanceResearchProducts In. ) AAdgietatitston @ 2W0e03i0sktgsInithiattion (6) 6Nm umberl nadnd6Sfees mxales Ilnddeinvdiidtuiaelantiuomnberedes 3g. 0) Husbandry @ HInoduisviidnugally, in suspended sinless see cages ' at 000264 !: -_ 0 cp omm-- enm 3| Com tio no Ts AFmues Fenn). mThoeufnaodf LisanbtoryaRaabiaDi yHhEe u532f6 (McI fo mlcopa a oronmen comm Ftweaiuorimalyrolm o3dtfosr icisteedm,iScampolesnof hsre dtrwe. orem omar. @ ThCroenearmwrinnnamonsas dcyon.tamina i thefodor wae atwold EnEtvneiirmroopmneemrennistlocfo1n9toCl 0o3r3thC.aneimaalreoommwiildlibesoft00%mi0ma%i.n nmdm5hduGUTSfeorwtcdksoc. The dk cl may oe NAciemarioan SSaiedncoenmeiasTnnsoAonfiumnarlss:weinghttsclrinpe0eCodvanc5e ShOPs. An olrorecaenadimmmetwointaodsigpsreoyudppspswoiorfnshsaythipiiandcieosndooonwei.thThreenadromizlsiwoinl be CFhiicobreciushe-- eoNfethw-- egseaanm-- souWnhteofablacikngraouind hafosrbmeaetniohneosnrtsi of ese. J 000265 --_--eee | -_-- compem--an Comme 3e3eo0tn0 B. Dose Administration Test Groups Goo (Como) 2 ToCoMnvuolcl l DMSO 684 (DposheeLenve)l oor s00 NuMmabsecofsoFiemzaulless 3 3 5 3 * Tobeadministered a3dose volume of 0.1 mk ofbody weight (2) OPnrtheedapy baeoffroErxeaptohtesiuniriteiaoAlrenesat nd convo material application. the a Back b awnidl.lni beeccelsitpspaerdyf(e0ebofihniewuintblheammiselheecdtrsikci<)l.pptehre.anTkhse osfhsacvehd aTrheawrielamceontnsiteis (aptparcoxsikmiant)elwyil2l0b%e ionfstpheectterdaorbovdeyrsfuerifancgelaecsai.ons. iTmhietatsinoni, losr dwielflecbtes tclhitppweodu3l3dnpereedceldudtehtrhoeuguhsoeutofhneysoifct.he nim. ) ADolseanAidmmailnsiwsitllracteioinve five consecutive daysofsdministsion ofthe epsppleiccaitvieocnonittrolsochdtaeys.t mTahteerieastpmliaetd eotihelsiammveiesrtehsrpeiecwtciilvlle bee bperdepeasriegfnraesthaeedxcDhaydaIy. oTfhaedcmoinnitsotlatmiaotne.riaTlhewirlstbedaapypolfiedreusunmdenitdila. awidtohsDevMoSluOm3e0odfapp0.l1 imedLt a dTohseevGorlouumpe2otefst.mamtLerkiga.l wiTlhlebedodsielsutfeodr {thaekaenoimnalDsayin|Gerofuoprs h1 eanfdis2wtialdlmibniestbraatisoanneatnhddeanppilimed'tobtohdeyawreeaigohft eaxnpdo2s)urweilinbaectohimnpalnetdenliyfcoorvmeraeydew, iTahte&-srphly o5f.a0p-plmicxa5ti0onc(mGgraouuzpe|s opavtecrh.wraAplplepdicwihtehsSwairlalnbeWrsaecpu"rneddwiEtlhasptaopeprasttapteapaero0unpdroavlideedgaens and o2c3c-lhuosuisvaeppdlreiscsaiingo.n Tpehreiodr.abbTihsewpirllepbaerecdoltleatremdidxutruirnegswoilclhbseppsriorxeidmaatte room temperature unl sminisration. i . i s 000266 . ar-------------------- i: | --_---------- CCoonnme-- es3o25w.2m00 aw2Hne ToRetsoon troRotee afmoemien xt ns, e A-- atesd-- c Samap nt amthrh osscent tmealspoinh,e TB a oet,r To G. Grrentomstarinss CBSloriteOovrurrvtagtos eouitine, thamrty35 a n Bo 5a may rion Do 39 Snri rei mB Rentengote iDernm om reaimoneintseeagn tyme bimSeanykmO0oh0138A Piltthreeo amhnP lsoeBet praatse, to oPBordyrDwroimiBeo orweeain noer crn ae ion ood Sample Clectins @ iFrqumneyeA E fogsIeno ei ens -- - 000267 Covance 6326-200 i i Comme 32r9e20n0 Paes {i (8) Method ofCollctonNoufmAnbimealrs i mBalrogoidnaslamcpalevsei(nsopfparllovainmimaatllsyatmtLhecianche)rawlilslpbreeccolslecttherdoufgrhom a. Day 22 bFcroobmshinepeodstferroimorevaecnhaanciamvaal, aspaprfoseimiantae2ml0oyrimbLuondfbcloondoidtiwoilnl (iffrpoomsesaicbhlec.onstpoplronxiimmatlelsyc2i0ic0e4d0onmLDoafy 2l9oo(dhewimlalxbiemaubtmined bvlooloudmewillpboesosibbaleiwnileldbeforboimesiancehd)Grnodupap2parnoixmiamlatsealcyri2f0icmeid onof Days. "cTehnetibfluogoedds,amapnldesthweislelpabreatseosreerdusmtaonodmcetelmpeerfaatucrteioannsdsttohreend in asfceicoaneosbetaitonemdaitntraoiung2h0DaCy +110wiCl.l beTsehe sefrurmozsedoncerlylacre 10 tsheerSupmoannsdorcoelnulos ftowcoiwoeneskosbpsriinoerdofiinceiDfaeymi1n5swtiilolnb.e sTehne fterromziennatoinond.ry TcheetSphonesoSrpoinsroerspwointshiibnleonfeorwteheekreatfenetrionniatned disposiion of he amples "The serum and cellar fsionssmpleswillbeshigped 0: JSaMmeEsTD..&JSohnson 9Bl3d5g.Bu2s3h6A.v0e9ne. SPaul MN 55106. JShaimpemsenD.toJfohtnhesosnaomrplheiss shtemae il be notified regarding the. D. Pathology (1) AUnnsycahneidmualledyiSnagcrdiufriicnegatnhdestDuedaythosr saciid in moribund condiion waiblnlorbmeasluibijeesctweidl0besnreacbobrrdeevd.iatAednigmraosl ninecromposrybeuxnadmicnoantdiiotnioannwdiallllbe 1 ." Co 000268 FE, H| --5Treg sete re ET . 000269 1i ceronn i mn i 8. puReport ------ D D Poceiere esosr be p stindoeomrn lmatls DoDTeosvotofornxipemsaomernaosyentbis enby srsnxdmedmitotaoiconne GTrebotsapioanntfoetTibnogdsy weighs br enw doe evel Ge reepaoa lo reogoefeinsenbdaheuSesy Dict 5. GoLocraStiiotnioofgRoorowcDorteosseReesecodtrdll, Reerserevle SaatmploneCf)osnndosFdfipeitRn.epeoWrtinng {nTitvienogndsasceommoedcoiut mwilhl rede Sl,hSvaorf Cgopvper34 rreeimodeofcnecepelhsolotoweinegmTSeeniSonpneoodfrsoreetpsorotlveOveensre fteSsrapcmihaofnd a oSeeCeoaonenei1ArhvSeoneoordrhosstPe v Covoi podeCoafthee ol rteterns E iSotoltsotrs-- greetamends SDaRodeawseavgmhtesiodnans SaAOmeprnasctliinpnstaoonoegresrt SSFookygapoermooreeos tnsindcon) a 000270 a" -~ 1 Covance 6329-200 Comme 119-200 Paez a"Trhcehifvoeldlowwiitnhgthseupspiodryidnasr.ecordswillbe renda Covance but wilnatbe WAantiemralaneaclyesiipssrceeciomrdastion records ARneifmrailgreoroamtnotdrempreerzaetrurteeamnpderhautmuirdeirteycorredcsords Instrument calibration and maintenance records " 000271 | J wa Celefrlrgiise, Fa oJ E(GrBtit 7 di,Lic, tbrk7 4-18-97 5nts 000272 - i. 3 : Covance 6329-200 Comme 119200 Attachment 1 Scoring SealeforAcute Dermal Reactions Erythe0m-a None 21-SMioidgehtrate 5-Severe Edema0- Nove 21. NSighlt boe(drlaisepedearrpcpreaopxittmiaotwbeeellyle|demfmi)ned by definite rising) 3 Severe (raised mre than | mm) Avonia0- None 21 .SNiigohdetrastleigshtowimpreutiem1ennofoerlmaaslt)icity) 3. Marked (noclastiit) Desqua0m-arNioonne 21. MoSldiegrhtatlieg(hstcaslceaslainngd)flakes) 5. Marked (pronounced kingwith denudedress) Corisce0o-usNnoeses 21.SMloidgehrtadteecreetahseeriyn ltiesxbtiulriet)y) 5. Marked (ough and brite) Fissuri0ng None 2.SlMiogdehrtadtee f(cnrackcsraicnkdserinmiesp)idermis) 5. Marked (cracks with bleeding) 50 000273 1 Cones 19:20 |i COVANCE errs a putST. ILE ii AMENDMENT No. 1 TO THE PROTOCOL #RotocoL Tree Daily Dose oDeTrmaal4Abmsa bToi Study Covance 6329-200 i `Sponsor Testing Facility i fwm ------ C$o3v0tKniensLmaanBboroird SNM dCoimerbepdagri7m0e25.02 Vadson wi 53708 2S0.m508s3s3313033220 Sponsor's Representative Study Director RogG. Prin. PD DAT 0 aeons Ths amendmen modish follwing prions of te prtce Efcive ay 20,1997 ChoenaebehreaDnecpo2f MedosroaOha clainidmes cwithsDyol2laoibnetsadsc.onTsoosiohne ing 0 easel 10 mova Dap 23 esd Oo 3 1 PCoalgecei,o.nEsx.p(erFirmeequnetnacyD.isMiganC,d.OibsseervcaotinonwoifthAhniemaFollsl,ow(i4ngBlsonndeSasmple ne SriemdioieisatnainmtaeDfryomouph Dfoaurydys15be.f2or4nhde isapledorsems eminnion 29) st 000274 --- Cove 329-200 Amendment. | Connce 63T2F9oa2e0m:0 2 FPCaoaglecrsi.oonsiE,hxi(p5es)riMemeetonhtnoadlwoDfaCisiogn,lColl.lOcobwsiienrgovwanntdieoNonsfiuoAnfAeminmibtmnae.glseM,o)tiBloootd Sooanle Bmlionodaslamplevsipnop0 nss mLh.ecaech)swillrbeecdoslltecufoomDa 23 PROTOCOL AMENDMENT APPROVAL SRpoensors onReerrpsrsmn5ieBRT 3 kl F2 FFH de1)wredone) Scwedy SDurdioers Covance Laoraors or521-97 a CQuoalintyeAsLsuirabncee rUnait. we52047 2 000275 | : | Cptoirnedssenwaishined15nepcdeone,urspi4nnoPdrnolehceerliod3n0coocNfmepywan Jos US Cmoodenstebwshiichhde msaFksiomnrgddmhhaemewpsoorrtCoTtvheaenpceriInnci fo sd bet bdo SReanms ssmmmcrne WU rsoamraansam hfoso:rGotmea Shaping Solutions 000276