AU2016269432A1 - Biodegradable introcular tyrosine kinase inhibitor implants - Google Patents

Abstract

H:\fm\Intrwovn\NRPortbl\DCC\FMT\I230729IL.docx- 12/6/2016 BIODEGRADABLE INTRAOCULAR TYROSINE KINASE INHIBITOR IMPLANTS Abstract Biocompatible intraocular implants include a tyrosine kinase inhibitor and a biodegradable polymer that is effective to facilitate release of the tyrosine kinase inhibitor into the vitreous of an eye for an extended period of time. The therapeutic agents of the implants may be associated with a biodegradable polymer matrix, such as a matrix that is substantially free of a polyvinyl alcohol. The implants can be placed in an eye to treat or reduce the occurrence of one or more ocular conditions, such as posterior ocular conditions.

Description

BIODEGRADABLE INTRAOCULAR TYROSINE KINASE INHIBITOR IMPLANTS by

Jeffrey L. Edelman, Patrick M. Hughes, Thomas C. Malone, Gerald DeVries,

Joan-En Chang-Lin, Jane Guo Shiah, Thierry Nivaggioli, Lon T. Spada and

Wendy M. Blanda

CROSS REFERENCE

This application is related to United States application number 10/837,361 filed April 30, 2004, the entire content of which is incorporated herein by reference.

This is a divisional of Australian Patent Application No. 2014203794, the entire contents of which are incorporated herein by reference.

BACKGROUND

The present invention generally relates to devices and methods to treat an eye of a patient, and more specifically to intraocular implants that provide extended release of a therapeutic agent to an eye In which the implant is placed, and to methods of making and using such implants, for example, to treat or reduce one or more symptoms of an ocular condition.

Delivery of drugs to the retina, vitreous and uveal tract is typically achieved by high systemic dosing, intra-ocular injections or other heroic measures. Penetration of systemically administered drugs Into the retina is severely restricted by the blood-retinal barriers (BRB) far most compounds. Although intraocular injection, such as intravitreal injections, resolves some constraints posed by the BRB and significantly reduces the risk of systemic toxicity, intraocular injection techniques may result in retinal detachment, physical damage to the lens, exogenous endophthalmitis, and also may result in high pulsed concentrations of drug at the lens and other intraocular tissues.

Compounds are eliminated from the vitreous by diffusion to the retrozonular space with clearance via the aqueous humor or by trans-retinal elimination. Most compounds utilize the former pathway while lipophilic compounds and those with trans-retinal transport mechanisms will utilize the fatter. Unfortunately, compounds that are eliminated across the retina have extremely short half-lives, Hence, for these compounds it is difficult to maintain therapeutic concentrations by direct intraocular Injection, and therefore, frequent injection Is often required,

Additionally, the rapid elimination of retinaly cleared compounds makes formulation of controlled delivery systems challenging. For example, tyrosine kinase Inhibitors {TKIs} may possess extremely short Intraocular half-lives, and thus, may pose a challenge to the formulation of controlled delivery systems,

The Inventors are unaware of any small molecule TKIs given by intraocular administration, let alone, intraocular Implants containing TKIs. 0,8. Patent No. 6,713,081 discloses ocular Implant devices made from polyvinyl alcohol and used for the delivery of a therapeutic agent to an eye in a controlled and sustained manner. The Implants may be placed subconjunotivally or intravitreally in an eye,

Blosompatlbl© Implants for placement in the eye have also been disclosed in a number of patents, such as U.S. Pat. Nos. 4,621,210; 4,853,224; 4,997,652; 5,164,188; 5,443,605; 5,501,858; 5,786,242; 5,824,072; 5,869,079; 6,074,881; 6,331,313; 6,369,116; and 8,699,493.

If would be advantageous to provide eye implantable drug delivery systems, such as intraocular implants, and methods of using such systems, that are capable of releasing a therapeutic agent at a sustained or controlled rate for extended periods of time and in amounts with few or no negative side effects.

SUMMARY

The present Invention provides new drug delivery systems, and methods of making and using such systems, for ©Mended or sustained drug release Into an eye, for example, to achieve one or more desired therapeutic effects. The drug delivery systems are in the form of implants or implant elements that may be placed In an eye. The present systems and methods advantageously provide for extended release times of one or more therapeutic agents. Thus, the patient In whose eye the Implant has been placed receives a therapeutic amount of an agent for a long or extended time period without requiring additional administrations of the agent. For example, the patient has a substantially consistent level of therapeutically active agent available for consistent treatment of the eye over a relatively long period of time, for example, on the order of at least about one week, such as between about one and about six months or even for more than one year after receiving an implant. Such extended release times facilitate obtaining suocassfu! treatment results. The implants allow for prolonged delivery of a therapeutic agent while reducing invasive procedures and reducing high transient concentrations associated with pulsed dosing.

Intraocular implants in accordance with tie disclosure herein comprise a therapeutic component and a drug release sustaining component associated with the therapeutic component. The implants may be solid, semisolid, ©r viscoelastic. In accordance with the present Invention, the therapeutic component comprises, consists essentially of, or consists of, a tyrosine kinase inhibitor (TKI), for example, an agent or compound that inhibits ©r reduces the activity of tyrosine kinase. The TKI may also be understood to be a small molecule TKI. The drug release sustaining component la associated with the therapeutic component to sustain release of an amount of the TKI into an eye in which the implant is placed. TKis may be released from the Implant by diffusion, erosion, dissolution or osmosis. The drug release sustaining component may comprise one or more biodegradable polymers or one or more non» biodegradable polymers. Examples of biodegradable polymers of the present implants may include pofy-iaciide-so-glyeolide (FLGA and PLA), polyesters, pcfy (ortho ester), poly(phcsphazlno), poly (phosphate ester), polycaprolaetone, natural polymers such as gelatin or collagen, or polymeric blends. The amount of the TKI Is released Into the eye for a period of time greater than about one week after the implant Is placed in the eye and Is effective in reducing or treating an ocular condition.

In one embodiment, the intraocular implants comprise a TKI and a biodegradable polymer matrix. The TKI Is associated with a biodegradable polymer matrix that degrades at a rate effective to sustain release of art amount of the TK! from the implant effective to treat an ocular condition. The intraocular implant is biodegradable or bioerodibfe and provides a sustained release of the TKI in an ©ye for extended periods of time, such as for mere than one week, for example for about one month or more and up to about six months or more, The Implants may be configured to provide release of the therapeutics agent In substantially on© direction, or the Implants may provide release of the therapeutic agent from all surfaces of the implant

The biodegradable polymer matrix of the foregoing Implants may be a mixture of biodegradable polymers or the matrix may comprise a single type of biodegradable polymer. For example, the matrix may comprise a polymer selected from the group oonsisting of poiyiaotides, poly (lactide-co-giyeoBdea), poiycaprofaoiones, and combinations thereof, in another embodiment, intraocular implants comprise a therapeutic component that comprises a TKI, and a polymeric outer layer covering the therapeutic component. The polymeric outer layer includes one or more orifices or openings or holes that are effective to allow a liquid to pass Into the implant, and to allow the TKI to pass out of the implant The therapeutic component Is provided in a core or interior portion of the implant, and the polymeric outer layer covers or coats the core. The polymeric outer layer may include one or more non-bfodegradabla portions. The implant can provide an extended release of th© TKI for more than about two months, and for more than about one year, and even for more than about five or about ten years. One example ©f such a polymeric outer layer covering Is disclosed in U,3. Patent No, 6,331,318,

Advantageously, the present implants provide a sustained or oontrolled delivery of therapeutic agents at a maintained level despite the rapid elimination of the TKls from the ©ya, For example, the present implants are capable of delivering therapeutic amounts of a TKI for a period of at feast about 30 days to about a year despite the short intraocular halMIves associated with TKls. Plasma TKI levels obtained after implantation are extremely low, thereby reducing issues or risks of systemic toxicity. The controlled delivery of the TKls from the present implants permits the TKIs to be administered into an eye with reduced toxicity or deterioration of the blood-aqueous and blood-retinal barriers,, which may b® associated with intraocular Injection ©f liquid formulations containing TKIs, A method of making the present implants involves combining or mixing the TKI with a biodegradable polymer or polymers. Hie mixture may then be extruded or compressed to form a single composition. The single composition may then be processed to'form Individual implants suitable for placement In an eye of a patient.

Another method ©f making the present Implants involves providing a polymeric coating around a ©ore portion containing a TKI, wherein the polymeric coating has one or more holes.

The implants may be plaoed in an ocular region to treat a variety of ocular conditions, such as treating, preventing, or reducing at least one symptom associated with non-exudative age related macular degeneration, exudative age related macular degeneration, choroidal neovascularization, acute macular neuroretinopathy, cystold macular edema, diabetic macular edema, Behcet’s disease, diabetic retinopathy, retinal arterial ocdusive disease, central retinal vein occlusion, uvettic retinal disease, retinal detachment, trauma, conditions caused by laser treatment, conditions mused by photodynamic therapy, photoeoagulation, radiation retinopathy, epiretinal membranes, proliferative diabetic retinopathy, branch retinal vein occlusion, anterior ischemic optic neuropathy, non-retinopathy diabetic retinal dysfunction, retinitis pigmentosa, ocular tumors, ocular neoplasms, and the tike.

Kits in accordance with the present invention may comprise one or more of the present Implants, and instructions for using the implants. For example, the Instructions may explain how to administer the implants to a patient, and types of conditions that may be treated with the Implants.

Our invention also encompasses a biodegradable intravitreal implant comprising a tyrosine kinase inhibitor (TKI) and a biodegradable polymer, Tha

Implant can release the TKi at a rat® effective to sustain release of the TKI from the implant for at least about one weak after the implant Is placed In the vitreous of an eye. The polymer can be a polylaotide, poly (faofld®*co»glyoolld©), polyeaprolaetona, or a derivative thereof, or a mixture thereof, Th© polymer can release the TKI at a rate effective to sustain release of an amount of the TKI from the Implant for more than one month from the time the Implant is placed In the vitreous of the eye, Th© TKI can be provided in an amount from about 30% by weight to about 70% by weight of the implant, and the biodegradable polymer matrix oan comprise a poly paotlde~oo-glyc©IIde} In an amount from about 30% by wight to about 70% by weight of the implant. More preferably, the TKI oan b© provided In an amount from about 40% by weight to about 60% by weight of the Implant, and the biodegradable polymer matrix can comprise a poly (lactlde-co-glycollde) in an amount from about 40% by weight to about 60% by weight of the Implant, The Implant oan be made by an extrusion process, A detailed embodiment of our invention oan be a method of making a biodegradable Intravitreal Implant, comprising the step of; extruding a mixture of a TKI and a biodegradable polymer to form a biodegradable Implant that degrades at a rate effective to sustain release of an amount of the TKI from th© Implant for at least about one weak after the Implant is placed In the vitreous of an eye. The mixture oan consist essentially of the TKI and the biodegradable polymer.

Our invention also encompasses a method of treating an ocular condition of an eye of a patient, comprising the step of placing a biodegradable intraocular Implant Into tii© vitreous of an eye of the patient, the implant comprising a TKI and a biodegradable polymer, wherein the Implant degrades at a rate effective to sustain release of an amount of the TKI from the Implant effective to treat the ocular condition. This method can be affective to treat, for example, a retinal ocular condition, glaucoma or a proliferative vltreoretinopathy.

The TKi used In the implant can have, for example, one of the following five possible structures;

Each and ©very feature described herein, and each and every combination of two or more of such features, la Included within the scope of the present invention provided that the features included in euoh a combination are not mutually inconsistent In addition, any feature or combination of features may be specifically excluded from any embodiment of the present invention.

Additional aspects and advantages of the present invention are set forth in the following description and claims, particularly when considered In conjunction with the accompanying drawings.

BBflMMBS

Figure 11s a graph showing the vitreous humor concentration of two TKis as a function of time,

Figure 2 Is a graph similar to Figure 1 for two different TKIs,

Figure 3 is a graph of the cumulative release profile for AGN 200954 as a function of time.

Figure 4 is a graph of the cumulative release profile for AGN 202314 as a function of time.

Figure 5 is a graph simitar to Figure 4 for different formulations of AGN 202314.

Figure 6 Is a graph of the TTL release for AGN 201634 as a function of time.

Figure 7 is a graph similar to Figure 8 with implants containing 30% AGN 201034.

Figure 8 is a graph similar to Figure 0 for AGN 201634 In different solutions.

Figure 9 is a graph of the percent of TKI released as a function of time in different a Tween 80/saline solution.

Figure 10 Is a graph similar to Figure 9 except in a phosphate buffer solution,

Figure 11 is a graph of the cumulative release profile for TKI AGN 201634 of Formulation 11n saline and PBS.

Figure 12 is a graph of the cumulative release profile for TKI AGN 201634 release of Formulation 3 in media of a pH of 6.0 {with 0.1 % GTAB), 7,4 (PBS) or 8.5 (with 0.6% SDS).

Figure 13 la a graph of the cumulative release profile for TKIAGN 201634 release of Formulation 4 in media of a pH of 6.0 (with 0.1% CTAB), 7.4 (PBS) or Θ.5 (with 0,5% SDS).

Figure 14 Is an lustration of a biodegradable implant comprising a drugreleasing active layer and a barrier layer.

Figure 1S is a graph of the cumulative release profile for a TKI-containing Implant and Dexamethasone-conteinlng Implants, in which the biodegradable polymer Is pofyoaprofactone.

Figure 16 Is a graph of the cumulative release profile for three different formulations of TKI (AGM2Q6639) containing implants in phosphate buffered saline release medium, pH 7,4,

Figure 17 Is a graph of the cumulative release profile for five further and different formulations of TKI (AGK206639) containing implants In phosphate buffered saline release medium, pH 7.4.

Figure 18 is a graph of the cumulative release profile for three different formulations of TKI (AGN2055SB) containing implants In phosphate buffered saline release medium, pH 7.4.

Figure 19 is a graph of the cumulative release profile for eight different formulations of TKI (AGN20632Q) containing Implants in phosphate buffered saline release medium, pH 7.4.

Figure 20 Is a graph of the cumulative release profile for eight different formulations of TKI (AQN206784) containing Implants In phosphate buffered saline release medium, pH 7,4,

Figure 21 is a graph of the cumulative release profile for seven different formulations of TKI (AGM206816) containing implants in phosphate buffered saline release medium, pH 7,4,

Figure 22 Is a bar graph showing on the X axis time {14 days and 28 days) after Intravltrea! implantation lot© rabbit eyes (N - 7) of either a RIGA placebo Implant or a TKI (AGN208639) PL8A Implant, and on the Y axis vitreoratlnal fluorescence (as determined by a scanning ocular fluorophotometry area under the curve fAUC] method after Intravenous Injection of sodium fluorescein). DESCRIPTION!

As described herein, controlled and sustained administration of a therapeutic agent through the use of one or more intraocular Implants may Improve treatment of undesirable ocular conditions. The implants comprise a pharmaceutically aooeptable polymeric composition and are formulated to release one or more pharmaceutically active agents, such as tyrosine kinase inhibitors (TKIs), over an extended period ©f lima. The Implants are effective to provide a therapeutically effective dosage ol die agent or agents directly to a region of the eye to treat, prevent, and/or reduce one or more symptoms of one or more undesirable ocular conditions. Thus, with a single administration, therapeutic agents will be made, available at the site where they are needed and will be maintained for an extended period of time, rather than subjecting the patient to repeated Injections or, In the case of self-administered drops, Ineffective treatment with only limited bursts of exposure to the active agent or agents.

An intraocular implant In accordance with the disclosure herein comprises a therapeutic component and a drug release sustaining component associated with the therapeutic component. In accordance with the present inventions the therapeutic component comprises, consists essentially of, ©r consists of, a TKI. The drug release sustaining component Is associated with the therapeutic component to sustain release of an effective amount of the therapeutic component into an eye In which the Implant is placed. The amount of the therapeutic component is released Into the eye for a period of time greater than about one week after the Implant is placed In the eye, and Is effective In treating and/or reducing at least one symptom of one or mors ©solar conditions, such as conditions wherein migration or proliferation of retina! pigment epithelium or glial cells causes or contributes to the cause of the condition. Some examples of ocular conditions that may be treated with the Implants of the present invention indude* without limitation, non-exudative age related macular degeneration, exudative age related macular degeneration, choroidal neovascularization* acute macular neuroretinopathy, cyetoid macular edema, diabetic macular edema, Behoofs disease, diabetic retinopathy, retinopathy of prematurity, retinal arterial occlusive disease, central retinal vein occlusion, uveltlo retinal disease, retinal detachment, trauma, conditions caused by laser treatment, conditions caused by photodynamic therapy, photocoagulatton, radiation retinopathy, epirettnal membranes, proliferative diabetic retinopathy, branch retinal vein occlusion, anterior ischemic optic neuropathy, non-retinopathy diabetic retinal dysfunction, retinitis pigmentosa, ocular tumors, ocular neoplasms, and the like.

Definitions

The following terms as defined as follows, unless the context of the word indicates a different meaning.

As used herein, an Intraocular implant" refers to a device or element that is structured, sized, or otherwise configured to be placed in an eye. Intraocular implants are generally biocompatibi© with physiological conditions of an eye and do not cause adverse side effects. Intraocular implants may be placed in an aye without disrupting vision of the eye.

As used herein, a “therapeutic component" refers to a portion of an Intraocular implant comprising one or more therapeutic agents or substances used to treat a medical condition of the aye. Th© therapeutic component may b© a discrete region of an Intraocular implant, or it may be homogenously distributed throughout th® implant. Th© therapeutic agents of toe therapeutic component are typically ophthalmicaliy acceptable, and are provided in a form that does not oaus® adverse reactions when the Implant Is placed In an aye.

As used herein, a "drug release sustaining component1' refers to a portion of the intraocular implant that is affective to provide a sustained release of the therapeutic agents of the implant, A drag release sustaining component may be a biodegradable polymer matrix, or it may be a coaling covering a core region of the implant that comprises a fherapeutio component.

As used herein, “associated with" means mixed with, dispersed within, coupled to, covering, or surrounding,

As used herein, an ’’ocular region" or "ocular site" refers generally to any area of the eyeball, including the anterior and posterior segment of the eye, and which generally includes, but is not limited to, any functional (e,g,s for vision) or structural tissues found In the eyeball, or tissues or cellular layers that partly or completely line the interior or exterior of the eyeball. Specific examples of areas of the eyeball in an ocular region include the anterior chamber, the posterior chamber, the vitreous cavity, the choroid, the suprachoroldal space, foe conjunctiva, the subconjunctival space, the episcleral space, the intracorneal space, the epicomea! apace, the sclera, the pars plana, surgically-induced avascular regions, the macula, and the retina.

As used herein, an “ocular condition" is a disease, ailment or condition which affects or involves the eye or on® of the parts or regions of the aye. Broadly speaking the eye includes the eyeball and foe tissues and fluids whioh constitute the eyeball, th® periocular muscles (such as foe oblique and rectus muscles) and the portion of the optic nerv® whioh Is within or adjacent to th© eyeball.

An "anterior ocular condition" Is a disease, aliment or condition which affects or which Involves an anterior (La, front of the eye) ocular region or site, such as a periocular muscle, an eye lid or an eye bail tissue or fluid which is located anterior to the posterior wall of th® lens capsule or clary muscles. Thus, an anterior ocular condition primarily affects or Involves foe conjunctiva, th© cornea, th® anterior chamber, th© iris, the posterior chamber (behind the retina but in front of the posterior wall of the lens capsule), the tens or the tens capsule and blood vassals and nerve which vascularize or innervate an anterior ocular region or site.

Thus, an anterior ocular condition can include a disease, ailment or condition, sueh as for example* aphakia; pseudophafda; astigmatism; blepharospasm; cataract; conjunctival diseases; conjunctivitis; corneal diseases;, corneal ulcer; dry eye syndromes; eyelid diseases; lacrimal apparatus diseases; lacrimal dust obstruction; myopia; presbyopia; pupil disorders; refractive disorders and strabismus. Glaucoma can also be considered to be an anterior ocular condition because a clinical goal of glaucoma treatment can be to reduce a hypertension of aqueous fluid In the anterior chamber of the eye (i.e. reduce intraocular pressure), A ’’posterior ocular condition1* Is a disease, aliment or condition which primarily affects or involves a posterior ocular region or site such as choroid or solera (in a position posterior to a plane through the posterior wall of the lens capsule), vitreous, vitreous chamber, retina, optic nerve (Le. the optic disc), and blood vessels and nerves which vascularize or Innervate a posterior ocular region or site.

Thus, a posterior ocular condition ran Include a disease, ailment or condition, such aa for example, aoute macular neuroretinopathy; Behcet's disease; choroidal neovascularization; diabetic uveitis; histoplasmosis; Infections, such as fungal or vlral-caused infections; macular degeneration, such as aoute macular degeneration, non-exudative age related macular degeneration and exudative age related macular degeneration; edema, such as macular edema, cystoid macular edema and diabetic macular edema; multifocal choroiditis; ocular trauma which affects a posterior ocular site or location; ocular tumors; retinal disorders, such as central retinal vein occlusion, diabetic retinopathy (including proliferative diabetic retinopathy), proliferative vitreoretinopathy (PVR), retinal arterial occlusive disease, retinal detachment* uvelc retinal disease; sympathetic opfhalmia; Vogt Koyanagl-Harada fVKH) syndrome; uveal diffusion; a posterior ocular condition caused by or Influenced by an ocular laser treatment; posterior ocular conditions caused by or influenced by a photodynamic therapy, photQoaagulation, radiation retinopathy, epi retinal membrane disorders, branch retina! vein occlusion, anterior Ischemic optic neuropathy, non-retinopathy diabolic retinal dysfunction, retinitis pigmentosa, and glaucoma. Glaucoma can be considered a posterior ocular condition because the therapeutic goal is to prevent the loss of or reduce the occurrence of loss of vision due to damage to or toss of retinal cells or optic nerve cells (i.e. rsauroproteoticn).

The term “biodegradable polymer® refers to a polymer or polymers which degrade in vivo, and wherein erosion of the polymer or polymers over time occurs concurrent with or subsequent to release of the therapeutic agent Specifically, hydrogels such as methyicelluloee which act to release drug through polymer swelling are specifically excluded from the term ’’biodegradable polymer". The terms “biodegradable" and "bioerodibfe" are equivalent and are used Interchangeably herein. A biodegradable polymer may be a homopolymer, a copolymer, or a polymer comprising more than two different polymeric units.

The term ’’treaf, ’’treating8, or "treatment8 as used herein, refers to reduction or resolution or prevention of an ocular condition, ooular Injury or damage, or to promote healing of injured or damaged ooular tissue.

The term "therapeutically effective amount" as used herein, refers to the level or amount of agent needed to treat an ooular condition, or reduce or prevent ocular injury or damage without causing significant negative or adverse side effeots to the eye or a region of the eye. intraocular implants have been developed which can release drug loads over various’ time periods. These implants, which when inserted inf© an aye, such as the vitreous of an eye, provide therapeutic levels of a TKi, for extended periods of time (e«g„ for about 1 week or more). The disclosed implants are effective In treating ocular conditions, such as non-exudative age related macular degeneration, exudative age related macular degeneration, choroidal neovascularization, acute macular neurorotinopathy, oystald macular edema, diabetic macular edema, Behcet’s disease, diabetic retinopathy, retinal arterial occlusive disease, central retinal vein occlusion, uveitic retinal disease, retinal detachment, trauma, conditions caused by laser treatment, conditions caused by photodynamic iherapys photocoagulation, radiation retinopathy, ©piratical membranes, proliferative diabetic retinopathy, branch retina! vein occlusion, anterior ischemic optic neuropathy, non-retinopathy diabetic retinal dysfunction, retinitis pigmentosa, ocular tumors,- ocular neoplasms, and the Ike,

In one embodiment of the present invention, an Intraocular implant comprises a biodegradable polymer matrix. The biodegradable polymer matrix is one type of a drug release sustaining component The biodegradable polymer matrix Is effective In forming a biodegradable Intraocular implant The biodegradable Intraocular implant comprises a TKI associated with the biodegradable polymer matrix. The matrix degrades at a rate effective to sustain release of an amount of the TKI for a time greater than about one week from the time in which the implant is placed in ocular region or ocular site, such as tie vitreous of an eye,

The TKI of the implant is typically an agent that inhibits or reduces the activity of a tyrosine kinase. The TKI may Inhibit tyrosine kinase activity by directly acting on a tyrosine kinase molecule, or it may cooperate with one or mom other factors or agents to achieve the desired Inhibition, Examples of TKIs useful in the present implants are described In US, Patent App Mos, 10/256,879 (U.S. Pub. No, 20030190478) and 10/259,703 (U.S. Pub. Mo. 20030225152).

In short, a TKI of the present implants include organic molecules capable of modulating, regulating and/or Inhibiting tyrosine kinase signal transduction. Some compounds useful in the present Implants are represented by tie following formula wherein Ff is selected

from the group consisting of halogen, NO& ON, Ct to C4 alkyl and aryl, e.g. phenyl; Bs is selected from the group consisting of hydrogen, C1 to C8 alkyl, GOCH3l CHsCHsOH.CHjCH&HsOH and phenyl; R is selected from the group consisting of P, halogen, 0¾ to Ce alkyl, CF3i OCF3, OCFgH, onm, CM, 3R2, (CR7Rs)sCCG)GR2, C(0)M{R2)a, (CR7 R3)cQR2f HNC(0)R2, HN *Cf0)0R2, (CR7R8)eN(Rs)2} SOg (CR7R%M(R2)a 0P(0)(0Ffy, OqO)OR2 OCHaO, HM-CH^CH, -N{CORa)CHgGH2i N^CH-S, 0{CR?R8)d"Re and (CR7R8)c~R€, -MRa(CR7R8)dR@ wherein R8 is selected from the group consisting of halogen, 3-fluoropyrrolIdinyf, 3-fluoroplperldlnyl, 2-pyridinyi, 3~ pyridlnyl, 4-pyritfnyl, 3-pyrnojinyf, pyrrofidirsyl, methyl isonipecotate, N~{2-methoxyeihyi)-M-motoyIamyl, 1s2,3,6”tetrahydropyridfnyl, morpholinyl, hexamethyleneiminyi, plperazinyl«2“©ne, piparaxinyl, N~(2~ m®tho«yethyl)ethylaminyI, thiomorphollnyl, hoptamethylanelminyl, 1-piperazinylcarhoxaldehyde, 2,3,6,74strahydrD>(1 H)-1,44ia»plnyM>{4H)-one, N-methyfhomopfperazinyl, {3-dlmethylamino)pyrroIldInyls N-(2-methoxyethyt)-N-propyiaminyi, fsoindolfnyl, nlpecotamldinyl, Isonlpecotamsdinyl, 1» aoetylplparazfnyl, 8-acetamIdopyrrolldfnyt, trans^decahydiOisoquinolinyl, cis-decahydroisoqulnoiinyl, N-acetylhomoplperazInyl, 3~(dl©thylamino)pyrrolidinyi5 1,4-dl©xa*S«azaapfrof4i>idecanlriyl, 1"(2-metho3cyefhyl}“pfperaisnyl, 2-pyrroiidfn~3-ylpyrldfnyl, 4^jyrrolidln-3-ylpyridinyI, 3-(methyfsu!fonyl)pyrrolldinyl, 3-pioolyimethylamlnyl, 2-(2“methylaminoethyl)pyr!dlnyli 1 "{2“pyrimidyl}»plp©rszlnyii 1 »(2-pyra2isiyf}-piperazInyl, 2-methylaminomeihy]-1 ,3~dloxolane,2-(M~methyI»2” aminoethyl}-1,3-dioxolane, 3”(N">acetyI~M«methylaniino)pyrr©Iidinyli 2» mathoxyethyfaminyl, tetrahydrofurfurylamlnyl, 4-amlnotetrahydropyran, 2-amfn©« 1 -methoxybutane, 2»methoxy!sopropySarriiny!, 1 "{S-aminoprcpyi) Imidazole, histamyl, NsN"diisopropytelliylen®dsamlnyl, 1 ~benzy!43-ami nopyr roildyl 2~ {aminomethyl)-5~methySpyrazsnyi, a^-dimethyH^dlOKolane^-methanaminyl, (R)“3~amino~1-N«80C-pyrro!idinyI, 4-amlno~1,2,2,6,e-pentamethylplperidinyf, 4* aminomethyliatrahyd ropyran, ethanolamlne and alkyl-substituted derivatives thereof and wherein when c is 1 said CHa may be

and CHaChfeCHg; provided aaid alley! or phenyl radicals may be substituted with on® or two halo, hydroxy or lower alkyl amino radicals wherein Ft7 and R8 may be selected from the group consisting of H, F and CrC4 alkyl or CR?R® may represent a oarbocyolio ring of from 3 to 6 carbons, preferably R? and R® are H or Cf-fe b Is 0 or an integer of from 1 to 3; a is 0 or an integer of from 1 to 5, preferably 1 to 3; o i® 0 or an integer of from 1 to 4, ' d is an Integer of from 2 to 5; the wavy line represents a £ or Z bond and pharmaceutically acceptable salts thereof.

In certain Implants, the TKI is a compound having the foregoing formula, wherein R1 is selected from the group consisting of H, La b is 0; OH3, F, Cl and phenyl.

Preferably, R is selected from the group consisting of CHSi CHgCHa, OCHa, OH, t-btstyl, F, ON, C(0)NH2j HM 0{O)CHS( CHgCfGlOH, SCfeNHs, 0(0)0H, OCFgH, Isopropyl, C2H5GH, C(0)OCHg, CHgOH, NH-CH=CH, HO-N-N-H, N^CH-S, 0{CR7R%R®, (CR7R«)0R8 and »NR®(CR7R8}dR8, wherein R8 is selected from the group consisting of 3-fiuoropyrrciidinyI, 3-fluoroplperldlnyl, 2~ pyrldinyl, 3-pyrldinyl, 4-pyridinyl, 3-pyrrolinyl, pyrroitdlnyl, methyl iionlpeootate, M~ (2~methoxyethy!)-N~methy!amyl, 1,2,3,6-tetrahydropyridinyi, morphollnyl, hexametbyleriefminy!, plperazfnyl-2-one, piperailnyl, N-(2* methoxyethyljethylaminyl, thlomorphollnyl, heptamethyieneiminyl, 1-piperazfnyicarboMldehyde, 2,3,6,7-tetrahydro-(1 H)~1f4~diazepinyl»g{4H)-one, N-methylhomopiperazinyl, (S^lrnethylaminolpyrolldinyl, N-(2-meihoxyeihy!)-N» propylaminyl, Isolndoflnyl, ntpeeofamldinyl, Isonlpeootamldiny!, 1» asetyipiperazinyi, S^acetamidopyrrolidinyl, tmns-deoahydrolsoquinaiiny], els-decahydiolsoquinoHnyl, N”aesfyfhoraoplperazinyl( 3-(dl6tiiylamino)pyirolidinyl, 1,4«dIoxa~8"aiaspirof4.g]decaninyl, H2-methOMyethy!)-piperazinyi, 2»pyrrolidin~3» ylpyridinyl, ^-pyrrolsdin^-ylpyrldiny!, a^rnethyisulfonyOpyrrolldinyl, 3- pioolylnMthylaminyll2-(2-methylamlnoett^)pyddiriyil1-(2-pyrimidyQ-piparezinyll H2~pyrazinyi)~piperazirsyf, a-rnetbyiarninomethyM ,3-dioxoiane, 2-(N-methyI-2-amlnoethyi)-1,3-dtoxoIaoas S-CN-aeefyFN-methyfaminolpyrrolidlnyl,, 2-methoxyethyiamlnyl, tetrahydrofurfuryiaminyl, 4-aminotetrahydr0pyran! 2-amino» 1 -methoxybufane, 2-fneihoxyisopropyIarninyI, 1 »(3-amInopropyI)imidazole, histamyl, N,R-dlisopropyfethytenediaminyI, l-benzyl-3-amlnopytroildyl 2-{aminomethylJ-S-methylpyrazmyl, 2!2»dirnetliyi-1(3-dioMoIane-4-mefhanam!ny!g {R)-3-amfno-1-^~BOG"pyaolldinyls 4-amino-1 f252,6!8-'pentanietiiylpiperldlnyi! 4-aminomethyitetrahydropyranyf, ethanolamine and alkyl-substituted derivatives thereof, e.g. R® is morphoiinyl or CH2N{CH3}2.

More preferably, R is selected from the group consisting of m-ethyi, p-methoMy, p-hydroxy8 m»hydroxy, p-cyano, m-C(0)NH2, p»HNC{0)GH3l ρ» GHgC(0)0Hs p-SOgNHg, p-CHgOH, m-methoxy, p-CHaCHaOH, HNCH-CH, HC-N-NH, p-morpholinyl, hfcGH-S, p-OCHFa, p«COOH, p-CH3i p-OCHa, m»F, m-CHSN{C2H3)2! (GR7R8)gR* , 0{CR7R%R®and NR2{CR?R%RS

It Is noted that R may represent a condensed ring that is attached to the above phenyl ring at two positions, For example, GHaCH2GH2 may be attached at the 3 and 4 (or m and p) positions of the phenyl ring.

Still more preferably, R Is selected from the group consisting of fluoro, methyl, (GR^^R8,0(CR7R8)dR8 and MR2(CR7Ra)£jR® wherein R® is selected from dsmethylamino, diethylamino, S-flooropyrrolldlnyl, 3-fIuoropiperldlnyl, 3-pyrldinyl, 4-pyridinyl, pyrrolidinyl, morphoiinyl, psperazinyl, heptamethyienelmlnyl, t©tiabydrafurfyrylaminyt, 4-amin0teirahydrapyrarsyif Ν,Ν» dlisopropytethylenedsamlnyl and 4-aminomefhyltetrahydropyran,

In particular, the compound® of fh© present Implants may be selected from the compounds of the tables below. TABLE 1

Unsubstituted 4-fifethyl & B-Ohhro 3-[(Sybsfityted Phtnylamlno)-m©!hyIane]-1!3»diliyiJra.ir!clo!-2~on0i,

Unsubsistuted, 4»mefhyl & 5-Chloro 3-|(Substifeted Phenylamino}-methylene]»1!l3"dihydfa»indol"2~on®s.

Unsubsfituied, 4-m®fsyl a S-Chloro S-HSuMuted Phenyiamino)» methy!en®J-'1i3"dlhydro-lndol'2-'®nes.

Unsobstltuted* 4~m©thyl & 5«Chtoro ^[{Substituted Phenylamino)» ηΐΘ!ΙινΊβίΐθ]-1(3^^ν€ΐΓθ"1π£ΐ0Ρ2·οπβ®.

UnsubsiUtuiad, 4-methyi, 5-Chloro &5-Ruoro S-fCSubstituted Phenylamino)» methyfeneJ-I.Sodihydro-Indoi^onas.

Unsubstituted, 4-methyl, 5-Chtora &5~Fluoro 3»[{8ubstitutecl Phenylamin©)-πιβΐΗν!βηθ}'Ίί3"εϋΗ)?ζίΓθ“5ηεΙοΙ~2-·οη©51 0" R1 4s i 1 >0

Unsybsfituted, 4-meihyl, 5-Chfora &5-Fluofo 3~[fSubstituted Pfienylamlno)-m©!hyIer.e]”1,3-dihydro-sndo!-2-ones, 5

WO 2O8S/M7708! PCT/USaOOS/HS]14

Uoiubitlfuted, 4-Fluo.ro, 4~m©fhy!, S-Chtoro, S-Cyano, 5~Ru©r©, 6-Nitro, 6~ Ruor© & 6-Aryl S-KSubstltuiad Phsnyiamlno)-m0ttiyten®]-1f3-elihytlr©-lndDl-2" ones.

Unsubstitutsd, 4-Fluoro, ^-methyl, S-Chloro, 5-Cyano, 5-Ruoro, 6-Nitrof a~Ruor© & 6-Aryl 3-KSubsiituted Phenylam!n0)~methylen©]-1,3« cHhydro-indoha-onas,

UrBubstltuted, 4-Fluoro, 4-m®thyljs 5-Chtoro, 5-Cyan©s S-Ruoro, S-IMItro, 6~ Huoro & 6-Aryl 3-[(Sub$t!tuted Phenylaminol-mathytenel-^S-cJifiydra-inctd-E" on®s.

Unsubstituted, 4-FIuoro, 4»mef!hyf( 5-Ch!oros S-Cyano, 5-Fluoro, 5-Nlfro, 8-Fluoro & β-Aryl 3~[{Substitutsd PhenylaminQl-mtthytenel-l^-dihydro-indoi-a» ones.

Unsubsilfuted, 4-Fluoro, ^-methyl, 5-Chloro, 5-Cyano, S~FSuoro, S-Mitro, 6-Fluor© & 6-Aryf 3*[{Subst!tutecJ Ph0ny[aminQ)"rnethyten®l“1s3"dfhydrO“ind@l“2“ ones,

Unsubstituted, 4-FIuoro, 4-methyl, 5-Chioro, S~Cyanos 5-Fluoro, 5-Nstro, 6» Ruoro & 6»Aryl 3“[{Substituted Phtny!amino}"m#thyten©]"1s3~dIhydrO"!!idDl-2-©n©s,

Unsubslfutadj 4-Fluom,, 4»m©thyl, 5-Chforo, S-Fluor© &8-Fluoro 8-[{Substituted Ph@nylamIno)”methylio©l“1!3"dihydrO“lnd©l-2-onas,

Unsufosflluted, 4-Ruoro, 4-methyi( 5-Chbro, S-Fiuoro &6»Ffuor© 3“[(Subslitutad Phenylamino}-m®ttiytene]-tf3“dihydro-ind©l-2"Onea,

The present implants may also comprise a TKi or a combination of TKIs represented by the following formulas

AGN 261684

Additional TKis that may be used In th© present Implants include those compounds disclosed In Goel of al., “Tyrosine Kinase Inhibitor®.· A Clinical Perspective*, Current ©neology Report®, 4:9-19 (2602); Haluska eta!., «Receptor tyrosine kinase Inhibitors", Current Opinion in Investigational Drugs, 212):280-286 (2001)* Hubbard ©t al., "Protein tyrosine kinase structure and function", Anno, Rev. Biochem,, 69:378-98 (2000); Basse et at, "Tyrosine kinase inhibitors: rationale, mechanisms of action, and Implications for drug resistance", Semin Oncol 28(suppl 16) 47-55 (2001): and Fabbra et al,, "Protein tyrosine kinase inhibitors: new treatment modalities?’, Current Opinion in Pharmacology, 2:374381 (2002).

Th© foregoing compound® may be synthesized using routine chemical technologies and methods including those disclosed in U.S. Patent App Nos. 16/256,079 (U.S. Pub, No, 20030199478) and 10/259,703 (U.S, Pub, No, 26080225152) and the other above-identified references.

The present Implants may also include salts of the TKIs, Pharmaceutically acceptable acid addition salts of th© compounds ©f the invention are those formed from acids which form non-toxic addition sails containing pharmaceutically acceptable anions, such as the hydrochloride, hydrobromide, hydroiodide, sulfate, or bisulfate, phosphate or add phosphate, acetate, maleate, fumarate, oxalate, lactate, tartrate, citrate, gluconate, saceharaie and p-teiuene sulphonate salts.

Thus, the Implant may comprise a therapeutic component which comprises, consists essentially of, or consists of a TKI, sate thereof, and mixtures thereof- The biodegradable polymer matrix of such implants may be substantially free of polyvinyl alcohol, or in other words, Includes no polyvinyl alcohol, .

Additional TKIs may be obtained or synthesized using conventional methods, such as by routine chemical syntheals methods known to parsons of ordinary skill In the art. Therapeutically effective TKIs may be screened and identified using conventional screening technologies used for the TKIs described herein.

The TKIs may be in a particulate or powder form and entrapped by the biodegradable polymer matrix. Usually, TKI particles in intraocular Implants will have an effective average size less than about 8000 nanometers. In certain implants, the particles may have an effectlv® average partiole size about an order of magnitude smaller than 3000 nanometers. For example, the particles may have an effective average particle size of less than about S00 nanometers. In additional Implants, the particles may have an effective average particle size of less than about 400 nanometers, and In sti further embodiments, a size less than about 200 nanometers.

The TKI of the Implant Is preferably from about 10% to 90% by weight of the Implant. More preferably, the TKI Is from about 20%® to about 80% by weight of the implant. In a preferred embodiment, the TKI comprises about 40% by weight of the Implant (e.g., 30%»S0%}, In another embodiment, the TKI comprises about 60%® by weight of the implant.

Suitable polymeric materials or compositions for us® In the Implant include those materials which are compatlbls, that is biocompatible, with the ey® so as to cause no substantial interference with the functioning or physiology of the eye,

Such materials preferably are at least partially and more preferably substantially completely biodegradable or bioerodible.

Examples of useful polymeric materials include, without limitation, such materials derived from and/or Including organic esters and organic ethers, which when degraded result in physiologically acceptable degradation products, Including the monomers. Also, polymeric materials derived from and/or including, anhydrides, amides, orthoesters and the like, by themselves or In combination with other monomers, may also find uss. The polymeric materials may be addition or condensation polymers, advantageously condensation polymess, The polymeric materials may be cross-linked or nan-cross-linked, for example not more than lightly cross-linked, such as less than about 5%, or less than about 1% of the polymeric material being cross-linked. For the most part, besides carbon and hydrogen, the polymers will include at least cne of oxygen and nitrogen, advantageously oxygen. The oxygen may be present as oxy, e.g. hydroxy or ether, carbonyl, e.g. non-oxo-oarbonyl, such as carboxylic acid ester, and the like. The nitrogen may be present as amide, cyan© and amino. The polymers set forth in Halier, Biodegradable Polymers In Controlled Drug Delivery, in: CRC Critioal Reviews in Therapeutic Drug Carrier Systems, Vol, 1, CRC Press, Boca Raton, FL1987, pp 39-90, which describes encapsulation for controlled drug delivery, may find use in the present implants.

Of additional internet are polymers of hydroxyaltphatic carboxylic adds, either homopolymers or copolymers, and polysaccharides. Polyesters of Interest Include polymers of EMactic acid, L-laetic acid, racemic lactic acid, glycolic acid, polyoaprolactone, and combinations thereof, Qenerally, by employing the L-lactate or D-lactate, a slowly eroding polymer or polymeric material is achieved, while erosion Is substantially enhanced with the lactate racemate.

Among the useful polysaccharides are, without limitation, calcium alginate, and function allied celluloses, particularly sariboxymethylcellutos© esters characterized by being water Insoluble, a molecular weight of about S kD to 500 kD, for example,

Other polymers of interest include, without limitation, polyesters, polyelhers and combinations thereof which art biooompatible and may be biodegradable and/or bioerodible.

Some preferred characteristics of the polymers or polymeric materials for use in the present invention may include biocompatibility, compatibility with the therapeutio component, ease of use of the polymer in making the drug delivery systems of the present Invention, a halMIfe In the physiological environment of at least about 6 hours, preferably greater than about on© day, not significantly Increasing the viscosity of the vitreous, and water insolubility.

The biodegradable polymeric materials which are included to form the matrix are desirably subject to enzymatic or hydrolytic Instability. Wafer soluble polymers may be cross-linked with hydrolytic or biodegradable unstable cross» links to provide useful water insoluble polymers. The degree of stability can be varied widely, depending upon the choice of monomer, whether a homopolymer or copolymer is employed, employing mixtures of polymers, and whether the polymer Includes terminal acid groups.

Equally important to controlling the biodegradation of the polymer and hence the extended release profile of the Implant is the relative average molecular weight of the polymeric composition employed in the Implant Different molecular weights of the same or different polymeric compositions may be Included in the implant to modulate the reiease profile. In certain implants, the relative average molecular weight of the polymer will rang© from about S to about 04 kD, usually from about 10 to about 64 kD, and more usually from about 12 to about 45 kD.

In some implants, copolymers of giyoolio acid and lactic add are used, where the rate of biodegradation la controlled by the ratio of glycolic acid to lactic acid. The most rapidly degraded copolymer has roughly equal amounts of glycolic acid and lactic acid. Homopolymer©, or copolymers having ratios other than equal, are more resistant to degradation. The ratio of glycolic acid to ladle acid will also affect the brittleness of the implant, where a more flexible implant Is desirable for larger geometries. The % of polylaofic add in the polylaotio acid polygiycollo aoid (PLOA) copolymer can be 0-100%, preferably about 15-85%, more preferably about 35-85%, In some implants, a 50/50 PLGA copolymer Is used. 5

The biodegradable polymer matrix of the intraocular implant may comprise a mixture of two or more biodegradable polymers. For example, the implant may comprise a mixture of a first biodegradable polymer and a different second biodegradable polymer. One or more of the biodegradable polymers may have o terminal add groups.

Release of a drug from an erodible polymer is the consequence of several mechanisms or combinations of mechanisms. Some of these mechanisms include desorption from the implants surface, dissolution, diffusion through 5 porous channels of the hydrated polymer and erosion. Erosion can be bulk or surface or a combination of both. As discussed herein, the matrix of the intraocular implant may release drug at a rate effective to sustain release of an amount ©? the TKI for more than one week after Implantation into an eye. In certain implants, therapeutic amounts of the TKI am released for more than about ϊ on© month, and even for about six months or more.

One example of the biodegradable intraocular implant comprises a TKI with a biodegradable polymer matrix that comprises a poly (factlde-co-glycoilde) or a poly (OjL-lactlde-co-glycolid©). The implant may have an amount of the TKI s from about 20% to about 80% by weight of the implant Such a mixture is effective In sustaining release of a therapeutically effective amount of the TK! for, a time period from about one month to about six months from the time the implant is placed In an eye.

t Another example of the biodegradable intraocular Implant comprises a TKI with a biodegradable polymer matrix that comprises a single type of polymer. For example, the biodegradable polymer matrix may consist essentially of a polycaprolactone. The poiycaprclactone may have a molecular weight between about 10 and about 20 kifodaltons, such as about 10 kifodaltons, These Implants are capable of providing a nearly linear release rate for at least about 70 days.

The release of the TKi(s) front the intraocular Implant comprising a biodegradable polymer matrix may include art initial burst of release followed by a gradual Increase lit the amount of the TK! released* or the release may Include an initial delay in release of the TK! followed by an Increase In release. When the implant Is substantially completely degraded, the percent of the TKI(s) that has been released Is about one hundred. Compared to existing Implants, the Implants disclosed herein do not oompietely release, or release about 100% of the TKIfs), until after about one week of being placed in an eye.

It may be desirable to provide a relatively constant rata of release of tbe TKI(s) from the Implant over the life of the implant, For example, it may be desirable for the TK!(s) to be released in amounts from about 0.01 pg to about 2 per day for the life of the implant. However, the release rate may change to either increase or decrease depending on the formulation of the biodegradable polymer matrix, In addition, the release profile of the TKI(s) may Include one or more linear portions and/or one or more non-linear portions, Preferably, the release rate Is greater than zero once the implant has begun to degrade or erode.

The implants may be monolithic, t®, having the active agent or agents homogenously distributed through the polymeric matrix, or encapsulated, where a reservoir of active agent is encapsulated by the polymeric matrix. Due to ease of manufacture, monolithic Implants are usually preferred over encapsulated forms. However, the greater control afforded by the encapsulated, reservoir-type Implant may be ©f benefit In some circumstances, where ths therapeutic level of the drug fails within a narrow window, in addition, the therapeutic component, including the TKi(s), may be distributed in a non-homogenous pattern In the matrix. For example, the implant may include a portion that has a greater concentration of the TKI(s) relative to a second portion of the implant.

On® such implant 100 is illustrated in FIG. 14. The implant 100 may b® understood to be a unidirectional drug delivery device. The implant 100 is characterized by comprising a first portion 110 and a second portion 120. First portion 110 comprises a mixture of a therapeutic agent, such as TKI, and a biodegradable polymer matrix, such as a matrix of RIGA, PLA, or a combination thereof. Second portion 120 oomprisas a polymer, such as a biodegradable polymer, and la substantially free of the therapeutic agent The polymeric component of the first portion 110 and the second portion 120 may comprise the same polymer material, e.g., both components may be made from a PLGA polymer, Although the therapeutic agent Is a TK!S other implants may include other therapeutic agents, including those described herein, First portion 110 may be understood to be an active layer, and second portion 120 may be understood to ba a barrier layer, which Is effective to prevent or reduce diffusion of the therapeutic agent from one side of the Implant The layers may be separately formed as films and pressed together using a Carver press, for example. Or the layers may be eo*extruded using conventional extrusion techniques or injection molded using injection molding techniques. The implant 110 Is effective to control the flow or release of a therapeutic agent In a specific direction, such as one direction. The implant can be applied to a diseased location, such as in an eye, that needs the release of the therapeutio agent in a specific and controlled manner, such as tor subconjunctival applications.

The present Implants may also comprise a combination of a TKI and polyoaprolactone, as described herein. Such implants may provide a single order release rate for about 70 days or more after placement In an eye. The polycaproiaetone may have a molecular weight of about 15 kilodaftons. Thus, one embodiment of the present implants, comprises a poorly soluble drug or therapeutic agent and a single polymeric component that releases the drug at a substantially linear release rate (e.g,, a zero order rats).

The present implants may also include a nan-biodegradable polymer component, as described herein. The release of a therapeutic agent, such as TKI, may be achieved by movement of th© therapeutic agent through one or more openings, orifices, ©r holes. An example of such an implant is disclosed In U.S'. Pat No. 6,331,313.

Ttie intraocular Implants disclosed herein may have a size of between about 6 pm and about 2 mm, or between about 10 pm and about 1 mm for administration with a needle, greater than 1 mm, or greater than 2 mm, such as 3 mm or up to 10 mm, for administration by surgical implantation, The vitreous ’ chamber In humans Is able to accommodate relatively large implants of varying geometries, having lengths of, for example, 1 to 10 mm. The implant may be a cylindrical peiief (e, g„ rod) wits dimensions of about 2 mm x 0,75 mm diameter. Or the implant may be a cylindrical pellet with a length of about 7 mm to about 10 mm, and a diameter of about 0.75 mm to about 1.5 mm.

The implants may also be at least somewhat flexible ao as to facilitate both Insertion of the implant in the eye, such as in the vitreous, and accommodation of - the Implant. The total weight of the implant is usually about 260-5000 more preferably about 500-1000/rg. For example, an implant may be about 500 pgs or about 1000 pg. For non-human individuals, the dimensions and fetal weight of the implants) may he larger or smaller, depending on the type of individual. For example, humans have a vitreous volume of approximately 3,8 ml, compared with approximately 30 ml for horses, and approximately 50-100 ml tor elephants. An Implant steed for use in a human may be scaled up or down accordingly for other animals, tor example, about 8 times larger for an implant for a horse, or about, for example, 26 times larger for an Implant for an elephant

Thus, implants can be prepared where the center may be of one material and the surface may have one or more layers of the same or a different composition, where the layers may be cross-linked, or of a different molecular weight, different density or porosity, or the tike. For example, where it is desirable to quickly release an Initial bolus of drug, the center may be a polylaotate coated with a polylacfate-polyglycolate copolymer, sc as to enhance the rate of Initial degradation. Alternatively, the center may be polyvinyl alcohol coated with polylaotate, so that upon degradation of the polylactate exterior the center would dissolve and be rapidly washed out of the eye.

The implants may be of any geometry including fibers, sheets, films, microsphares, spheres, circular discs, plaques and the like. The upper limit for the Implant size will be determined by factors such as toleration for the implant, size limitations m Insertion, ©as® of handling, etc. Where sheets or films are employed, the sheets or films will be In the rang© of at least about 0.5 mm x OJ mm, usually about 3-10 mm x 5-10 mm with a thickness of about 0.1-1.0 mm for ease of handling. Where fibers art employed, the fiber diameter will generally be In the range of about 0,05 to 3 mm and the fiber length will generally be In the range of about 0.5-10 mm. Spheres may be in the range of about 0.5 pm to 4 mm in diameter, with comparable volumes for other shaped particles.

The size and form of the Implant oan also be used to control the rate of release, period of treatment, and drug concentration at the ate of implantation, Larger implants will deliver a proportionately larger dose, but depending on the surface to mass ratio, may have a slower release rate. The particular size and geometry cl the Implant are chosen to suit the site of implantation.

The proportions of TXI{s), polymer, and any other modifiers may be empirically determined by formulating several implants with varying proportions. A USP approved method for dissolution or reiaasa test can be used to measure the rate of release (USP 23; IMF 18 {1995} pp. 1790-1798). For example, using the infinite sink method, a weighed sample of the Implant is added to a measured volume el a solution containing 0,9% NaCI in water, when the solution volume will be suoh that the drug concentration is after release is less than S% of saturation. The mixture is maintained at 37®C and stirred slowly to maintain the Implants in suspension. The appearance of the dissolved drug as a function of time may be followed by various methods known In the art, such as speotrophctometrloally, HPLC, mass spectroscopy, ©to. until the absorbance becomes constant or unfit greater than 90% of the drug has been released.

In addition to the TKI(s) included In the intraocular implants disclosed herein, the intraocular implants may also include one or more additional cphthalmloally acceptable therapeutic agents. For example, the implant may include one or more antihistamines, one or more antibiotics, one or more beta blockers, one or more steroids, one or more antineoplaetlc agents, on© or more immunosuppressive agents, on© or more antiviral agents, on© or more antioxidant agents, and mixture© thereof.

Pharmacologic or therapeutic agents which may find use In the present systems, include, without limitation, those disclosed In U.S. Pat, Nos. 4,474,451, columns 4-6 and 4,327,725, columns 7-3.

Examples of antihistamine© Include, and are not limited to, loradatlne, hydroxyzine, diphenhydramine, chlorpheniramine, brompheniramine, cyproheptadine, terfenadlne, clemastine, triproildine, carblnoxamlne, dipbenylpyrallne, phenlndamine, azatadine, trlpelennamine, dexohlorpheniramine, dexbrompheniramsne, methdiazine, and trimprazlne doxylamine, pheniramlne, pyriiamlne, ehlorcyclizlne, thonzyiamin®, and derivatives thereof.

Examples of antibiotics include without limitation, eefazclin, csphradine, eefador, cephapirin, cefiizoxime, ©efoperazona, cefotetan, oefufoxime, cefotaxime, eafadroxi, ceftazidime, cephalexin, caphaiofhin,, cefamandole, cefoxitin, cefonicid, caforanide, ceftriaxone, cefadroxil, cephradine, cefurexime, cyclosporine, ampioillin, amoxicillin, cyclacillin, amplciln, penicillin <3, penicillin V potassium, piperacillin, oxacillin, bacampiclin, cioxaciilin, ticamlin, azlociliin, carbentelliin, methisiin, nafoiliin, erythromycin, tetracycline, doxycyetine, minocycline, azfreonam, chloramphenicol, ciprofloxacin hydrochloride, clindamycin, metronidazole, gentamicin, lincomydn, tobramycin, vancomycin, polymyxin B sulfate, colistimethate, colistin, azithromycin, augmentin, sulfamethoxazole, trimethoprim, gatiftoxacln, ofloxacin, and derivatives thereof.

Examples of beta blockers Include aoebutoloi, atenolol, labataloi, metoprolol, propranolol, timolol, and derivatives thereof. i

Examples of steroids include corticosteroids, such as cortisone, prednisolone, fiurometholone, dexamethasone, medrysone, loteprednol, ffuazacost, hydrocortisone, prednisone, betamethasone, prednisone, methylprednleoione, riamcinoione hexacatonide, paramethasone acetate, difiorasone, fluocinonide, ffuQcinotene, triamcinolone, derivatives thereof, and mixtures thereof.

Examples of antineoplastio agents include adriamycin, cyclophosphamide, actinomycin, bleomycin, duanorubicin, doxorubicin, epirubicin, mitomycin, methotrexate, fluorouracll, carbopiatin, carmustlne (BCMU), methyi-CCNU, cispiatln, etoposide, interferons, camptotheoin and derivatives thereof, phenesterina, taxol and derivatives thereof, faxotere and derivatives thereof, vinblastine, vincristine, tamoxifen, etoposide, piposulfan, cyclophosphamide, and flufamide, and derivatives thereof.

Examples of Immunosuppressive agents include cyclosporine, azathloprine, tacrolimus, and derivatives thereof.

Examples of antiviral agents include Interferon gamma, zidovudine, amantadine hydrochloride, ribavirin, acyclovir, vaiciclovfr, dideoxycytldine, phosphonoformlc acid, ganciclovir and derivatives thereof.

Examples of antioxidant agents Include ascorbate, alpha-tocopherol, mannitol, reduced glutathione, various carotenoids, cysteine, uric acid, taurine, tyrosine, superoxsd® dismufase, lutein, zeaxanthln, cryotpxanthin, astazanthin, lycopene, hl-acetyl-cysteine, sarncsine, garnma-glutamyicysteirte, quercitin, laotcferrin, dshydrolipoic acid, citrate, Ginkgo Blioba extract, tea cateohlns, bilberry extract, vitamins E or esters of vitamin i, retinyl palmltate, and derivatives thereof.

Other therapeutic agents includs squalamine, carbonio anhydmse inhibitors, alpha agonists, prostamides, prostaglandins, antiparasitics, anffungals, and derivatives thereof.

The amount of active agent or agents employed in the Implant, Individually or In combination, will vary widely depending on the affective dosage required and the desired rate of release from the implant As indicated hareln, the agent will be at least about 1, more usually at least about 10 weight percent of the implant, and usually not more than about 60, more usually not more than about 40 weight percent of the Implant.

In addition to the therapeutic component, the intraocular implants disclosed herein may Include effective amounts of buffering agents, preservatives and th© Ilk©. Suitable water soluble buffering agents Include, without limitation, alfcaii and alkaline earth carbonates, phosphates, bicarbonate®, citrates, borates, acetates, succinates and the like, such as sodium phosphate, citrate, borate, acetate, bicarbonate, carbonate and the like. These agents advantageously present in amounts sufficient to maintain a pH of th© system of between about 2 to about 9 and more preferably about 4 to about 8. As such the buffering agent may be as much as about 5% by weight of the total implant. Suitable water soluble preservatives Include sodium bisulfite, sodium bisuifate, sodium thiosulfate, ascorbate, benzaikonlum chloride, ohlorobutanol, thimarosal, phenylmercuric acetate, phenylnrereuric borate, phenylmercuric nitrate, parabens, methylparaben, polyvinyl alcohol, benzyl alcohol, phenylethanol and the like and mixtures thereof. These agents may be present in amounts of from 0.001 to about 5% by weight and preferably 0.01 to about 2% by weight.

In addition, the implants may include a solubility enhancing component provided in an amount effective to enhance the solubility of the TKI{s) relative to substantially identical implants without the solubility enhancing component. For example, an Implant may Include a β-oyciodextrin, which is effective in enhancing th© solubility of the TKi, The p-cyclodextrfn may be provided In an amount from about 0.5% (w/w) to about 25% (w/w) of the Implant, In certain implants, the β~ oyclodextrin Is provided in an amount from about S% (w/w) to about 15% (w/w) of the implant in some situations mixtures of implants may b© utilized employing the same or different pharmacological agents. In this way, a cocktail of release profiles, giving a biphaslc or triphasic release with a single administration is achieved, where the pattern of release may be greatly varied. Th© implant© may also have a sigmoidal release profile.

Additionally, release modulators such as those described in U, S. Patent No, 05869,079 may be included In the implants. The amount of release modulator employed will be dependent on the desired release profile, the activity of the modulator, and on tie release profile of the TKf In the absence of i modulator. Electrolytes such as sodium chloride and potassium chloride may also be included in the Implant. Where the buffering agent or enhancer is hydrophilic, It may also act as a release accelerator. Hydrephie additives act to increase the release rates through faster dissolution of the materlai surrounding the drug particles, which increases the surface area of the drug exposed, thereby s increasing the rate of drug bloerosien. Similarly, a hydrophobic buffering agent or enhancer dissolve more slowly, slowing the exposure of drug particles, and thereby slowing the rate of drug bioerosion.

Various techniques may be employed to produce the implants described herein. Useful techniques Include, but are not necessarily limited to, solvent evaporation methods, phase separation methods, fnterfadal methods, molding methods, injection molding methods, extrusion methods, co-extrusion methods, carver pres® method, die cutting methods, heat compression, combinations thereof and the liks.

Specific methods are discussed In U.S. Pat, No. 4,997,652, Extrusion methods may be used to avoid the need for solvents to manufacturing. When using extrusion methods, the polymer and drug are chosen a© as to be stable at the temperatures required for manufacturing, usually at least about 85 degrees Celsius. Extrusion methods use temperatures of about 25 degrees C to about 150 degrees C, more preferably about 65 degrees C to about 180 degrees C. An Implant may ba produced by bringing the temperature to about 80 degrees C to about 150 degrees O for drug/pofymer mixing, such as about 130 degrees C, for a time period of about 0 to 1 hour, 0 to 80 minutes, or 5-15 minutes. For example, a time period may b© about 10 minutes, preferably about 0 to 5 min.

The implant® are then extruded at a temperature of about 60 degrees C to about 130 degrees C, such as about 75 degrees C,

In addition,, the implant may be ceextruded so that a coating is formed me? >a core region during the manufacture of the implant.

Compression methods may be used to make the implants, and typically yield Implants with faster release rates than extrusion methods. Compression methods may use pressures of about 50-1 SO psi, more preferably about 70-80 psl, even more preferably about 76 psi, aid us© temperatures of about 0 degrees C to about 115 degrees C, more preferably about 25 degrees C.

The implants of the present invention may be inserted Into the eye, for example the vitreous chamber of tie eye, by a variety of methods, including placement by forceps or by trooar following making a 2-3 mm Irtslsion In the solera, One example of a device that may be used to Insert the implants into an eye is disclosed in U.S. Patent Publication Mo. 2004/0054374, The method of placement may Influence the therapeutic component or drug release kfnetlos,

For example, delivering the implant with a trooar may result in placement of the implant deeper within the vitreous than placement by forceps, which may result In the implant being closer to the edge of the vitreous. The location of the Implant may influence the concentration gradients of therapeutic component or drug surrounding the element, and thus influence the release rates fag», an element placed closer to the edge of the vitreous may result In a slower release rate},

The present implants are configured to release an amount of the TKI|s) effective to treat or reduce a symptom of an ocular condition, such as a posterior ocular condition.

The Implants disclosed herein may also be configured to release the TK! or additional therapeutic agents, as described above, which to prevent diseases or conditions, such as the following: MACULOPATHIES/RETIMAL DEGENERATION; Mon-Exudative Age Related Macular Degeneration (ARMD), Exudative Age Related Macular Degeneration (ARMD), Choroidal Neovascularization, Diabetic Retinopathy,

Acute Macular Neuroretinopathy, Central Serous Chorioretinopathy, Cystold Macular Edema, Diabetic Macular Edema, UVEITIS/RETINITIS/CHOROIDITIS: Acute Multifocal Ptacoid Pigment Epitheliopathy, Behcet's Disease, Blrdshe! Retsnoehoroidopathy, Infectious (Syphilis, Lyme, Tuberculosis, Toxoplasmosis), Intermediate Uveitis (Pars Planiiis), Multifocal Choroiditis, Multiple Evanescent White Dot Syndrome (MEWDS), Ocular Sarcoidosis, Posterior Sciarftis, Serpignous Choroiditis, Subretinal Fibrosis and Uveitis Syndrome, Vogt-Koyanagl-Harada Syndrome. VASCULAR DISEASES/EXUDAT1VE DISEASES: Goafs Disease, Parafoveal Telangiectasia, Papillophlebitls, Frosted Branch Angitis, Sickle Cell Retinopathy and other Hemoglobinopathies, Angioid Streaks, Familial Exudative Vitreoretlnopathy. TRAUMATIC/SURGIGAL: Sympathetic Ophthalmia, Uveitic Retinal Disease, Retinal Detachment, Trauma, Laser, POT, Photocoagulation, Hypoperfusion During Surgery, Radiation Retinopathy, Bone Marrow Transplant Retinopathy. PROLIFERATIVE DISORDERS: Proliferative Vitreal Retinopathy and Epiretinal Membranes, Proliferative Diabetic Retinopathy, Retinopathy of Prematurity (reirolenta! fibroplastic). INFECTIOUS DISORDERS: Ocular Histoplasmosis, Ocular Toxocariasis, Presumed Ocular Histoplasmosis Syndrome (PGHS), Endophthalmitis, Toxoplasmosis, Retinal Diseases Associated with HIV infection, Choroidal Disease Associated with HIV Infection, Uveitic Disease Associated with HIV Infection, Viral Retinitis, Acute Retinal Necrosis, Progressive Outer Retinal Necrosis, Fungal Retinal Diseases, Ocular Syphilis, Ocular Tuberculosis, Diffuse Unilateral Subacute Neororetinffis, Myiasis. GENETIC DISORDERS: Systemic Disorders with Associated Retinal Dystrophies, Congenital Stationary Night Blindness, Cone Dystrophies, Fundus

Fiavimacuiatus, Best's Disease, Pattern Dystrophy of the Retina! Pigmented Epithelium, X-Linkad Retinosohfsis, Sorsby’s Fundus Dystrophy, Benign Concentric Maoutepathy, Bietti’s Crystalline Dystrophy, pseudoxanthoma elastieum, Osier Weber syndrome. RETINAL TEAR8/11GLES; Retinal Detachment, Macuiar Hole, Giant ‘ i

Retinal Tear. TUMORS: Retinal Disease Associated with Tumors, Solid Tumors, Tumor Metastasis, Benign Tumors, for example, hemangiomas, neurofibromas, trachomas, and pyogenie granulomas, Congenital Hypertrophy of the RPE, Posterior Uveal Melanoma, Choroidal Hemangioma, Choroidal Osteoma, Choroidal Metastasis, Combined Hamartoma of the Retina and Retinal Pigmented Epithelium, Retinoblastoma, Vasoproliferatfve Tumors of the Ocular Fundus, Retina! Astrocytoma, Intraocular Lymphoid Tumors. MISCELLANEOUS: Punctate Inner Choroidopathy, Acute Posterior Multifocal Piaooid Pigment Epfthellopathy, Myopic Retinal Degeneration, Acute Retinal Pigment Epitheiitis, Ocular Inflammatory and immune disorders, ocular vascular malfunctions, Comeal Graft Rejection, Neovascular Glaucoma and the like. in one embodiment, an Implant, such as the Implants disclosed herein, Is administered to a posterior segment of an eye of a human or animal patient, and preferably, a living human or animal. In at least one embodiment, an Implant is administered without accessing the subretlnal space of the eye. For example, a method ©f treating a patient may Include placing the implant directly Into the posterior chamber of the eye. in other embodiments, a method of treating a patient may comprise administering an implant to the patient by at least one of intravitmai injection, subconjuotival injection, sub-tenon injections, retrobulbar injection, and supraehoroidal injection.

In at least one embodiment, a method of improving vision or maintaining vision In a patient comprises administering one or more implants containing one or more TKIs, as disclosed herein to a patient by at least one of intravltreal injection, subconjuefivaf injection, sub-tenon injection, retrobulbar Injection, and suprachoroldaf infection. A syringe apparatus Including an appropriately steed needle, for example, a 22 gauge needle, a 27 gauge needle or a 30 gauge needle, can be effectively used to inject the composition with the posterior segment of an ®ye of a human or animal. Repeat injections are often not necessary due to the extended release of the TKI from the Implants.

In another aspect of the invention, kits for treating an ocular condition cf the eye are provided, comprising: a) a container comprising an extended release implant comprising a therapeutic component Including a TKI, and a drug release sustaining component; and b) instructions for use. Instructions may include steps of how to handle the implants, how to insert the implants info an ocular region, and what to expect from using the implants,

Example 1 infravitraal Pharmacokinetics of TKIs In Fluid Compositions

The ocular pharmacokinetics of AGN 199689, AGN £00954, AGN 201088 and AGN 201666 following single intravltreal injections into female albino rabbit eyas was determined. The animals were dosed with a 50 pL Intravltreal injection of 242 ng AGN 201088,128 ng AGN 201866,114 ng AGN 199659 or 222 ng of AGN 200954 per eye. Vitreous humor samples (n ~ 4 eyes per timepoint) were oollaoted at 0.5,1,2,4S 8, and 12 hr postdosa. The TKI concentration In the vitreous humor was determined using a liquid chromatography tandem mass spectrometry method (LG-MS/MS).

All compounds were eliminated fairly rapidly from the rabbit eye. This indicates a transretinal route of elimination. Thera was no bias to compound nucleus. However, even though elimination was extremely rapid it was determined that local sustained delivery was feasible. Based on the vitreal clearance determined In this study for 3"[C4”Morphofin«4»yi-plienyIamino)-mefhyleneJ-198-dihydro«sndoi*2"one, S-Ce-Amsno^H-isobeniofuran-l -ylldenej-S-οΝαΓθ-1,3-0%Ρίο~ΙηΰοΙ-2-οπ®, AGN 201088 and AGN 201666, and assuming steady state efficacious concentration at twice the EG50 values (determined by In vitro receptor binding and Intracellular C@2+ assay) all the tyrosine kinase inhibitors tasted could be formulated Into 1 mg Implants that would maintain the desired steady state drug vltreai concentrations for a duration of about six months. This data is summarfeed in Table 1 and Figures 1 and 2.

Tbbl® 1. TKI Pharmacokinetic Parameters after a Single Intravitreai Injection

Example Z • TKI Biodegradable Implants

Tyrosine kinase Inhibitors were Incorporated into PISA or PLA implants by extrusion. The TKIs wars milled with the polymers at certain ratios then extruded Into filaments. These filaments were subsequently cut Into Implants weighing approximately 1 mg. Several TKIs were formulated in the PISA and PLA implants based on their potencies and physicochemical properties as shown In Table 2.

Table 2, Tyrosine Kinase Inhibitors Formulated in PLGA Implants

TKI release from the implants was assessed In vitro, implants were Placed Into vials containing release medium and shaken at 37 SC. At the appropriate time points a sample was taken from the release medium for analysis and the medium totally replaced to maintain sink conditions. Drug In the sample was assayed by HPLC and the cumulative percent release of drug from the Implant noted as a function of time. The in vitro release prates of AQN 200954, AGN 202314* ΑΘΝ 201635 and AON 202564 are depicted in Figures 3 through 10, respectively.

From the formulation release data depicted in Figures 3 through 10 It is evident that TKIs over a wide range of physicochemical properties ran be engineered to release drug in vitro over a period of weeks to a year.

Encamp!© 3 in Vivo Pharmacokinetic Properties of TKi-Contalning Implants

Implants containing AGN 202314 war® placed intravitrealiy or suboonjunctivally in an ©ye. The implants released AON 202314 in-vitro over a 14 day period (Figure 3.). The Intent of this study was to achieve an Intravttreai in-vivo/ In-vitro correlation with the Intravitreal implants and assess the feasibility of periocular delivery. intravitreal Implants, PLGA (400 μg AGN 202314 dose, 1 mg total implant weight), were Implanted by surgical incision into the mid vitreous of albino rabbits. At days 8,15,31 and ©1 rabbits were sacrificed and the vitreous humor, lens, aqueous humor and plasma assayed for AGN 202314.

Subconjunctival Implants, RIGA (1200 pg AGN 202314 dose; three implants) and PLA microspheres (300 pg AGN 202314) were Implanted subconjunctlvally. At days 8,15,31 and 61 rabbits were sacrificed and the vitreous humor, lens, aqueous humor and plasma assayed for AGN 202314.

The data are summarized In Tables 3 through 5

Table 3. PK Results from 2 Month Intravitreal Implantation AGN 202314 implant (400 pg rod)

Table 4, PK Results from 2 Mo Subconjunctival Implantation of AON 202314 Microspheres

BLQ = Retina {<5 ng/g), VH («30 ng/g), tens f<5 rsg/g), AH (<Q.S ng/mt), plasma («0-5 ng/mL)

Table 5. Month Subconjunctival Implantation (3 rods with a total of 1,2 mg AGN 202314)

The data from this study indicates that a good In vitro In vivo correlation was established for AGN 202314. The AGN 202314 implant released drug over a two week pelted both in vitro and In vivo. It Is also important that plasma levels remain BLQ or extremely low for all time points. This shows that even in a worst case scenario if Intravitreal delivery over two waste systemic exposure la negligible, it was also noted that periocular delivery was unsuccessful at delivering ASM 202314 to the vitreous and retina A foilow-on two-month ocular pharmacokinetic study of AGN 202814 following a single Intravitreal implantation Into albino rabbit eyes was initiated.

The formulations delivered AGN 202314 over a period olfour months in-vitro.

The following 1 mg implants were evaluated; 30% AGN 202314/ 70% Purac PLA; Lot# JS493028 (FIG. 4), 50% AGN 201634/50% Purac PLA; Lot # JS493034 (RG 6.). Two rabbits (4 eyes and 2 plasma) were used per timepoint. Implants were administered by a bilateral surgical intravitreal placement by sclerotomy without vitrectomy. The vitreous humor and retina AGN 202314 concentrations were assayed at dap 8» 15,31 and 61. The data are displayed in Table 6.

Table 6. One Month Date from the AGN 202314 intravitreal Study

The retinal levels achieved from this study approach therapeutic levels by the first week and are maintained over the first thirty days. This date shows that actual in vivo sustained delivery of a TKI locally Is feasible, A six month pharmacokinetic study was Initiated with Intravitreal and subconjunctival AGN 200954 implants. The implants released AGN 200954 In-vitro over a 180 day period {Figure 3), Intravitreal Implants, PLGA (500 pg AGN 200954 dose, 1 mg total Implant weight, Purac polymer) and PLGA {500 pg AGN 200954 dose, 1 mg total Implant weight, RG503H polymer), were implanted by surgteal Incision Into the mid vitreous of albino rabbits. At days 8, IS, 81 and 61 rabbits war© sacrificed and the vitreous humor, lens, aqueous humor and plasma assayed for AQN 200954 Subconjunctival implants, PLGA Implant {500 pg AGM 200954 dose, 1 mg total implant weight, Purae polymer) and PLGA mferospheres (370 pg and 740 pg AGM 200954), were administered. At days 8,15,31 and 61 rabbits were sacrificed and the vitreous humor, lens, aqueous humor and plasma assayed for ASH 200954, AGN 2009S4 Pharmacokinetics after Intravitreal Administration

BLQ “ Retina (5 ngfa), ¥H (30 ng/g), tens (5 ng/g), AH (0.05 ng/mL), plasma (0.0S ng/mL) it is evident from the data that a considerable in-vivo fag time exists for the first formulation not seen In vitro. Neither formulation exhibits measurable plasma concentrations.

Example 4

In vitro release of a TKI (AON 201634) from an implant TKI release was examined for Implants made from poly {D,L-iaGtide-QQ~ glyoeilde) (PDLG) or poly {D.L-laofide) (PDL) In different media with or without addition of detergent at 37¾ in a shaking water bath. AGN 201634 was obtained from Allergan, and Its chemical structure Is shown below. It was used as received without further purification. PDtG/PDL polymer materials were obtained from Purac Amerloa Ino.

TKI release was examined in various medium, including saline, phosphate buffer saline of pH 7.4, SOmM bicarbonate buffer of pH 6.0 ± 0.1 with 0.1 % cetyltrimethylammonfurn bromide (CTAB), and 50my borate buffer of pH 8.S ± 0.1 with 0.6% sodium dodecyl sulfate |3DS) in a shaking water bath {Precision} at 37¾. Sample was incubated In 10 mL of medium, and was totally replaced with fresh medium at each sampling time. Drug concentration was determined by HPLC using a Waters 2690 Separation Module equipped with a Waters XTerra RP8 column (3.9x150 mm, 5pm, equilibrated at ambient) and a Wafers 896 photodiode array detector (set at 238 nm) using 0.1% acetic acid In acetonitrile/water (40/60 by volume} as the mobile phase under a flow rate of 1.2 ml/mln. The column was equilibrated with mobile phase for at least 30 min before Initiating any sample injection.

Th@ characteristics of formulations, including formulation Identification, Lot number, drug loading, inherent viscosity of polymer, and extrusion temperature are summarized In the following table, Tha drug load Is from 20 to 50%. The formulations were extruded from a 750 μπι nozzle to form cylindrical DOS,

Table a Characteristics of TKI formulations.

The stability of AGN 201834 standard solution in deionized watsr/aoetonftrile (75%/25%) was examined at 4®C, and the results are summarized In tha following table. The concentration of standard solution was from 0,0695 jugfonL to 8,683 /rg/ml, and was analyzed on day 14, 21, and 35. The results show that the recovery was all greater than 95%, indicating a good stability of AGN 201634 in .- deionized water/acetonitrite (75%/25%) at 4°C for up to 35 days even the concentration was up to 8.7 ^rg/mL

Stability of AGN 201634 standard solution of various concentrations In D! water/aoetonftrite (75%/25%) at 4 X (Table 8).

Stability of AQN 201634 standard solution of various eonearstratfons In Dl water/asetonitrile (7S%/26%) at 4 ®C (Table 9),

To examine the stability ©f TKI in formulation, Formulations 3 and 4 ware prepared to various concentration in a medium of pH 8,0* 7.4 or 8,S, respeotively, and subjected to an Incubation condition of either 7 days under ambient condition i or 14 days at 4°C, and the results are summarized in the following table. Hie results show that th© recovery was all better than 98%, indicating that AGH 201634 was stable in media of pH 6.0,7,4 and 8,5, and lasted for 7 days In ambient or 14 days at 4aG, i Table 10: Stability ©f TKI in Formulations 3 and 4 In madia under various incubation conditions.

TKI releases of Formulation 11n 20 mL of saline or 20-30 mL of PBS are demonstrated In Figure 11, The DOS was incubated In a vial of either 40 or 20 ml, and 10 ml sample solution was replaced by §arn© volume of fresh medium, respectively, at each sampling time, The release profiles in saline and PBS were obviously different. Less than 5% of TKI was released in saline during the first 70 days. In contrast, loss than 5% of AQN 201034 was released at the first 3 weeks when DOS was Incubated in PBS, the same as In saline, but more than 80% of AGN 201034 was released after TO days. However, no significantly difference In release profile was found when DOS was incubated In 20 or-30 ml of PBS in a 20 or 40 mL vial It seems that release medium plans an important role In the variation of release profile Instead of incubation volume. Due to this slow and diverged release profile, the release profile of Formulation 2 was not performed since it© formulation was based on the same polymer with a lower drug loading, TKI releases of Formulations 3 and 4 in 10 mL media of a pH of 8,0 (with 0.1% CTAB}, 7.4 (PBS) or 8,5 (with 0.5% SDS} at 37¾ are demonstrated in Figures 12 and 13, respectively. For F3, more than 50%, 45%, and 75% TKI was released at the first 3S 7, and 2 weeks, when DOS was Incubated in a medium of pH 6.0 (with 0.1% CTAB), 7.4 (PBS) and 8.5 (with 0.5% SDS), respectively. On the other hand, approximately 47%, 6%, and 68% of TKI was released from F4 when DOS was incubated in media as described above. It seems that TKI release in different pH medium Is pH 8.5 > pH 6.0 > pH 7.4, with or without the assistant from detergent in the medium. No large standard deviations are found In all media for both formulations.

To monitor the appearance of DOS during dissolution, the images of F3 and F4 formulations incubated in 10 mL media of a pH of 8.0 (with 0,1% CTAB), 7.4 (PBS) or 8.5 (with 0.6% SDS) at 37¾ ware. All formulations experienced swelling followed by matrix degradation, resulting in drug release. No complete disintegration of formulation matrix was observed within 153 days at 37 ®C,

In summary, tyrosine kinase inhibitor (AGH 201834) DDS were formulated using various RIGA ©r PLA at various drug loading. The stability of AGN 201834 solution in Dl wafer/acetonitrile (75%/25%) at 4°C was more than 35 days, and DDS solution in various pH medium was more than 7 days under ambient condition or 14 days at 4°C. Different drug release profiles were found when DDS was tested in PBS or saline. Drug buret effect was found only in Formulation 3 when incubating In a medium of pH 6.0. Controlled AGN 201834 release in vitro was more than 4 weeks in a medium of pH 8.5, and more than 5 months in media of pH 7.4 and pH 8.0.

Example 5

Biodegradable implants with a linear release profile

Biodegradable implants are made by combining a TKi with a biodegradable polymer composition in a stainless steel mortar. The biodegradable polymer composition comprises a single type of biodegradable polymer, The combination is mixed via a Turbula shaker set at 98 RPM for 15 minute®. The powder blend is scraped elf the wall of the mortar and then remixed for an additional 15 minutes. The mixed powder blend is heated to a seml-molten state at specified temperature for a total of 30 minutes, forming a poiymer/drug melt.

Rods are manufactured by pelletizing the polymer/drug melt using a 9 gauge polytetralluoroefhylene (PTFE) tubing, loading the pellet Into the barrel and extruding the material at the specified core extrusion temperature Into filaments. The filaments are then out into about 1 mg siae implant® or drug delivery systems. The rods have dimensions of about 2 mm long x 0,72 mm diameter. The rod implants weigh between about 900 pg and 1100 pg,

Wafers are formed by flattening the polymer melt with a Carver pres® at a specified temperature and cutting the flattened material into wafers, each weighing about 1 mg, The wafers have a diameter of about 2,5 mm and a thickness of about 0.13 mm. The wafer implants weigh between about 800 pg and 1100 pg. in-vitro release testing can be performed on each tot of Implant (rod or wafer). Each Implant may be placed into a 24 mL screw cap vial with 10 ml of Phosphate Buffered Saline solution at 37¾ and 1 mL aliquots are removed and replaced with equal volume of fresh medium on day 1,4,7» 14,28, and every two weeks thereafter. .

Drug assays may be performed by HPLC, which consists of a Waters 2590 Separation Module (or 2698), and a Waters 2998 Photodiode Array Detector. An Uitrasphere, C-18 (2), 5 pm; 4,6 x 156 mm column heated at 80 s C can be used for separation and the detector can be set at 264 nm. The mobile phase can be (10:90) MeOH - buffered mobile phase with a flow rate of t mL/min and a total run time of 12 min per sample. The buffered mobile phase may comprise (08:0.75:0,25:31) 13 mM 1-Heptane Sulfonic Acid, sodium salt - glacial acetio acid - trfethylamine - Methanol, The release rates cars be determined by calculating the amount of drug being released In a given volume of medium over time In pg/day,

The single polymer chosen for the implant was poly(caproiactone). Rod and wafer Implants were formulated at a ratio of 50:50 (poly(caprolactone):TKi). Thus, a 1 mg implant comprises about 500 pg poiy{capro!aefene) and SG0 /jg TKL AON 200954 was used as the TKL

As shown In FIG, 15, Implants formed from a poorly soluble drug (TKI) and a single type of a biodegradable polymer (pcly(caprolactcne)) released TKI at nearly zero-order rate for at least about 70 days. The particular poly(caproiactone) had a molecular weight erf about 15 kitodaltons. The nearly linear release rate is extremely hard to achieve with other biodegradable implants based on a single polymeric component, as shewn for the dexamathasone containing implants in FIG, 15,

Example 6

Manufacture and testing of Implants containing an TKI and a biodegradable polymer matrix

Additional biodegradable Implants art read© by combining a TKI with a biodegradable polymer composition as described In Example 5. The polymers chosen for the implants oan be obtatned from Boehrfngtr Ingelheim or Purac America, for example. Examples of polymers include; RG502, RC3752, R2Q2H, R2G3 and R208, and Purac PD LG (50/50), RGSG2 is (50:50) poIy(D,L-!aetid®-cO” giycollde), RG752 is (75:25) poIyCD^Iaetldt-co-glyoolida), R202H is 100% poiy(D, L-lactide) with acid end group or terminal acid groups, R203 and R208 ar© both 100% po!y(D, L-Iactide). Purac PDLG (50/50) is (50:50) pofy(D,L-lactide-co-glyeollde), The Inherent viscosity of RG5Q2, RG752, R202H, R203, R206, and Purac PDLG are 0.2,0.2, 0.2,0.3,1.0, and 0.2 dl/g, respectively.

The average molecular weight of RG502, RG752, R2Q2H. R203, R208, and Purac PDLG are, 11700,11200,6500,14000,83300, and 9700 daltons, respectively.

Example 7 inyitroM ’

Summary TKls can inhibit the Intrinsic tyrosine kinase activity necessary for activation of vascular endothelial growth factor receptors (VEGFR). ¥EGF and VEGF signaling pathways can induce angiogenesis and increase vascular permeability, activities required for neovascularization. Thus, TKls oan have utility to prevent or to treat choroidal neovascularization (CNV), auoh as CN¥ that can result from or be a symptom of, for example, age-related macular degeneration (AMD) and diabetic retinopathy (PRO).

In this experiment Implants containing one of five different TKls (receptor-mediated tyrosine kinase inhibitors) with antlangiogenic activity were made and evaluated, Th® implants w®r© formulated as TKI, sustained release, biodegradable polymer implants with different polyp^aotida-eo-glyoelide) and polyCD-L-lasiida) polymers, made by a melt extrusion process. These Implants are syllable for Intraocular (such as Mravireai) us® to treat on® or more ocular disorders. Specifically, we made and evaluated controlled release Intravitreal implants for the TKls AQN208689, AGN205558, AGN2O6320, AGN206784, and AGN206S18, showing that such implants can consistently release a TKi over a period of from about thre® to about six months. Th® implant formulations were evaluated fit vitro in two different release media (phosphate buffered saline and dtrate phosphate buffer with 0,1 % oetyltrimethylammonium bromide). The effects of the elevated temperature storage and of gemma sterilization on potency were also examined.

Although the implants were made by melt extrusion with poly (lactide) or poly (iactlde-oo-giycollde) polymers various implant formulations were mad® with or under different drug (TKI) load, laeilde-glycolld® ratio, intrinsic viscosity, and extrusion temperature. Th® polymer implant drug delivery systems (DDSs) mad© were assayed by HPLG for potency initially, post-sterilization, and after exposure to accelerated conditions. The TKI release from th® DOS was assayed by HPLC after Incubation in two different release media at 37°C; (1) phosphate buffered saline (pH 7.4), and; (2} oltrat® phosphate buffer with 0.1 % oetyltrimethylammonium bromide (pH S.4), Generally, release rates war® higher for similar polymer systems with higher drug (TKI) loading.

Table 11 sets forth the five different TKIs formulated into drug delivery systems (i.e. implants).

Table 11: Chemical properties of the TKIs AGN206639, AGR20555®, AGN2G6320, AQM2QS784, and AGN2Q6316.

The polymers used to formulate the TKi drug delivery systems were:

Purasorh PDLS PolyCDsL-taofde)» Purac Corp. tot #DQ6?6GA (Inherent viscosity [tv] Is up to 6 df/g, molecular weight [mw In Daltons] Is up to TOOK).

Resomer RG5G2,50:50 PolyCD.L-laotfde-oo-glycolicle], Boehringer Ingelhelm Corp. Lot #R02MQQ2(iv is 0,16 to 0,24 df/g).

Resomer RG502S, 50:50 Poly(DfL-lactlde-oo-glycolids)f Boehrlnger logalhelm Corp, Lot #Res-Q354 (Iv Is 0.1Θ to 0,24 dl/g).

Resomer 13(3504, SOSO PolyP.L-laclide-co-glycolide), Boehringer Ingelhelm Corp. Lot #1009731 (Iv is 0.45 to 0.60 di/g),

Resomer RG505,50:50 PoiyiD.L-lactide-co-giycollde}, Boehringer Ingelhelm Corp. Lot #228799 (Iv Is 0.61 to 0.74 di/g),

Resomer RG50S, 50:50 Poly^L-laotide-co-glyeollde), Boehringer ingelhelm Corp. Lot #34034 (iv fs 0.75 to 0.95 dt/g).

Resomer RG752,75:25 PotyCDjL-lactlde-oo-giycoIide), Boehringer Ingelhafm Corp, Lot #RG2A005 (hr Is 0.18 to 0.24 dtyg).

Resomer RG755, 75:25 Poly(D,L-lactlde-eo~glyaolido), Boehringer Ingelhelm Corp. Lot #1009232 (iv Is 0,50 to 0.70 dl/g).

Resomer R104, Poly(D(L-lactlde), Boehringer Ingelhelm Corp. Lot #290588 (mw Is determined by the presence in the polymer of between about 1,500 and about 2,250 of the repeating monomer unit C3H402).

Resomer R207, Poly^L-lactide), Boehringer Ingelhelm Corp. Lot #280911 (Iv is 1.3 to 1.7 dl/g).

Citrate phosphate buffer (CTAB) solution used was prepared by adding 27.5© g sodium dibasic phosphate heptahydrate, 9.32 g citric acid, and 2 g (1%) oatyltrimethylammonlum bromide (CTAB, JT Baker) to a 2-L volumetric flask and filling with deionized water.

Phosphate buffered saline (PBS) solution used was prepared by adding two packets of PBS (Sigma catalog #P-3818) granules to a 2-L volumetric flask and adding deionized water. i

Release Profile Standards

For stock standard preparation for compounds other than AGM206784,5 mg was added Into a 50-mL volumetric flask and acetonitrile was added to the mark. Working standards were prepared by adding 5 mL of stock standard to a 50-mL i volumetric flask and adding a blend of 60:4Q acetonifrile:water. For AGN208764 stock standard preparation, 5 mg of compound was added Into a 50«mL volumetric flask and a solution of 80% acetonitrile and 20% water was added until full. For working standard preparation, 5 mL of stock standard was added to a 50»mL volumetric flask and a solution of 40:60 aeetonitr!!e:waier was added until full.

Release profile mobile phase

Acetonitrile (ACM) was manufactured by Burdick and Jackson, Trifliscroacetic acid (TFA) was manufactured by Burdick and Jackson, A blend of 75:24.5:0,5 ACftwatenacatlc acid with δ mM hexanesuifbnkj acid was used for alt analyses except for AGN206748 formulations where a blend ©f 60:40:1 AON:Watsr:TFA was used as the mobile phase.

Equipment:

Powder blending: a Glenn Mis inc. Turbuia shaker type T2F, ID number 990720 was used, in addition, an F. Kurt Retech GmbHfi, Co model MM200 ball mill was used.

Powder compaction: A modified Janesville Tool and Manufacturing Inc, pneumatic drive powder compactor, model A-1624 was used.

Piston Extrusion: A custom built piston extruder produced by APS Engineering Inc. was used with a Wallow 93 temperature controller and thermocouple.

Weighing; A Mahler Toledo MT6 balance, S/N 11 f 8481643 was used.

Sample Incubation: A Precision Inc. Reciprocal Shaking Bath with water was used. HPLC: A Waters LC module 1 plus, S/N M38LCJ242h1 with a Supefoo HSF5 μπι, 4.6x150mm column and Waters 2487 dual wavelength absorbance detector was used. Data was analyzed using Peak Pro software, version 9,1b,

Powder blending

The drug (TKI) was stored at room temperature with minimal light exposure, and polymers wer© stored at 5eC and allowed to equilibrate to room temperature prior to use. Both materials ware used as received, Formulations, listed in Table 12, were blended In a stainless steel mixing capsula with two stainless steel balls and placed in a Retselh mi at 30 cps or Turbuia blender at 86 rpm for S to IS minutes. Depending on tie starting materials, formulations underwent four to six blending cycles at five to fifteen minutes each. Between blending cycles, a stainless steel spatula was used to dislodge material from the inside eurfaoae of thi mixing vassal. Formulation ratios and extrusion temperatures far all formulations are listed in Table 2.

Powder compaction A die with a 720 pm opening was attached to a stainless steel barrel. The powder compactor was set to 50 psf. The barrel was inserted Into the powder compactor assembly. A stainless steel powder funnel was used to add a small amount of powder Into the barrel and then the pneumatic compactor was actuated. This process was repeated until the barrel was full or no more powder remained.

Extrusion A piston extruder was set to temperature and allowed to equilibrate, The extrusion temperature was chosen based on drug load and polymer. The extrusion temperature was adjusted for each formulation to produce smooth, uniform looking filaments. After the extruder temperature equilibrated, the piston extrusion barrel was inserted Into the extruder, and a thermocouple wee inserted to measure the temperature at the surface of the barrel. After the barrel temperature equilibrated, the piston was inserted into the barrel and the piston speed was set at 0.0025 in/mln. The first 2-4 inches of extrudat® was discarded. Afterwards, 3-6-inch pieces ware cut directly Into a centrifuge tube. Samples were labeled and stored in a seated foil pouch containing desiccant.

Formulations with higher drug load required higher extrusion temperatures, Polymers with higher Intrinsic viscosities required higher extrusion temperatures than polymers with tower Intrinsic viscosities, Lactfde-gSycoiide co-polymers with a higher iaotide percentage (75:25) required a lower processing temperature than polymers with a lower lacffda percentage (50:50). Formulation information and extrusion temperatures are listed in Table 12.

Content uniformity analysts

Ten samples of 1mg (+/-10%) were cut from each formulation, Each was weighted and placed Individually Into 50-mL volumetric flasks. For AGM206784, a 40:00 ACNrWater or 100% acetonitrile was added and samples were sonicated. Samples were analyzed according to the HPLO method used for release profile analysis, below, For the other TKIs, a 60:40 ACftWater was added to each 50-mL volumetric flask. Flasks were sonicated and samples were tested according to the same HPLC method that la used for in-vivo release (below).

Gamma sterilisation

Samples were weighed and packaged in vials, and each vial was sealed in a foil pouch with desiccant and labeled. All samples were sterilized with 25-40 kGy of gamma radiation.

Stability testing:

Filaments were cut into 1 mg (+/-10%) samples and packaged together in screw-top vials. Formulations were then placed in an oven at 40¾ and ambient humidity, After 14 days, samples warn tested for percent TKi content

In Vitro Release Profile Analysts

Twelve samples of 1 mg (+/-10%) were out from each formulation. Each sample was then weighed and placed Individually Into 60-mL sample vials. Fifty milliters of citrate phosphate buffer solution was added to she vials and fifty milliters of phosphate buffered saline release media was added to six vials. All vials were placed into a shaking water bath set at 37¾ and 50 RPM. At each time point 2 ml was taken from each vial for analysis, the remaining solution was disposed of, and 50 mL of new release media was added to the vial.

Table 12: Formulation conditions tor TKfs AGN206839, ΑΘΝ205558, AC3N206320, AGN20S784, and AGN2Q8316 implants.

In Table 12 the API (active pharmaceutical ingredient [i.e. the TKI] valu© is a weight percent value, HPLC Assay

The HPLC was conditioned until stable at 1 mL per minute flow rate at 2S0nm, Samples were transferred to auto sampler vials with added samples for system suitability and standardization. Total run time was 10 minutes, temperature was ambient and injection volume was 20 pL Samples were taken on day 1 s 4,7, and at 7 day intervals after that until the studies were ended or 100% release was achieved. The total TKi present was calculated from the height of the peak at 280 ran compared to the height of the standard peak. Percent of drag released, total microgtams released, and standard deviations within formulations were calculated from the amount of drug detested.

Results

The content uniformity analysis carried showed that most formulations tested at 100% label strength plus or minus 20%.

Figures 16 to 21 are graphs which provide examples of in vitm release data {in either pH 6.4 citrate phosphate buffer release medium or in pH 7.4 phosphate buffer saline release medium) for the five TKI used in implants with varying polymer formulations. Figures 161© 21 show that TKIs with higher solubilities had the tendency to release at a faster rate than TKIs with lower solubilities. Additionally, Figures 16 to 21 show that TKI Implant formulations with lower drug (IK!) loading released at a slower rate than those with a higher drug loading.

Thus, a number of different sustained release biodegradable polymeric formulations {implants) were made for live different tyrosine kinase Inhibitor compounds. The results show that the release of TKIs from PLGA polymer implants oars be modified by changing polymer matrices and extrusion conditions. Notably, sustained release was achieved for all five TKI compounds in phosphate buffered saline release medium (pH 7.4) and CTAB media and the results show that TKI release from the polymer matrix can last from about one to over six months, depending on the formulation. Significantly, linear, consistent drug release profiles were obtained achieved from for each of the five TKIs in phosphate buffered saline (see formulations 7409-024,7409-009,7409-022, 7409-032, and 7408-071).

Example 8

In Ww Evaluation.Ma.TKl Implant

An experiment was carried out to evaluate effects of an intravltreat TKI implant upon mammalian vision. Thus, either a PLGA placebo implant or a TKI RIGA implant was Inserted into the vitreous of rabbit eyes, ft was determined that an intravitreal TKI PLGA implant can be usad to effectively Improve and maintain vision by iff actively treating conditions such as, or common to, for example, retinal vasodilation, retinal vessel tortuosity (blood flow Increase), blood-retinal barrier breakdown, retinal edama and macula edema,

We used as a model system for determination of vision improvement, rabbits with VEGF Induced retinal damage. It is known that infravitrea! Injeotion of recombinant human MBGFm in rabbits can cause robust vasodilation and vessel tortuosity (blood flow increase), blood-retinal barrier breakdown, and retinal edema. See ©,g. Edelman, J.L et ak, Corticosteroids inhibit VEGFdrsduc&amp;d vascular leakage in a rabbit model of blood-retinal and blood-aqueous harder breakdown, Experimental Eye Research, 80:249-258 (2005) These VEGF Induced ocular effects are believed to be mediated by activation of VEGF reoeptor-1 and VEGF receptor-2.

In this experiment we found that such VEGF Induced retina! injuries can be treated (that is inhibited, alleviated, raversed and/or prevented) by use of a TK! intravftreai implant, Presumably, the tyrosine kinase inhibitor released (In therapeutic levels from the Intravltreai implant) inhibits receptor phosphorylation at the target tissue, thereby blocking the VEGF-mediated responses. Thus, we determined that a 2 mg AGN-206639 (TK!) PLGA can for at least about one month after vltreal implantation provide significant inhibition of the VEGF induced effects, such as blood-retinal barrier breakdown, in this experiment seven female Dutch Belt rabbits (2.1 to 2.6 kg body weight) were used. A 1 mg PLGA placebo implant (made as set forth In Example 7) wae inserted into either the right or left eye (the control) ol six rabbits (N~@). in the remaining eight eyes, a 2 mg TKIFLGA implant (made as set forth in Example 7. See Table 12; Formulation 7409-041 was used as the TKI Implant) was Inserted (N ™ 8), Table 1 sate forth characteristics of the implants used T&amp;bfe 1, Implant composition, size, and In vitro release rales

The implants were intravltrealiy inserted as follows. At T0 {day zero} the rabbits (H =7) were anesthetized with ketamine/xyiazine (subcutaneous), andth© orajlar surface was anesthetized with 2 drops of 1% proparacaine, Irrigated with ophthalmic grade Betadine, and the conjunctiva was incised and refracted from the underlying sclera In the superotemporal quadrant of the ®y®, A 20Θ yVR knife was used to perforata the sclera at the pars plana (about 3 mm posterior to th® limbus) adjacent to the temporal insertion of th® superior rectus muscle, On© implant (placebo or active) was inserted through tha sclerotomy Into th® vitreous with sterile forceps. The scleral and conjunctival Incisions were closed with 8»0 coated Vicryl suture. The implanted eyes was then Instilled with topical kefarofac (Acular) and gentamicin ti.d. Immediately following surgery and continuing for at least 3 days.

At days 12 and 26, the implanted rabbits (N = 7} were anesthetized with intravenous ketemine/fyhxdna and f % proparacafne and Betadine were topically applied to the ocular surface. Immediately thereafter (on days 12 and 28} SQQ ng VEGF186 was Injected intravltrealiy in 100 uL of sterile PBS via a 28Q needle Inserted about 8 mm posterior to the limbus into each eye of each rabbit which had received an implant (placebo or active),

The Implanted eyes were evaluated at several time points after implantation by: (1) fundus evaluation and angiography, and; (2) fluoraphotometric measurement,

Fundus evaluation and angiography was earned out at 14 days and at 20 days post-implant insertion. Thus, using a Zeiss retinal camera,, late phase angiograms were acquired S to 10 min, after intravenous infection of sodium fluorescein (10 mg/kg) via the margin ear vein. All images were masked and than graded by three examiners for severity of retinal fluorescein leakage (normal ” 1; sever© ~ 5). Statistical significance was determined using the Kruskal-Waliis non-parametrio ANGVA test, the Dunn's multiple comparisons test, and the Mann-Whstney test (two-tailed). It was determined, lor example, that at 14 days post implantation a placebo implant ay© scored as high as a 4.6 grade, while a TKt implant eye scored as low as 1.0. Additionally, It was determined, for example, that at 28 days post implantation multiple placebo eyes scored as high as a 5.0 grade, while multiple TKt implant eyes scored as low as about 1,0.

These results ofearly demonstrate that compared to placebo, the 2 mg TKI (ΑβΝ2Ο6630) implant significantly inhibited VEGF-induced angiographic fluorescein leakage at both 14 days and 28 days after surgical insertion o! the TKt implant in rabbit eyes.

Fluorophotometric measurement was also carried out at 14 days and at 28 days post-implant Insertion, Thus, fifty minutes after Intravenous Injection of sodium fluorescein, blood-retinal barrier integrity was assessed using scanning ocular fluorophotometry. The area under the curve (AUC) for vitrecratina! fluorescence was calculated using Kafaidagraph software. Statistical significance was determined using the single factor parametric ANOVA test and the two-tailed unpaired Meet. As shown by Figure 22, compared to placebo, the 2 mg AGN-208839 implant significantly inhibited VEGF-Induced blood-retinal harrier breakdown, as measured by scanning ocular fiuorophotomstry at 14 days and at 28 days after surgical insertion of the Implant In rabbits,

This experiment demonstrated that for at least about one month after vitrea! implantation, a TKI PLGA Intravltreal implant can be used to provide significant vision Improvement, as determined by, for example, Inhibition of experimental blood-retina! barrier breakdown In rabbits. These results therefore show that an intravitreal TKI implant can be used to treat a variety of ocular conditions, including ocular vascuiopathies Including diabetic macuiar edema, proliferative diabetic retinopathy, and choroidal neovascularization associated with age-related macular degeneration.

All references, articles, publications and patents and patent applications oiled herein are Incorporated by reference in their entireties»

While this Invention has been described with respect to various specific examples and embodiments, It is to be understood that the Invention is not limited thereto and that it can be variously practiced within the scope of the following claims.

Throughout this specification and the claims which follow, unless th© context requires otherwise, the word "comprise", and variations such as "comprises” or "comprising", will b© understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

The reference in this specification to any prior publication {or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates. -

Claims (41)

1. The claims defining the invention are as follows:

   1. A biodegradable intraocular implant comprising: a tyrosine kinase inhibitor and a biodegradable polymer matrix that releases drug at a rate effective to sustain release of an amount of the tyrosine kinase inhibitor from the implant for at least about one week after the implant is placed In an eye.
2. 2. The implant of claim 1, wherein the tyrosine kinase inhibitor is a compound having the formula
3. 3. The implant of claim 1, wherein the tyrosine kinase inhibitor is a compound having the formula .
4. 4. The Implant of claim 1, wherein the tyrosine kinase inhibitor is compound having the formula
5. 5. The implant of claim 1, wherein the tyrosine kinase inhibitor is a compound having the formula
6. 6. The implant of claim 1, further comprising an additional ophthaimically acceptable therapeutic agent.
7. 7. The implant of claim t, wherein the tyrosine kinase inhibitor is dispersed within the biodegradable polymer matrix.
8. 8. The implant of claim 1, wherein the matrix comprises at least one polymer selected from the group consisting of polylactides, poly (lactide-co-glycolides), polycaprolactones, derivatives thereof, and mixtures thereof.
9. 9. The implant of claim 1, wherein the implant comprises a first portion comprising a mixture of the tyrosine kinase inhibitor and a biodegradable polymer, and a different second portion comprising a biodegradable polymer substantially free of the tyrosine kinase inhibitor.
10. 10. The implant of claim 1, wherein the matrix comprises a single type of polymer, and the implant releases the tyrosine kinase inhibitor for about 70 days at a substantially linear rate.
11. 11. The Implant of claim 1, wherein the matrix releases drug at a rate effective to sustain release of an amount of the tyrosine kinase inhibitor from the implant for more than one month from the time the implant is placed in the vitreous of the eye.
12. 12. The implant of claim 1, wherein the implant is structured to be placed in the vitreous of the eye.
13. 13. The implant of claim 1, wherein the tyrosine kinase inhibitor is provided in an amount from about 40% by weight to about 70% by weight of the Implant, and the biodegradable polymer matrix comprises a poly (lactide-co-glycolide) in an amount from about 30% by weight to about 60% by weight of the implant.
14. 14. The implant of claim 1 formed as a rod, a wafer, or a particle.
15. 15. The implant of claim 1 which is formed by an extrusion process.
16. 16. A method of making a biodegradable intraocular implant, comprising the step of: extruding a mixture of an tyrosine kinase inhibitor and a biodegradable polymer component to form a biodegradable material that degrades at a rate effective to sustain release of an amount of the tyrosine kinase inhibitor from the implant for at least about one week after the implant is placed in an eye.
17. 17. The method of claim 16, wherein mixture consists essentially of a tyrosine kinase inhibitor and a biodegradable polymer.
18. 18. The method of claim 16, further comprising a step of mixing the tyrosine kinase inhibitor with the polymer component before the extrusion step.
19. 19. The method of claim 16, wherein the tyrosine kinase inhibitor and the polymer component are in a powder form.
20. 20. The method of claim 16, wherein the polymer component comprises a polymer selected from the group consisting of poiyiactides, poly (lactide-co-glycolides), polycaprolactones, and combinations thereof.
21. 21. A medicament, which is a biodegradable Intraocular implant, for treating an ocular condition of an eye of a patient by placing the biodegradable intraocular implant in an eye of the patient, the implant comprising a tyrosine kinase inhibitor and a biodegradable polymer matrix, wherein the implant degrades at a rate effective to sustain release of an amount of the tyrosine kinase inhibitor from the implant effective to treat the ocular condition.
22. 22. The medicament of claim 21, wherein the medicament is effective to treat a retinal ocular condition.
23. 23. The medicament of claim 21, wherein the ocular condition is glaucoma.
24. 24. The medicament of claim 21, wherein the ocular condition is proliferative vitreoretinopathy.
25. 25. The medicament of claim 21, wherein the implant is placed in the posterior of the eye.
26. 26. The medicament of claim 21, wherein the implant is placed in the eye with a trocar.
27. 27. The medicament of claim 21, wherein the implant is placed in the eye with a syringe.
28. 28. The medicament of claim 21, wherein a therapeutic agent is administered in addition to the tyrosine kinase inhibitor to the patient.
29. 29. A biodegradable intravitreal implant comprising: a tyrosine kinase inhibitor (TKi) and a biodegradable polymer, the implant releasing the TKI at a rate effective to sustain release of the TKI from the implant for at least about one week after the implant is placed in the vitreous of an eye.
30. 30. The implant of claim 29, wherein the polymer is selected from the group consisting of polylactides, poly (lactide-co-glycolides), polycaprolaclones, derivatives thereof, and mixtures thereof.
31. 31. The implant of claim 29, wherein the matrix releases the TKI at a rate effective to sustain release of an amount of the TKI from the implant for more than one month from the time the implant is placed in the vitreous of the eye.
32. 32. The implant of claim 29, wherein the tyrosine kinase inhibitor is provided in an amount from about 30% by weight to about 70% by weight of the implant, and the biodegradable polymer matrix comprises a poly (lactide-co-glycolide) In an amount from about 30% by weight to about 70% by weight of the implant.
33. 33. The implant of claim 29, wherein the implant is made by an extrusion process.
34. 34. A method of making a biodegradable intravitreal implant, comprising the step of: extruding a mixture of a TKI and a biodegradable polymer to form a biodegradable implant that degrades at a rate effective to sustain release of an amount of the TKI from the implant for at least about one week after the implant is placed in the vitreous of an eye.
35. 35. The method of claim 34, wherein mixture consists essentially of the TKI and the biodegradable polymer.
36. 36. The method of claim 34, wherein the polymer comprises a polymer selected from the group consisting of polylactides, poly (lactide-co-glycolides), polycaprolactones, and combinations thereof.
37. 37. A medicament, which is a biodegradable intraocular implant, for treating an ocular condition of an eye of a patient by placing the biodegradable intraocular implant into the vitreous of an eye of the patient, the implant comprising a TKI and a biodegradable polymer, wherein the implant degrades at a rate effective to sustain release of an amount of the TKI from the implant effective to treat the ocular condition.
38. 38. The medicament of claim 37, wherein the medicament is effective to treat a retinal ocular condition.
39. 39. The medicament of claim 37, wherein the ocular condition is glaucoma.
40. 40. The medicament of claim 37, wherein the ocular condition is proliferative vitreoretinopathy.
41. 41. The implant of claim 37, wherein the TKI is a compound selected from the group of compounds consisting of compounds having the structures: