JOURNAL OF OCULAR PHARMACOLOGY AND THERAPEUTICS
`Voiume 24, Number 4, 2008
`© Mary Ann Lieberh Inc.
`D01: 1€1.1089/;op.21)07.0082
`
`24-Haw Evaiuaiéan of the Qcufiar Qisiribuiiorz 01
`‘4C-Labeied Bmmfenac Faificwing Tcpicai instiiiation
`into the Eyes 61 New Zeaiand White Rabbits
`
`George A. Bakiayan, Hai
`
`iv. Patterson, Ciara K. Song, James A Gow,
`
`Timothy R. McNamara
`
`Abstract
`
`Purpose: The aim of this study was to determine the diséribution and cc.r:\Centratio:1:5 of bromfenac ophthalmic
`solution in ocular tissues following topical irzstiiiation in New Zeaiand, White (NZW) rabbits.
`Design: Two animal shidiess were conducted.
`Methods: A Sihgie 50111. ]4(§.'~bmmfenac ophthalmic 2;oiL1ti~z)ra (204.3 ;J.Ci or O.()':?%) was administered into the
`right eyes of 14-18 randomly assigned NZ‘:/V rabbits. At \/'aI'i()L1S time points, ocular tissues were collected and
`atmiyzed for 1”~‘C~br0mfenaC contems. Ocular tissues were combusted and the amount of radioactivity was de-
`termixaed by liquid 3(:intiiia’:ion «tounting (LSC). Aquec)us~hum0r sampies were directly transferred to LSC Viais.
`Resufis: Peak concemrations of '4C—bmmfenac were observed in the aque0L1s humor and most ocuiar tissues
`at or before 2—h0urs. The highest concennaiiorxs were in the cornea, conjunctiva, and sciera. Similar amounts
`were detected in the aqueous humor, iris--ciliary body, choroid, and, to a siightiy lesser degree, in the retina.
`i\/ieasurahie arnounts of bmmfena»: were detected in all samples; at the 2/£~h0ur3 time point (T/:O.{3{}i Mg equiv-
`alent,/'g).
`Cmzciusimzs: Significant jpenetmticm and measurabie ammuwss of ”C—’:*.~mmfenac were detected in ai} ocuiar tis-
`sues over 24 h, inciuding the sciera, Choroici, and retina. These resuits stmngiy sugges? the uti my of bromfenac
`ophthaimic s(3h1’ci0n 0.09% in treating infiammatiotz of both the anterior and p0steri()r (sc:L-Jar segments.
`
`introduction
`
`
`Ro1\.iFr.N.\C OPHTT T11/HC SOUJTION 0.09% (><ibmm"»“; JSTA
`
`Pharnxacetiticai
`’
`inc, Irvine, CA)
`is a {apical mm-
`steroidai anti-inflammatory drug (NSAID) for the treatment
`of postfiperative inflammation and the reduction -;)f ucular
`pain in patients who have undergone a cataract extracti-3:1.‘
`Bromfen-ac sadium n}*.1'nt11air:1ic SOiLlti(3l\ 0.1% was first ap-
`proved in _Faparx in 2000 as Br<mm;‘k‘3’ (Senju _P1mrma::eutica.1
`Company Md... Osaka, iapan) for the irea :1nem of postoper-
`ative inflammation, bk>.pha1'itis,. coniunctivitis, and Scleritirsf
`in 2005, the same fmrmulation was; approved in the United
`States as bmmfenac sodium ophthalmic solution 0.09% (Xi-
`brom) for the ’r1‘ea‘cme11t of post:-pera%:ive inflammation ful-
`iowing cataract surgery. In 2006, the Food and Drug Ad»
`
`n1inist1'ati0n (FDA) expanded the indicatic:-11 of bromfclmc
`f).09"/u to in-dude 61¢ reduction of pain following ca‘:amctsu1‘-
`g_;er_<,-2
`For 5: topics} drug to be effective, in addition 10 potency,
`the drug should penetrate the affected tissue(s). For zzxampie,
`an Gphthalmic tr.-pica] NS/-UD would be effective in the pre-
`vention or treatment of cystoid macular edema (CME) if it
`penetrates the ocular tissues ta reach the retina. Ocular instii»
`lation of various topical NSAIDS provides angular tissues and
`<1qL:,e0Lzs humor with leveis adequate in iuhihii pr'osl’agia*rx,d‘rn
`synthesis and, thus, the ami--inflammatory role. Ilowever, the
`pe1':etrat:'on of the various NSAIDS Varies considerably among
`agents, as dc-as the drug potency and efficacy.
`In recent years, there have been VE.\1‘iO1lS mudeis proposed
`describilxg ‘The binding: of NSAEDS to the cycle-ox}:gena5e en-
`
`ISTA :"iuarr::a<ceut£calsi“, Ema, Irvine, CA.
`Y
`ies were coxiducted b ESTA Phr1r111aceu‘:icais‘"i, Inc.
`,
`Em:
`
`’ of Cafarargf and, Re-§rac'rive Silrgery (ASCRS) Amwuai M.eet'mg, San Frzmciscgo, CA,
`paper was presented a‘: the funericar
`March 17-22, 2.006, as Ahstrao‘. i’I‘.27, and aim at the A fiuciairizm for Research in Vision and Oph's}'u:—:E;]wIng§./ (ARVO)‘Anrn,sai Met-1ing,,
`E.aL1<ierda1e,. FL, April 30-May 4, 2806, as Abstract A5086.
`’fXibr0xn Package insert. L‘5TA 1’harm::ceuti;a' ‘ Inc, irvine, CA 7.006.
`Téronucirz. pa«:k3ge ins‘:-ri, Senju i’ham13Ceut:;ca1 -:,Company, Ltd" Osaka, Japan, 2005.
`392
`
`.
`.
`lrxiixm, CA . ‘Iht: axsticrirs are emr)im'ecs and 5|.'()Ci(i1()idt‘Z'S of ISTA 1/’1mr—
`
`CONFIDENTIAL
`
`PROLO167067
`
`Petitioner |nnoPharma EX 1151
`
`|PR2015-00902
`
`|PR2015-00903
`
`Page 1
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`Page 1 of 7
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`
`OCULAR DlSTRlBUTEON OF “C-LABELED BROMFENAQ
`
`393
`
`ISTA Pharmaceuticals. The protocol, materials, methods, and
`zyme, their role in the inhibition of the enzyme, and, conse-
`data analysis were identical for the two studies. The first
`quently, their potency as anti—inflammatory agents. These
`study had 14 NZW rat‘-bits randomized into seven groups of
`piiarmacol<inetic properties are dependent. to a great degree,
`on the structure of the molecule itself. ‘v’\1aish Qt ail siildied
`2 rabli-its each. ‘Hie second study had 18 .\lZ‘/‘»" rablaits ran-
`domized into :3i:< groups of 3 1':'1l’}i')il'S each. Tlie‘: studies, com-
`the physicochemical properties of ainfenac and l‘) other de«
`rivatives and found that the addition of a suiostituent to the
`plied with the animal welfare, policies of 1‘3‘i”C and were ap»
`proved by the institutional Animal Care and Use Committee.
`molecule decreased the antieinflammatory activity, whereas
`the addition of a group to the aromatic ring had a pro-
`The first study was designed to ensure that enough mea-
`1
`.
`_
`e. ' xd
`
`The researchers reported that the most potent compounds
`study tested comn1erciai—strengti1 brom tenac ophthalmic so»
`lution 0.09"/o.
`were those with a halogen in the 4"~posi1ion (Br ~ I ,> Cl
`‘>
`F :> H). Bromfenac has
`cheniical struc‘:ure very similar to
`that of anifenac {the active form of the prod rug nepafenac),
`except for the critical addition of a bromine atom in the xl—
`position of the benzoyl ring (Figure 1A and ] it’-‘;.‘ Preclinical
`data dermmstrated that this lialogenaticm of the molecule not
`only pron"! uced greater in vim) and in viva potency,2’5 but also
`enhanced bi‘on1E:31iac absorption across the cornea and pen
`etmtion in ocular tissues.”
`‘lihe <:--:tanol,/water (O/ W) partition coefficient (Chg p) and
`the quantitative structur-evactivity relationship (QSAR) of a
`drug is another factor that influences its penL=t.ration prop-
`erties and, consequently, its potency. C1,)?’ p estimates the w.1~
`tor solubility and the level of lipophilicity of a compound {a
`key determinant of ‘the pharmacoldnetics pamnieter). it is
`commoniy used in drtigdesign studies, since this property
`is related to drug absorption. bioavailability, metabolism,
`and toxicity. The higher the value, the better the penetration,
`with a 1.0-unit differeiice in the coefficient representing a
`tenfold. cl,il’ferersc'e in penet1'ati.on. R Lli.”~', er, al.“ reported that
`the C1,,“ p of l7mn1fer:ac (2.2.7>‘- is higher than other NSNDS,
`such as amferiac -(1.23) and ketorotac (188)-. This difference
`in (C-m|,;1> explains the higher iipophilicity of bromfenac that
`provides a more rapid saturation of the epithelium and a
`minimal lag time before the drug croases the cornea and,
`thus, the fast arzalgesic action.“
`The aim of this preclinical investigation was to evaluate
`the penetration of ”C~»bromfenac following a single ocuiar
`instillation in New Zealand White (NZW) rabbit
`Methods
`
`Test article
`
`The “C-lar-Limfenac sodium (Daii-ahi Pure Chemicals Co,
`Ltd, Tokyo, Japan) was arialyzed to cleteiniine the radioactiv-
`ity purity prior to its use in the study and was fouiitl to be 100%
`pure (3.13 Mttq/mg, 83.97 ,wCi,"mg). Stability testing of the
`proclnct was not conducted in this study. Staliiiity t sting was
`previously conducted by Senju Pharmaceutical Co, L.TD.
`
`Dosing-soilztion preparation and anaij/sis
`
`The dosing soiution was prepared by adding either ”C~
`bromfenac to brornfenac ophthalmic; solution 0.09‘?/E»
`to
`achieve a
`target concentration of zapproximateiy 500
`;.LCi/mL or to target a concen tration of 0.09%. Three (3) all»
`qua-ts (l€l0 /.-LL) of the dosing solutions were weighed and
`iaruught to a volume of 25 ml, with saline. Duplicate all»
`quois (100 iii.) of the diluted solutions were quantltated
`
`{or radio;+ctiVi.ty by LSC. Tine Cit,
`ing solution was prepared
`imn1ed,iately prior to do:si.n;-g; therefore no refrigzzratiim
`was needed. After dosing and prior to hi,gl':~performance
`liquid chromatography (1 IFLC), the solution was refriger~
`ated. The radiocheinlcal purity of the dosing solution was
`tested again by i"il’LC foliowing dosing and was fotrnd to
`be ltli. % pure.
`
`Animals
`Female NZW rah‘l3i,ts were obtained from The Rab‘oit
`Source éhzttnona, CA) or Covance (Denver, PA). The aniniais
`
`were at least l2 weeks old and weighed 218- .32 kg at the
`time of dosing. The animals were housed in individual cages
`and were identified with ear tags and cage cards.
`
`Two animal studies were conducted at the Biological Test
`Center (ETC, lrvine, CA} as per—study pmtoctols provldwl by
`
`Amtenac
`
`Bromtenac
`
` B
`
`+:
`
`NC
`
`IG. 1.
`Chemical. structure of newer generation of mmsteroidai 2mti—irnflanm1ato1*y drugs, (A) amtenac (active form of pm-
`drug nepafenac) and (B) bromfenac.
`
`A F
`
`CONFIDENTIAL
`
`PROL0167068
`
`Page 2 of 7
`
`Page 2
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`Page 2
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`

`
`394
`
`BAKLAYAN ET AL.
`
`TABIJ3. 1. Tm: T:'HS1.IF. SAMIMI-cc; Tm]: AFIER L\:s1'31,.r,A1':oN
`AND my AM<)U;\”‘.' Eilzov NAC AND RADIO Acnvrry
`
`EACH TmzA.rMFNr GROUP
`
`Average BI-" dose Average RA dose
`______
`
`0.276
`0.284
`
`22.14
`22.81
`
`0.306
`0.236
`0.282
`
`24.57
`18.93
`22.66
`
`1 h 1‘: 5 min
`2 h
`15 min
`4 h 3: 1:» mm
`8 h .1‘: 15 min
`12 h 3: 15 mm
`1 in
`
`A
`B
`C
`D
`E
`‘
`
`E>mxnfe,-xwac; RA, ra<ii«_>a<:tivi'ry.
`
`Pretreafrrtem‘ axarrunafior: and dosirwg procedyrs
`Prior to placement in the study, each animal underwent a
`f.1=.or0u gh pretteam1ent Qplxthaimic examination with sht
`lamp. Acceptance criteria for placement in the study were
`scores of <1 for Confiunciival congestion and rawcliing and
`scores of zero for ail ether observation variables. Prior to dosr
`ring, animals were \-veighed arm‘, randomly aseigiled to six
`study groups. (gmups A—F in Table 1') of 2 {study 1) or 3 rab-
`bits each (study 2).
`On day 1, 50 M. of the freshly prepared dosing solutien
`was administered into the right eye of eavzh animal, ursing a
`calfhratrad pipette, and the time 0f dose administration was
`reccrded. The actual cime the animals received rangs.>::( from
`13.3 to 24.6 ,LoCi (stuciv 1} er 0.09% bromfenac (studv 2). Ac-
`ma! dosing Values in mg and MCI were used in all su,hse~
`quem calcltiatiorss. Animais were obswived for nmriahfy
`and/01' mmbidity duling the course of the study.
`
`Samples processing
`
`The animais were eiltlzajlizetl with an i1‘ctra\/enuus injec-
`tion of euthallasia scduiion, and the ocuiar fissues we1'.‘: COI-
`lected fmnl the dosed eye at 1-, 2, 4-, 3-, 1}.‘--,. and 24-h time
`points following d-ssing (Table '}_). The 36-h iimepoint, grcsup
`(3 in study 1, was rendered unusable due to c(:-ntamination.
`The c0njLmCtiva, csmea, lens, iris-ciiia1'y body, sciera, cho-
`roid, and retina were weighed intu combustion cones and
`Combuzited. Combusted sa.m,p1e.~'. were trapped in Caflmm-14
`
`TABLE 2.
`
`Cocktail (R. J. Ha1'v»3y, Hillsdaie, N3) in LSC Viale, and the
`anmunt of radioactivity was determin€:d by ISC.
`Approximately 25-'}.l.L dupiicate aliquots (1-fcach aqueous-~
`hunmr sample were transferred t0 LS-C viales, usirxg Instmfiel
`as sc§_r1tiHz1tion fluid, and the amgtmt of radioactivity was de-
`
`homog-
`tcrmhwd by LSC. Each vitreous humor sample :
`enized, and duplicate aliquots (100 ;:.1. each) were traxxsferred
`to [SC vials, using {n:;ta-Gel (P(zrZ<1'n Elmer Life and Ana-
`},
`Hm, W:r}H1m°n, )'. ..mfi&dd,
`and the amount 0f radiaactivity was determined by LSC.
`
`F?ad.ioacziv:'ty measurements
`
`Radioactiviiy mea.<.us"ems2nts were periermal by using, a
`Eeckmam Liquid Sztintiliation Spectrometer {BQCRHIRIE C0111-
`O
`ter 1115., Fullerton, CA . Cmmtino tinw W88 to a statisticai ac-
`curacy of
`0//0 or a maximum of 10 min, whiche\«'rer came
`first. Backgrmmd noise, appro>:imateE.y 100 ziisimegrations
`per minute (dpm}, was subtracted mttomaticaliy. in additiim,
`.,.~..,,s.,L
`',
`2. J:
`...,c i<'..(
`.
`‘cc
`¢':<_
`*9’
`the " 7P"*I‘“)fK’1€‘lk’E‘\‘J1<
`r ygmmmei E) 11;! wmau 10 ! mv rt
`counts per minute (rpm) kc» dpnx.
`
`Calculations and statistical anarfysis
`
`The foliovvmg are the formulas used in this study. For the
`specific activity (dpnz,/Mg) of ”C—brmnfs3nas::
`Specific activity 2
`
`
`For the 'ppm (/;.g,/5,3} of dosed 1‘:‘C—br(.=111femac:
`
`ppm 1
`
`>
`(if-n“.,/g of sampie
`Specific activity of dosing soluti-an
`(dpnn/ug’)
`
`PM the percem‘. of dose:
`
`‘"25 of dose *
`
`Raciiwzstix/ikv in sample (dpm)
`Tomi racli-:3ac:i\‘/ity
`.adn'n'.nistered (dpm)
`
`X ‘J00
`
`When applicable, the mean and standard deviation were
`used to characterize the data.
`
`“C-BRoMFE.\:Ac RESIDUES xv OCLILAR TESSUE AT DIP RENT '1‘:‘~.4E—PoLm3 FOLLOWENG A
`
`
`
`‘
`ST LA? N 1N'Ir:> we REC.
`‘E OF F\I2:‘w ZFALAND
`E31
`51. T
`
`~.
`Gr‘01.¢{? L.’ (4 Ir)
`h)
`Gran}; 8 (.: fa’)
`(,}rr;up E) (8 /1)
`(‘I‘“ 51) me 1 ‘mm *' 9}’
`‘
`
`
`‘cm;
`*‘ 9:’
`
`
`
`
`/3.,
`G701./g;
`W2
`'1
`‘NW1
`
`
`
`5.632 .
`3.889
`..
`.
`.
`
`.2". 2.189
`1‘; 7.3"
`373*" .1: 8/"'Z.’:'
`LJ1 1.
`0.012 : 0.002
`0.027 : 0.004
`0.022 : 0.004
`0.016 : 0.007
`0.441 t 0.123
`0.280 i 0.026
`0.113 f 0.025
`0.064 : 0.002
`0.4.95 :*: (3.191
`0.3.1‘: : '0 09]
`0.099 : 0.016
`0.039 t
`.012
`13.370 1' 0.143
`0.272
`.106
`0.131 : 8.074
`0.074 :' 0.060
`0.007 t 0.008
`0.002 _. 0.000
`0.001 f 0.000
`0.004 _ 0.005
`0.118 + 0.072
`0.096 “ 0.044
`0.080 3 0.082
`0.038
`0.028
`2.500 : 1.037
`1.223 : 0.421
`1.505 : 1.316
`6.212 E 0.352
`
`
`
`10.693 2 0.993
`
`9.7.
`V: “.929
`0.005 : 0.00]
`: 0.059
`3; 0.0.52
`: 0.094
`: 0.000
`.
`0.081 "; 0.015
`4.511 i 0.301
`
`
`
`0.835 1’ L‘.
`'
`
`t.
`0.012 : 0.001
`0.033 i 0.013
`0.004 3; 0.001
`0.019 : 1‘.-.00]
`0.000 : 0.000
`0.009 3 0.003
`13.4143 : 0.095
`
`Conjuncfiva
`Cam-ea
`Lens
`Iris-cifiary body
`Aqueous humor
`Chore-id
`Vitreous hunirsr
`Retina
`Sclera
`
`ppm, parts per mi11i0n;SD, standard dexriaiixuxl; Boldface, peak tissue Coxmelltration.
`
`CONFIDENTIAL
`
`Page 3 of7
`
`Page 3
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`PROL0167069
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`Page 3
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`

`
`OCULAR DlSTR§BUTEON OF “C-LABELED BROMFENAQ
`
`395
`
`TABLE 3.
`
`‘“‘C—BI<oMF2NAc 0.09% RESIDUES IN OCULAR TISSUT"
`
`'
`’
`mm 11-11:. R:
` S1\
`' 3? AL lN.'.%T
`
`A1" DEZFFERENT Tnvle-POINTS FOLLOWD\TC. A
`I«‘.~/F. OF N’/.W RABBITS
`
`
`
`
`G‘/rvup /‘s (I I7)
`mezm [mm J: Si? mam ;7;)ru
`
`. Sf)
`
`1mw: zlpm t D
`
`
`Gmup E ' " it)
`Grmga F (24 F1)
`Group 0 (#3 E?)
`
`
`SD rriean pf)”: ;“ Si}? m .m ;7;.vm ..
`.91’)
`
`Slitfifllt?
`
`Cc-niumttiva
`Cornea
`Lens
`Eris-~ciliary body
`Aque0s.1;~7’nL1mor
`Choroici
`'\/’itre(:-us humor
`Retina
`Sclera
`
`0.437 : 0.133
`‘L069 1 0.053
`0.001 : 0.001
`0.087 : 0.029
`0.030 1; 0.008
`0.040 : 0.013
`0.000 : 0.000
`0.008 t 0.003
`0.238 : 0.070
`
`0.6i2 : 0.2€)2
`0.735 :; 0.207
`0.006 : 0.008
`0.044 : 0.005
`0.026
`0.007
`0.039 : 0.003
`
`0.000 . 0.000
`0.011 .; 0.005
`0.186 : 0.015
`
`0.484 : 0.451
`0.887’ 1: 0.302
`0.002
`0.000
`0.057 : 0.001
`0.039 : 0.012
`0.048 i 0.018
`0.000 : 0.000
`0.007 4; 0.005
`0.267 : 0.19.7
`
`0.126 : 0.085
`0.269 :; 0..F76
`0.00’! : 0.000
`0.027 : 0.006
`
`0.010 *‘ 0.0 "
`0.013 A‘ 0.002
`0.000
`0.000
`
`0.002 ’_ 0.001
`0.031 : 0.012
`
`0.226 : 0.338
`032'! i 0.057
`0.001 : 0.000
`0.022 : 0.004
`0.008 "V 0.007
`0.005 : 0.002
`0.000 : 0.000
`0.00] 1: 0.001
`0.038 i 0.041
`
`0.397 : 0.132
`0.046
`0.042
`0.002 : 0.002
`0.013 : 0.007
`0.003 V’ 0.003
`0.003 : 0.005
`0.000 : 0.000
`0.003 “* 0.003
`0.394 : 0.52.
`
`ppm, parts per million; SD, stand-3rd deviati(m.
`
`Results
`
`Dosing and sampie harzd/ing
`
`The raclioactivity present in the conjunctiva (sf 2 dosed
`eyes in group G {firststL1dy)\raried greatly, indicating a pos-
`silt-le comaminatlon. Co:1sec[ue11tly, the samples were ex»
`
`cluded. E.
`ve tearing ¢;:ecw'red in 1 am'ma'i in group
`of the sank": . tudy, following dosing. Excessive tearing was
`abscwtved with absorbent eye wipe(s‘). For each eye~wipe
`san1p1e, the sample was placed in a vial, E5 ml. of distilled
`water was added to it, and each Vial was put on a Wrist-ac-
`{ion 5hal<e1*for aE'p1'0x;'matelV‘ 1 hour. Duplicate aliquotrs (25
`M.) of each sample were a'nai.y:cecl by L‘:3C. The 1'm'li0activ«
`ity pre:‘~.ent in each wipe was considered as radioactivity lost
`during dosing, and actual administmed dosee Vvere calcu»-
`lated accorctmgly.
`
`Data analysis
`
`After the admiztistratirm of a single 50—,y'..l.. dose of “C—lw1'0m—
`fenae ophthalmic solution to achieve a target dose af 20-25 ,<cCi
`into the right eye of randomly ::_~:s‘ig11erJ l‘\V'7.W rabbits, mea»
`surable amounts of radioactivity were detected in all tissues
`but the vi.trer)us l‘1‘.H’110I’. Peak comtentratians 0f radi.0lal*-eled
`bmmfenac were observed in all (scalar tissues in the first study
`at 2 ll, with the exceptiorn of the mrtjttnctiva, whicla showed
`peak tissue eoncentraticm at 3 h, and the lens, which slmweri
`peak tisstte ::0ncentra.tior: at .3 h (Table 2). In the rsecoxnd study,
`peak comtentra tions of radiniabeled bromfenac were 0bse1'Ved
`in all ocular tissues at 1---2 h, with the exception of the lens and
`vitreous humor (Table 3). The b1‘on1fenas: concentraticms were
`highest in the cornea. Siniilm‘ amounts of radialabeled br(:-m»
`fenac were measured in the aqtteous humor, iris-xjliziry body,
`Cl‘.OXOlCl, and, to
`sliglw tly lesser degree. the retina. Furtl*:er, ;,'a~
`L«l'l'C'lH'U(.l§K.l b'!'U1'TY.‘fE'JzdT.‘ Vclb detegtett
`in all E-JTllP'l.to
`ft Ril-
`lowing topicai adn1inistra’don in the first and second SlILldl9fS,
`with the exception of the vitrec-L15 humor. Althc-ugh the clini»
`cal significance of these animal studies is u.nl<m>wn, the data
`suggest that the extensive penetrati<..m profile and the sustaixwri
`cuculamissue concentration of bromfenac in the anterior and
`poster§<_=:.' segments of the eye may support the use mi brom-
`fenas: in treating inflammatory cii:=.0rde:‘s in other ocular 05--
`suea, such as CME (Fig. 2).
`
`Discussien
`
`In addition to pct-tency, aI1oth~;>.1' important critericm of
`topical anti~inflamm;1t<:-ry drugs is the ability to penetrate
`the tissues ‘co reach their targeda) in .3 timely m.a1me1' and
`appropriate c;011cent1'ation.. Thus, the ability to penetrate
`ocular t‘155ues is an impmtalit d.eter1‘ninm'xr. of the Safety anrl
`effiuiacy of ophthalmic NS.MI}s. The two animal sstudies
`§>1‘e:‘-.ented in this report dem0nss‘:1‘ate that a single topical
`dose 0f Commercial-strength but-mfenac ophthalmic solu—
`tion 0.09% rapidly, within 2 h, a-thieved measurable levels
`in all ocular tissues and detectable levels were sustained
`
`other ocular tis.«:ue:3, 1*adic-labetecl bromfenac was um‘le--
`tec.tabl,e in the vitreous humor after '1
`ts {€0.00} pg (.>qu§v--
`alem/gm’). This was most
`likely due to the enhanced
`113;.-ophil§v:it_y of l3r(Jl’!'1fé3lIt‘.C, which could impart a rnore
`
`DYHBZ
`%§6!éIris-Cliary Body
`°1*R‘PwAqueous Humor
`“=°EWRetina
`Wdfléwchoraid
`
`
`
`Concentration(pgequiv./g)
`
`Time (h)
`
`FIG. 2. Radioactive C<\.m3ent‘cation.~; of “C-bromfenac in or.-
`ular tissues f@3l0wi31g a single 0.09% topical dose of com-
`n1ercial~stre11g’th bmmfenac to the iglrst eye of New Zealand
`white rabbits. Detr-(table levels of varixnus ocular tissueze
`were seen tlwmuglw 24 la and bE‘.V‘V’(‘.-fld.
`
`CONFIDENTIAL
`
`Page 4 of 7
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`
`396
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`BAKLAYAN ET AL.
`
`ihrz
`tissues and,/or
`t}1i'oLigh
`time
`rapid rirug—tx'ansit
`choiioidai blood flow E0 pos,terios'-segiiient tissues. A sep-
`arate, but similar, p€.1‘l€‘I]”€U.‘i01‘|
`study of “C--iiepafzinac (pre-
`cursor of zimfenac), using three times the commercially
`available dose, found no Significant ievela of nepafeiiac/"
`amfenac in the aqueous humor and choroid after 12 h and
`in the retina afte1' 6 in foilowing ii}?-i.'iiiai’i01‘:.3 However, the
`cornea aim‘, iris-ciiiary body‘ _~:howed detectabie drug levels
`
`1 /1C—Bmrrrfenac
`
`
`
`FIG. 4. Comparison of the estimated aqueoLzs—humor con»
`ceutration after the instiilzitimi of B1'oima:k® (bromferiac so-~
`dium 0.1% ophthalmic solution; Seriiu Pharmaceutical Com-
`pany‘, Ltd., Osaka, Japan) to the human eye with the {C53
`Value of recombinzmt human cycio—o:<yge1iase—2.
`
`of nepafa-mac/amfenac at 24.1 h f0I.10w'mg the topical ad-
`rni1‘:isri‘ari0n of ”C—::epafenz1c" {_Fig,. 38).
`The ami—infiam112atory and analgesic effects of a1} NS/UB5;
`are due to their inhibitory activity (311 cyclwoxygeiiase err
`zymes, mainly cycl0—oxygenase—2 (CC>X~2). I-Ia:-wever, the rel—
`ative potencv of NSALDS against COX-2 varies among the
`different NSAID moiecuies. The unique chemical simchire
`of bronifeaiac, wifh a {drum ins: atom at the 4’—position of the
`aromatic ring, has Ci(‘.n10]’lSt1‘:‘:'tQd its pronounced effects on
`bi‘0mfcnac's potency, absorption across tine cornea, and pen»
`etratir:-rs into ocular tissues where it maintained detectable
`levels for up to 24 h following topical acfmiinistratioii, mak-
`ing it the most potent ophthalmic NSAID in inhibiting the
`
`CO‘<—" erizyme.2”5'1" Kida et (11.5 coiripai‘e3d COXV2 inhibitory
`activities of the four coniniercialiy avaiinble ophthalmic
`NSAIDS: amfenac (arrive metaboiite of iiepafeaiac}, b1‘Cn1-
`Eenac, diciofenac, and ketc-rolac. Bra:-mfenac was eipproxk
`mately three to four times more poterit ihzm the other three
`NSAJD muiecules in inhibiting the COX-2 enzyme. This po-
`{ency may explain why bromfenac op-hthaimic solution
`0.89
`5 fhe fir$t and only ophtliainiic NSAID approved for
`tw’
`-daily dosage.‘ Furthe1', Qgawa et 5].” evaluated the
`pharmacokiraetic profile of bromfenac sodium 0.1% in 54
`subjects undergoing cataract surgery. One {1} <:i1'(:-p was ad»
`ministered at various time points 3-0-345 min prior to sur-
`gery. Tiie coricenkration of hi‘0mfei'iac in a 'l{)()v,mL sampie of
`ihe aqueous i’lL1l‘{‘aO3,‘ was determined by using HPLC. The
`peak aqueous-humor concentration of bmmfenac was
`ar_..
`1
`athim-red2.S—3.€r-E1
`.
`'
`3.
`'.
`‘
`'.
`.
`"
`remained at a therapeutic ccmcentration above the EC5g value
`for COX-2 for 12 ii. in arldition, a computer simulation pm-
`jecied a rneasu_:'a‘ni,e cm1c.emr,‘atio,n at or beyond 24 11 (Fig. 4).
`‘C:.m'erst1y_, the four pi'incE.pe11 roies O}’)i1ii”lEii1fi§fl NS/\,iDS
`play in ophfssalmi-: surgery are thepreVex1ti.on of n"eio$isd1.u*--
`ing atataract s;urgei:y, management of }')0E%i2\“:}I>€‘,1‘{ii£i\/"t": pain and
`inflammation foliowing cataract and 1'efrac’tiVe surgery, and
`the prevention and treatrnenf of CME after cataract sur-
`
`
`
`Page 5 of 7
`
`Page 5
`
`PROL0167071
`
`
`
`
`
`r
`
`1
`
`r
`18
`
`1
`24
`
`-9- Cornea
`-‘E- |r5s—C.‘-iiiary Body
`-—~£n—- Aqueous Humor
`—%-— Choroid
`--£3-— Retina
`.
`1
`6
`
`1
`
`,
`‘i2
`Tmre Ch;
`
`Concentration(pgequivjg}
`
`A
`
`0-07
`
`0001
`
`14C-Nr:paienac
`
`-9- Comea
`---B---
`€ris—CiEiary Body
`--.fir- Aqueous Humor
`—-%~ Choroid
`-$- Retina
`
`
`
`79?
`i
`‘
`I
`3
`1 is
`E
`U‘
`3
`3
`I
`4*
`i
`9
`Z 01 ~,
`.9
`3
`G.‘
`5
`5
`i
`o
`U
`3 0.011
`
`. E
`
`1E’3 i
`
`O
`
`I
`
`1
`‘S
`
`I
`
`I
`18
`
`t
`94
`
`2
`‘.2
`Time (h)
`
`B F
`
`IG‘ 35 Comparison of ocI,iia1"—€issue coiiceiitrations of “C-
`bromfenac (A) or 14C—i1epafenac (B) following the adminis-
`t1'az‘.i<.m of ii singie topical dose, three times the coznmercial
`st1'eng‘rh, to rabbits. (A) Detectabie levels in all ocular tiasues
`through the /Awh time—poi,nt. (PS) Retina not detectable at 6 h
`and aqueous. humcxr ami choroid mi? deiecia bie at 12 h,
`
`CONFIDENTIAL
`
`Page 5
`
`

`
`OCULAR DlSTR§BUTEON OF “C-LABELED BROMFENAQ
`
`397
`
`potency as an anti—irifiarim1ato1y agent. Tlie highly lipophilic
`molecule allows for a rapid peI‘1€:ti‘Al“lO1‘i of ocular tissues, re:
`sulting in both rapicl and sustained d(‘,l:z”:L‘.lZ€-.l’JlC drug levels in
`all relevant ocular tissues, including the retina for over 21 h
`following a single topical administration. Alfliough these
`were preclinical studies, these results strongly suggest that
`Xibrom may also be effective in treratliig inflammation of the
`sclem, choroid, and retina. including CME because of en-
`:«
`c;
`.l.
`
`.
`.
`.
`.,
`:
`H’
`.. O;
`c7£r(, v‘\*fi’l-
`ranted to explore these therapeutic facets of bromfenac oph»
`thal mic solution.
`
`References
`
`Conclusions
`
`The unique chemical structure of bromfenac ophthalmic
`solution 0.09% {Xib1'oin) plays a major role in enliaricing its
`
`V~J E‘-‘
`
`CONFIDENTIAL
`
`Page 6 of 7
`
`Page 6
`
`PROL0167072
`
`l. ‘Nalsh, D.A., Moran, l-{.‘W Slminblee, DA., -22’: al. Pmti-in-
`flammatory agents. Synthesis and pharmacological evaluav
`tion of 2-amino-3~lve: '
`cefir; acid and analogues.
`,7.
`./Med, Ciiem. 1l:E3'/*9
`
`2. (Joe):
`osier,Y.Nonstcroidalantfirsfiamma
`rory‘ drugs and
`
`
`305-
`ocular mllammation. Klin. Mormeezi’.
`zlzigrzrz/z:"z/iciil. 21
`308, 2001.
`Yaimi. }.M., Gralf, G., and Helihergg, MR. Topicaliy acl:‘m’*i—
` «e
`istereii ctmiposition conrair
`1;, 3-benzoyiphenyiacetic acid
`derivatives for treaiinent of or-h iiialmic ‘mi'Eam:11atory disor~
`(Jcrs. Alcoa Labomtors'r:s, lnc., assigziee. US. Patenf 5,-'l75,(l34.
`December 12, 1995.
`4. Waie1'l'n.xry, i..D., Siilimaii, 0., and _Io‘.as;, T. Comparison of
`c\'clo—o><_Vgenase inliibir(>i')/ activity and ocular anti-inflam-
`matory effects of lzetorolac tromethaniine and bromfenac
`5
`Sodium. Curr. Moi. R
`.
`H3 5114:’), 2006.
`
`Kida, T., Ogawa, T., McNamara, T.
`at al. Evaluaiion of the
`li.,i-n1an-C89é-
`L. L, bro. h‘-ezna ,, cl-i lofcrmr,
`and ketorolac‘. Proceedings of the 3.3t‘d Symposium on Ca?-
`aract, IOL, and Refractive Surgery of the American Society
`
`of Cataract
`tid Rcfracfive Surgery, April 27—Mzajv 2, 20-’.)7,
`San Diego. CA.
`6. R uiz, ]., Lopes, EVL, Mila, ,I., et ai. QSAR and conformational
`analysis of the an'ri-inflammatory agent amfenar: and am»
`logues. I. Cor/;pt.—;‘li:ie¢i Mafcc. Des. 7:i83~l9'8, 1993.
`Sancilio,
`Nolan, ].C., Wagner, L.E., or ill. The analgaasic
`and anti—infiamma‘:ory activity and pliarmacologic proper-
`ties of iironitenac, %‘:m’z'rHii1e{fi!r:;!7h2m_
`‘ 19, 1987.
`
`ES. Acosta, J\/l.{.‘., Lima, 2%., Gratf, (3,, er ai. C<.>mpara'rive eitects
`of the nonsteroidal anti—inflainmatory drug nepafenac on
`co teal sensory nerve fil)crs responding to chemical irrita-
`tion. Invest. Ophtlzalnzal. Vis. Sci. -'l81l82—188, 2007.
`
`9. Helberg, M.R,, and Nixon, _I,( Use of no:
`teroidal anti-i'rs~
`fiammafr_>ry agenis in combimstion with comp.o1.mds that
`have PE prostaglandiii agonist activity to treat glaucoma and
`ocular hypertension. Alcon i.al1oratoi'§r:s, l1‘.L'.., assigrioo. US.
`Pat-'-.>:it 6,342,524 Bl. fanuary 29, 2002.
`O. Donnenfeid, E.l.')., and Donnenfeld, AB. Global experience
`wirh, Xibmm (bromtenac zjplithalrxiir scluti,on) 0.09%: The
`
`first iwic «daily ophthalmic nonsieroidal antidnflarnmaiory
`drug. Int. Opl5.th.v7.x'.V.'znI. C.’z'n. 46:2} 39, 2006.
`,_.\ ,_. . Ogawa, I. Miyake, K, Mci\lamara, TR, et al. i’harmacol»:i~
`netic profile of topically applied bromfenac sociium oplv
`tlfiialmic: solution 0.1% in fsitiwgects unriergoing cataract sur-
`gery. Proceedings of the; 78th Association for Research in
`
`Vision and Ophtiialim‘;logy {ARK/O) Annual i\/lee rig, Ft.
`Lauderdale, Fl... April 30»l\/lay ll, 20%, as A687.
`
`O’Bri»2. , TI’. Emerging guidelines
`vr the use of NSAZD the!»
`
`apy to optimize ca
`act surgery and parierst care, Om . A/lm’.
`
`.. Gpin. 2l:1l3'i~'l137, 2905.
`
`
`
`
`
`go
`
`{J7
`
`fl
`
`ge1‘y.‘2’” Several stuclies corrx,§~.'ared the efficacy Mlwromfenac
`to dicl<,>fenac in reducing in fiam mation after C£3la1‘:'1CiSL§]’g¢Z‘rV‘ .
`Bronifenac demonstrated a faster onset of antidriflzimmatory
`activity than diclofenacln’; l‘1(.‘-WEVSI‘, the effect on postoperm
`rive iriflaniiiiation was comparable between the ‘two formu-
`lationsu‘ When compared wlfli vehicle. hromfenac was
`statisticaily significanilv niore effective in conl*rolli,ng in-
`flammation following cataract surgeiy and was more potent
`:
`U9
`l).t:ll;lTr?'l,;
`fl'Tcl'R
`1'
`—
`ass
`i_ L
`10,17
`compared to 41 days, following cataract surgery.
`Whereas the exact etiology of CME remains unclear, most
`investigators agree that increzised prostagiandiri synthesis
`secondary to inflammation is a major etiologic factor. The
`primary use of NSAIDS, although not indicated by the FDA,
`is the prevention and treatnwnt of postoperative, CME. Rho
`et al.“‘ compared twice—ciai.iy bromfenaa: with fou1'—times—
`daily dictlofenac and ketorolau: for the treatnteni of acute
`pseudopliakir: CME, and concluded that rwis:e—daily brom-
`fenac was siafisti,Caliy
`effective as diclofenac or l<eto1'olac
`closed four times daily. Although ihe Clli'|l.C:'!l slgiiificance of
`these aninial studies are unknown, the data in these pre-
`clinical investigations suggest that the extensive penetration
`profile and the sustained ocular ‘tissue concenuatirr-n of
`brc-nifenac in the anterior and posterior segments of the eye
`may support the use of bromfenac in treating inflamrnatory
`disorders in other ocular tissues, such as ClVll€
`(Fig. 2). Fur-
`tlier, bromfenac was reported to l‘.'€‘ as effective as Cliclofenac
`in the prevention of miosis during cataract surge.ry,1"
`ef~~
`fective as pemirolasi potassium for the treatment of seasonal
`allergic (;‘{)I1jL1i'U,‘llVilCi§,2U as effective as ophthalmic steroids,
`though safer than steroids.
`in the treatment of anterior
`uveit§s,2l and better tolerateri and more c<3mfoz‘table than ke—
`torolaagz
`O«:ula1' safety" assessments for subjects treated with brom-
`fenac were generally equivalent or better than those for sub»
`jects treated with the vehicle. One of the serious side effects
`of cataract surgeiy and the po&.‘roperati\/e use of NSAH35 is
`corneal cyifhelial damage. Two studies reported that al-
`though there was mild damage to the corneal epithelium fol»
`lowing cataract surge-17,’ and siihsequent use of eitl‘xer brorn«
`fenao: or diciofenac, there was no difference between the two
`formulations on corneal epithelial functions, as measured by
`fluoropliotometery.154“ Likewise, systemic safety assess-
`ments for bromfenac demonstrated that there have been no
`systemic side effects associated with the brom fenac 0.1% for»
`m.ulation.2“ The plasma concentration following 2 drops four
`times a day for 28 1 ays (two times the presci'il:ved frequency
`and duration) was below the detection level of 50 ng/m§..
`Shifimzm et ail?“ contirznecl the systemic safety of bromfenac
`by siuclyiiig its effect on liver functions, which were within
`normal range limits for more than 90% of subjects, suggest-
`ing no clinically significalit systemic adverse cvents,24 ln ad~
`
`dition, 3 postniarkcting SEW? lance study (julv 2000--janu~
`ifiyzfifififioithefirfcfyarmiefficacy zafnoiirfharnnc
`solution in more than 3006 subiects concluded that this prod-
`uct is safe and effective in treating inflammation of the an-
`terior segment of the eye and postoperative in{lam1natiora.35
`
`Page 6
`
`

`
`398
`
`BAKLAYAN ET AL.
`
`21.
`
`22.
`
`24.
`
`Y’.
`Us-.ui, M., and Massuda, K. Effect 0fb1‘mni'e11a
`ll.
`
`
`107 2B) eye d1'0_r)s on anterior uveitis. f. R:
`.
`max. 91:39——44», 1997.
`Perry, H.D., Chou, T,,. Dimnenfelcl, EA‘—34I 0+ al. fxnesthesia
`and ocular t<>lera,bility of topical monster-‘niclai 2mtx'~infEam-
`matory Limgs: A direct Compmison ‘c»2‘n/vevzn bromfenac and
`l<r-.'tr.>1‘olac. l’r(;r.'eedirsg*s of the Pflth Annuai Op§‘,1t‘rmlmic Anes-
`thesia Society {OAS}; Scientific Meeting, Chicago, IL, Dctov
`bar l3~15 2006.
`ei ai. Inves-
`. Shiffman, ML, Dunnenfeld, E11, Holland
`tigation cf liver toxicity ff-H-swing topicai treaim-znt with Xi-
`br0mT"‘ 0.1%, an l‘\lS/KID for‘ p05: cataract sm'gE.-ry i;‘.t2am~
`mation. Proceedmgs (if the 31st Sympoflum on Cataract,
`E01,, and Refractive Surgery of the Amerhsax‘. Society of Cai-
`aract and Refractixte Surgery, April 15—20, 2005, Washmg-
`ton, DC.
`Kitao, N, Shimoji, H., and Fukucla, M. P05tmarl<r ‘mg star»
`veillance of bmmfenar;
`sodium {BR<3NUCI(®).
`I. Eye
`22:1299—«13'Ll8, 2005.
`
`Received: fuly '18, 2007
`Accepted: March 24, 2008
`
`(‘marge A Baklnf;/.a.r1
`Reprint Raquel
`,:
`{STA Pizarrnncezmcnls, Inc.
`£5295 Alton Parkway
`Irvine, CA 92618
`
`l€‘~umi.?: gbaklayan@i_smvisioamonx
`
`14-3.
`
`18.
`
`topical
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`ment of acute ps::>..ldophal<ic Cysztuszzl maCul.3r edema. 0,1711
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`«'h:”II':‘I'I:Jf(?(|_{]/I ]07:203«l~‘?.( , ,, ’
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`200-1.
`. Damnenfeld, F..E?., t
`lmliaml, E-._l,, Stewart, R.H., et al. Bromfenac
`
`ophthalmic solution
`% (Xiblwn) for po:~tCwp&mtiVe ocular
`pair: and inflamrntwtimx. O,7J.*12‘I:m'n1.:7}r)gy ll4:16'3.3 1662,. 2307.
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