123
`
`24-HC>ur Evaluation of the Ocular 1Pharmacokine1tics of 14C-Labeled Low-Concentration, Modified Bromfenac
`Ophthalmic Solution Following Topical Instillation into the Eyes of New Zealand White Rabbits
`
`~,:;~~~~~A:~:-:-;e: To evaluate the 24-hour ocular distribution
`and concentrations of 14C-Iabeled
`low-concentration,
`modified bromfenac ophthalmic solution following topical
`instillation 1n New Zealand White rabbit eyes.
`
`~.:~~::U·:o::j~;;: Eighteen New Zealand white rabbits were
`randomly assigned to 6 treatment groups (3 animals per
`treatment group). A single dose of 50~L of the dosing
`solution was topically administered mto the conjunctival
`sac of both eyes of each animal. Animals were
`euthanized and the aqueous humor was collected at 1
`hour ± 5 m1nutes, 2 hours ± 15 m1nutes, 4 hour~. ± 15
`minutes, 8 hours ± 15 minutes, 12 hours ± 15 minutes,
`or 24 hours ± 15 m1nutes
`following dosing. The
`iris/ciliary body, lens, vitreous, retina, choroid, sclera,
`conJunctiva, and cornea
`(target
`tissues) wem also
`collected from each eye for analysis. Dosing solutions
`were analyzed
`to
`confirm
`radiochemical
`punty;
`radioactive concentration of the dosing solutions was
`calculated us1ng liquid scintillation count1ng (LSC).
`
`14C-Iabeled
`of
`residues
`Radioactive
`R~::~;;:}~t:"?~
`bromfenac, expressed as mean parts per million [(ppm)
`~g/g] was seen 1n all target t1ssues of the eyes, with the
`highest concentrations found in the cornea, conjunctiva,
`and sclera. The conCE:mtrat1ons 1n the t1ssues diminished
`to varying degrees over the 24 hour study period, with
`the exception of the lens, which increased insignificantly
`from the 1 hour tim<= point. The levels detected in the
`lens and VItreous humor were
`low and close
`to
`background levels.
`
`C::;..n::;~:~~:-:-;:::_;n: Significant penetration and measurable
`amounts of 14C-Iabe1Ed bromfenac were detected in most
`ocular target t1ssues over 24 hours, with highest levels
`the cornea, conJunctiva, and sclera. The 14C-Iow
`1n
`concentration bromfenac residues in ocular tissues were
`similar to those previously reported with 0.09% 14((cid:173)
`bromfenac,
`the currently available concentration of
`bromfenac formulation.
`
`Bromfenac is a non-steroidal anti inflammatory drug
`(NSAID) with an extens1ve history of clinical efficacy; 1t
`acts by blocking prostaglandin synthesis by inhibiting
`cyclooxygenase 1 and 2 1n
`the arachidonic acid
`pathway.
`
`Baklayan G.A.
`Bausch & Lomb Inc.
`
`Bromfenac, like most of the NSA!Ds, are weakly acidic
`drugs, which ioniz" at the pH of the lachrymal fluid and
`therefore have limited permeability through the an1on1c
`cornea, which has an isoelectric point (pi) of 3.2.
`Reducing
`the pH of the
`formulation 111creases
`the
`unionized
`fraction of
`the drug which enhances
`permeation. 4
`
`(bromfemac ophthalmic solution) 0.09%,
`Xibrom '"
`administered twim daily, was approved by the Food and
`Drug Administration (FDA) on March 24, 2005 for the
`treatment of patients with post-cataract ocular
`inflammation, and 1n January 2006 for the treatment of
`ocular pain following cataract surgery. 5
`
`(brornfenac ophthalmic solution) 0.09%
`Bromday'"
`administered once daily, was approved by the FDA on
`October 16, 2010 for the treatment of postoperative
`inflammation and reduction of ocular pa1n 1n pat1ents
`who have undergone cataract extraction. 6
`
`An advanced formulation of bromfenac (PROLENSA™
`[bromfenac ophthalmic solution] 0.07%) has been
`developed which contains a lower pH (7.8) compared to
`earlier formulations. The lower pH facilitates corneal
`penetration. PROLI::NSA also has a lower concentration
`compared
`to
`other
`available
`formulations
`of
`bromfenac 7
`
`14C-Iabeled bromfenac 0.07% was analyzed to verify the
`radioactive purity prior to its use in the study.
`
`Prior to treatment, 18 animals were weighed and randomly
`assigned to 6 treatment groups.
`A physical exam1nat1on was performed on each animal
`including a pre-treatment ophthalmic examination (slit lamp
`and
`indirect ophthalmoscopy). Acceptance cntena
`for
`placement on study were as follows:
`< Scores of ,; 1 for conjunctival swelling
`~ Scores of 0 for all other observation variables
`Each rabbit received topical ocular doses of a 50~L test agent
`dose into the conjunctival sac using a calibrated pipette and
`the eyelid was held closed for 5-10 seconds following the
`dose.
`
`3 rabbits (both eyes) were assessed per time point over a 24
`hour period
`
`Animals were euthanized at the following 6 t1me po111ts: 1 hr
`± 5 min, 2 hrs ± 15 min, 4 hrs ± 15 min, 8 hrs ± 15 min, 12
`hrs ± 15 m111, or 24 hrs ± 15 m1n following dosing.
`
`The iris/ciliary body, lens, VItreous, ret1na, choroid, sclera,
`conjunctiva, and cornea were collected from each eye and
`weighed.
`
`The ocular pharmacokinetics of 14C-Iabeled bromfenac
`0.07%, pH=7.8 was assessed at 6 time points over a 24
`hour t1me interval.
`
`.·.·::.·--Aqueous Hurnor
`
`W»">Iris/Ci:i~ry Body
`
`·.·:·:·.·.Ler.s
`
`,...Ret!na
`
`.·.·.·.··Sclera
`
`The mean ~g/g of drug-derived radioactivity following
`administration of 14C-bromfenac was seen in all tissues of
`the eyes at low levels, with the highest concentrations
`found in the cornea, conjunctiva, and sclera (Figure 1).
`
`The concentrations in the tissues diminished to varying
`degrees over the 24 hour study period. Levels 1n the lens
`were very low and remained essentially unchanged. The
`radioactivity detected by LSC 1n both the l"ns and the
`vitreous humor was very low and close to background
`values.
`
`The bromfenac 0.07% formulation has been shown to
`Improve the
`penetration 111to ocular t1ssues
`thereby
`allowing for a lower concentration with comparable tissue
`concentrations to those seen with BROMDAY.
`
`the advanced
`and BROMDAY,
`to XIBROM
`Similar
`formulation of bromfenac 0.07% provides
`therapeutic
`concentrations throughout ocular t1ssues for 24 hours
`which allows for once daily dosing.
`
`PROLENSA (bromfenac ophthalmic solution) 0.07% was
`FDA approved on April 5th, 2013 for once daily dosing for
`the treatment of postoperative inflammation and reduction
`of ocular pa111 1n pat1ents who have undergone cataract
`surgery.
`
`Significant penetration and measurable amounts of
`14C-Iabeled bromfenac were detected in most ocular
`target tissues over 24 hours, with highest levels in the
`cornea, conjunctiva, and sclera. The 0.07Dfo
`14C(cid:173)
`bromfenac residues in ocular tissues were similar to
`those previously reported with 0.09% 14C-bromfenac.
`
`1. Baklayan GA, Patterson HM, Song CK, Gow JA, McNamara TR. J Ocular
`Pharmacal Ther 2008;24(4):392-398.
`2. Donnenfeld, E.D., Holland, E.J., Stewart, R.H., et.al. Ophthalmology.
`2007; 114:1653-1662.
`~~~: S.J., Flach, A.J., Jampol, L.M. Surv Ophthalmol. 2010; 55(2):108-
`
`3.
`
`4. Ahuja M, Dhake AS, Sharma SK, Majumdar DK. AAPS J 2008;10:229-41.
`XIBROMTM Prescribing Information. ISTA Pharmaceuticals, Inc. 2010
`5.
`BROMDAYTM Prescribing Information. Bausch and Lomb, Inc. 2011.
`6.
`PROLENSATM Prescribing Information, Bausch and Lomb, Inc. 2013
`7.
`
`The chemical structure of bromfenac conta1ns a
`bromine atom at the C4 pos1t1on, which
`Imparts
`distinguishing characteristics
`from other ophthalmic
`NSA!Ds
`including
`improved potency and enhanced
`lipophilicity
`of
`the molecule, which
`facilitates
`~~1?~!:2~~~~!!5?.~~~~~5?.~5?.~~~1 ~~~-~ ~ ~~ ~:.~: .. 1. -.:. • • • • • • • • • • • • • • • • • • • • • • • • • •••• -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~--------- --------------------------------------------- -~~~~~~~~~~~~~~~~~~~~~- •••••
`Animal use for this study was approved by the facility's Institutional Animal Care and Use Committee (IACUC) and conformed to the ARVO "'Statement on the Use ~~-~~;~~~·;·i·~-~~-~~~~~-~i·~-~~d-vi·;~~~-R~-;~~~~;;_:··~~~~~~~~~~~~~--------------------------------------------------------------------------~~~~---················-···················
`Presented at the 2013 Association for Research in Vision and Ophthalmology (ARVO) Annuall Meeting; May 5-9, 2013; Seattle,. WA
`
`0.0001 L . ._ - - - - : - · - - - - - - - - - - - - - - - - - - - - - - - - - - -
`
`Hours
`
`Conjunctfva
`
`Financial support: Bausch & Lomb Inc., Irvine, CA, USA
`Financial disclosures: GA Baklayan is an employee of Bausch & Lomb, Inc
`
`PROL0280755
`
`SENJU EXHIBIT 2223
`INNOPHARMA v SENJU
`IPR2015-00903
`
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