Document oeD5en1RObGGBYKbr3oEVdy9X

K JfH'BNA* Pf fb>LKiR At f V i|,N. li.lV-rilwWi ftnl.i . V.SJ. t Metabolism of Ri phenyl and 4-CIilorobiphenyl in the Rabbit* AY.U.Ttll D. BLOOi AND IIebbkrt II. Counku From. the Dejtarttutat of Dermatology, Medicol School, The Unipcreily ef Michigan, Ann Arbor, Michigan (Urod^Td (or publication, August 24,1930) TV ability of the rabbit tonv-taholi*e nnphtliak-neandelilorinitrd ncphlhaknc wa* markedly influenced by tlic drjrw of baloQruutian of Urn compound aitortling to Cornish and Hlock (2). Xaphthaknc is related structurally 10 biphenyl, in whieh s cua\ial linkage connects two phenyl molecules. WhclluT Uic presence of a ebloriix* atom in biphenyl al o alters the mrtaliolifm of this compound as it <k*.s in the ease ef naphtliakne has not been Trj-orUd. Stroud (3) and W't-M ct al. (4) have rtudkd the nrtnbolkio of iin$ubstitut<d 'nipl.t-nyl rerpvctivdy in rabbits and bi rat.*. The promt investigation is a 'Comparative melabohc ttadj of the urinary end-produets of biplietiji and 4-ehloroliipinyl in the rabbit. EKITJIIMUXTAL T -rooip-xinds biphenyl1 and 4-elilonibipltenyP wi-n- recrya- talh from aleoltol until the mc-Kini; points were constant, 60-10' and 76-77', respectively. Sis male albino rahhilr (2 kg in -vcight), fed a diet of oo<s and cablugc, were r*d for each coii.|-ound studied. Twcnty-four- Irnur iiri-ie samples were eollect<*d ior a 4-<lay period both before and alt- r administration of either 1 g of biplicuyl or ) g of 4-clilo- mhiplienyl in eoni oil giivn bv ftone"eh tube. Each animal served us bis own control. The urine samples were cnalynd for their content of the fol lowing inctaljnliUs: Fhiiijskluroi.ie acid-*, phenolic eoni|ioiinils, ettieTcal sulfates. and incrtaplr.rie :uids (neutral sulfur), ('ronti- ni'ie ci-ntcnt wa also det- mured. Ghiroshhirnnie aeiiK wen- assayed by tlw procedure of llanson H at. (5). 1 Mh-i!*-lit* i nnipniiii-ls ware determined with the i'olin- Ciocnlleu rvu> ut (I'd, with l ii\ ln-\\bipl:i nyl as a standard, tsilfiu part it ion* wire bv tl.e 1 >. i him miI fate precipitation procedure of K-)'in (71. I 'r-.-at;: e eoni rut was detennimil by a )4>riloni-tii'". ii.islita :.iioa -if Fi.iinV picrl- arid method (N). Tie- amount of the \:- t.ili>hl> iM.irb-d by i-nrli rabbit during the 4 days wlii n m-itlii i l-ipla nyl nor 4-r!'.l:iruliiplionyl was f<d, wrr* fubtra- t.il f.oin the are-nn: - \. u li il during the 4 iLiy-. after the conipounil wa* fed. Tin im n mi nt, which was pn.-iiin-d to br can't i! by fit din- of t!>c !n d. -nrlioii, wl ah'ul.'tnJ in terms id milligrams of eitlur l-ijile nyl i.r -l-ili).-t..biphenyl. For ftudi-'S in whieh mo era I oi tin- exert tory products derived from Inpl.eiiyl 4--l.lori-l-iphi nyl w.-p- i-nlatcd from urine, 1 of i-i-a |--.iiitl w i> fid (*i ral-l.iis feil ivieh eonipoiin-li and ll*e i: work was M!;.p..r:,-.| in part bv I'nilcd Stales I'tihlii: lie: till s'-fvire tlralit IP! ' PM. A pi-ri|Hri lias lss-n pr- r.l (|). * lt;|'!iinyl was i.l-taid. .1 fru.ii KaMiiiaii Organic Cliiinir.ils, IVo-iliaiion JV.-l-ii o ladu-tu.-'. Ib r. New Vi-rk. 4- thlo'i-iiipbciiy I v.;ts i*tii .iin >1 i!.;'.-ti:-li lie- i`ourie..v r,f NIiMiiaiilo (la :.ii- sl ('oinpanv, Si l.n-n-. Mi--ini. urine coHccled far 4S hours Urine collected from animals fed the taioc compound was pooled and frozen until analyzed. An atirprot of the urine was adjusted to pH 7.2 to 74 with 10% NaOII and continuously extracted with ctlicr far 24 hours. The ether extract was evaporated to dryness and lire residue taken up in hot ab-silute alcohol and water added until Use nlulioo hrenme. turtiid. The mixture was placed in the raid (15r) far 24 hours and a nyMaDinn phenolic eompiiund obtained. After several reeryt-talliutions this ciHiipouiul gave a rooutant indting point. Tlic elhtr-extracUd urine was eonemtrated to one-half its original vohimo under reduced ireerun-, acidified to pll 2.0 with 10% 1IC1, and placed in the cold (15*) for 24 hrra. A yellow precipitate higli in ghienskluronv' acid coutcnt w.-is se cured. This material was n-vrystalliscd from hot absolute alco hol until a constant inching point was ditalned. umn 9 McMtUic ffidia Talde I shows average 4-day exeretinn valuer of various uri nary metabolites both before and after insertion of biplienyl ami of 4-chlorobiplmyl. lk-eaurc normal daily cxrrctiou of these metabolite* is variible, ebingi-s in exendinn of hr* than 20% are uni rowkWred im-aniusfui. Amounts of ingest id bpliciiyl ami 4-cIiIuroliiplii-nyl nceounltd for by the four urinary metaUdiU-s determined were c*i*e:itially tlc time, 030 mg and 047 mg. Approximately 20% or 202 mg of the biphenyl was exrrrtid as tlic ghn-OEkluruuir nekl ili-rivative, Imt 50% or 505 mg of tin* ingi-slcd J-i-bloroliipl*'iiy|.was rxi rvtori in thi;: maum-r. Free jdienolie mmpcnrwl-' in tin- urim: aprountrd for 2-13 mg of the inpo tsl bijilienyl a-- eimtraytul to 30 mg of tin- Tchlorobi|ilicnyl. Idlu-tval sulfate cxercliun accounted for 13 ami 11 %, rir|Kvtivi4y, of tin*, ingi-slcd Inplmriyl and 4-ehk>nibiphenyl. XeitliiT liiphinyl rmr 4-ililuroliiphi'nyl n-emed to he cxrn-tiil as nwriapturir at-kl derivative*. 1fetation Sludir* Bipt cnyl .l/ilitbolilci--l-llydro\yliipl.nyl (in.p. 105*) was iso lated from the etlH-r-solublc fraetkni of uriin- i xrn-tiil by rnlihll* fid Itiftltarty 1. The nivtute and Uur.ixite ik-rivatives of this eum|xnintl Wi-te ptvpnd ail franwl to have melting points of SS* and 151*, nvja-ctiwly, which an* cuti-ist-'iil with vaha - n |x.r!cd in tlic literatnn: (0;. P.iplMnylgltuivi-lurntiie nekl (m.p. ISiJ1S-IC) w.-i-' L olafid fnnn tla- ellH r-hiSuluble fr:n-|k*n of tlx- mini-. Hydrolysis of the Mnn|wiuia| with aekl or d ghn iiiiinid:i-e y k klul 4diydro\ytiipliiiiyl (m.p. I*."'*). .\ilditkm of know n l-byilr\ybiphenyl dkl nut ik-|n-*s tl*- melting |mint. A-ei.it-- ami ls-nZiKili- derivutixes of tlx* i-ulaleii -I-hydnixylnplii uy 1 had melting |min(h nf K7-Sv>* and 1 !!", rcsjuvtivrly. 3301 DSW 255034 STLCOPCB4060725 Sift? Alrltilxifisiii nj fby>/i'-// owl i'hUtni`ifJirntd vl XJI.X,. \: , i r>f IHJ-- .'.i-n f{ t'f urinary Tmi.i: 1 (iJ {-fW4i/'/,riii/ on eicrctimn by Ike T,:l-bil ('14KI134 ;r t| Mi Jfv I| Cbcaji>U Mttutf lu:* I i'l trr !* noii> rvii*>mwU i"l; I 5* "_V-5 ivTMMl l nr><o- tl . i c?t Ifj s | lli 3 |t: Fa!I -- ^ | p^ad frtl - | H I * - Control.. tol ... 4 (lilontbi* llruyl... X "t | ri 7! ev m' lo ii 41 IIS Ii is 112' SO' ftI "I ml f % 510 311 Sol! 262 3st 213 630 r.t 512! 231 1 IRJj 505 50Gj 30 or 65 The values represent the total 1 d.vy excretion. Each v:iluc h the average of result-, obtained on six different tiuinuli. lltilmljsn In- & of tin- plue*'i-h,riiiiiif. : lalvd fmm tie* urines of auimul-s fed cither rompannl itt<It. a l-tu linb^i' till I In- phi'-irdriuronir acid innirty. AllNaijIt li)>lciiyl is ivlnlcd structurally to in pi.that h, . tied Imtli are ronijxised of lun |4>cnyl Knni(K bid with dilT.-r. . linkages, (lie |ctthwny fur uu-t.-dioli-ni, by the r-'!>Vit, of tin- , com|itiinls arc rl'i.-similir. Cornish nnd block (it r.|.i t d t`> [ tlx* ingi-Atrd u.-iiditliukmr wa* rxcreh-d ns mere 'ptnrir :t.,; (Irrivutivcs,al*d 2',|.ns free jJielmtie com|>ound;. In the pr, , study, no tuen-ment in nvrraiitiiric mid excretion r.ut-.-.l feeding tdjihrnyl could la- rlclretrd, and, n.< niontioni-d l.f..,. free |ihcnnli<- ikiivativo* were an ini|>rtnnt cxcielt.ty p-i-h., Cornish and block (2) did find that morr f-cht'irmtaphth i!,-n and dichlonmaptilliak'iie Ilian naphthalene we* ewn ted a.-. ii.- plucosiduroti'nle. The prr~enl study would roiTnu th. Imi;. . Ihxit ghicosiduronide formation Ik-nnur* him.- im|>ort-iiit e> mctalmlism of tin- ingestiat Imlmcrrlfoti by the rabbit wl.-c e. com|iutmd U luilogruated, nnd Hint apparently (lie rnhlnt ;,, - a mechanism for llir. ddialopcnation in tiro of sin-h cohiimmi (2)- 4-Chloroiiphi nyl MctpMit'S--4-(/>-C'hloroplK.-iiyl)plicitol (m.p. I-Mi1) as isolated from thr rther-.vilulrle fraction of urine ex creted by rabbits fed 4-chlorobiphviivt. Acetate nnd In-nioat derivatives of this compound hail melting points of 110* and IS'2V, r>-^]s.vtively. These values arc consistent with tho.v n-(>urled in the literature (10). 4-Chlorobiplienyl giuensiduromde (ru.p. l.vV; was isolated from tlie ctlirr-insohible fraction of tire urirs\ llydroUfis of iIk- compouml with acid or ^-ghK-uronulase yielded !-(/ clitoroi*t:ciiyl)phcnol (m.p. l-15a) whose acetate and Ix'inoafc ik-rivatives lent nu-lting points of 110--1 ll0ar>d 181, respectively. The isolation procedures used in this study do not rule out llic pre-cnee of otln-r inefalxrlitcs of biphenyl and 4-cldorobiphenyl in the urine of rabbits fed these eoni|>outids. uisccsstox The results of this study indicate that 4-chlorobiplienyl is inejalir.lireil as readily as hiphetiyl by the raWiit. Tire presence of m chlorine atom in the 4-jiositinii of biphenyl, however, iky-a alter the didiilnttion of the urinary metalMditis of the eom|SHind. Kvs-iitially twice as mil' ll l-<ldicohiph. iiyl ns hiplienyl was excn-l\t a-1 the i-.hieosHliiroiiii: m ic! deiii ative. 'flic isrJation stud ies iiidiralid that the gltico-idiironir a-ids of tmtli 4-<-tihnitii1 'i- i:yI ai:<l hip'ienyl were foruieit by con;iie,ati<m in the 4- nr /urn-podium. Oxidation of the 4' |msil'on of ttn- 4-thIonl>iphenyl to a phenol group must time taken plane in order for th'- ghi< I'.-iduionidi- to b.1 forna-d. and l-(/M-hloropln nyll|ili. ii"l wa- i-otab d from (he e(her->oli'Me fnietion of urilM' rolhvtiil fr.an ralibits fid 4-i tilornliiplu iiyl. Nevcrtlieli'ss, tlie aniiiiat pn-f.n-ntLilly form'd the glm o iduroiiide derivative for eXtretHHl tciaii-.- only a small fraction of the inge.-di-d 1-chhirnbiplieiiyl (l';i was i-M'icted as ttie fn-e ph uolic eoin|Kiiind. Hliis |iaftiway was fi major oik* for the excretion of biphenyl (21 ` i) au<l was i-ijuatlv as iin|>or1.ni( as tin- fnrnmtioii of ghi'-osidiironiiks in bip!i'-u> I ii'rt.ilxilism. tUUMARV Twcnty-four-tiiKir urine sanipk-a were rolhvUd for a lsi>. period fioth ln-forr am! after oral administration of cither 1 r biplicnyl or 1 j; of 4-ctiloroluplwnyl to rabbits. The urine i, lik^s were analyzed for tln-ir content of glucOPidnronic ncids e.v pliritolie compouiKls, mcrrapturic ncids, and ctlmnail r>ulf.-.:- - These mctalxiHtes in tlie urine accounted for Clh or th. gested bijjietiyl or 4-chlorobiplienyl. 4-Cldorobiphcny! we errted mainly as the ghtc<*iduti>a>de (50^); free phenohr . pound.; (24^i) and gliiifisiiluroniitr* (26^i) in the m ine aceo; u1 ' for most of the biphenyl. Tims tlie presence of a chlorine ; in Uh* biphenyl moleeuk' altensl the metabolic jeithway ia t; rafibit. Isolatkzii studies indicated Ibat the ghicoskluronic acid dcrr.atives of both compounds were fornuat by conjugation in t!"- 4or pnru-posilion. HEFF.lt I'XCJvS 1. Cokmsii, H. II., Axr Hixck, W. 13., Federation. Proc., IS (ItTfl). . 2. Cornish, It. 11., and Dus k, \V. D., J. Biol. ChemS3!, . - :t. Stkouu, S'. \V., J. Endocrinol., 1. 55 (19t0). t. \Vt>T. It. I)., I.AWSOX, J. It , MlI.LKR, I- H-, AMI M-ITlie:'-' (i. Jt., .trrk. Hiorhrm. Iliophya., 60, 11 (1`3-Vi). 5. IIanmxs, S. W. F., Mii.lw, (h T., axo Wim.iams, H. T., / rkm. J., 38. 271 (19M). . 0. F>US,<t., AMI ClOeAl.TKV, V.,/. Bird. Ckrm13, f>?< (t I 7. Foi.ix, O., .I. J. rkijninl., 13. 45 (1905). ti. Kolin, ()., J. Iliel. f'krm., 17, 469 (tiilt). 9. Clll-WOMS, X. !>., AM> EnTIUKIK, J. It., ScnlnricrC qn '.l't'i > orfanir audfysM, 2nd edition, Inlcrscicncu I'uhlislicrs. 1 - Xew York. 1957. 10. S.woy, C. M.t ami Aiickxatht, 3. I.., J. ,lw. Chem. .. 1 1219 (1912). DSW 255035 v/ V' STLCOPCB4060726