Document JrwEQ0Zb1Vb8XjqdKnvxMj7nr

AD BIOSEARCH, iNC. p.o.box 8598 philadelphiap,ennsylvania19101 t e Ie p h o n e 2 1 5 848-4499 Submitted to: Material: Sample Received: Date of Report: Test: Obiectof Test: Procedure: Restilts: Conclusion: 3M Company 3M Center St. Paul, Minnesota 55101 3M Company - T-1388 November 28, 1975 March 4, 1976 Acute Oral Toxicity- Rats To study the acuteoral toxicityin rats of the subjectmaterial. Four groups of 5 male & 5 femalealbino rats of the Sherman-Wistar Strain weighing between 200 and 300 gm were employed in this study. The rats were deprived of food but not water for 24 hours prior to dosing. Each animal was weighed and dosed by direct administrationof the experimentalmaterial into the stomach by means of a syringe and dosing needle. The sample was dosed as a 25% w/v suspension in corn oil. The following dosage levelswere administered: 0.63 gm/kg 1.25 gm/kg 2.5 gm/kg 5.0 gm/kg Followingadministrationthe animals were allowed food and water ad libitumfor the 14 day observationperiod during which time the rats were observed for signs of toxicityand mortalities. See Table 1. The subject material when studied in male & female albino rats has an acute oral LD,,OF less than 0.63 gm/kg. I-Inia KaLrk ab iel, V.M.D.,Ph.D. Director TABLE I Acute Oral Toxicity Material: 3M Company - T-1388, as a 25% w/v suspension in corn oil. Dosage Number Mortalities Total T Level of Animals Days Dead s gm/kg Dosed 1 2 3 4 5 6 7 8 9 10 11 12 13 14 14 Days 0.63 5 males I I 1 0 000000 0 0 0 0 3 0.63 5 females 1 2 0 0 0 0 0. 0 0 0 0 0 0 0 3 1.25 5 males 22 1 0 0 00000 0 0 0 0 5 1.25 5 females 200 30 0 0000 0 0 0 0 5 2.5 5 males 3 20 0 00000 0 0 0 0 0 5 2.5 5 females 4 1 0 0 00 0000 0 0 0 0 5 5.0 5 males 5000000000 0 0 0 0 5 5.0 5 females 5000 0000 00 0 0 0 0 5 The LD50 is less than 0.63 gm/kg. DraftReport 5-DailyDose Dermal Absorption/Toxicity Study of T-6684 inRabbits I PP,EPARED FOR: 3M COVANCE STUDY NUMBEI;L: 6329-200 ISSUE DATE: July 11, 1997 Sponsor: 3M St.Paul,Minnesota DRAFT FINAL REPORT Study Title: 5-DailyDose Dermal Absorption/ToxicitSytudy ofT-6684 inRabbits Author: F.Bud W. McDonald Study Completion Date: July 11,1997 TestingFacility: Covance LaboratoriesInc. 3301 Kinsman Boulevard Madison,Wisconsin 53704 TestingFacilitPyrojectIdentification: Covance 6329-200 Page I of 52 QUALITY ASSURANCE STATEMENT (To be providedinthefinalreport) Covance 6329-200 2 STUDY IDENTIFICATION Covance 6329-200 5-DailyDose Dermal Absorption/ToxicitSytudy of T-6684 inRabbits TestMaterial Sponsor Sponsor'sRepresentative Study Director TestingFacility Study Timetable Study InitiatiDoante Experimental(In-lifSet)artDate In-lifEend Date ExperimentalTerminationDate Study CompletionDate T-6684 3M ToxicologyServices MedicalDepartment 3M Center,Bldg.220-2E-02 P.O. Box 33220 St.Paul,MN 55133-3220 Roger G. Perkins,PhD, DABT 3M ToxicologyServices MedicalDepartment 3M Center,Bldg.220-2E-02 P.O.Box 33220 St.Paul,MN 55133-3220 (612)733-3222 F. Bud W. McDonald Covance LaboratoriesInc. P.O. Box 7545 Madison,Wl 53707-7545 608.242.7901 Covance LaboratoriesInc. 3301 Kinsman Boulevard Madison,WI 53704 April18,1997 May 5,1997 June 2, 1997 (Datefinalreportissigned) (Datefmalreportissigned) 3 Acute Studies F. Bud W. McDonald Study Director JeffxeyB. Hicks In-lifSeupervisor Rose M. Bridge AdministrativSeupervisor ToxicologySupport Kathy Myers Manager CalvinL. Horton Supervisor KEY PERSONNEL Covance 6329-200 LaboratoryAnimal Medicine Cindy J.Cary,DVM Diplomate,ACLAM Supervisor Anatomical Pathology Thomas E. Palmer,PhD AnatomicalPathologist Deborah L. Pirkel/ Jack Serfort Supervisors Necropsy QualityAssurance SherryR. W. Petsel Manager 4 CONTENTS Covance 6329-200 QUALITY ASSURANCE STATEMENT .......................................2........... STUDY IDENTIFICATION ..................................................3........... KEY PERSONNEL .........................................................4........... SUMMARY ...............................................................7.-.......... OBJECTIVE ..............................................................9........... REGULATORY COMPLIANCE ..............................................9.......... TEST AND CONTROL MATERIALS .........................................9........... Identificati..o.n.........................................................9........... Purityand Stabilit.y...................t..................................1.0........... Storageand Retention....................................................1.0........... SafetyPrecaution.s......................................................1.0........... TEST SYSTEM ...........................................................1.0........... TestAnimal .............................................................1.0.......... Housing................................................................1.0.......... Animal Diet.............................................................11........... SelectionofTest Animals .................................................1.1........... Study Design............................................................I.I.......... JustificatifoonrSpeciesSelectio.n..........................................I.I.......... PROCEDURES ...........................................................1.2........... PreparationofExposure Area ..............................................1.2........... Dose Administratio.n....................................................1.2........... Reason forRoute ofAdministratio.n........................................1.3........... ObservationsofAnimals ..................................................1.3.......... Blood Sample Collections/Shipme.n.t......................................1.3........... Pathology..............................................................1.4........... Shipment of Bileand Tissues..............................................1.4........... StatisticAanlalyses......................................................1.4........... Locationof Raw Data,Records,and FinalReport..............................1.4.......... 5 Covance6329-200 RESULTS ................................................................1.5.......... Body Weights ...........................................................15........... ClinicalObservation.s....................................................1.5.......... Dermal Irritati.o.n......................................................1.5........... Pathology..............................................................1.5........... DISCUSSION .............................................................1.6.......... SIGNATURE .............................................................1.7........... REFERENCE .............................................................1.7........... TABLE I IndividuaalndMean Body Weights(g)....................................1.8.......... 2 IndividuaCllinicaSligns...............................................1.9........... 3 IndividuaDlermal IrritatiSocnores.......................................2.0.......... Group I(Control-)DimethylSulfoxid(e0.0mg/kg/day)....................2.0........... 4 IndividuaDlermal IrritatiSocnores.......................................2.1.......... Group 2 -T-6684 (3.0mg/kg/day).......................................2.1........... APPENDIX A ............................................................2.2........... IndividuaAlnimal PathologyData..........................................2.3........... APPENDIX B ............................................................3.5........... ProtocolDeviation.......................................................3.6.......... ProtocolTP6798 ........................................................3.7........... Amendment No. I To The Protoco.l........................................5.1........... 6 SUMMARY Covance 6329-200 This study was done to assess the systemic absorption/toxicityand relativeskin irritancy of T-6684 when appliedto the skin of rabbitsforfiveconsecutivedays. The study was conducted using threemale and threefemale acclimated rabbitsof the Hra:(NZW)SPF strainforeach treatmentgroup. Group Control/ Test Material Dose Level (mg/kg/day) Number of Animals Males Females I (Control) Dimethyl 0.0a 3 3 Sulfoxide 2 T-6684 3.Oa 3 3 a Administered ata dose volume of 0.1 mL/kg of body weight. The back of each rabbitwas clipped freeof hairand a singledose of the respective controlor testmaterialwas administeredto the skinof the rabbitsfor 5 consecutivedays. The treatmentsitesremained intact.The area of applicationwas covered with a gauze patch secured with paper tape around alledges and overwrapped with Saran Wrap's and Elastoplaststape to provide an occlusivedressingforeach approximate 23-hour exposure period. Clinicalobservationswere conducted predose on Day I only and atapproximately 1,2.5, and 4 hours aftereach controlor testmaterialadministrationon Days I through 5. Additionalclinicaolbservationswere conducted dailythereaftetrhrough Day 29. Mortalitychecks were conducted twice dailyon Days 2 through 29 (am. mortalitycheck only on Day 29). Body weights were determined on Day -5 forrandomizationpurposes, beforethe firsctontrolor testmaterialadministration(Day 1),and atthe scheduled sacrificienterval(Day 29). The initiadlermal irritatiorneadingwas made before the first controlor testmaterialadministration(recordedas the Day I reading).Subsequent readingsof dermal irritatiownere made aftereach patch removal on Days 2 through 6 and on Day 13. A blood sample (approximately4 mL) was collectedfrom each animal on 7 Covance 6329-200 Days -3,8, 15,and 23. In additiona,tthetimeof necropsy(Day 29),approximately20 to 40 mL of bloodwas obtainedfrom each controlanimaland approximately20 niL of blood was obtainedfrom eachGroup 2 animal.Allsampleswere centrifugeadnd separatesamplesof senun and cellulafrractionwsere obtainedand sentfrozenon dry ice totheSponsor.On Day 29,theanimalswere anesthetizewdithsodium pentobarbital, bledviatheposteriovrena cava,exsanguinatedand necropsied.The whole liverb,ile,an approximate1-cm x 1-cm sectionof thedermalapplicatiosnite,and both kidneys (collecteadsone swnple)were collectefdrom allanimals,weighed (volumeonly determinedforbile)a,nd sentfrozentotheSponsor. ApplicationofT-6684 didnotresulitn any testmaterial-relatcehdangesinbody weight gainormacroscopicfindingsatnecropsy.Allanimalsappearednormal throughoutthe studywiththeexceptionof one Group I animaland two Group 2 animalswhich had decreasedfeedconsumptionforone tothreedays withinthefirsntinedays of thestudy. Slightdermalirritatiwoans observedinone controlanimalwhileslightomoderate dermal irritatiwoans observedintheotherfive.contraonlimalstreatedwithdimethyl tulfoxid(eDMSO). IntheanimalstreatewdithT-6684,slighdtermalirritatiwoans observedinone animalwhile slighto moderatedermalirritatiwoans observedinthe otherfiveanimals.No macroscopiclesionswere seen inany oftheanimalsatnecropsy. 8 OBJECTIVE Covance 6329-200 The objectiveof thisstudy was to assessthe systemic absorption/toxicitaynd relative skin irritancoyf testmaterialswhen appliedto theskinof rabbitsforfiveconsecutive days. REGULATORY COMYLL4NCE This study was conducted in accordance with the United StatesFood and Drug AdministrationGood Laboratory PracticeRegulationsforNonclinicalLaboratory Studies,21 CFR 58,with the exceptionthatanalysisof thetestmaterialmixtures for concentrationh,omogeneity/solubflitya,nd stabilitwyas not conducted. All procedures used in thisstudywere in compliance with the Animal Welfare Act Regulations.In the opinion of theSponsor and study directort,hestudy did not unnecessarilyduplicateany previous work. AU proceduraltimes presentedinthisreportfallwithinthe acceptable ranges as specifiedin the Covance LaboratoriesInc.(Covance) StandardOperating Procedures (SOPs). TEST AND CONTROL MATERIALS Identification The materialswere identifiedand describedas follows: Identification PhysicalDescription T-6684 (test) off-whiteliquid DMSO (control) Clear colorlessliquid Dimethyl sulfoxidewas manufactured by MallinclcrodtChemical. 9 Covance 6329-200 Purityand Stability The Sponsorassumesresponsibiliftoyrtestmaterialpurityand stabilitdyeterminations (includinugndertestconditions)A.nalysisofthetestmaterialmixturesfor concentrationh,omogeneity/solubiliatnyd, stabiliwtays notconducted.The purityand stabilitoyf thecontrolmaterialwere consideredto be adequateforthepurposesof this study. Storage and Retention The controland testmaterialswere storedatroom temperature.A reservesample of each thecontroland testmaterialswere takenand storedina freezersettomaintaina temperatureof -20'C 10*C. The controlmaterialreservesample willbe'retaineadt Covance forone year.The testmaterialreservesample willbe sentto theSponsor.Any unused testmaterialwillbe returnedtotheSponsor.Any remainingcontrolmaterialmay be used forothertestingand vrilnlotbe discardedafterissuanceof thefmal report. SafetyPrecautions The controland testmaterialhandlingprocedureswere accordingtoCovance SOPs and policies. TEST SYSTEM Test Animal Adult albinorabbitsoftheHm:(NZW)SPF strainwere receivedfrom Covance Research ProductsInc.,Kalamazoo, Michigan on April16,1997. Housing Afterreceiptt,heanimalswere acclimatedfora periodof atleast7 days. During acclimationand throughoutthestudy,theanimalswere individuallhyoused insuspended stainlesssteelcages.Enviro=ental controlsfortheanimalroom were settomaintaina 10 Covance 6329-200 temperatureof 16 to 22'C, a relativheumidityof 50% 20%, and a 12-hourlight/12-hour darklightincgycle.In caseswhere variationfsrom therequiredtemperatureand humidityconditionsexistedt,heywere documented and consideredto have had no adverseeffecton thestudyoutcome. Animal Diet The animalswere providedaccesstowaterad libitumand a measured amount of LaboratoryRabbitDietHF #5326,PNE Feeds,Inc.The feedisroutinelaynalyzedby the manufacturerfornutritionaclomponents and environmentalcontaminants.Samples of thewater areperiodicallaynalyzed.There were no known contaminantsinthefeedor wateratlevelsthatcouldbe expectedto interferweith or affectheresultosf thestudy. SelectionofTest Animals The animalswere identifiebdy animalnumber -andcorrespondingeartagand were placed intostudygroupsusinga stratifibeoddy weightrandomizationprogram. The randomizationbody weightswere obtainedon Day -5. Study Design Animals weighingfrom 2,067to2,500g and approximately15 weeks ofage atinitiation oftreatmentwere placedintothefollowingstudygroups: Group Control/ Dose Level Dose Volume Number ofAnimals TestMaterial (mg/kg/day)(mL/kg/day) Males Females I (Control) Dimethyl 0.0 0.1 Sulfoxide 2 T-6684 3.0 0.1 3 3 3 3 Justification for Species Selection Historically, the New Zealand VNte albino rabbit has been the animal of choice because ofthelargeamount ofbackgroundinformationon thisspecies. PROCEDURES Covance 6329-200 Preparation ofExposure Area On theday beforethefirsctontrolortestmaterialapplicationt,heback and,ifnecessary (toobtainunblemishedskin),theflanksofeachrabbitwas clippedfreeof hairwithan electricclipper.The clippedareamade up approximately20% of thetotalbody surface area.The testsites(intacstkin)were inspectedforinterferilnegsionsi,rritatioonr, defectsthatwould precludetheuse of any oftheanimals.The animalswere clippedas needed throughoutthestudy(Days 3,6, 13 and 29). Dose Administration Allanimalsreceivedfiveconsecutivedaysof administrationftherespectivceontrolor testmaterial.The firsdtay oftreatmentwas designatedasDay 1. Group 1. An individuadlose was calculateadnd measured based on each animal's body weighton thefirsdtay oftreatment'.Me controlmateria(ldimethylsulfoxide) was appliedtothetestsiteata dose volume of 0.1 mL/kg ina thinand uniform layer. Group 2. An individuadlose of thetestmaterialmixture(T-6684and dimethyl sulfoxidea)ta concentratioonf 30 mg/mL was calculateadnd measured based on each animal'sbody weighton thefirsdtay oftreatment.The testmaterialmixture was appliedtothetestsiteata dosevolume of 0.1 mL/kg of body weight ina thin and uniformlayer. Each areaofapplicatioinn Groups I and 2 was coveredwitha 4-ply5-cm x 5-cm gauze patch.Each gauze patchwas securedwith papertapearoundalledges and overwrapped with Saran WrapQ)and Elastoplastt'aspetoprovidean occlusivedressing.Collarswere used torestraitnheanimalsduringeach approximate23-hourexposureperiod.One Group 2 male was found withoutitscollaron atthea.m.mortalitcyheck on Day 3. The collarwas placedback on theanimalatthistime(seeprotocoldeviationspage). Approximately23 hoursaftereach controlortestmaterialapplicationt,herestraining collarasnd patcheswere removed and any residuamlaterialremoved from theapplication sitesusingtapwaterand disposablepapertowels. 12 Reason forRoute ofAdministration The dermalrouteisa potentiarlouteof exposureinhumans. Covance 6329-200 Observations ofAnimals Clinicalobservationwsere conductedpredoseon Day 1 only and atapproximately1,2.5, and 4 hoursaftereach controlortestmaterialapplicatioonn Days 1 through5. Additionalclinicaolbservationwsere.conducteddailythereaftetrhroughDay 29. Mortalitycheckswere conductedtwicedailyon Days 2 through29 (am. mortalitycheck onlyon Day 29). Body weightswere determinedon Day -5 forrandomizationpurposes,beforethefirst controlor testmaterialadministratio(nDay 1),and atthescheduledsacrificienterval (Day 29). The initiadlermalirritatiroenadingwas made beforethefirsctontrolortestmaterial applicatioanccordingto the I Draize technique(recordedas the Day I reading). Subsequentreadingsof dermalirritatiwoenre made within30 to37 minutesaftereach patchremoval(Days 2 through6) and on Day 13. Blood Sample Collections/Shipment A blobdsample(approximatel4y mL) was collectefdrom amarginalearveinof each animalon Days -3,8,15,and 23. In additiona,tthetimeof necropsy(Day 29), approximately20 to 40 mL ofblood was obtainedfrom eachcontrolanimal and approximately20 mL of bloodwas obtainedfrom each Group 2 and 3 animalviathe posteriovrena cava. Allsampleswere storedatroom temperatureuntilcentrifuged. Aftercentrifugatiosne,paratesamplesof serum and cellulafrractionwsere obtainedand storedina freezersettomaintaina temperatureof -20*C 10*C untilshippedto the Sponsor. The senan and cellulafrractionsamplesobtainedon Days -3,8,a'nd 15 were shippedfrozen(ondry ice)totheSponsor(JamesD. Johnson,3M E.T.& S,Bldg.2-3E09,935 Bush Avenue,St.Paul,MN, 55106)on Day 16. The serum and cellulafrraction samplesobtainedon Days 23 and 29 were shippedtotheSponsortheday afterthe experimental(in-lifdea)teof terminatioinnthesame manner asthesamplesobtained priorto Day 23. The Sponsorisresponsiblfeortheretentioannd dispositioonfthe 13 Covance 6329-200 samples. Covance does notacceptany responsibiliftoyrtheanalysisof thesamples collecteidnthisstudynor aretheseresultpsresentedinthisreport. Pathology At terminatioonfthein-lifeexperimentaplhase(Day 29),animalswere anesthetizewdith sodium pentobarbitabll,ed viatheposteriovrena cava,exsanguinated,and necropsiedin random order.The sitesofcontrolortestmaterialapplicatiownere washed withwarm tapwater beforethenecropsyprocedure.Allanimalswere subjectedto an abbreviated grossnecropsyexaminationand any abnormalitiewsere recorded.Tle whole liverb,ile, an approximateI-cm x I-cm sectionof thedermalapplicatiosnitea,nd bothkidneys (collecteads one sample)were collectefdrom allanimals,weighed (volume only determinedforbile)a,nd placedina freezersettomaintaina temperatureof -20'C 10'. Afternecropsy,theanimalswere discarded. Shipment of Bile and Tissues One week afterin-lifeexperimentalterminationt,hetissue(swhole liversd,ermal applicatiosnitesa,nd kidneys)and bilewere sentfrozen(ondry ice)totheSponsor (JamesD. Johnson,3M E.T.& S, Bldg.2-3E-09,935 Bush Avenue, St.Paul,MN, 55106),alongwithdocumentationoftheircorrespondingweightsor volumes. The Sponsorisresponsiblfeortheretentioannd dispositioonfthesamples.Covance does not acceptany responsibiliftoyrtheanalysisofthesamplescollectedinthisstudynor are theseresultspresentedinthisreport. StatisticaAlnalyses No statisticaanlalyseswere requiredby theprotocol. Location of Raw Data, Records, and FinalReport The raw data,records,and an originaslignedcopy ofthefinalreportwillbe retainedin thearchivesofCovance inaccordancewith Covance SOP. 14 RESULTS Covance 6329-200 Body Weights Individualand mean body weightsareinTable 1. Allanimals,withtheexceptionof one controlanimal,exhibitead weightlossfrom randomizatiotnoinitiati(oinn-lifweh)ich canbe attributetdotheacclimationoftheanimalstorestrainincgollarcsonductedfor 3 days (approximatel2y1-23 hours/day)beforetheinitiaclontroland testmaterial administrationA.llanimalsgainedweightfrom Day ItoDay 29. ClinicalObservations IndividuacllinicaslignsareinTable2. Allanimalsappearednormalthroughoutthe studywiththeexceptionof one Group 1 animaland two Group 2 animalswhich had decreasedfeedconsumption forone tothreedays withinthefirsntine days ofthestudy. Dermal Irritation Individuadlermal irritatisocnoresareinTables3 and 4. Slighterythema,edema and fissurinrgeactionwsere observedinone controlanimalwhileslighto moderate erythema and edema and slightdesquamationreactionwsere observedintheotherfive controlanimalstreatedwithdimethylsulfoxide.Of thesefivecontrolanimals,two animalsalsoexhibitedslightcoriaceousnesrseactionsI.n theanimalstreatedwith T-6684,one exhibitesdlighteryffiemae,dema and fissurinrgeactionso,ne exhibited slighterythemaand slighto moderate edema reactionso,ne exhibitedslighto moderate erythema and slightedema and fissurinrgeactionso,ne exhibitedslighto moderate erythema and slightfissurinrgeactionso,ne exhibitedslighto moderateerythema and edema and slightdesquamationand coriaceousnesrseactionsa,nd one exhibitedslighto moderate erythemaand edema and slightdesquamation,coriaceousnessa,nd fissuring reactions. 15 DISCUSSION Covance 6329-200 The acute systeniic absorption/toxicity and relative skin irritancyof T-6684 was evaluatedin male and female albinorabbitswhen administereddennally forfive consecutivedays. Applicationof thismaterialdid not resultin any testmaterial-related changes in body weight gainor macroscopic findingsatnecropsy. All animals appeared normal throughoutthe study with the exceptionof one Group I animal and two Group 2 animals which had decreasedfeed consumption for one to threedays within the firstnine days of thestudy. Slightdermal irritatiownas observed in one controlanimal while slightto moderate dermal irritatiownas observed in the otherfivecontrolanimals treated with dimethyl sulfoxide.In theanimals treatedwith T-6684, slightdermal irritatiownas observed inone animal while slightto moderate dermal irritatiownas observed in the other five animals. 16 SIGNATURE Covance 6329-200 F.Bud W. McDonald Date StudyDirector Acute Studies REFERENCE 1. Draize,J.H.,"AcuteDermal Toxicity(SingleExposure),I"n:AppraisaloftheSafety of@ChemicalisnFoods,Drugs and Cosmetics-Dermal ToxicityA,ssociatioonfFood and Drug OfficialosftheU.S.,pp.54-56(1959). 17 Table 1 Individualand Mean Body Weights (g) Covance 6329-200 Males Animal Random Number -ization Day 1 29 Females Animal Random Number -ization Day 1 29 Group 1 (Control-)DimethylSulfoxide(0.0mglkg/day) F63011 F63012 F63013 2,315 2,430 2,447 2,277 2,529 2,500 2,766 2,388 2,708 F63017 F63018 F63019 2,424 . 2,380 2,263 2,333 2,755 2,277 2,783 2,244 2,619 Mean 2,397 2,388 2,668 Mean 2,356 2,285 2,719 Group 2 -T-6684 (3.0mg/kglday) F63014 F63015 F63016 2,404 2,485 2,447 2,402 2,722 2,444 2,754 2,390 2,805 -F63020 F63021 F63022 2,262 2,438 2,395 2,067 2,429 2,321 2,506 2,804 2,735 Mean 2,445 2,412 2,760 Mean 2,365 2,272 2,682 Table 2 IndividuaCllinicaSligns Covance 6329-200 Animal Number Sex Observation Day I Day 2 --ifour- Hour 1 2.5 4 1 2.5 4 Day 3 Hour 1 2.5 4 Day 4 Hour 1 2.5 4 Day 5 Hour 1 2.5 4 67 Day(s) 8 9 10-1516-29 Group I (Control-) DimethylSulfoxide(0.0mgfkgtday) F63011 M Appeared normal VV V V Decreasedfood consumption F63012 M Appeared normal V F63013 M Appeared normal V F63017 F Appeared normal V F63019 F Appeared normal F63019 F Appeared normal V of ol or of r Group 2 -T-6694 (3.0mg/kglday) F63014 M F63015M F63016 M Appeared normal / V Appeared normal / / Decreafsoeodd consumption Appeared normal --f -- V .1 --- V VV oe t of F63020. F Appeared normal Decreasedfood - - - - - - - - - - F63021 consumption F Appeared normal .1 Z .1 V V V or F63022 F Appeared normal .1 - of ol V Conditionexisted. Conditionnotevidenl 19 Covance 6329-200 Table 3 IndividualDermal IrritatioSncoresGroup I (Control)-Dimethyl Sulfoxide(0.0mg/kg/day) Dermal Reaction Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Males Study Day 1 2 3 4 5 6 13 Animal No. F63011 000 1222 000 1222 0 000000 0 0 00001 00 00000 0 0 00 00 0 Animal No. F63012 0111 111 000001 1 00 00 00 0 0 000 00 0 0 000 00 0 000000 1 Animal No. F63013 011 1121 000 1 22 1 0 00 00 00 000000 1 000 0 00 0 00 00 00 0 Females Study Day 1 2 3 4 5 6 13 Animal No. F63017 0 01 222 1 0001 22 1 0 0 00 0 0 0 000000 1 0000 1 00 00 00 0 0 0 Animal No. F63018 01 1 1122 0 1 1 1 2.2 1 00 0 0 00 0 000000 1 0 0 0 0 0 1 ol 0 00 0 00 0 Animal No. F63019 0 1 1 1 222 01 1 1 1 22 00 0 00 0 0 000000 1 00 0 00 0 0 00 0 00 0 0 20 Table 4 Individual Dermal IrritationScores Group 2 -T-6684 (3.0mglkglday) Covance 6329-200 Dermal Reaction Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissufing Erythema Edema Atonia Desquamation Coriaceousness Fissuring Males Study Day 12345 1-3 Animal No. F63014 00 1 1 1 1 1 00 1 1220 00 000 00 00 000 00 00 000 00 00 00 00 0 Animal No. F63015 0111111 0011 11 1 0 00000 0 0 00000 0 0 00000 0 0 00000 1 Animal No. F63016 01 11 12 1 0 000 1 1 1 0000000 0 00000 1 0000000 0000000 Females Study Day 1 2 3 4 5 6 .1,,) Animal No. F63020 00 12221 000122 1 0 000000 0 0 00 00 1 0 0 000 1 0 00000 1 0 Animal No. F63021 01 22222 01 1 122 1 0000000 000000 1 00 000 1 0 0000000 Animal No. F63022 00 1 122 1 00 1 1 1 2 1 0 000000 0000000 00 00000 0000001 21 APPENDIX A IndividualAnimal Pathology Data Covance 6329-200 22 Appendix A Individual Animal Pathology Data ----------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: F63011 SEX: MALE DOSE GROUP: I SACRIFICE STATUS: SCHEDULED, TERMINAL SACRI DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT.- NOT DATE AND TIME OF NECROPSY: 06/02/97 8:59 PROSECTOR: DAVID SCHUETTE RECORDER: KEVIN BILLI POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALF ----------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- ST ---- DERMAL APP. SITE (IS) KIDNEY (KD) LIVER (LI) BILE (VOLUME) (BI) 1.2216 15.3938 67.8896 0.5000 --------------- --------------- ---------------------- -------- WEIG WEIG WEIG WRIG ----------------------------------------------------------------------------------------------------------------- * ** G R 0S S PAT H0 L0 GY 0 BS ERVA T 10N S ORGAN NA14E SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ------------------------------------------------------------------------------------ GENERAL COMMENT (GC) ... .. .. .. .. .. .. .. ... .. ... .. .. ... .. ... .. .. .. -NO MACROSCOPIC LESIONS -ABBR NBC PERFORM. ui ----------------------------------------------------------------------------------------------------------------- Appendix A individual Animal Pathology Data ----------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: F63012 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUSI SCHEDULED, TERMINAL SACR DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: N(Y DATE AND TIME OF NECROPSY: 06/02/97 8:35 PROSECTOR: DAVID SCHUETTE RECORDER: KEVIN BILLI POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALF ---------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0 ORGAN NAME (GRAMS) TO BODY WEIGHT (ft) WEIGHT RATIO ST ---------------------- -------- -------- -------- --- DERMAL APP. SITE (IS) 1.1614 -------- -------- WEI KIDNEY (KD) 16.1562 --------------- -------- WEI -------- WEI LIVER (LI) BILE (VOLUME) (BI) 61.6643 0.5000 -------- -------- WEI ----------------------------------------------------------------------------------------------------------------- SEVERI;Y,* r-R PA TH 0L 0GY 0 8S ERV AT 10N S ORGAN NAME KEYWORD(0S)q SOR PHRASE FREE -TEXT COMMENTS AND NOTES ---------------------- ----------------------------------------------------------------------------------- GENERAL COMMENT (GC) .. .. . .. ... .. .. .. ... .. .. ... .. .. ..I .. .. ... ... -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM --------------------------------------------------------------------------------------------------------------- Appendix A Individual Animal Pathology Data ------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: F63013 SEX: MALE DOSE GROUP: I SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIF DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT* NOT DATE AND TIME OF NECROPSY3 06/02/97 9:OS PROSECTOR: JILL PAUS RECORDER: KEVIN BILLING POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFO ------------------------------------------------------------------------------------------------------------------ ABSOLUTE ORGAN WRIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME ------- ----- ---------- (GRAMS) ------ TO BODY WEIGHT -------- WEIGHT RATIO -------- STA -- -- - DERMAL APP. SITE KIDNEY (KD) (IS) 0.7738 15.1059 ---------------------- --------------- -------- WEIGH WEIG WEIGH LIVER (LI) BILE (VOLUME) (BI) 73.3509 0.6000 -------- -------- WEIG --------------------------------------------------------------------------------------------------------- * * * G R 0 S S P A T H 0 L 0 G Y 0 B S E R V AT 1 0 N S ORGAN NAME ---------------------- SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ------------------------------------------------------------------------------------- GENERAL COMMENT (GC) .. .. .. .. ... .. .. ... .. .. .. ... ... .. .. .. .. .. ... . -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. --------- -------------------------------------------------------------------------------------------------------- Appendix A Individual Animal Pathology Data ----------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: F63014 SEX- MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIF DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT DATE AND TIME.OF NECROPSY: 06/02/97 8:17 PROSECTOR: JILL PAUS RECORDER: KEVIN BILLING POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFO ------------------------------------------------------------------------------------------------------------------ ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (W) -------- WEIGHT RATIO -------- ST ----- DERMAL APP. SITE (IS) 0.4833 -------- -------- WEIG KIDNEY (KD) 12.5487 -------- -------- WEIG LIVER (LI) 68.0339 -------- -------- WEIG BILE (VOLUME) (BI) 0.9000 -------- -------- WEIG ----------------------------------------------------------------------------------------------------------------- * * * G R 0 S S P A T H 0 L 0 G Y 0 B S E R V A T 10 N S ORGAN NAME SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ------------------------------------------------------------------------------------ GENERAL COMMENT (GC) ... .. .. ... .. ... . ... .. .. ... .. .. ... .. .. ... ... -No MACROSCOPICLESIONS -ABBR NEC PERFORM ---------------------------------------------------------------------------------------------------------------- Appendix A individual Animal Pathology Data Corning Hazleton Inc. Madison, Wisconsin USA ------------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: F63015 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFI DATE OF DEATH: 06102/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT I DATE AND TIME OF NECROPSY: 06/02/97 8:25 PROSECTOR: DAVID SCHUETTE RECORDER: KEVIN BILLINGI POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST- DR. TOM PALMER WEIGHER: JOHN S. HALFORI ------------------------------------------------------------------------------------------------------------------ ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- STA ----- DERMAL APP. SITE (IS) 1.0044 -------- -------- WEIGW, KIDNEY (KD) 1S.9025 -------- -------- WEIGH LIVER (LI) 74.7248 -------- -------- WEIGH BILE (VOLUME) (DI) 1.9000 -------- -------- waiatr ------------------------------------------------------------------------------------------------------------------ *** G R0 S S PAT H 0L0 GY 0 BSE RV AT 10N S O-R-G-A-N--N-A-M-E------------- S-E-V-ER-I-T-Y-, ---KE-Y-W-O-RD-(-S-)---O-R--P-H-RA-S-E------------F-R-E-E--T-E-XT----C-OM-M-E-NT-S----AN-D--N-O-TE-S--------------------- GENERAL COMMENT (GC) ... . .. ... .. . ... .. .: . .. . ... .. ... ... . ... .. . -NO MACRO-SC6PiC LESIONS -ABER NEC PERFORM. ------------------------------------------------------------------------------------------------------------------ Appendix A Individual Animal Pathology Data Corning Hazleton Inc. Madison, Wisconsin USA ------------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: F63016 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFI DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH- 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT DATE AND TIME OF NECROPSY: 06/02/97 6:56 PROSECTOR: JILL PAUS RECORDER: KEVIN BILLINGI POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORI ------------------------------------------------------------------------------------------------------------------ ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- STA DERMAL APP. SITE (IS) 0.6073 -------- -------- WE10tr, KIDNEY (KD) 24.9690 -------- -------- WEIGER. LIVER .(LI) 61.8641 -------- -------- WEIGW BILE (VOLUME) (BI) 0.6000 -------- -------- WE10tr ------------------------------------------------------------------------------------------------------------------ *** G R 0 SS PA TH 0L0 GY 0 BS BRV AT 10N S ORGAN NAME ---------------------- SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ------------------------------------------------------------------------------------- GENERAL COMMENT (GC) .. .. 00 -NO MACROiC'OPiC*LiSiOiS* * . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -ABBR NBC PERFORM. ------------------------------------------------------------------------------------------------------------------ Appendix A individual Animal Pathology Data Corning Hazleton Inc. Madison, Wisconsin USA ----------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: P63017 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRI DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT DATE AND TIME OF NECROPSY: 06/02/97 8:47 PROSECTOR- DAVID SCHUETTE RECORDER: KEVIN BILLIN POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFO ----------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WRIGHT (t) -------- WEIGHT RATIO -------- ST ---- DERMAL APP. SITE (IS) 0.9965 -------- -------- WEIG KIDNEY (KD) 14.5047 -------- -------- WRIG LIVER (LI) 55.4966 -------- -------- weir. BILE (VOLUME) (BI) 1.4000 -------- -------- WEIG ----------------------------------------------------------------------------------------------------------------- ** * G R0 9S PA TH 0L0 GY 0 8S BRV AT 10N S O-R-G-A-N--N-A-M-E------------- S-E-V-E-RI-T-Y-,---K-EY-W-O-RD-(-S-)---O-R--P-HR-A-S-E------------F-R-E-E--T-E-XT----CO-M-M-EN-T-S---A-N-D--N-OT-E-S-------------------- GENERAL COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . bi -NO 14ACROSCOPIC LESIONS -ABBR NEC PERFORM. ----------------------------------------------------------------------------------------------------------------- Appendix A Individual Animal Pathology Data Corning Hazleton Inc. Madison, Wisconsin USA ------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: F63018 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIF DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT DATE AND TIME OF NECROPSY: 06/02/97 8:14 PROSECTOR: DAVID SCHUETTE RECORDER.- KEVIN BILLING POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFOR --------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WRIGHT RATIO -------- ST A ----- DERMAL APP. SITE (IS) 0.8229 -------- -------- WEIGH KIDNEY (KD) 13.4273 -------- -------- WEIGH LIVER (LI) BILE (VOLUME) (BI) 63.0821 1.6000 --------------- --------------- WEIGH WEIGH ------------------------------------------------------------------------------------------------------------------ * ** G R0 SS PA TH 0L 0GY 0 B SERVA T 10N S ORGAN NAME SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ------------------------------------------------------------------------------------- GENERAL COMMENT (GC) . ... .. .. ... .. . ... .. .. ... .. ... .. ... .. .. .. ... . -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. ----------------------------------------------------------------------------------------------------------------- Appendix A Individual Animal Pathology Data Corning Hazleton Inc. Madison, Wisconsin USA ------------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: P63019 DATE OF DEATH: 06/02/97 DATE AND TIME OF NECROPSY: SEX: FEMALE DOSE GROUP: I STUDY DAY OF DEATH: 29 STUDY WEEK 06/02/97 8:23 PROSECTOR: JILL PAUS SACRIFICE STATUS: OF DEATH: 5 SCHEDULED, TERMINAL SACRIPI TERMINAL BODY WEIGHT: NOT E RECORDER: KEVIN BILLINGS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFORD ------------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WRIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0RG ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- STA ------ DERMAL APP. SITE (IS) 0.6150 -------- -------- WEIGM KIDNEY (KD) 14.0486 -------- -------- WEIGM LIVER (LI) 51.5804 -------- -------- WEIGHI BILE (VOLUME) (81) 0.6000 -------- -------- WEIGHI ------------------------------------------------------------------------------------------------------------------- ** * 0R 0 SS PAT H0 L0G Y 0 BSE RVA T 10N S ORGAN NAME SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ----------------------------------------------------------------------------------------------------------- GENERAL COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -NO MACROSCOPIC LESIONS -ABER NEC PERFORM. ------------------------------------------------------------------------------------------------------------------- Appendix A Individual Animal Pathology Data Corning Hazleton Inc. Madison, Wisconsin USA --------- ---------------------------------------------------- 7 --------------------------------------------------- ANIMAL NUMBER: F63020 DATE OF DEATH: 06/02/97 SZX: FEMALE STUDY DAY OF DEATH: DOSE GROUP: 2 29 STUDY WEEK SACRIFICE OF DEATH: STATUS: 5 SCHEDULED, TERMINAL SACRIF TERMINAL BODY WEIGHT: NOT DATE AND TIME OF NECROPSYI 06/02/97 8:49 PROSECTOR: JILL PAUS RECORDER: KEVIN BILLINC POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFO --------- ------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME .(GRAMS) TO BODY WEIGHT (1k) WEIGHT RATIO -------- ST ---- ---------------- -------- -------- DERMAL APP. SITE (IS) 0.6370 --------------- --------------- WEIG WEIG KIDNEY (KD) 13.4440 -------- --.!------ WEIG LIVER (LI) BILE (VOLUME) (BI) 52.3928 0.8000 ------ -------- WEIG --------------------------------------------------------------------------------------------------------------- *** GR 0S 9 PAT H0 L0 GY 0 8S ERV AT 10N S ORGAN NAME ---------------------- SEVERITY, KEYWORD(S) OR PHRASE ---------------:-----------r. ----------- FREE-TEXT COMME14TS AND NOTES --------------------------------------------- GENERAL COMMENT (GC), . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . L@j -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. --------------------------------------------------------------------------------------------------------------- Appendix A Individual Animal Pathology Data Corning Hazleton Inc. Madison, Wisconsin USA ------------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: P63021 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFI DATE OF DEATH: 06/02/97 STUDY DAY OF DEATH.- 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT E 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 ------------------------------------------------------------------------------------------------------------------- A13SOLUTS ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0 R0 ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- STA ------ DERMAL APP. SITE (IS) 0.8344 -------- -------- WHIGHI KIDNEY (KD) 17.0195 -------- -------- WEIGHI LIVER (LI) S3.0880 -------- -------- WEIGHI BILE (VOLUME) (BI) 1.5000 -------- -------- WEIGHI ------------------------------------------------------------------------------------------------------------------ * * * G R 0 S S P A T H 0 L 0 G Y 0 B S E RV A T 10 N S ORGAN N-A-M-E------------ S-E-V-E-RI-T-Y-,---K-EY-W-O-R-D(-S-)---O-R--P-H-RA-S-E------------F-R-E-E--T-E-XT----C-OM-M-E-NT-S---A-N-D--N-O-TE-S--------------------GENERAL COMMENT (OC) . . . . . . . . . . . . . . . . . . . . . . . ... I . . . . . . . . . . . . . . . . . . -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM. ------------------------------------------------------------------------------------------------------------------ Appendix A Individual Animal Pathology Data ------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: F63022 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIF STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: NOT DATE OF DEATH: 06/02/97 DATE AND TIME 00 NECROPSY: STUDY DAY OF DEATH: 29 06/02/97 9:07 PROSECTOR: DAVID SCHUETTE RECORDER: KEVIN BILLING POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: JOHN S. HALFOF ------------------------------------------------------------------------------------------------------------------ ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN To BRAIN 0R ORGAN NAME (GRAMS) -------- TO BODY WEIGHT (t) -------- WRIGHT RATIO -------- ST ---- ---------------------- DERMAL APP. SITE (IS) 1.0944 -------- -------- WEIG KIDNEY (KD) 14.2647 --------------- - -------- -------- WEIG WRIG LIVER (LI) BILE (VOLUME) (BI) 60.3910 1.6000 -------- -------- WBIG ---------------------------------------------------------------------------------------------------------------- ** * G R0 SS PA T H0 L00 Y 0 BS ERVA T 10N B O-R-G-A-N--N-A-M-E------------- S-E-V-ER-I-T-Y-, ---KE-Y-W-O-RD-(-S-)---O-R--P-HR-A-S-E------------F-R-E-E--T-EX-T----CO-M-M-EN-T-S---A-N-D --N-OT-E-S-------------------- GENERAL COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . I. . . . . . . . . . . . . . . . . . . . . . -NO MACROSCOPIC LESIONS 4@- -ABBR NBC PERFORM. ----------------------------------------------------------------------------------------------------------------- APPENDIX B ProtocolDeviation ProtocolT?6798 Amendment No. 1 To The Protocol Covance 6329-200 @5 Protocol Deviation Covance 6'@29-200 Protocol Actual Procedure Page 7, 7. Experimental Design, B. Dose Administration,(3)Dose Administration,Ninth Sentence: The rabbitswillbe collaredduring each approximate 23-hour applicationperiod. Animal No. F63016 (Group 2 Male) was found without itscollaron atthe a.m. mortalitycheck on Day 3. The collar was placed back on the animal atthis time. The patch appeared to have remained inplace on the applicationsite This deviationisnot consideredto have had an adverse effecton the outcome of the study. 36 Covance 6329-200 c 0 V A-N--CE@ TME DEVELOOMENT SERVICES COMPANY Covence Laboratories Inc. P.O. Box 7546 Madison. Viisconsin 53?07-7345 Packages: 3301 Kinsman Bouieverd Madison. Vrosconsin 537'04 Tel:608/241-4471 Fax: 903/241-7227 Sponsor: 3M St.Paul,Minnesota PROTOCOL TP6798 Study Title: 5-DailyDose DermalAbsorption/ToxiciSttyudy of T-6684 inR@abbits Date: April18,1997 TestingFacility: Covance LabomtoriesInc. 3301 Kinsman Boulevard Madison,Wisconsin53704 TestingFacilitPyrojectIdentification: Covance 6329-200 THE A#AENTCAS EUROPE ASWPACIFIC AFRICA 37 Covance 6329-200 Covance 6329-200 TP6799 Page 2 STUDY IDENTIFICATION 5-Day Dose Dermal Absorption/Toxicity Study ofT-6684 inRabbits Covance No. Test Material Sponsor 6329-200 T-6684 3M Toxicology Services Medical Department 3M Center,Bldg. 220-2E-02 P.O. Box 33220 St.Paul,MN 55133-3220 Sponsor'sRepresentative Roger G. PerL-insP,hD, DABT 3M Toxicology Services Medical Department 3M Center,Bldg. 220-2E-02 P.O. Box 33220 St.Paul,MN 55133-3220 (612)733-3222 Study Director F. Bud W. McDonald Covance LaboratoriesInc. P.O. Box 7545 Madison, WI 53707-7545 608.242.7901 TestingFacility Covance LaboratoriesInc. 3301 Kinsman Boulevard Madison, WI 53704 Proposed Study Timetable Experimental StartDate ExperimentalTermination Date DraftReport Date Week ofMay 5,1997 Week ofJune 2, 1997 Week ofJuly 14,1997 38 Covance 6329-200 Covance 6329-200 TP6799 Page 3 1. Study 5-DailyDose DerrnalAbsorption/ToxicitSytudy inRabbits 2. Purpose To assessthesystemicabsorptionand toxicityand relativeskinirritancoyf a test materialwhen appliedtotheskinof rabbitsforfiveconsecutivedays 3. Regulatory Compliance This studywillbe conducted inaccordancewith thefollowingGood Laboratory PracticeRegulationstStandards/Guidelinweisth theexceptionthatanalysisof the testmaterialmixturesforconcentrations,olubilityh,omogeneity, and stabilit%yVill notbe conducted: [ ] Conduct asa Noaregulated Study [X] 21 CFR 58 (FDA) 40 CFR 160 (EPA-FIFRA) 40 CFR 792 (EPA-TSCA) C(S 1)30(Final()OECD) 59 NohSan No. 3850 (JapaneseMAFF) NotificatioNnos. 313 and 870 (JapaneseMOHW) All proceduresinthisprotocolareincompliancewith theAnimal Welfare Act Regulations.In theopinionof theSponsor and study directort,he study does not unnecessarilyduplicateany previouswork. 4. QualityAssurance The protocol,study conduct,and the finalreportwillbe auditedby the Quality Assurance Unit inaccordance with Covance LaboratoriesInc.(Covance) Standard Operating Procedures(SOPS) and policies. 5. Test Material A. Identification T-6694 B. PhysicalDescription Off-whiteliquid 39 Covance 6329-200 Covance 6329-200 TP6799 Page 4 C. Purity and Stability The Sponsor assumes responsibilitfyorpurityand stabilitdyeterminations (includinugnder tw conditions)S.amples of testmaterial/vehicmliexture(s) forconcentrationh,omogeneity/solubilitya,nd stabilitaynalyseswillnot be taken. D. Storage Room temperature E. Reserve Samples Reserve sample(s)of each batchtlotoftestmaterialwillbe taken forthisstudy. The testmaterialreservesample(s)willbe storedatCovance ina freezersetto maintaina temperatureof -20*C 10*C untilreturnedto theSponsor after completion ofthe in-lifpehase of thestudy. F. Retention Any unused testmaterialwillbe returnedtothe Sponsor. G. SafetyPrecautions As requiredby Covance SOPs and policies 6. ControlMaterial A. Identification Dimethyl Sulfoxide(DMSO) B. PhysicalDescription Clear,colorlessliquid C. Purityand Stability To be documented by Covance (informationfrom the supplier) D. Storage Conditions Room temperature 40 Covance 6329-200 Covance 6329-200 TP6799 Page 5 E. Reserve Samples Reserve sample(s)of each batch/lotof controlmaterialwillbe taken. The controlmaterialreservesample(s)willbe storedatCovance ina freezerset tomaintaina temperatureof -20*C 10*C. F. Retention Any remaining controlmaterialmay be used forothertestingand willnot be discardedafterissuanceof thefinalreport. G. SafetyPrecautions As requiredby Covance SOPs and policies 7. Experimental Design A. Animals (1) Species Rabbit (2) Strain/Source Hra:(NZW)SPF/Covance Research ProductsInc. (3) Age at Initiation Adult (4) Weight at Initiation 2.0to 3.0kg (5) Number and Sex 6 males and 6 females (6) Identirication Individualnumbered ear tag (7) Husbandry (a) Housing Individuallyi,nsuspended stainlessteelcages 41 Covance 6329-200 Covance 6329-200 TP6798 Page 6 (b) Food A measured amount of LaboratoryRabbitDiet HF #5326 (PMI Feeds,Inc.)T.he food isroutinelaynalyzedby themanufacturer fornutiitionaclomponents and envirorunentaclontaminants. (C) NVater Ad libititfmroman automatic system. Samples of the water are periodicallaynalyzedforspecifiedmicroorganisms and environmentalcontaminants. (d) Contaminants There areno known contaminantsin thefood or water thatwould interferweith thisstudy. (e) Environment Environmental controlsfortheanimal room willbe setto maintain a temperatureof 19*C to23*C, a relativheumidity of 50% 20%, and a 12-hour light/12-hourdark-cycle. The dark-cyclemay be interruptedue toin-lifperocedures. (f) Acclimation At least7 days (8) Selectionof Test Animals Based on healthand body weight accordingtoCovance SOPS. An adequate number of extraanimals willbe purchased so that no animal inobviouslypoor healthisplacedon test.The animals willbe placedintostudygroups using a stratifiebdody weight randomization program withinnine days of study initiation. (9) JustificatiofnorSpecies Selection Historicallyt,heNew Zealand White albinorabbithas been the animal of choice becauseof the largeamount of background informationon this species. 42 Covance 6329-200 Covance 6329-200 TP6798 Page 7 B. Dose Administration (1) Test Groups ControV Test Material Dose Level (my/kglday) Number of Animals Mal= Females I (Control) DMSO 0.0* 3 3 2 T-6684 3.0* 3 3 To be administeredata dose volume of 0.1 mL/kg of body weight (2) Preparation ofExposure Area On theday beforetheinitiatlestand controlmaterialapplicationt,he back and, ifnecessary(toobtainunblemished skin),theflanksof each rabbit%%illbe clippedfreeof hairwith an electriclipper.The shaved areawillconstitutaepproximately200/oofthe totalbody surfacearea. The treatmentsites(intacstkin)willbe inspectedforinterferinlgesions, irritatioonr,defectsthatwould precludethe use of any of the animals. The animalswillbe clippedas needed throughoutthestudy. (3) Dose Administration All animalswillreceivefiveconsecutivedays of administrationof the respectivecontrolortestmaterialappliedtothesame respective applicatiosniteeach day. The testmateriavvehiclemixture willbe preparedfresheach day of administrationT.he firstday of treatmentwill be designatedas Day 1. The controlmaterialwillbe appliedundilutedat a dose volume of 0.1 mL/k-g. The Group 2 testmaterialwillbe diluted %vithDMSO and appliedata dose volume of 0.1 mL/kg. The doses for theanimals inGroups I and 2 willbe based on theanimal'sbody weight taken on Day I beforethe firsatdministrationand appliedtothearea of exposure ina thinand uniform layer.The area ofapplication(Groups I and 2)willbe completelycoveredwith a 4-ply 5.0-cm x 5.0-cm gauze patch. Allpatcheswillbe securedwith paper tapearound alledges and over%NTapped with Saran Wrap@ and Elastoplasetape to provide an occlusivedressing.The rabbitswillbe collaredduring each approximate 23-hour applicationperiod.The preparedtestmixtureswillbe storedat room temperatureuntiladministration. 43 Covance 6329-200 Covance 6329-200 TP6799 Page 8 (4) Reason for Route of Administration The demal routeisa potentiarlouteof exposure inhumans. (5) Removal of Test Material Appmximately 23 hours aftereach testor controlmaterialapplication, thepatchesand collars%%ilble removed and theresidualmaterialwillbe removed using water or an appropriatesubstance,ifnecessary.Ile sites of testand controlmaterialapplicatiownillbe washed with warm tap water on Day 29 priortothetissuecollectionprocedure. C Observationof Animals (1) ClinicalObservations Before controlor testmaterialadministrationa,tapproximately 1,2.5, and 4 hours aftereach administratioonf controland testmaterialon Days 1-5 forclinicasligns,dailythereaftefrorclinicasligns,and twice daily(a.m.and p.m.)formortalitystartingon Day 2 and continuingto Day 29 (am. mortalityonly on Day 29). Observationsmay be extended %vhen directedby thestudy director. (2) Reading of Dermal Irritation Before theinitiaclontrolor testmaterialadministrationt,hedermal irritatiorneadingwillbe made and recordedas theDay I reading (Attachment1).Additionaldermal irritatiroenadingswillbe made %vithi3n0 to 60 minutes aftereach patch removal (Days 2-6)and on Study Day 13. Individualdermal irritatiroencordswillbe maintainedfor each animal. (3) Body We ights For mndomization, beforethe initiatlestor controlmaterialapplication (Day 1),on Day 29, or atdeath(when survivalexceeds I day) (4) Blood Sample Collections (a) Frequency Predose (anytimefrom up tofourdays beforetheinitiadlose administrationto Day 1),on Days 8, 15,22, and atexperimental terminatio(nDay 29) 44 Covance 6329-200 Covance 6329-200 TP6799 Page 9 (b) Method of Collection/Numberof Animals Blood samples (approximately4 mL each)willbe collectedfrom a marginal carvainof allanimals atthe intervalpsredosethrough Day 22. From the posteriorvena cava,approximately20 mL of blood will be obtainedfrom each animal sacrificeidna moribund condition (ifpossible)a,pproximately20 to40 mL of blood willbe obtained from each controlanimalsacrificeodn Day 29 (themaximum volume possible%vilble obtained),and approximately20 mL of blood willbe obtainedfrom each Group 2 animal sacrificedon Day 29. The blood samples %villbe storedatroom temperatureand then centrifugeda,nd theseparateserum and cellularfractionstoredin a freezersetto maintain-20*C 10*C. The serum and cellular fractionsobtainedthroughDay 15 willbe sentfrozenon dry iceto the Sponsor one to two weeks priortoin-lifteermination.The serum and cellulafrractionosbtainedafterDay 15 willbe sent frozenon dry iceto theSponsor withinone week afterin-life termination.The Sponsor isresponsibleforthe retentionand dispositionof thesamples. The serum and cellulafrractionsamples willbe shipped to: James D. Johnson 3M E.T. & S Bldg.2-3E-09 935 Bush Avenue St.Paul,MN 55106 James D. Johnson or hisalternatewillbe notifiedregardingthe shipment of thesamples. D. Pathology (1) Unscheduled Sacrificesand Deaths Any animal dying duringthe study or sacrificeidna moribund condition willbe subjectedtoan abbreviatedgross necropsy examination and all abnormalitiewsillbe recorded.Animals ina moribund conditionwillbe 45 Covance 6329-200 Covance 6329-200 TP6798 Page 10 anesthetizewdithsodiumpentobarbit(avliainjectioinnthemarginalear vein),bledvia thevena cava,and exsanguinated.The whole liver,bile (allavailablea)n,approximateI-cm x I-cm sectioonfthedermal applicatiosniteand both kidneys(collecteads one sample) willbe collectedfrom allanimalsdying duringthestudy or sacrificeidna moribund condition,weighed (volume only determinedforbile),and placed intoa freezersettomaintain a temperatureof-20*C ,10*C. After necropsy,the animals %%ilble.discarded. (2) Scheduled Sacrifice On Day 29,the animalswillbe anesthetizedwith sodium pentobarbital (viainjectioninthemarginalcar vein),bled viathe vena cava, exsanguinated,and subjectedto an abbreviatedgrossnecropsy examination.The animals willbe necropsiedinrandom order(via computer-genemted random numbers) and allabnormalitieswillbe recorded.The whole liverb,ile(allavailable)a,n approximate 1-cm x 1-cm sectionof the dermal applicationsite,and both kidneys(collected as one sample) %%ilble collectedw,eighed (volume only determined for bile),and placed intoa freezersettomaintaina temperatureof -20*C :tIOOC. Afternecropsy,theanimals willbe discarded. (3) Tissue Sample Shipment The samples (liverb,ile,demial applicatiosnite,and kidneys)collected atthescheduled sacrificweillbe sentfrozenon dry icetothe Sponsor within one %veckaftercollection.Samples collectedatunscheduled sacrificeasnd deaths(ifapplicablew)illbe sentwith and inthe same manner as thesamples from thescheduled sacririceT.he samples and theircorrespondingweights orvolumes willbe shipped tothe person listedin'Sectio7n.C.(4).(b)T.he Sponsor isresponsiblfeorthe retention and dispositionof thesamples. E. StatisticaAlnalyses No statisticaanlalysesarerequired. 46 Covance 6329-200 Covance 6329-200 TP6799 Page II 8. Report A finalreport including those items listedbelow will be submitted. Descriptionof the testand controlmaterials Descriptionof the testsystem Procedures Dates of experimentalinitiatioannd termination Tabula6on of mortalitydata by sex and dose level Descriptionof any toxiceffects/dermalirritation Tabulationof mean body weights by sex and dose level Gross pathology findings Gross pathology report(ifrequestedby theStudy Director) Individualanimal tissue%veightsand bilevolumes 9. Location of Raw Data, Records, Reserve Sample(s),and FinalReport Originaldata,or copiesthereof,willbe availableatCovance tofacilitataeuditing the studyduring itsprogressand before acceptanceof the finalreport.When the finalreportiscompleted,controlmaterialreservesainple(s)a,lloriginalpaper data, includingthose items listedbelow willbe retainedinthearchivesofCovance fora periodofone year followingsigningof the finalreport.One yearaftersigningof the finalreport,allof theaforementioned materialswillbe sentto theSponsor and a returnfeewillbe charged. The Sponsor may electtohave thematerialsretainedin the Covance Archives foran additionalperiodof time and Covance willcharge a storagefee.Ifthe Sponsor chooses to have Covance disposeof thematerials,a disposalfeewillbe charged. Protocoland protocolamendments Dose preparationrecords In-lifreecords Body weights Randomization data Dose administration Observations Anatomical pathology records Samp!e collectionrecords Shipping records Study correspondence Finalreport(originaslignedcopy) 47 Covance 6329-200 Covance 6329-200 TP6798 Page 12 The followingsupportingrecordswillbe retainedatCovance but willnot be archivedwith the studydata. Animal receipt/acclimatiroencords Water analysisrecords Animal room temperatureand humidity records Refrigeratorand freezertemperaturerecords Instrumentcalibratioannd maintenance records 48 PROTOCOL APPROVAL Covance 6329-200 Covance 6329-200 TP6799 Page 13 Rog'er'U.Perk-insP,hD, DABT SponsoesRepresentative 3M F.Bud W. McDonald Study Director Acute Studies Covance LaboratorieIsnc. Representative QualityAssurance Unit Covance LaboratoriesInc. Date Date Date 49 Covance 6329-200 Attachment I Scoring Scalefor Acute Dermal Reactions Covance 6329-200 TP6798 Page 14 Erythema 0 - None I -Slight 2 -Moderate 3 - Severe Edema 0 - None I - Slight(barelyperceptibleto %%,eldlefinedby definiteraising) 2 - Moderate (raisedapproximatelyI mm) 3 - Severe(raisedmore than I mm) Atonisk 0 - None I -Slight(slighitmpairmentof elasticity) 2 -Moderate (slowreturntonormal) 3 -Marked (no elasticity) Desquamation 0 -None I -Slight(slighstcaling) 2 -Moderate (scalesand flakes) 3 -Marked (pronounced flakingwith denuded areas) Coriactousness 0 - None I -Slight(decreaseinpliability) 2 - Moderate (leatherytexture) 3 - Marked (toughand brittle) Fissuring 0 - None I- Slight(definitceracksin epidermis) 2 - Moderate (cracksin dermis) 3 - Marked (cracks%vithbleeding) 50 Covance 6329-200 c 0 V A-9-C--E@ THEDEVELOPMKP(T &["as COMPANY Covence Laborstofi*s Inc. P.O. box 7545 Madison, VA*consin 53707-7S45 Packages: 3301 Kinsman Boulevard Madison, Vfisconsin 63704 Tat: 608/241-4471 Fax:lOa/241-7227 AAIENDMENT NO. I TO THE PROTOCOL PROTOCOL TP6798 5-DailyDose DermalAbsorptionrroxiciStyudy ofT-6684 inRabbits Covance 6329-200 Sponsor 3M ToxicologyServices MedicalDepartment 3M Center,Bldg.220-2E-02 P.O.Box 33220 SL Paul,NiN 55133-3220 Sponsor'sRepresentative Roger G. PerkinsP,hD, DABT TestingFacility Covance LaboratorieIsnc. 3301 Kinsman Boulevard Madison,Wl 53704 StudyDirector F.Bud W. McDonald Thisamendment modifiesthefollo%vinpgortionsoftheprotocol: EffectivMeay 20,1997 The observanceofMemorial Day coincideswithDay 22 ofthestudy.To avoidtiavintgo conducttheDay 22 bleedson a holiday,modifythefollowingtwo sectionsofthe protocoltomove theDay 22 bleedstoDay 23. I. Page 8,7.ExperimentalDesign,C. ObservationofAnimals,(4)Blood Sample Colicction(sa,)Frequency.Modifythissectiowniththefollowingunderlined change: Predose(anytimefrom up tofourdaysbeforetheinitiadlose administmtiotnoDay 1),on Days 8,15,2.1,and atexperimental terminatio(nDay 29) THE AMINICAS EUROPE ASWPACIFIC AFRICA Covance 6329-200 Amendment No. I Co%,ancc6329-200 TP6798 Page 2 2. Page 9,7.ExperimentalDesign,C. ObservationofAnimals,(4)Blood Sample Collection(sb,)Method ofCollection/NumbeorfAnimals.Modifythefirst paragraphinthissectionwiththefollowingunderlinecdhange: Bloodsamples(approximatel4ymL each)willbe collectefdrom a marginalearveinofallanimalsattheintervalpsredosethroughDay 21. PROTOCOL AMENDMENT APPROVAL Roger DPerk-ins,PhD, DABT Date Sponsoes Representative 3M F.Bud W. McDonald' Study Director Acute Studies Co%,anccLaboratorieIsnc. f Representative QualityAssuranceUnit Covancc LaboratorieIsnc. Datc 5,-2 Date 52