Document RJVjv1pLpx7BzLKJaBKZMy298

July 30, 1998 OTs atid! Fax to: Dr. Jack Moore Dr. Joe Rodricks Fax # 202-289-8530 Fax # 703.516.2393 From: John L. Butenhoff Fax #651.733-1773 Tel #651.733-1962 1 have attached our rationale for dose setting on the six month chronic oral study of 'PFOS in Cynomolgus Monkeys. Youwillalso find attachedthereport fromtheenvironmental laboratoryofserumlevels for thefourweekrangefinderanimals. Serum leav ree repl orts edfordays -7, 2, 3, 14 and 29. We have doneananalysisof the slopesofthe cumulative dose vs. serum concentration for these range finder animaattlhse lowdosofe 0.02 mg per kgperday. This is also attached. Bythe endoftheday,wewillhaveananalysisoftheslopesfor thetwohighdoseanimals. 1willfaxthat toyouwhenavailable. "The interesting thing to know from this analysis is the extreme linearityof the cumulative dose vs. serum concentration. `We willneedto produce new charts becauseof a slight error inthedataanalysis. For example: If a serum value is reported for day two,therehas onlybeen one dose since collection ofserum is alwayspriorto dosing on the dayofserum collection. `Thevalue that I stateintherationale (5.8ppm PFOS in serum per 1 mgper kg of PEOS. dose), has taken that fact into consideration. 4:20 Contd. `We look forward to discussing this dose-setting exercise with you on August 3, at 5:30 pm (Eeasten) during our conference call (DialIn # (303)633-6052). ongral fo LZ 03576 Dose-Setting Rationale for Six-Month Chronic Oral Study in Cynomolgus Monkeys Prepared by John Butenhoff July 29, 1998 Recommended Dose Groups: 0.0, 0.02, 0.15 and 0.75 mg/kg/day Rationale: Resultsofanalyses for serum concentrationsofPFOS from the four-week rangefinder (attached) suggesta highly linear relationship between total accumulated dose and serum concentration. A significant male/female difference is not apparent. The consistency of the ratioofserum [PFOS] to total accumulated dose at either dose level and all time. points gives confidence in using this preliminary data to set dose levels for the six-month definitive study based on estimated serum concentrations. `The data provided by the Environmental Lab suggest that serum PFOS concentration increases incrementally with each daily dose of PFOS in direct proportion to the `magnitudeofthe dose. The data give us a good degree of confidence in deriving a factor relating accumulated dose in mg/kg to ppm PFOS serum. This relationship suggests that 1 mg/kg results in a serum PFOS concentration increaseof approximately 5.8% 0.39 `ppm. Assuming that this zero-order accumulation kinetic will holdatthehigherdose over the course ofa six-month study, this allows us to predict serum concentrations that `wouldbeattained with chronic dosing. The apparent precisionofthis is invaluable in setting dose levels that will result in serum PFOS concentrations which can be compared directly to known human serum PFOS concentrations for risk characterization. In addition, we now have the data to estimate the serum concentrations that were associated with effects observed in the previous 90-day rhesus studies. Going back to the original 90-day rhesus studies, the LOEL, which we believe to be close to a NOEL, of 0.5 `mg/kg/day would have resulted in an estimated serum PFOS concentrationof261 ppm. Atthe 1.5 mg/kg/day dose, 783 ppm would have been attained after 90 days. Death resulted within seven weeksofdosing at 4.5 mg/kg/day, which is now estimated to have produced serum PFOS concentrationsof approximately 1305 ppm. We have the following primary objectives for the six-month study: 1) to establish a N10OpAroEvLidefoar cfhrraomneiwcoerxkpofsorurien;te2r)spteocdieestecrommipnaeritshorens;haonldd,ef4f)etcotsprfoorvihdeealethresfuerrveeinlcleadnocsee;:3) for human risk characterization. It is not our objective to study higher levelsofexposure which would result in serious illness in the test monkeys. Dose levels for the six-month study should be set to allow us to achieve the stated objectives: 03577 tLhoewraDnogseef:indTehrestluodwy,dwoseeenxepeedcst ttoo baechaiecvoenfaiprpmreodxiNmOatEeLl.yI21fwpepumsePFtOheSlionwtdhoesseerfurmomof `tahneilmaolws dwoisle baeniamtaalnsdatabtohveeentdheoafvseirxagmeonstehsr.um Acfotnecrentthreaftiirostnotfwoouwreewkorsokffordcoes.ingA,t tshteudy `temramg,nitthuedseeaabnoivmealasvewirlalgehawvoerkseerrusmercuonmcceonntcreanttiroantsiotnhsat. aWreeapaprerosxuigmgaetsetliyngatnhaotrdweer okfeep 0.02 mg/kg/day dose as our low dose. tMhiedrhDeossues:stTuhdye,mwied-kdnoosewitshatthemimnoosrt epfrfoebcltesm(aqtuiecstfiroonmabaledossieg-nsieftitcianngcep)eroscpceucrtrievde.aFtr0o.5m Imfgw/ekgu/sdeaythoivsesre9r0umdacyosn.cenWterateisotnimaasteartehfaetrseenrcuemfcoornsceetnttirngatdioosnes aotntaesrimx-wmeornet2h61stpudpy,. `we would assume that similar serum concentrations would be reached with 0.25 mg/kg/day. From a risk characterization perspective, 261 ppm serum PFOS is two orders otfhismalgevneiltoufdeexpaboosvuereavweorualged wboerdkeesirrsabelreu;mho[wPeFvOeSr],ocfon2-f3idpepnmc.e tAhacthi0e.2v5inmgg/akNgO/dAaEyL at `wouldbe a NOAEL is not as high as confidence that 0.15 mg/kg/day would be a sNeOrAuEmLt.rigTlyhceerSi.deCs.iJnomhanlseosnaWt a0.x3 9m0g-/dkagy/ddaryi.nkiFnegmwaalteesrhsatdudiynicnreraastesdyliievledrewdeirgehdtusceddown 100.06 mg/kg/day; however, we know that the rat liver responds more than the monkey 10 PFOS. Setting the mid-dose at 0.15 mg/kg/day would achieve an estimated serum PFOS concentration of 156 ppm at the endof six months. This serum level is still reasonably close to two ordersofmagnitude above average employee serum levels and close to four ordersof magnitude above currently understood general population values. Therefore, the 0.15 mg/kg/day dose seems to be the best fit for the mid-dose. High Dose: The high dose should be designed to produce threshold effects. After one month of dosing at 2.0 mg/kg/day (resulting inanaverageof 306 ppm PFOS in serum), themaleand female rangefinder monkeys (one per sex) demonstrated the expected reduction in serum cholesterol and triglycerides, which appears from rat and monkey studies to bean early marker ofbiologic effect. There were no remarkable histopathologic effects. Even though there were no major disturbances aftr one month at 2.0 mg/kg/day, six-monthsofexposure at this dose is now predicted to result in lethal body burdens. The rangefinder data tell us that threshold effects begin to occur at serum levels approximating 300 ppm. Using the serum concentrations estimated to have been produced in the 90-day thesus study by 1.5 mg/kg/day (783 ppm) as areference, 2 0.75 mg/kg/day high dose would seem appropriate 0 achieve marginal effects on body weight `and general energy level of the animals. Raising the dose to 1.0 mg/kg/day would result in serum levelsof approximately 1044 ppm by term, which would approach fatal serum concentrations. Therefore, 0.75 mg/kg/day is the suggested high dose that will clearly take us beyond threshold effects of serum cholesterol and triglygeride reduction. Comment: tis interesting to note that the consistent linearity of increasing serum [PFOS] and apparent lackofsignificant elimination may make it possible to give much 03578 less frequent doses or even to give a single dose atthe beginningofthe study and adjust that dose periodically to maintain a relatively constant serum level. We will not reach a steady state in the study as it is currently designed and will be studying the effect of increasing body burden over time. Is there also value in attempting to hold animals at a relatively constant serum level over time? Also, it would be interestitnog look at serum concentration at extremely low level exposure over time; e.g, 0.1 to 1.0 ug/kg/day. 03579 amo . in 32 ren Esramcrae -- em S S --iHE =H --== sn | 238 a@2 rat AMDT 041596.1 oo so 300 00 I 2 100 o 8aa2 o1 02 03 os 05 AccuniatvDose page 08 1A05c3c4D4oMse mgkg000 000046 014 002s8 [serum] __ugiml 0010517200 0014744900 2173870000 105347M AcmcgDkogse 080 000046 001248 058 u[gserum] 00140 0.1470 0072788700 2153660000 10A5c3c4D8oMse mgkg000 004 000164 002588 [serum] ugh 00100 01610 0028879400 3154760000 105364F AcmcgDkogse 0%0 000046 014 005288 [usemrulm] 00161 0.1580 0027832600 5157860000 Sheatt ssleop(selope) Fsvqaulauree 55 (reg) a02s12 54010188 5.749 -000068 se (yyiinntt) 0104 so (osto)f 0043 ss (resid) slope. 56(slope) Forvsaqluauree 55 (reg) 401473 667.074599 4671 000054 so(yyienin)t 008 4 se (yest) of 0028 ss (resia) slope 56 (slope) Fosvqaulauree 55 (reg) 613 024 642078495 8944 -006 yint 007 012 ssoe((yyeisnt)) 00564 ss (resiodf) slopo. 56 (slope) Fsvqaulauree 55 (reg) 085 038 yint 106 029 se (yin) 86.01299 0524 se (yesto)f 23.092 1072 ss (resid) Page 1 03583 10A5c3c6D5oFse mg030 000046 002184 058 [usergum] 0010114780 0062268700 122625000 10A5c3c7D0oFse mag000 000046 002184 058 suergum] 000110T0H0 0062814300 3125060000 snoott ~~ s5h0op(slope) Fsaqvuaalvree ss (0g) a0s0s8 683.160802 4989 000022 so (yyiinn)t 0064 so (esld)f 0005 ss (resid) s5i0o(psolope) rFsvqaulauree 55 (09) s01e3 1686.190808 7623 000084 so (iyinn)t 0064 so (yesto)f 0016 ss (rosid) Page? 03584 `Dose-Setting Rationale for Six-Month Chroaic Oral Study in Cynomolgus Monkeys RecordofAugust 3, 1998 Conference Call Participants: DDrr.. JMoahrnvBCuatseen,b3ofM, 3M DDrr.. JJeofhfnMaMonodreel,,E3MHR DDrr.. GJoeearRyodOilcskens,,3EMNVIRON Dr. Andrew Seacat, 3M Outcome: Dccuaomtnucalefanrttoriamvtetidohonespreaeanrgnmedgf/isnkedgreurPwmFPaOFsSdOidSsoccsouens.cseeDnditisrncacutlsiusoidnoinn(vgtegrti,hfeiftedr$oJun2lg7ry+e/l2-a9t0,i6o1n19s9ph8ippwmbreistteewrnuemeraPntFioOnaSl.e f0aoc.rc0e2spetmtetgdi/.nkggdT/ohdseaesystwtuoidt0y.hw0oi3lnlmepgcr/hokacgne/gededa.oyDntroA.kuMegeouposrdteo1s2se%us,gagt1e9sm9utl8etdwitiphtlahettsthhoeeffl.oowTlhdeolsseuobdggoeewsrseatiliiseoevnndewflgasrs:om CLoonwtrDoolse 00..0030 mmga/kkggl/ddaayy MHiigdhDdoossee 00.7155 mmgy//kkgg/lddaayy Copies: PDra.rtLiacrirpyanZtosbel SDtr.udByiFlilWeep(Tp-r6e2r95.7) : 03585 6-month toxicity study with PFOS in Cynomolgus Monkeys Date: Ir-6295.7 Covance 6320-223 316/99 urobilinogen (UB) Analysis Results from Dr Joseph P. McConnell., Porphyrins [and nutritional Biochemistry lab, Dept of Laboratory Medicine and Pathology. [Mayo Clinic, Rochester MN. [Average IInso Female [asvoerage n7 rest Male asvoerage n |asvoerage nr-rest CmongtroUlB/g 00007187 6000 075 magi0a112 0075 60030507 mCogntrUolB/g 00.008317 6000 075 mar0i064 05.001080 0.059 (n=) 603586 T2057 mont toxic study in Gymomogus Monkeys CcDaowtveea:necze nnUurieorbteionnaolgeBnocAnhaelmysiirsyRaebsu,lDtsepftroomf,LOarboJroasteorpyhMP.edMicoCnoanned,PaPhoorlpohgyyri.nMsaaynod Cin, Rochestor Mi. Annuimmbaelr DLeovseel oossszs9 TiFF ssssas IIFF lloosssseess TIFF A0verage n sossas a4fF o05s5s306 44FF Oosssssi1e 4afF Avarage nTSDest o0s5s107 11mM o08s52t0 m1MM s055z27 1iMM Asnvoerage mg UB per water Stool eight AWadtdeerd WToetiaglht sgpaecmiiod _o wnt fi 1e00 01010 210800 o01o1r 00d10 11800 000078 B6o0 fdoo aisoo 000068 72s2 1o0 w2s2 000082 Go sso 6% 8500 1100 21800 ootose W2o0 1100 220200 00028 Gs0o 1100 11500 00005s 52 10 102 on 3eo2 0& 3so2 060.00307 000 1100 210600 00018s 0e001010 21000 00110 7G00 1d0o 11700 000077 812s 100 e1zs 001003 eo 6 eo eo } 603587 105506 4M 105507 4M 108511 4M l0s512 4M 105522 4M Average so n T-TEST 80 10 180 008 30 10 130 0.04 100 20 300 006 60 10 160 006 70 10 170 007 68 2 188 008 28 4 65 002 50 5 50 5.00 0.059492800 (n=6) 603588