7;.!. /:ran:
`
`DCp~lrlmt’lll of D~21-lll~l,Oll.;~_v. I.[lliVgl-.~il\
`i’ll AI’k;IIISlI>, I’Of I~,Icclicatl ,%cic’nces.
`IJItlu" I/llck. i\l’k., lISA
`
`Absorption of Amorolfine
`through Human Nail
`
`Key Words
`A,noroll!inc
`Nail
`
`Pl2rcl, t~_in0OLl.~; tlbF.ort]tio,]
`
`Pilal-macokin¢lic~
`
`Abstract
`Tho percutanoous abso,l~tion (it a nc\v anl.ifl.ingal agon[, amorolfine, has been
`,no~lStll’0d through hoth skin :rod nail L,~ing an in vitro technique. Application of
`~1 5% conctHltrtition i,l either an ethanol or methylene chloride lacclticzr l’u’stiltcd
`in pcrmmai.ion ,~,|¢h through n~li] in the; r~t[lgm o[ 2()-11)11 nglclw’th and somewhat
`higher through skin. Absorl)tion was gr0lltor from tile lno[hvlcllg chh)ridc lac-
`qu0r th~in lh0 c:thatio] ]acqucr.
`oot lllloooololallo*oa
`
`Introduction
`
`Topical ihcrapy for fungal infccikms of lhc human nail
`has been I~t,’<7ol.%.’ incffectho, and this failure" nlay’ ill [’~ai’i be
`duo to poor ponolraiion of the thei’at~0utic agent into the
`nail plale. The thickness of the nail ~,,lt! its relatively co,ll-
`pact cOtl~trLiC!ion make it ~i forillid~ib]¢ barrier to lilt
`on\,’)’ of topically applied :_lTgiltS;. In this .gltidv ihc po,lol,’S.l-
`lion of amoroltinc l-i-13-[p-( 1. !-dhllcthylp,-ol~yl)l~henyl]-2-
`n~c’ fl ly]-l~ rotly I i -2.6-ci.~-d hl~ofl~ y I m l)rl~hol h~c h vdrochlo-
`rid~l into Ilt1111.3,1 nail has boc~,l ox~ullincd using ~_l,1 in vitro
`p~2 Nll~,lio,1 moctud.
`
`Materials and Methods
`
`r\vl) LOlllllL]~llil]ll~ o1~1!1"1Ol-O11{11c’ \\el-t_. Cx~ii~]illCtl in thin s, udv. l],mh
`
`\~c’]’c’ ~u!)plic’d I’~) t ](l[lt’ili~il’lli-[ .~i R(~u’hc (NU, Icy. N ..!. ;.llltl t’l~nsin, cd of
`5% (cid:128)trug in ciihc’r :~ nlmlh\ic’llc’ chl~wictc 11t’ c’,h~lliol lacqumi. 1"o c’;ic’h
`
`~\~ln ~ltltlctl ~ ~r’accr atnotiiil of ’ll-an~(~loll’hlc (spc~cilJu" ac’l{\ilV 1(1ii.7
`ft(’illng). ~i\’iT’lg ~.1 IJilal ~lC’i\ iIV Of a[~l~]’OXh,l~’c’i)’ I(lll l~(.’i/iI~l.
`[[Lln~Jin i~;iiL~ w~rc’ ob[ainc’d ill ~lLlt(ll’~S) I]’o1,1 hw~ll hospi’als and
`
`~hliod in ~,Cl-C\%’-t.’{Ip glass vlal~ :it -](! (’. Pri~ir ,o Llb;c [fit.’ nail~ wu’ru
`cJc+;Iti~¢d t)l’ iidhc’rt)ll, hlootl lit ,iSSllt." u~in.,., ~1 MI]~I[] t.’l.llt’t,t-’ ~l[]f.tft)l ,~lrl
`wii,c.Y. [hcw \vt..D2 ,h0!] r’nol.lll,~2tl hi H~cci~d gl~.~ t[i fl’Ll.’,ion charnh~2ry, l’i,-
`
`Ic’d \vilh a plaslic lop ]]avhlg ,ht-’ ~]llc’ c’ur\~llLH’c’ ~l.~ that of tt~o II~li].
`
`Wht.’n tl~.ill7 ,hu~ gl-~it, hl~liiiil or ,hLInlbni~il. il \\ils Iirsi sl~lil in half so
`,h~, t~.lt~h I¢~.I I’Ol’lnl.ll~l,iOll c~)uld be Itlll in dLil~licalt.’ ~ill piec-es t)l nail
`hiL\<[ll~ Iht.’ .~.Llllt.’ ,IIIc’LT]t.’S~ lind obi[~h~t.’d I’l%)tl, lhu" same donor. The
`rc’ct.’p,or i~ol-lic)n of ili~ dilfu~io]~ c’li~lllll~c’l~. \va~. fi]lc’d ~ilh i~ho~,f~h~ll~>
`
`bLLfl’c’rcd ]~c)l(II~it-" ~llinc (pll ft.0);tnc] n~iinlltint-’c] ~1I 37 ’(" b\ C’il-Cul~l,-
`
`in[ ,u’n’tl)~2r;l,llrc:-i;on~rollt.x] \\’;ilt.’i’ ~hrotl~h fin Ol=llCl" .i~ck~l..~lil-l-h]~ ol’
`
`,lie" Tc’t’c’p,~<ir pha.~c \~ a.s achimvu’d h\ ~l t c’floll c’o\ u’Fmtl II’I~.i~TI.I..’, :let i\ ~t,t-’(I
`
`b~. ~.t (~()(I-i-r~m i-0.olor p];~c-~¢l t’~t_’llk.’~lt]l ,lira c’h~ll~lbL’l’. lhu’ tllj!CI< p~lrliim ol
`
`,lie n~iil \\~ts 1¢1’, ~rl?n 1~ ~imh{on, lal~or~i,or\ coiidili~ins. ~il~l~l-oxin]atcl_t
`22-23 ’:’(’ ~.tl,tl 3.~-.~5<f’i, rchdhc: humidh\:
`])L’lmtl[ ~IIIUOLIS; ~.lh~;.)rll,Jo[1 ’,~’~.ls I1,L’i.LSllrt.’d tl$]ll7 ~l [11oi.[ll’lt:’t,i(] rl of ,he:
`i]lt.’tllotl of Fl~ln~ I] I- lq)tlo\~ [l~ ~l ]4-f’iollf CtiltlilihIHtiOII periled, t-.ii~.-h:
`[’(~l’Mlda,itll, ~.\iiy, tll~l~lic’(I lo ,ht.’ n~i] ~) rl,it;r’tlsvl’hlg? ~ll iL (fOb;L> Of II.(ll
`ndlc’ni’. lhc" t’a,c :.llltl c’xlc’lll of llh~.oipIiOll \~~lb. 1,tc_.~l~,tli’mtl bv It-’ll,O~ i1~7
`
`,ht-’ l~t-’t.’l~[Cli-~ol,~iion ~11 inlc’i-\ ;d’+ ovc’r the’ liL",;I I’u’\V cl~i\n il!ill ~i~iX ili7 ti
`
`IIl~iil;i~ I. I’l.ill/. ~+111
`
`, ]ll~l~ I~.:ll’~.’c’l \(, IL~’,,:
`
`lhlk" R~,~’k. All 7.~114 t!:h.\l
`
`~7~
`cs~
`
`ARGENTUM EX1018
`
`Page 1
`
`

`

`[-ml aliquot Ior ra(IJo;_it:tJv~., £’Ol’it@l’il bv Hqliid scintillati<m spectros-
`
`copy, In some c×pcrJnlt_’lltS, additional do~¢s ,a,f alllOioIliliC \v£’r~_’
`applied ~lf~gl" I’il’Sl rcmo\:kng the prior dose wJ(ll acetone-soaked c[)1[Oll
`
`swab,s.
`
`For COlllt’~al’~llivt.’ ptlll~tlscs, scvcla[ c×pcl’illlelllS \V~’lC Ctllltltlcicd [11
`
`which amoro[lille ab~orplion lhmugh htLrnan ~kill wa~ a[~o TIICaSLII’C(t,
`
`I hl111i.ln IFU, ilk skin wus obtained l.iI ~ltllOl)s_v. sgclil.llled wilh a del-t!l~l-
`IOllll,." 1(I ~l tllickllc,Ss of aprirox.iltlatcl_v 25(I ~HI1 i.II1(.I ,,;lored at -20 "(.’. JHY, t
`
`prior hi rise it was tllawcd by placing {Jl 37 ~’(" Ior 5 illi11, till illlo small
`s~2cTioIIS and motmied on t)-i111ll I:l’~lll/. coiN. All other aspects o1 ~.hc
`
`L’XpCl’illlQIlt~, \vt~r¢ Silllital to those dcscl’ibcd I’or tile IllL’;.l%tllCmrdllt of
`
`nail pcrlllealiOll.
`The tlpl[ikg oJ" anlol’tdlhl~: al eqilJlJbriUlll bv ~lll~ll pJcct:s of n~lJl
`plate \%~lS also tlcterillJllcd, N,"d{~ from 2 don(]r,s were CU(Jilt() nluhiplc
`21)- lO 40-1rig scctiom and placed in sm,’lll screw-cap vials to which 1 ml
`
`o1" oil.her the ethanol or mctll,,-Iclle chloride lacqt.cr was added. After
`
`4g h l.it I()Olll tcl’tlperaturc, tht? ’S.C,L’t R)IlS WCl"C I’CTllOV~d fill[] placed lol- 1
`
`nl{tl in RIO nil el ethallol. \\’hid] was Then vigorously M{rf’¢d, Followhl~
`(his tile ~.’ctions were remo\¢tl, blotted dry. and tlJssoJ\cd ill 3 nil
`
`,~.(.)ILICI]t’ {[)~.lCkilld h~strtlmCnl Compall’)’). An (I.l-m[ a[i¢lUOl was
`
`Ct,)ttTlt£’t] t(’r dCt¢lItllitlJ~ radioactive {+I(llltl21l[,
`
`Results
`
`The rate of absorpikm of an]orolfinc through hunlatl
`thulnl~tlail following a single applicatiot~ of the ethai]ol alltl
`methylene chloride lacquer is shown in figure 1, The flux
`roaches a peak belv¢Cell 5 and 2..’7’ h. lhcll appears to slowly
`decline. Following an acetone wash of the surface at 68 h,
`there is a further decline in the flux. Amoix)lfinc absorption
`from the methylene chloride lacquer ix somewhat greater
`lhan that from the ethanol lacquer over the entire time
`cotu’se of the experiment. At the peak, the flttx approxi-
`mates Ill0 ngtcm-’th from the methylene chloride vehicle.
`Reapplication of lhe lacquers to the sanle tlaJl sections
`shown in figure 1 were nlade 3 times over the suhseqLtel~t P,
`days. The results are shown in figure 2. With each rcappli-
`cation the rate of absorption profile was found to be similar
`to that seen with the first application. Following a peak
`reached after 10-20 h the rate slowly declined until the time
`of the next acetone wash ~llld rcapplication. The variable
`nature of the all,el-el fine peak on each reapplication of tim
`ethanol lacquer is puzzling and its significanec is not known
`tit this time,
`FOI" eonlparison, alnorolfine absorption through tt’unk
`skin is shown in figure 3. As with nail an initial peak is seen.
`al appl-oximately 2 I1, followed by a steady slate over most
`of the subsequent 48 h, The Ilux is greater from both lac-
`quers through skin lhalt throttgh nail. l"angillg froth 100 to
`2(RI 1]gtcm~/h. but is again seen to be higher from the methy-
`lene chloride \chicle.
`Amorolfine absorption through nail k)llowing pretleal-
`lnenl with the penelralioil enhancer dinlclhvI sulR)xidc
`
`0,15
`
`0,12
`
`0,09
`
`0,03
`
`Fig. I, ItatC of absorrJt[oi; (llg/(_,lll-’/’tl) I’ollo\vin# a single applk’,’i-
`
`~1o1~ t)l amoroll’i0,c To d0,plicalC hah’cs o1 both thumbnails [mTn a shlglc
`t]OllOl’, t-)ilia \vel-c [l\cragcd I’or 1211gh vehicle, Vorlical line [L]dicatcs
`
`stlrl’ai2c wash, ¯ = Meth\-I¢ilo ch]ol’ittt:: ..’% = ethanol,
`
`0.18
`
`0.15
`
`0.12
`
`0.09 -
`
`0,06
`
`0.03
`
`0.00
`
`I
`120
`
`I I
`145 170 195 220
`
`1
`245
`
`I
`270
`
`I
`295
`
`95
`
`Time, h
`
`Fig, 2. Rate of ahsol-ption (!l~¢nl:lh) following i’eapl)l[carioi;
`
`(vel-ticztl lines) of arnorolfin¢ to the sr.lilli2 nail sections slIOWll il’t figure
`
`l, D{tta wcrc avci’aged I’of hoth vehicles, ¯ = Methylene chloride:
`"-’ = ¢’d]arl,ol,
`
`kL.
`
`0.20
`
`0,15
`
`0,10
`
`0,05
`
`0,00
`
`0
`
`110
`
`210
`
`3%
`
`40
`
`510
`
`Time. h
`
`Fig, 3. Rate ot anlol-(t|[iliL’ absorptkul (!!~Crll-~!h) thlough hulllilfl
`tlLIt]k skin. Each vehicle \vas applied to triplicate s¢ctiOllS lroill lhc
`~alllC’ tlo~]or and the dat.a a\eragt~d. ¯ = Metl~vlellO chlorlti¢: {) = etha-
`nol,
`
`~5
`
`2
`
`~g
`
`Page 2
`
`

`

`0.5
`
`0.4
`
`0.3
`
`x
`
`0.2
`
`0,1
`
`0
`
`Table 1. Nail uptake of a.lfl(n-ol[ille following 48-hour soak
`
`Vehicle
`
`Drug, lag./mg nail
`
`Methylene chloride
`Et hallol
`
`2.9 +_ 0.6
`1.2 _+ 0.4
`
`Values are mean+gD.
`
`n
`
`4
`4
`
`0 20 40 60 80 100 120 140 160 180 200
`
`Time, h
`
`Fig. 4. Rate of amorolfioc absorption [% of dose) folEowing
`application fl’om mh,cnc. ])MSO was applied to one of tl]i’ce nail sec-
`t{oils from tllu S;iLITl~2 dont)r 5 Ill[ll prit~[ to drug application.
`O "= |)l~[gO: L~I I I10 D~/[S{): ~:= Ilf,) D~,~(),
`
`(DMSO) is shown in figure 4. DMSO was applied to one
`section of a single donor’s nail for 5 rain. then removed and
`the surfi, ce carefully dried. A tracer amotLnt of ~H-amorol-
`fine in toluene was then applied and the rate of absorption
`measured. For comparison, the rate of absorption through
`two nail sections fl’om the same donor not pretreated with
`DMSO is also shown. The results are similar to those seen
`following use of I)MSO on skin [2]. A large increase in
`drug penetration is noted in the pretrcated as opposed to
`untreated nail sections, and this increase persists over the
`entire 8 days of the experiment.
`The uptake of amorolfinc into the nail plate from both
`of the lacquer formulations following a 48-hour eqtLilibra-
`
`tion is given in table I. Uptake is greater from the methy-
`lene chloride lacquer than the ethanol lacquer (2.9 vs. 1.2
`~g drug/rag nail, respectively), and this in keeping with the
`flux data.
`
`Conclusions
`
`The absorption of amorolfine through human nail and
`trunk skin has been measured using an in vitro technique.
`When applied from either a methylene chloride or ethanol
`lacquer the drug is found to penetrate bolh nail and skin. In
`nail the rate of absorption profile is characterized by a peak
`followed by a slow dectine. Rates of permeation through
`nail are in the range 2(!-100 n~cm-~/h, and somewhat higher
`through skin, Absorption was found to be consistently
`higher from the methylene chloride lacquer than the etha-
`nol lacquer. Pretreatment of nail with the penetration
`enhancer DMSO resulted in a large increase in amorolfine
`penetration.
`
`~0
`
`Fl’all/
`
`AlllOl’oll’itlc PcrlllealiOl;
`
`2
`
`=7:
`
`Page 3
`
`