Document EdZVrjrQZk2paqq29B80rqXYn

PRARMACOKINETIC STUDY OF POSF IN RATS - STUDY OUTLINE 06/22/00 T-7098.1 POSF PK Study Objective: The objectiveof thisstudy isto assessthepotentialfororal absorption,urinai@v and fecalclearanceand biologicaplersistencoefperfluorooctanseulfonylfluoride(POSF) in male Sprague Dawley ratsaftera singleoraldose. The POSF compound isthe startinmgaterial forthesynthesisofa wide varietyofperfluorooctanseulfonate(PFOS) basedmaterials.The purpose ofthisstudyisto understandtherateof metabolism ofPOSF toPFOS by the liver.This studvwillprovidedataforproperriskcharacterizatiofn POSF. GLP Status:This studywillbe performed inthe3M StrategiTcoxicology Laboratoryundera definedprotocoland classifieads a "Class B Study" as explainedin TOX SOP 0950, Strategic Toxicology Lab GLP Program Procedure. Test Animals: Male, Sprague-Dawley rats,150-250g,obtainedfrom Harlan. Dose Groups: 0 and 5 mg/kg accordingtothefollowingschedule: Group I Dose N Euthanasia 1 0 mg/kg 5 day I postdose 2 0 mg/kg 5 day 4 postdose 3 0 mg/kg 5 day 29 postdose 4 5 mg/kg 5 dav I postdose 5 5 mg/ka 5 ay 4 postdose 6 5 mg/k- 5 day 29 postdose Test Article:Dan Hakes, ProductResponsibilitLyiaison3M Chemicals Division,willfurnish high-purityPOSF. Compound Administration:Oralgavage. ClinicalObservations:Once daily. Sample Collection:Urineand feceswillbe collectedthreespecifircatsfrom groups 3 and 6 on days 1,2, 4, 15 and 29-postdose. These animalswillbe placed in metabolism cages for24 hours foreach foreach collectionN.ecropsieswilloccur on days 1.4 and 29 post-dose.Liver, kidneys.sera,subcutaneousfatand whole carcasscollectiownilloccur at necropsy. Sample Analysis:The 3M EnvironmentalLaboratorywillmanage theextractioannd anaivslsof sera,liveru,rineand fecesfortheparentcompound, POSF, and itspresumed metabolite.PFOS. Alltissuesamples willbe retainedforpossiblefutureanalysisoftotalorganicfluorine(TOF). 3M Environmental Laboratoryor itsdesigneewould perform such an analysisifdeemed necessaryby theStudy Director. Data Analysis:Data collectedon tissuelevelsof parentcompound and identifiablmetabolites willbe analyzedfortoxicokinetipcarametersand forstatisticalsliygnificandtifferencesbetween groups usingStudentsT-testand/orANOVA. Estimated Timeline (Assuming EHS&R approvalon Jan.5Ih.): In-LifSetartJ:anuarIyIth1,999 In-LifEend:Februar9yth,1999 AnalyticCaolmpletionM:arch22nd,1999 FinaRleportA:pri]l9th1,999 StudyDirector: AndrewM SeacaPth.D. 3M MEDICAL DEPARTMENT, CORPORATE TOXICOLOGY Protocol for Study No. T-7098.1 PHARMACOKINETIC STUDY OF POSF IN RATS 12/30/98 T-7098.1 POSF PK Study Objective: The objective of this study isto assess the potentialfor oral absorption, urinary and fecal clearanceand biologicalpersistencoef perfluorooctanesulfonylfluoride(POSF) in male Sprague Dawley ratsaftera singleoraldose. The POSF compound isthe starting materialforthe synthesisof a wide varietyof perfluorooetanesulfonate(PFOS) based materials.The purpose of thisstudy isto understandthe rateof metabolism of POSF to PFOS by the liver.This study willprovide dataforproperriskcharacterizatioonf POSF. ResearcitClient: 3M SpecialtyChemicals Division 3M Center,Building236 SaintPaul,MN 55133-3220 Sponsor: 3M SpecialtyChemicals Division 3M Center,Building236 SaintPaul,MN 55133-3220 Study Location: 3M StrategicToxicology Laboratory 3M Center.Building270-3S-06 room SB314 Saint Paul,N4N 55133-3220 Study Director: Andrew M. Seacat,Ph.D. Sr. Research Toxicologist 3M Medical Dept. / CorporateToxicology 3M Center,Building220-2E-02 SaintPaul,MN 5513')-)'220 Ph.:651-575-3161 FAX: 651-73)-1773 Study Toxicologist: Deanna Nabbefeld,MS Advanced Research Toxicologist 3M Medical Dept. / CorporateToxicology 3M Center,Building 220-2E-02 Saint Paul, NIN 5513 @)'-)-2'20 Ph: 651-7')7-l)74FAX: 651-733-1773 Ih.): Proposed Study Timeline(Assuming EHS&R approvalon Jan.5 In-LifSetartDate:JanuarIyIth,1999 In-LifEend Date:Februar9yth,1999 AnalyticCaolmpletioDnate:March22nd,1999 FinalReportCompletioDnate:April19th1,999 12/30/98 POSF PK Regulatory Compliance: This study will be performed in the 3M Strategic Toxicology Laboratory under a defined protocoland classifiedas a "Class B Study" as explained in TOX SOP 0950, Strategic Toxicology Lab GLP Program Procedure. Test Material: Dan Hakes, Product ResponsibilityLiaison 3M Chemicals Division,will fumish highpurityPOSF. Identification: Name: Perfluorooctane Sulfonyl Fluoride Molecular Formula: To be provided. Lot Number: The lotnumbers willbe maintained in the raw data. Purity: Documentation willbe kept in on file. Stability: Documentation willbe kept on file. Storage Conditions: Upon receipt,testmaterialwill be storedtightlysealed at room temperature. Characteristics: Information on synthesis methods, composition or other characteristics thatdefinethe testmaterialwill be kept on file. Animals: Species: Rat Strain: Sprague Dawley Source: Harlan Age at initiationof treatment: Weight at initiationof treatment: Number and sex: 30 males 6-8 weeks approximately 150-250g Table Group *1 *2 *3 4 5 6 * Rats I - Dose Groups Dose 0 mg/kg 0 mg/kg 0 mg/kg 5 mg/kg 5 mg/kg 5 mg/kg in groups 1-3 will be used N 5 5 5 5 5 5 concomitantly Euthanasia day I post dose day 4 post dose day 29 post dose day 1 post dose day 4 post dose day 29 post dose as control animals in studies T-70 71.2, Pharmacokinetic Study of M556 in Rats, and T- 7099. 1, Pharmacokinetic Study of FX-845 in Rats. Identification: ear tag with animal number or unique tailmark. 2 12/30/98 POSFPK AUA Number: 2154 Husbandty: Housing: Three specificratsfrom groups 3 and 6 willbe housed individuallyin metabolism cages forportionsof the study(seeTable 2). When not in metabolism cages,theseratswillbe group housed in standardcages. All otherratswillbe group housed in standardcages throughoutthe study. Diet[Water: Harlan Teklad LM-485 Mouse/Rat SterilizablDeiet,suppliedby Harlan Teklad,Madison, WI, and tap water willbe provided to allratsad libitum throughoutthe study. Environment: Environmental controlsforthe animal room willbe setto maintain a temperatureof 72 3'F, humidity of 30-70%, a minimum of 10 exchanges of room airper hour and a 12 hour light/darkcycle. Dose and Dosing Procedures: Method of administration/Dose preparation: A single5mg/kg dose of POSF willbe administeredvia oral gavage to rats in groups 4-6 on day zero of the study. The POSF willbe prepared as a I% (I mg/ml) uniform suspensionin 2% Tween 80 using a 15 ml tissue grinder.A volume of 5 ml suspension/kg body weight willbe administeredto each rat.Re-suspension of solidswillbe performed with 5 strokesof thetissuegrinderpestelbeforeeach sample isdrawn-up in the syringefordosing. A single5 ml /kg body weight dose of 2% Tween 80 willbe administered via oralgavage toratsin groups 1-3 on day zero of the study. Observation ofanimals: ClinicalObservations: Each animal willbe observed dailyformortalityand morbidityand notablefindingswillbe recorded.Additionalfindingswillbe recordedas they areobserved. Body Weights: Each animal willbe weighed immediately priorto dosing,weekly thereafterand immediately priorto euthanasia. Specimen Collection: Frequency (seeTable 2): Urine and fecescollectionwsillbe made on days 1,2,4, 14 and 29 post dose. Necropsies willbe performed on days 1,4 and 29 post dose. 3 12/30/98 POSF PK Table 2 - Schedule Jan 10 Jan 11 day 0 DOSING Jan 17 day 6 PD Jan 24 day 13 PD Jan 18 day 7 PD Jan 25 day 14 PD Switchto met cages. Jan 31 day 20 PD Feb 7 dav 27 PD Feb I dav 21 PD Feb 8 day 28 PD switchto met cages Jan 12 Jan 13 Jan 14 day I PD day 2 PD day 3 PD Collection Collection Switchto Dy I PD Switchto met cages. sac reg cages. Jan 19 Jan 20 day 8 PD day 9 PD Jan 26 Jan 27 day 15 PD day 16 PD Collection Switchto reg.cages. Feb 2 Feb 3 day 22 PD day 23 PD Feb 9 day 29 PD collection Dy29PD sac Jan 21 day 10 PD Jan 28 day 17 PD Feb 4 day 24 PD Jan 15 Jan 16 day 4 PD day 5 PD Collection Switchto reg cages. Dy 4 PD sac. Jan 22 Jan 23 day 11 PD day 12 PD Jan 29 Jan 30 day 18 PD day 19 PD Feb 5 Feb 6 day 25 PD day 26 PD Method of Specimen Collection: Urine and feces will be collected from each metabolism cage at the designatedtimes. The initiavlolume of urinewillbe recorded,thesidesof the urinecollectionapparatuswillbe washed with 10-20 ml deionized water and the finalvolume of urinewillbe brought to 45 ml with additionaldeionizedwater. Daily fecesweight willbe recordedforeach animal. At the designatedtimes,animals willbe euthanizedby CO, and grossnecropsy performed. During necropsy,blood (;:6:@ml) willbe collectedvia the abdominal aortaand transferretdo blood collectiontubes withoutanticoagulant.Blood samples willbe allowed to clotfora period of 15 to 30 minutes atroom temperature,and the clotwillbe spun down in a centrifugeat I100 x g for5 minutes. The serum willbe transferredto labeled1.5mi microfuge tubesand centrifugedagainat 2000 x g to remove any remaining red blood cells.Each serasample willthen be transferretdo a separatelabeledpolypropylene microftigetube and flashfrozenin liquidnitrogen.Liver,kidneys and subcutaneous fatfrom each animal willbe removed, weighed, flashfrozen inliquidnitrogenand placed individuallyintolabeledsterilseample bags. The remainder of each carcasswillbe placedin a labeledziplockbag and frozen(-70 'C) untilanalysis. Specimen Handling: 4 12/30/98 POSF PK Specimens willtemporarilybe storedin a freezerset to maintain -60 to 80'C. For metabolite analysis,thesespecimens will be packed in dry ice and shipped to: Kris Hansen, Ph.D. 3M Envirorunental Technology and Safety Services 935 Bush Avenue St.Paul, MN 5513-'1-3331 Ph: 612-778-6081, FAX: 612-778-6176. The 3M Envirorunental Laboratory will manage the extractionand analysisof sera,liver,urine and feces for the parent compound, POSF, and itspresumed metabolite,PFOS. All tissuesamples will be retainedfor possibleftituraenalysisof totalorganic fluorine(TOF) ifdeemed necessary by the Study Director. 3M Environmental Laboratory or its designee would perform thisanalysis.All resultswillbe provided for inclusionin the finalreport. The number, type and date of collectionof specimens to be generated for analysisare as follows: Table 3 - Specimens Specimens day I post dose Serum 10 Liver 10 Kidneys 10 Subcutaneous fat 10 Carcass 10 Urine 6 Feces 6 day 2 post dose 6 6 day 4 post dose 10 10 10 10 10 6 6 day 15 post dose 6 6 day 29 post dose 10 10 10 10 10 6 6 Total 30 30 30 30 30 30 30 Data Analysis: Data collected on tissue levelsof parent compound and identifiablemetabolites will be analyzed for toxicokineticparameters and for statisticalsliygnificantdifferencesbetween groups using Students T-testand/or ANOVA. Responsibilities: Deanna Nabbefeld and Andrew Seacat will be responsiblefor dosing the animals, collectingin-lifespecimens, performing the necropsy and collectingand sending tissuesamples for analysis. Kris Hansen, 3M Environmental, willbe responsiblefor analyticalanalysis. 5 12/30/98 POSFPK Andrew Seacat will drafta finalreportand ensure the reportreceives appropn'ate 3M review before a finalreportisissued. Signatures: Dr. Andrew Seacat Senior Research Toxicologist Study Director Deanna Nabbefeld, MS Advanced Toxicologist Study Toxicologist Sponsor Representative Date Date Date 6