Document a44GXG8g0Z890X161255KmD0y

ARI 6-005Ly) Synthesis and Characterization of FC-95-1%C 0 = mr Report by: . Yut, in ls oF nncmacoriaias ~ Do + 2 Bet "175 chemical Characterization Section Reviewed by: E(Wey wig CLO 7 reeterebartgreatine 000019 2. Sumary Tihsedessycnrtihbeesdi.s ofThea s2p0ec.i0gficlotacotfiviFtCy-95i-s140C.45(9ca&rb0o.n0-0184 ulCaib/enlg.a toThisnu-llfauyreratom) acnhdemiccoallulmyn cpuhrreo.matoThgerapFhCy-95s-h1o4wCedwatshe fFoCu-n9d5-s1u4iCtabtloebeforatmelteaasbtol9is9m% rsatduidoie-s. 000020 3. Introduction A(asesruilefsonoifc eaxcpiedrismaeltn)tsanids tphleapnonsesdibtioliitnyveosftigFaCt-e807thebemientgabbiooltirsamnosffoFrCn-e9d5 tcoarbFoCn--9154anlda/boerletdheFC-a9l5cohwoals s(yFnMt-h34e2s2i)z.ed aTnodfcahcairlaicttaetreiztehde.se Tehxepersiymnetnhtest,ic pathuay illustrated in Figure 1 is described. 000021 a A. Synthesis of pc-95-14c I. Fractional Distillation of C.F,"CF ,S0F (step 1) TGahlseahsescdruwdoteowlicceetlolwirdterhmaoivcneoildnt,ghse difcleruloltmectaesrlaslt.urruanTtheeRd-3KEH2lC5uO6o,-r3ocawhneedrmeitcwaiflcielptrweoirdteuhdcatthtwreaorsu;gh wtahsehperdodcuecltl vdarsaindirnigesd "nalysis shoved TL.14 owC.F,avseSrOobsFtial.iinceadgOenfler.ohmucnAedlyrlieedlrdusnioxfRt-y3229g53r6.a-7mB9s. ooff GbLtaChsee-base- w(asseheedTabclelel1d).rainAinnaglsyswiiss fpourripfeixecdenbyt CfrFa.ctSiOonFalwdaisstdiolnleatbiyognas chroma- 5to3g0r3a6phyOF (parroedaucpterwcheincth).vasFrfaocutnidonfso c1o1n2t1a8i7nwef9e8.1c5o%mbiCGnFe,d StOoF.give II. Preparation of FC-951-4Mc (xsoec.recF Vwaetreerad(d3e1d.19)0 been equipped aandS0p0omtlasstihurnes-hyndercokxeidderopuenldl-ebtost,tom(e8d5% falsassaky,whi1c5.h53h3a)d with a reflux condenser, thermometer, an air-driven meeqcuhaalniizceadlasdtdiirtrieorn, fTurnnue-lB.oredThestifrlraesrk acsosnetrebnltys Fractional CF, fCFS0.F (27.0, 96.15% assay) avnedreahesmaatleld, tpore7s8s-u8r0eC-. was added dropwise Cttiuhorrneo.ubgehtUwptehoeennAcdo80dm-fp8tl3ie0otCfi.onfinGonfuemltyh,aetwahd4idtiretaitoesnolsiuodffsfitwcheeireenrtadftiooormamecadtiindvtuearipinenrgftihteuhoertoea-mddpie-ra- 3oicnttaotnhees-msaeulnlldfeoronfyslothleifdlutophririeedceee,sh.outsheWrametiaexcrttui(ro1en0.w8taismmel,)hevattahesedgaadutdmemdy8.5mCasTshfeorhapid3 bhoorfuorktseh.ne Sanodluttihoenswoalsid~s w1e3.r5e.waTshheedupspuecrceasqsuievoeulsy wpihtashewwaatserre(m5o4v.e0dml()asaptirator) 2s5t-i5r0r0eCd. atThaehisgohlidasgiwtearteionwasrhaeted,onacned mtohreeuwpiptehr waaqtueerous(1p7h.a8smel)w,as dSioslciadrdperdo.ductI,soapnrdopatnhoelmi(x1t9u.r15e5)wasandhewaatteedrat(1r7.e0fgl)uxwetreempaedrdaetduretofothreone ohnouer.hourA.L Tohfethfelasskolicdosntheandtsdiwesrseolpveodureafdteirnthoeaatignlgasstheevmaipxotruarteingfordish. TChoencefnltasrkatwiaosn woafshtehde wpirtohducitsowparsopadnoonle o(n24a.7smtle)a-nvbaattehr. (1A7.0fimnla)l.drying oifn tahevapcoutuanssdeisuimcepaetrofrl.uorTohoectadnrsy psouwldfoenrawtaeswatsritdounreataetd o6n0c9eC wfiorth3 hours FSrteiornrer113lutborirceamnotve; Tshmealldriterdacepsrodofuctsil(i2c7o.n89)oilvasimapsusriitgynedusetdheasfollowing number: Fe-95-1%c, 20.05, 14544. sCu-l9f5onynlormfalluloyridies p(raosdsuayced66-f6r7o4m).one-ThpelahtieghdisatsisalyledC.Fp)erffClFuo,rSoOoFcta(n9e8-.15%) will also be used to prepare FC-807-14c. 000022 5. 5. specific Activity Determination of Fe-95-}14ic TwehrreeeprsetpaanrdeadrdbysowluetiigohnisnogfI,C2-59.50-0%CmgE; (I1I4,54246,.84seneg;Synantdh,esi1s11,Sec2t6i.o7n9)mg mientthoantohlreeand25smollutciloansss AIfvoalnudmeTtIricwerfelaspkrse.pareSdolwuittihonaI1w:1asmpirxetpuarreed with w(ivtohlumteh:evoalpupnreo)priosftewatseorlvaenndt maentdhamnioxle.d bTyheinvvoelrutmiensg wbeyrehanadd.jusCtaeldibtroat2e5dml meiacchropsiolpuettitoonrsdtiwreercetlyuseidntotosacliinqtuioltlatsiioxn1c0okulntianngd sviiaxls5.0 iToaltihrqeueotsofofthe VEiraolnscaccohntapirniimnagry1s0oluultiaolni,quo1tBslanodf twoatetrhraenedco15ntmalinionfgAq5u0a4s1olealdiwmeuroetsadded. The remaining vials vere prepared with 2.5 ml of methanol and 7.5 ml of wMeIiqStuShiESl,aibPrTaahctekearsidnamMptolhedeseldawre3kr3e6b5ecfoLooirlqeeudidtthoSecyrienweftrrieilgleacrtoaiutonontredSpteetcmwtporeortmaietmtueersre.atandT5heamilnlcuootwueensdtientagoch efficiency as determined for each sample by the internal standard method. `The averaged data were reduced to dpm and the uCi/mg was calculated for each vvAieqaiulga.hsionlgTeshaenvdadsaMtIaSwSitahrweienresh2ow4wintofhiinneacT1ha.b2lo7ef a2t.nhed TotthhheeemrmeeamaennasnusbC.eit/wmTegheenfomu1en0adnuslfoabrentedwaecSehonuolf the vreerpeliwciattehsin (112.3%f.romTheeachoveprrailmlarayvewreaigsheinsgp)ecwiafsic0a.c4t5i9vi+ty0.b0a0s8e0dKCoin/ntgh.e 36 C. Radiochemical Analysis of re-95-Y4c I. Thin-layer Chromatography Systems and Carbon-14 Analysis Tthhein-alnaayleyrsischorfomartaodgiroacphheymicsaylstpeumrsituysiwnagseictahrerriedSGoFut25w0itmhicaronvarpireet-yscoofred UPnliaptleastewsotr.eorropurtei-naedlsyodrbeevnetlopSeGdF 2in501m0icxr12o"nxp"re-tshcionr-eldayUenriptlaantkeesft.fitted wdSicetrvhaepligonlpgas1s5uesclmidascancdoanmwdpelrilesihneesddcrwawipitethdh aslaacttueusrrtaaoltmli-yomnaipdnaed0s.t5e.mpcmlPawltiaedteeasndsweegsrmheeanrtapsle.lnoewdeTdheto Stainless steel spatula ground to exactly 0.5 cm width. 2 Riker Isotope Number 442. 2 WbeailgahnicnegswhiwcerhehaacdcobmepelnisrheecdenotnlyacaSl-ipblraacteeMdetbtyle3r HMoedtroelloegcyt.ronic 1/1 Micropipettor, Lab Industries, Berkeley, California. < New England Nuclear, Boston, Massachusetts. Modified TSS:25.2 PPO, 1.0lg Dimethyl POPOP and 3.8 liters toluene. analtech, 75 Blue Hen Drive, Newark, Delaware. 2 supelco Inc., Bellefonte, Pennsylvania. 000023 .. TCchaoetnetsas"icnriaTnphgeed.2.ss5ialmaiplclaeosgfveemlertesheagcnmooeulnn.ttsedwTeoarnedthcetohlmeeltechcoatmneeodsl,iPne7r.s5cMiinnnlttivloelfatM(iIcoSgnmS) vwviaeasrlss TCSoeemrmreecncottmse.wdesrcfeorrapnbeoadtskcfaorTroormuencbdteelduosifnToghre aefofrsiiecsiiitenanbcioyen. bshleaTnhkeplac(taeezrbmbolenlilyndgSawcesosm0ae.rn5etdo.omf "ach segment was expressed as percenotf total carbon-1 on the place: SSmpemeog nnm sege menn t t 100. = 4casrbboonn-1-4L4ccoonntteennttinin segsmeequnetnt. TmShebetiavpiretrycTpeehnaitkssc.aprrcbooovrnir-de1es4dponcadointnthgeinnt-ilooafyseeerpaacrrhaatdesideoqdmhceoinmotpmoawntaeosnstEpsalnoitnsthetodhweinvmgeartstuehsreiassleagdmieont Seriied to the plate. II. Solutions Used for Rasiochemical Purity Analysis SmToehltiuhstainTooenlpodr1:esscwTrahsiebeddFeCsi-io9nn5ta-ht1ee%dCsmeaacadteSioolnfurotonimoSn2p5e.1.c0i0fmigc oAfctirvci-t9y5-4Decterinmin2a5t.i0onm,1 MoSpeouemlbutseterio5dn4543e2):0nsvu1ar02se8..01pm)2ilnawicminegtgdh.ofmirentteho-a9na5ol-1.0Ycw,hTheLv-o4ls5uo4ml4eu,ttriio(cnRivkfaelsraskiIsnaovntedorpteetdheInsvveeovnleturomarely III. Column Chromatography ofFe-95-14c - ASSi1u'rseieylicoif"Acs1ai2cl0iimdclic(aGalnciiisqidu1oti)notfecoaSloulLmuitmwiaosn1.0p3.rveapgsalraesadsppibciyoelpduomuntgobinttgoheaatchoehpilgoohrftoftoohfren C5"oo5tliu0mmmne.);of"FTCrhahiceestki,osnucFcr3eascstaiinvoden203170002mmL00omffLrmaecotfthiaconhnoislc,rwoefrFoerrancc-tomileoltnehcat3n.eodlEaaf11ct1ehro(afvpopltulhmeyei:ng G3f20o0'nami11t2rorsmesecnt,ciooannnrsdifwruaegsceoenvtsuatbpiaot,ruatteeevddapwowiritathhte0da,2r0soomtnlaetaoirfngdcerhvylanoperosorsfaotwroinrt-,hmetathraasnntcsrifeearm1r:e1d aCnodluanneasluyosleidmea)s.deFsicfrtiyvemdipcrreovliiotuesrisy.wis spplisd to thin-layer plates tv. Results ang Discussion Since the chemical identity of the re-95-19C is well established by synthesis (see synthesis of FC-95-19c, this report) unlabeled FC-95 was not co-chromatographed with the labeled FC-95-14c. The results oCfhrothmeatorgardaipohcyhemaivcealShpouwrnitLyn Taanballeys3isanodfinFC-F9i5g-u1r4eCs b3y.5.thinT-hleasyeeranalyses dhT(iaeedeteeinnooFatiCcgoiudlvriuiesmtcye6rF)n.rraecactnoFiyvroeanrcsi.etmdipouFnrriiontm3iFera(sahcsatitiilnhoainncFoiCdc-3)95a(-ccL1ioi4d]nct.scaniolnloTeurhdmoifn0o-.srl8hmao-pywmeeeerrdtha3cane3nan.olt1lys)soiofsFtohtfehe 2 nChiesmiic,alacCotmipvaantyesd snieli.ciWcilAlciiad,msp1o0r0e-,20P0ommmesehy.ivanCilaa.rkson 2 Column was fitted with a sintered-glass base and stopcock. 000024 7. recovered radioactivity (see Pigure 7). The impurity contained in Fraction 3 has a different same solvent systems. The Ri,fpu(r0i.t40y) wafsronmotthaitdeonftifFiCe-d9.5 (0.27) in the overall, the radiochenical purity of FC-95-1%C is at least 90%. The FC-95-14c is suitable for metabolism studies. Acknowledgement Tohfethaeuthcoarrsbong-r1a4teflualbleyleadcCkFn,owlCeEd,geStOh,eF gfarsaccthiroonmsdaotnoegrbayphiMcr.anTaoldydsis Mathisen of Commercial CheAibals Division and the assistance of John C. Hansen and larry Headrick during the electrochemical fluorination of labeled C.F) CF,SOF. 000025 5. List of Tables and Figures Table 1: DCa4ta8fry57oCFm, FSrOacFtional Distillation of Base-washed NB 50942 p. 34,35. Table 2: SNpBec5i1f3i1c2 Ap.cti3v3.ity Determination of re-95-Yc. Table 3: NThBin5-1L6a0y7erp. Ch21r.omatography Systems forre-95-2c. Figure 1. Figure 2: Figure 3: Figure 4: Figure 5: PNBath5w0a94y2 fpo.r S36y.nthesis of FC-95-1dc. NThBin5-1L8a0y7erp.Rad4.iochromatogram Plate 1. XThBin5-1L8a0y7erp.Ra4d.iochromatogran Plate 2. TNhBin5-1l8a0y7erp.Ra5d.iochromatogran Plate 3. TXhBin5-1L8a0y7erp.Rad5.iochromatogran Plate 4. Figure 6: STihliinc-iLcayAecridRaCdioolcuhmroEmlauteongtr,amFroafctFiCo-n95-214c N(BCh5l1o8r0o7forpn.-e1t8h.anol) . Figure 7: sTihliinc-iLcayeArcidRaCdioolcuhmroEmlauteongtr,amFrofactFiCo-n953-i(cethanol) NB 51807 p. 18. 000026 5 mate 1 Data from Fractional Distillation of Base-Washed CF,Cr 50. itel ERmeEw Fraction bE C (@ weiof cg CcgoFi1i3Ss0o2lF CasoF ms15tS 13 5 ccwe waa ss wa Go-s3 a7 ass 2e0e i: 5Sa2as as1l mGsem 1a2sse :; sSdaors 413a saoae emsass 05 isolss 2L5e mdaedrs 115.%03 110 iLeesz o2a7 s72u3 msaazs 312 1 loea s7laa ik ome oaos 1I9F3 = B1ot16eBons seoo domeae o ==. Blend FR 1-16 oolms 72.2a0 --= 12d1se7 12e3om3s ==sdae msias == ssem eeems == so4is% seslnsesss oj=uos Vosz wseilss oollss olfoase sSahm ee2 08.15 2 Area percent was determined by gas chromatographic analysis. Lox hotiing non-functional Fiuerocasbons below Gof. siginhg,-unibdenotifiied Fluorocarbons boiling above CgF, S0,F. 000027 Table 2 Specific Activity Determination of re-95-14c (25S.o0l0u5ti9o/n251. m1) Ving 00..44555385 o.0a.4s6e528 00..44871366 ol0.a4s623l7 00..44568397 00..4460553 45.0. 0.4634 + 0.0085 comparison of primary Solutions (26.S84olmuat/io2n5.011m1) ci /mg 0.04.4563100 00..4655325 00..4651017 o0.a4s4e2s2 00..4450460 00l.4a5s1766 0.4554 +0.0063 10. (26S.o79lutmigo/n25.1011m1) img 00..44865407 00..4455392 00..44565444 00..4452756 00..44566275 0o.u4a4s510a 0.4565 + 0.0071 Comparison of Scintillation Solvents \MCiS/Smg 00..44685186 00..44762367 00..44668553 00..44665027 o0l.6451610 00..44551786 ol04.4s53524 00..44662454 00..44541530 3 5.0. 0.4614 + 0.0084 aquVeais/onrs 00..44553558 oo.lasse)2 00..44653879 00.44651300 o.0a.4s5e3s5 0ol.s4a4s2e2 00..44666407 00..44452958 00..4455767 0.4558 + 0.0067 Comparison of 10 4 and 50 ul Aliguots Te1i0znpsl 00..44553558 o.0.a4s6e528 00..4403266 00.44563100 ol0a.s635s2 00..44661017 00..44664670 00..4553528 ol0a.s6s4a8 0.4615 + 0.0076 \5i0/ngul 00..44562317 00..44568397 00..44665533 o0..a4s6e5s 0o.l4a4d2s26 00..4551786 00..44452756 00..44566275 0o.l4a4s910a 0.4557 + 0.0075 000028 aes a oEmS he BFe= Ea = Thin-tayer Chromatography Systems for p-95-'c. . we m1 acmn . we 1rmym wr . we pBaEm cite . a reine oEBoaE ae wor gg. . Semestnnres pessmtniyv-- o tnmeo rcs 2Solvents were prepared voluse:voluse; a 100 ml aliquot of solvent 12. Figure 1 Pathway for Synthesis of FC-95-1%c C,H, C'H SO,F2a EBCgEFg CF, EF,S0,F R-3256-B * CA7sFsya5yCF7,1S.O1,%F Fractional : Distillati=on ACs7sFa1y5F982.5105F% 1 1+ on H,M20 " Foor KOSSCE,CF Fc-95-14ck * Denotes position of Carbon-14. 2C,H CH,S0,F was prepared by Pathfinder Laboratories. b pc-95-14c, potassium perfluorooctanesulfonate. 000030 60 50 8 =4 E o 30 &8 s 2 2 & 10 13. Figure 2 Thin-layer Radiochromatogram of FC-95-14c, Plate No. 1 SGF Uniplate: 110000 cacheltoornoeform Total CPM on Plate = 39770 3 0 0 1 2 3 4 s 6 7 8 9 10 11 12 13 14 1s Distance from Origin mn 000031 Figure 3 1a. Thin-layer Radiochromatogram of FC-95-14c, Plate No. 2 SGF Uniplate: 110000 cmehtlhoarnooflorm 2 acetic acid 60. Total CPM on Plate = 41058 50 = 40 5 E8 7 30 3 E 20 2 10 0 0123 T 7 8 Tou 12 Is 1a 1s Distance from Origin (0 000032 15. Figure 4 Thin-lFacy-e9r5-1R4acd,iocPhlartoematNoo.gra3m of SGF Uniplate: 10100 bwuattaenrol 10 acetic acid 60 Total CPM on Plate = 38549 50 = 40 E5 o 30. 2 S = 20. 8 & 10 0 c 1 z 3 4 5 6 7 8 Ilo 11 12 1s 14 1s Distance from Origin (on 000033 16. Figure 5 Thin-lFaCy.e9r5-1R4aCd,iocPhlartoematNoo.gra4m of 60 SGF Uniplate: 15500 mcehtlhoarnooflorm 5 ammonium hydroxide Total CPM on Plate = 40842 50 == 40 5 E5 o 30 &5 s 20 25 10 0 6 1 235 & 5 6 7 8 9 10 11 12 13 14 1s Distance from Origin (cM) 000034 17. Figure 6 Fe-95-1144,Thsiinl-ilciacyerAciRdadCioolcuhmrnomaEtluoegnrta,m oFfraction 2 (Chloroforn-Methanol) Pre-adsorbent SGF Uniplate: 10305 mcehtlhoarnooflorm 60. S ammonium hydroxide Total CPM on Plate = 429,720 50 Zao =5 Ez5 o 30 S e 2 20. 2 10 o 0 1 2 3 4 5 6 7 8 9 10 12 11 3 11 4 15 Distance from Origin ) 000035 18. Figure 7 FC-95-1144Thisni-lilcaiycerAcRiaddioCcolhurmonmatElougernatm, ofFraction 3 (Methanol) 60. Pre-adsorbent SGF Uniplate: 10305 mcehtlhoarnooflorm S ammonium hydroxide Total CPM on Plate = 3565 50 S = 40 =5 E5 3 3 FY) 10 0 0 1.3 5&5 & o 1 Iz Iz I Is Distance(cMf)ron Origin 000036