AU774495B2 - The use of a protein tyrosine kinase pathway inhibitor in the treatment of ocular disorders - Google Patents

Description

WO 00/67738 PCT/US00/1 2339 THE USE OF A PROTEIN TYROSINE KINASE PATHWAY INHIBITOR IN THE TREATMENT OF OCULAR DISORDERS TECHNICAL FIELD OF THE INVENTION The present invention relates to a method for the prophylactic and therapeutic treatment of age-related macular degeneration, as well as methods for the prophylactic and therapeutic treatment of degeneration of the retina, degeneration of the choroid, and thickening of Bruch's membrane.

BACKGROUND OF THE INVENTION As the average human life span is continually lengthened by improvements in medical technology, there is an increasing need to address medical issues associated with age. The aging process results in physical and chemical changes in the eye, which lead to loss of visual acuity, decreased contrast sensitivity and, ultimately, complete vision loss. Blindness is perhaps the leading debilitating condition afflicting the elderly population. Age-related macular degeneration is the leading cause of blindness in patients over 65 years of age. In fact, vision loss attributed to age-related macular degeneration was found in approximately 10% of the United States population over 65 years of age (Gerster et al. Age Ageing 20: 60-69 (1991)). As the elderly population of the world increases, the incidence of age-related macular degeneration is expected to increase dramatically, reaching a predicted 7.5 million cases in the United States alone by the year 2030 (Hyman et al. Am J Epidemiol 118: 213-227 (1983)).

Age-related macular degeneration is a progressive, degenerative disorder of the eye resulting in loss of L r ~rll 'Wilt,~ B~i~L' r l l. r~lr l~~.YhrR~l I IInl~~ IJFI r~llY Y1! WO 00/67738 PCT/US00/12339 2 visual acuity. Symptoms of age-related macular degeneration include blurred vision, decreased ability to read, especially in dim light, trouble with dark adaptation and, in relatively few cases, abrupt vision loss. Complications associated with advanced age-related macular degeneration are divided into two categories, atrophic and exudative. Atrophic complications are associated with retinal pigment epithelial cell loss resulting in atrophy of the retinal pigment epithelium (RPE). Exudative complications, which appear in approximately 10% of age-related macular degeneration cases, include disciform scars scarring involving fibrous elements) and neovascularization. Ultimately, blindness from age-related macular degeneration stems from degeneration of the RPE and the subsequent death of photoreceptors.

Risk factors associated with age-related macular degeneration include age, heredity, systemic disease, environmental factors, such as smoking and light exposure (see Chesapeake Bay Waterman Study, Taylor et al., Arch. Ophthalmol. 110: 99-104 (1992)), and nutritional deficiency. Hyperopia and iris color also have been linked to the disease. Age-related macular degeneration has been correlated to light-colored irises, perhaps due to chronic exposure to damaging light which is normally absorbed by dark-colored irises. The disease also appears more often in women than men.

The best documented mechanism for the development of age-related macular degeneration involves molecular degradation in the RPE. Incomplete digestion of abnormal molecules, most likely altered post-synthetically, results in the formation of pockets of waste in RPE cells which eventually interfere with the normal metabolism of U WiAMm'r 'A WO 00/67738 PCT/US00/12339 3 the cell. Consequently, aberrant excretions from RPE cells aggregate within Bruch's membrane as basal laminar deposits, drusen and debris. It is believed that such deposits invoke neovascularization and/or the death of RPE cells (Young, Survey of Ophthalmology 31(5): 291-306 (1987)) There is currently no known prophylactic or therapeutic treatment for age-related macular degeneration (AMD). While not a cure for AMD, laser photocoagulation is used to treat choroidal neovascularization associated with age-related macular degeneration. Such treatment has been proven to reduce the risk of severe vision loss. Laser photocoagulation also has been used to treat drusen. However, laser treatment may cause permanent blind spots corresponding to the treated areas, leading to a decrease in visual acuity. Laser treatment may also cause persistent or recurrent hemorrhage and increase the risk of sensory retinal detachment. Many patients eventually experience severe vision loss in spite of treatment. Likewise, there currently is no method available for the prophylactic or therapeutic treatment of physical and chemical changes in the eye associated with the aging process, in general.

Given the prevalence of age-related macular degeneration, there remains a need for an effective prophylactic and therapeutic treatment of age-related macular degeneration. Accordingly, it is a principal object of the present invention to provide a method of prophylactically and therapeutically treating age-related macular degeneration, including treatment of the atrophic and exudative complications associated with the disease.

A need also exists in the art for an effective L n~ WO 00/67738 PCT/US00/12339 4 prophylactic and therapeutic treatment for deterioration of the eye, such as degeneration of the retina, degeneration of the choroid and thickening of Bruch's membrane. The present invention provides such methods of treatment. These and other objects of the present invention, as well as additional inventive features, will become apparent from the detailed description provided herein.

BRIEF SUMMARY OF THE INVENTION The present invention is directed to a method for the prophylactic and therapeutic treatment of age-related macular degeneration, including treatment of the atrophic and exudative complications associated with the disease.

The present invention is also directed to a method for the prophylactic and therapeutic treatment of an animal for degeneration of the retina. A method of prophylactically or therapeutically treating an animal for degeneration of the choroid is also provided, as is a method of prophylactically or therapeutically treating an animal for thickening of Bruch's membrane. The methods involve the administration of an inhibitor of the protein tyrosine kinase pathway. Preferably, the inhibitor of the protein tyrosine kinase pathway is a compound of formula: x ]Um WO 00/67738 PCT/US00/12339 wherein V, W and X are selected from the group consisting of hydro, hydroxyl, alkoxy, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, and Y is selected from the group consisting of oxygen, sulfur, C(OH), and C=O, and Z is selected from the group consisting of hydro and C(O)ORI, wherein RI is an alkyl. Preferably, the alkoxy is a Ci-Cs alkoxy. Preferably, the halo is fluorine, chlorine or bromine. Preferably, the ester is a Ci-C 6 ester. Preferably, the ether is a Ci-Cs ether. Preferred pharmaceutically acceptable salts of the carboxylic acid group include sodium and potassium salts. Preferably, the alkyl groups are Ci-C 6 alkyl groups. Desirably, the protein tyrosine kinase pathway inhibitor is genistein.

DETAILED DESCRIPTION OF THE INVENTION The present invention is predicated on the discovery that an inhibitor of the protein tyrosine kinase pathway, specifically genistein, is effective in treating prophylactically and therapeutically age-related macular degeneration, including the exudative and atrophic complications associated with age-related macular degeneration, and ocular disorders, such as degeneration of the retina, degeneration of the choroid and thickening of Bruch's membrane. Accordingly, the present invention provides a method for the prophylactic and therapeutic treatment of age-related macular degeneration. By "prophylactic" is meant the protection, in whole or in part, against age-related macular degeneration, in particular choroidal neovascularization and retinal pigment epithelium atrophy. By "therapeutic" is meant a 21u2a r ALo! r. K r WO 00/67738 PCT/US00/12339 6 the amelioration of age-related macular degeneration, itself, and the protection, in whole or in part, against further age-related macular degeneration, in particular choroidal neovascularization and retinal pigment epithelium atrophy. Preferably, age-related macular degeneration is that which results from age, hyperopia, systemic disease, such as cardiovascular disease or hypertension, smoking, light exposure or nutritional deficiency.

The method comprises the administration of an inhibitor of the protein tyrosine kinase pathway in an amount sufficient to treat the macula for age-related macular degeneration prophylactically or therapeutically.

Any inhibitor of the protein tyrosine kinase (PTK) pathway can be used in the method of the present invention as long as it is safe and efficacious.

The present invention additionally provides a method for the prophylactic and therapeutic treatment of both atrophic and exudative complications associated with agerelated macular degeneration. Atrophic complications include pigmentary disturbance of the retinal pigment epithelium, hard, soft and/or confluent drusen, basal laminar deposits, a thickening of Bruch's membrane, and RPE atrophy. Drusen are yellow or white excresences between the basement membrane of the RPE and Bruch's membrane which are often precursors for neovascularization. Exudative complications include choroidal neovascularization, RPE detachment, RPE tears, disciform scarring, vitreous hemorrhage and subretinal hemorrhage. Prophylactic and therapeutic treatment of exudative complications can be effected by prevention or inhibition of disruption of the integrity of the basement membrane.

ani~~~uiur 7a". W WO 00/67738 PCT/US00/12339 7 The method comprises the administration of an inhibitor of the PTK pathway in an amount sufficient to treat an exudative or atrophic complication of agerelated macular degeneration prophylactically or therapeutically. Any safe and efficacious PTK inhibitor can be used.

The present invention further provides a method for the prophylactic and therapeutic treatment of an animal for degeneration of the retina, in particular that which is due to aging. As used herein, degeneration of the retina, in particular that which is due to aging, includes, for example, pigment epithelial abnormalities and ganglion cell loss. Degeneration of the retina also includes loss of photoreceptor cells, which can result in decreased contrast sensitivity and vision loss. By "prophylactic" is meant the protection, in whole or in part, against further degeneration of the retina. By "therapeutic" is meant the amelioration of degeneration of the retina, itself, and the protection, in whole or in part, against further degeneration of the retina.

The method comprises the administration of an inhibitor of the PTK pathway in an amount sufficient to treat degeneration of the retina, in particular that which is due to aging, prophylactically or therapeutically. As indicated above, any PTK pathway inhibitor can be used in this method as long as it is safe and efficacious.

In addition, the present invention provides a method for the prophylactic and therapeutic treatment of an animal for degeneration of the choroid, in particular that which is due to aging. Degeneration of the choroid, in particular that which is due to aging, includes, for example, flattening of the choroid. Degeneration of the XiEE 4.4A WO 00/67738 PCT/US00/12339 8 choroid due to aging also includes atrophy of choriocapillaries. By "prophylactic" is meant the protection, in whole or in part, against degeneration of the choroid. By "therapeutic" is meant the amelioration of degeneration of the choroid, itself, and the protection, in whole or in part, against further degeneration of the choroid.

The method comprises the administration of an inhibitor of the PTK pathway in an amount sufficient to treat degeneration of the choroid, in particular that which is due to aging, prophylactically or therapeutically. Any safe and efficacious PTK pathway inhibitor can be used.

Ocular disorders further include thickening of Bruch's membrane, which is located between the retina and the choroid, in particular thickening of Bruch's membrane as a result of aging. Thickening of Bruch's membrane can interfere with vision by separating photoreceptor cells from the vascular-rich choroid and causing abnormalities in the retina. Accordingly, the present invention provides a method for the prophylactic and therapeutic treatment of an animal for thickening of Bruch's membrane. By "prophylactic" is meant the protection, in whole or in part, against thickening of Bruch's membrane.

By "therapeutic" is meant the amelioration of thickening of Bruch's membrane, itself, and the protection, in whole or in part, against further thickening of Bruch's membrane.

The method comprises the administration of an inhibitor of the PTK pathway in an amount sufficient to treat Bruch's membrane for thickening prophylactically or therapeutically. Any safe and efficacious inhibitor of the PTK pathway can be used.

%3A.iE WO 00/67738 PCT/USOO/12339 9 Herein, "PTK inhibitor" will be used to refer to such compounds and is intended to encompass any and all compounds that inhibit the PTK pathway, irrespective of at which point in the pathway the compound effects inhibition. PTK inhibitors are known in the art; others can be identified in accordance with assays known in the art. One of ordinary skill in the art will appreciate that, while complete inhibition of the PTK pathway by the PTK inhibitor is preferred, partial inhibition of the PTK pathway can be sufficient to achieve a prophylactic or therapeutic effect in the context of the present invention.

Preferably, the PTK inhibitor is genistein (5,7dihydroxy-3-(4-hydroxyphenyl)-4H-l-benzopyran-4-one) or a pharmaceutically acceptable, PTK pathway-inhibiting analogue or prodrug thereof or a pharmaceutically acceptable salt of any of the foregoing. Accordingly, the PTK inhibitor can be a compound of the following formula: x X 7

V

wherein V, W and X are selected from the group consisting of hydro, hydroxyl, an alkoxy, halo-, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, and Y is selected from the group consisting of oxygen, sulfur, C(OH), and C=O, and Z .~IAtWBAM!!bjt~tt,.~ WO 00/67738 PCT/US00/12339 is selected from the group consisting of hydro and C(O)ORI, wherein Ri is an alkyl. Preferably, the alkoxy is a Ci-Cs alkoxy. Preferably, the halo is fluorine, chlorine or bromine. Preferably, the ester is a CI-Cs ester. Preferably, the ether is a CI-C 6 ether. Preferred pharmaceutically acceptable salts of the carboxylic acid group include sodium and potassium salts. Preferably, the alkyl groups are Ci-Cs alkyl groups. Desirably, the PTK pathway inhibitor is genistein.

The prodrug can be any pharmaceutically acceptable prodrug of genistein, a PTK pathway-inhibiting analogue of genistein, or a pharmaceutically acceptable salt of either of the foregoing. One of ordinary skill in the art will appreciate, however, that the prodrug used must be one that can be converted to an active PTK inhibitor in or around the macula, retina, choroid or Bruch's membrane. A preferred prodrug is a prodrug that increases the lipid solubility of genistein, a PTK pathway-inhibiting analogue of genistein, or a pharmaceutically acceptable salt of either of the foregoing. A preferred prodrug is one in which one or more of V, W and X are independently derivatized with an ester, such as pivalic acid.

Compounds of the above formula are widely available commercially. For example, genistein is available from LC Laboratories (Woburn, MA). Those compounds that are not commercially available can be readily prepared using organic synthesis methods known in the art.

Whether or not a particular analogue, prodrug or pharmaceutically acceptable salt of a compound in accordance with the present invention can treat macular degeneration, degeneration of the retina, degeneration of the choroid, or thickening of Bruch's membrane, any one innm-l .ri mrm rrn ni~-n~-ni ~Ri~ni~Riu+,;r v~*nriinn r~I~on~ llilri~i~ WO 00/67738 PCT/US00/12339 11 of which can be due to aging, prophylactically or therapeutically can be determined by its effect in the mouse model used in Examples 2 and 4-6. Alternatively, analogues, prodrugs and pharmaceutically acceptable salts of inhibitors of the PTK pathway can be tested by in vitro assays such as, for example, the method set forth in Example i.

In addition, color Doppler imaging can be used to evaluate the action of a drug in ocular pathology (Valli et al., Ophthalmologica 209(13): 115-121 (1995)). Color Doppler imaging is a recent advance in ultrasonography, allowing simultaneous two-dimensional imaging of structures and the evaluation of blood flow.

Accordingly, atrophic complications, such as retinal pigment epithelium atrophy, and exudative complications, such as choroidal neovascularization, can be analyzed using such technology. Similarly, complications associated with degeneration of the retina, such as photoreceptor loss and ganglion cell loss, as well as complications associated with degeneration of the choroid, such as atrophy of the choriocapillaries and flattening of the choroid, can be analyzed using color Doppler imaging.

A PTK inhibitor can be bound to a suitable matrix, such as a polymeric matrix, if desired, for use in the present inventive method. Any of a wide range of polymers can be used in the context of the present invention provided that, if the polymer-bound compound is to be used in vivo, the polymer is biologically acceptable (see, U.S. Patent Nos. 5,384,333 and 5,164,188) An advantage of genistein is that it is very safe and efficacious. For example, when genistein was orally .N~UV*I~L~R~m'"-~~UUf~s'LIIYl~~~i~nlL~~h WO 00/67738 PCT/US00/12339 12 administered to Zucker diabetic fatty rats, genistein was found to be nontoxic to the retina at dosages ranging from 75 mg/kg/day to 300 mg/kg/day over a period of six months as measured by electroretinography. In addition, oral administration of genistein was found to have no effect on food intake and body weight for male and female rats. Also, no effect of orally administered genistein was found with respect to the weight of the ovaries and the uterus in female rats.

The PTK inhibitor, which is preferably genistein, a PTK pathway-inhibiting analogue of genistein, a PTK pathway-inhibiting prodrug of genistein, or a pharmaceutically acceptable salt of any of the foregoing, can be administered in accordance with the present inventive methods by any suitable route. Suitable routes of administration include systemic, such as orally or by injection, topical, intraocular, periocular subTenon's), subconjunctival, subretinal, suprachoroidal and retrobulbar. The manner in which the PTK inhibitor is administered is dependent, in part, upon whether the treatment of age-related macular degeneration, degeneration of the retina, degeneration of the choroid, or thickening of Bruch's membrane is prophylactic or therapeutic. For instance, the manner in which the PTK inhibitor is administered for therapeutic treatment of age-related macular degeneration is dependent, in part, upon the cause of the disease.

For example, given that the appearance of drusen and RPE hyperpigmentation is often the first indication of age-related macular degeneration, the PTK inhibitor can be administered prophylactically as soon as drusen and RPE hyperpigmentation are detected. For the prophylactic treatment of age-related macular degeneration, the PTK L 4' 4 4 .24k 4~ WO 00/67738 PCT/US00/12339 13 inhibitor is preferably administered systemically, e.g., orally or by injection. For the therapeutic treatment of age-related macular degeneration, treatment of atrophic and/or exudative complications, the PTK inhibitor can be administered systemically, orally or by injection, intraocularly, topically, subconjunctivally or periocularly subTenon's), for example.

The PTK inhibitor is preferably administered as soon as possible after it has been determined that an animal, such as a mammal, specifically a human, is at risk for age-related macular degeneration (prophylactic treatment) or has begun to develop age-related macular degeneration (therapeutic treatment). Treatment will depend, in part, upon the particular PTK inhibitor used, the amount of the PTK inhibitor administered, the route of administration, and the cause and extent, if any, of macular degeneration realized. Likewise, the PTK inhibitor is preferably administered as soon as possible after it has been determined that an animal, specifically a human, is at risk for retinal or choroidal degeneration or thickening of Bruch's membrane (prophylactic treatment) or has begun to develop degeneration of the retina or choroid or thickening of Bruch's membrane (therapeutic treatment) One skilled in the art will appreciate that suitable methods of administering a PTK inhibitor, which is useful in the present inventive method, are available. Although more than one route can be used to administer a particular PTK inhibitor, a particular route can provide a more immediate and more effective reaction than another route. Accordingly, the described routes of administration are merely exemplary and are in no way limiting.

a a Tan WO 00/67738 PCT/USO/12339 14 The dose administered to an animal, particularly a human, in accordance with the present invention should be sufficient to effect the desired response in the animal over a reasonable time frame. One skilled in the art will recognize that dosage will depend upon a variety of factors, including the strength of the particular PTK inhibitor employed, the age, species, condition or disease state, and body weight of the animal, as well as the amount of the macula, retina or choroid about to be affected or actually affected by degeneration or the amount of Bruch's membrane about to be affected or actually affected by thickening. The size of the dose also will be determined by the route, timing and frequency of administration as well as the existence, nature, and extent of any adverse side effects that might accompany the administration of a particular PTK inhibitor and the desired physiological effect. It will be appreciated by one of ordinary skill in the art that various conditions or disease states, in particular, chronic conditions or disease states, may require prolonged treatment involving multiple administrations.

Suitable doses and dosage regimens can be determined by conventional range-finding techniques known to those of ordinary skill in the art. Generally, treatment is initiated with smaller dosages, which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. The present inventive method will typically involve the administration of from about 1 mg/kg/day to about 100 mg/kg/day, preferably from about 15 mg/kg/day to about mg/kg/day, if administered systemically. Intraocular administration typically will involve the administration i t I WO 00/67738 PCT/US00/12339 of from about 0.1 mg total to about 5 mg total, preferably from about 0.5 mg total to about 1 mg total.

A preferred concentration for topical administration is 100 LM.

Compositions for use in the present inventive method preferably comprise a pharmaceutically acceptable carrier and an amount of a PTK inhibitor sufficient to treat agerelated macular degeneration and/or an atrophic or exudative complication thereof prophylactically or therapeutically. Alternatively, compositions for use in the present inventive method of treating degeneration of the retina, such as that which is due to aging, degeneration of the choroid, such as that which is due to aging, or thickening of Bruch's membrane, such as that which is due to aging, preferably comprise a pharmaceutically acceptable carrier and an amount of a PTK inhibitor sufficient to treat degeneration of the retina, degeneration of the choroid or thickening of Bruch's membrane, respectively, prophylactically or therapeutically. The carrier can be any of those conventionally used and is limited only by chemicophysical considerations, such as solubility and lack of reactivity with the compound, and by the route of administration. It will be appreciated by one of ordinary skill in the art that, in addition to the following described pharmaceutical compositions, the PTK inhibitor can be formulated as polymeric compositions, inclusion complexes, such as cyclodextrin inclusion complexes, liposomes, microspheres, microcapsules and the like (see, U.S. Patent Nos. 4,997,652, 5,185,152 and 5,718,922).

The PTK inhibitor can be formulated as a pharmaceutically acceptable acid addition salt. Examples F1P~ur~rrr-~inrawn ui~i l~~lari n rM T-1-FEWM. L-11-11-a~ :m~ WO 00/67738 PCT/US00/12339 16 of pharmaceutically acceptable acid addition salts for use in the pharmaceutical composition include those derived from mineral acids, such as hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric and sulfuric acids, and organic acids, such as tartaric, acetic, citric, malic, lactic, fumaric, benzoic, glycolic, gluconic, succinic, and arylsulphonic, for example p-toluenesulphonic, acids.

The pharmaceutically acceptable excipients described herein, for example, vehicles, adjuvants, carriers or diluents, are well-known to those who are skilled in the art and are readily available to the public. It is preferred that the pharmaceutically acceptable carrier be one which is chemically inert to the PTK inhibitor and one which has no detrimental side effects or toxicity under the conditions of use.

The choice of excipient will be determined in part by the particular PTK inhibitor, as well as by the particular method used to administer the composition.

Accordingly, there is a wide variety of suitable formulations of the pharmaceutical composition of the present invention. The following formulations are merely exemplary and are in no way limiting.

Injectable formulations are among those that are preferred in accordance with the present inventive method. The requirements for pharmaceutically effective carriers for injectable compositions are well-known to those of ordinary skill in the art (see Pharmaceutics and Pharmacy Practice, J.B. Lippincott Co., Philadelphia, PA, Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel, 4 th ed., pages 622- 630 (1986)). It is preferred that such injectable Y r~ n"2Z*AflL22iW ThV~., MYWt~' iNiU4&i .t' WO 00/67738 PCT/US00/12339 17 compositions be administered intramuscularly, intravenously, or intraperitoneally.

Topical formulations are well-known to those of skill in the art. Such formulations are suitable in the context of the present invention for application to the skin. The use of patches, corneal shields (see, e.g., U.S. Patent No. 5,185,152), and ophthalmic solutions (see, U.S. Patent No. 5,710,182) and ointments, eye drops, is also within the skill in the art.

Formulations suitable for oral administration can consist of liquid solutions, such as an effective amount of the compound dissolved in diluents, such as water, saline, or orange juice; capsules, sachets, tablets, lozenges, and troches, each containing a predetermined amount of the active ingredient, as solids or granules; powders; suspensions in an appropriate liquid; and suitable emulsions. Liquid formulations may include diluents, such as water and alcohols, for example, ethanol, benzyl alcohol, and the polyethylene alcohols, either with or without the addition of a pharmaceutically acceptable surfactant, suspending agent, or emulsifying agent. Capsule forms can be of the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers, such as lactose, sucrose, calcium phosphate, and corn starch. Tablet forms can include one or more of lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, and other excipients, colorants, diluents, buffering agents, disintegrating agents, moistening agents, preservatives, AM RIM IRRs- a-Ai-Sl l~l*i "~rir m ~nnl~~m~n~ WO 00/67738 PCT/US00/12339 18 flavoring agents, and pharmacologically compatible excipients. Lozenge forms can comprise the active ingredient in a flavor, usually sucrose and acacia or tragacanth, as well as pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acacia, emulsions, gels, and the like containing, in addition to the active ingredient, such excipients as are known in the art.

Formulations suitable for parenteral administration include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain anti-oxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The inhibitor can be administered in a physiologically acceptable diluent in a pharmaceutical carrier, such as a sterile liquid or mixture of liquids, including water, saline, aqueous dextrose and related sugar solutions, an alcohol, such as ethanol, isopropanol, or hexadecyl alcohol, glycols, such as propylene glycol or polyethylene glycol, dimethylsulfoxide, glycerol ketals, such as 2,2-dimethyl- 1,3-dioxolane-4-methanol, ethers, such as poly(ethyleneglycol) 400, an oil, a fatty acid, a fatty acid ester or glyceride, or an acetylated fatty acid glyceride, with or without the addition of a pharmaceutically acceptable surfactant, such as a soap or a detergent, suspending agent, such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agents and other pharmaceutical adjuvants.

"r~r;~i;WYr ~ullr r~LF S i i(~I P WO 00/67738 PCT/US00/12339 19 Oils, which can be used in parenteral formulations include petroleum, animal, vegetable, or synthetic oils.

Specific examples of oils include peanut, soybean, sesame, cottonseed, corn, olive, petrolatum, and mineral.

Suitable fatty acids for use in parenteral formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

Suitable soaps for use in parenteral formulations include fatty alkali metals, ammonium, and triethanolamine salts, and suitable detergents include cationic detergents such as, for example, dimethyl dialkyl ammonium halides, and alkyl pyridinium halides, anionic detergents such as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates, nonionic detergents such as, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylenepolypropylene copolymers, amphoteric detergents such as, for example, alkyl-paminopropionates, and 2-alkyl-imidazoline quaternary ammonium salts, and mixtures thereof.

The parenteral formulations will typically contain from about 0.5 to about 25% by weight of the active ingredient in solution. Preservatives and buffers may be used. In order to minimize or eliminate irritation at the site of injection, such compositions may contain one or more nonionic surfactants having a hydrophilelipophile balance (HLB) of from about 12 to about 17.

The quantity of surfactant in such formulations will typically range from about 5 to about 15% by weight.

Suitable surfactants include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate and the high -i~rrr.~nM=-V~nW r -1 4 S 1 I 0I .I WO 00/67738 PC/USO/12339 molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol. The parenteral formulations can be presented in unit-dose or multi-dose sealed containers, such as ampules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid excipient, for example, water, for injections, immediately prior to use.

Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described.

Such compositions can be formulated as intraocular formulations, sustained-release formulations or devices (see, U.S. Patent No. 5,378,475). For example, gelantin, chondroitin sulfate, a polyphosphoester, such as bis-2-hydroxyethyl-terephthalate (BHET), or a polylactic-glycolic acid (in various proportions) can be used to formulate sustained-release formulations.

Implants (see, U.S. Patent Nos. 5,443,505, 4,853,224 and 4,997,652), devices (see, U.S. Patent Nos. 5,554,187, 4,863,457, 5,098,443 and 5,725,493), such as an implantable device, a mechanical reservoir, an intraocular device or an extraocular device with an intraocular conduit 100 t 1 mm in diameter), or an implant or a device comprised of a polymeric composition as described above, can be used.

The present inventive method also can involve the co-administration of other pharmaceutically active compounds. By "co-administration" is meant administration before, concurrently with, in combination with the PTK inhibitor in the same formulation or in separate formulations, or after administration of a PTK inhibitor as described above.

unu~- -l WO 00/67738 PCT/USOO/12339 21 For example, corticosteroids, prednisone, methylprednisolone, dexamethasone, or triamcinalone acetinide, or noncorticosteroid anti-inflammatory compounds, such as ibuprofen or flubiproben, can be coadministered. Similarly, vitamins and minerals, e.g., zinc, anti-oxidants, carotenoids (such as a xanthophyll carotenoid like zeaxanthin or lutein), and micronutrients can be co-administered. In addition, other types of inhibitors of the protein tyrosine kinase pathway, which include natural protein tyrosine kinase inhibitors like quercetin, lavendustin A, erbstatin and herbimycin A, and synthetic protein tyrosine kinase inhibitors like tyrphostins AG490, AG17, AG213 (RG50864), AG18, AG82, AG494, AG825, AG879, AG1112, AG1296, AG1478, AG126, RG13022, RG14620 and AG555), dihydroxy- and dimethoxybenzylidene malononitrile, analogs of lavendustin A AG814 and AG957), quinazolines AG1478), and thiazolidinediones, can be co-administered with genistein or an analogue, prodrug or pharmaceutically acceptable salt thereof (see Levitzki et al., Science 267: 1782-1788 (1995); and Cunningham et al., Anti-Cancer Drug Design 7: 365-384 (1992)). In this regard, potentially useful derivatives of genistein include those set forth in Mazurek et al., U.S. Patent No. 5,637,703.

Neutralizing proteins to growth factors, such as a monoclonal antibody that is specific for a given growth factor, VEGF (for an example, see Aiello et al., PNAS USA 92: 10457-10461 (1995)), or phosphotyrosine (Dhar et al., Mol. Pharmacol. 37: 519-525 (1990)), can be co-administered. Other various compounds that can be coadministered include inhibitors of protein kinase C (see, U.S. Patent Nos. 5,719,175 and 5,710,145), cytokine i~YiJI~ rE'4iL Jrm1Zh M .Li ~Ur' ~i WO 00/67738 PCT/US00/12339 22 modulators, an endothelial cell-specific inhibitor of proliferation, thrombospondins, an endothelial cell-specific inhibitory growth factor, TNF(, an anti-proliferative peptide, SPARC and prolferinlike peptides, a glutamate receptor antagonist, aminoguanidine, an angiotensin-converting enzyme inhibitor, angiotensin II, calcium channel blockers, y-tectorigenin, ST638, somatostatin analogues, SMS 201-995, monosialoganglioside GM1, ticlopidine, neurotrophic growth factors, dihydroxycinnamate, an angiogenesis inhibitor, e.g., recombinant EPO, a sulphonylurea oral hypoglycemic agent, gliclazide (non-insulin-dependent diabetes), ST638 (Asahi et al., FEBS Letter 309: 10-14 (1992)), thalidomide, nicardipine hydrochloride, aspirin, piceatannol, staurosporine, adriamycin, epiderstatin, (+)-aeroplysinin-1, phenazocine, halomethyl ketones, anti-lipidemic agents, etofibrate, chlorpromazine and spinghosines, aldose reductase inhibitors, such as tolrestat, SPR-210, sorbinil or oxygen, and retinoic acid and analogues thereof (Burke et al., Drugs of the Future 17(2): 119-131 (1992); and Tomlinson et al., Pharmac.

Ther. 54: 151-194 (1992)). Selenoindoles (2-thioindoles) and related disulfide selenides, such as those described in Dobrusin et al., U.S. Patent No. 5,464,961, are useful PTK inhibitors. Those patients that exhibit systemic fluid retention, such as that due to cardiovascular or renal disease and severe systemic hypertension, can be additionally treated with diuresis, dialysis, cardiac drugs and antihypertensive agents.

5M-W_-JWWA'_W4MAVRW NIK J 2 W VM AU "QJT' WO 00/67738 PCT/US00/12339 23

EXAMPLES

The following examples further illustrate the present invention but, of course, should not be construed as in any way limiting its scope.

EXAMPLE 1 This example demonstrates that genistein is effective in inhibiting disruption of the integrity of basement membrane and subsequent invasion of basement membrane by retinal endothelial cells.

Retinal capillary endothelial cells were cultured on Matrigel (Collaborative Research, Bedford, MA) in the presence of bFGF (1-10 ng/ml) and in the presence or absence of genistein (0.2 mg/100 ml). Matrigel is derived from basement membrane-producing tumor tissue and is comprised of type IV collagen and laminin as is the retinal basement membrane, differing only in the relative proportions of the components.

Retinal capillary endothelial cells cultured in the presence of bFGF alone invaded Matrigel. The presence of genistein reduced retinal capillary endothelial cell invasion into Matrigel by .8 mm, genistein inhibited invasion by approximately 32%.

This example illustrates the ability of genistein to inhibit disruption of the integrity of and the subsequent retinal capillary endothelial cell invasion of a basement membrane.

EXAMPLE 2 This example demonstrates the ability of genistein to inhibit photoreceptor loss as associated with agerelated macular degeneration and degeneration of the retina associated with aging.

C~S~.K&32LkV& I~A% ~YW2~A~~ S*!IYJ' A3,~Mfl'2A~rAi.USAA~V Its JeawA~!MIt A*ItLattLKs~ It ALSIt!tSAIS JA~)A4ItWIjIt~ttt *~~#ftt0~ibIj, ii, WO 00/67738 PCT/USOO/12339 24 Two strains of mice, senile accelerated mice strain 9 (SAM9) and resistant mice strain 4 (R4) (both strains were gifts from Kyoto University, Japan and are currently maintained at Johns Hopkins University), were exposed to a light source in order to determine the effect of genistein on light-induced damage of the macula. SAM9 mice are prone to accelerated age-associated changes in the skin and eye. R4 mice are identical to SAM9, except that R4 mice are resistant to physical changes associated with accelerated aging.

A total of about 20-25 mice of each strain were divided into control and treatment groups. The treatment groups received genistein (150 mg/kg food) by oral lavage once daily up to about 4 weeks. Mice were sacrificed at i, 2 and 3 weeks following light exposure. Eyes were enucleated and cut serially for histological study.

Specifically, the photoreceptor nuclei were counted to determine the degree of photoreceptor loss over specific areas of the posterior and peripheral retina.

At week 1, an average of six layers of photoreceptor nuclei were observed in both the treated and control groups for both strains of mice. There was no significant difference in the amount of light-induced damage between the control and treatment groups. At week 2, genistein had significantly reduced the amount of light-induced damage in the treatment groups of both strains. An average of two to three layers of photoreceptor nuclei were observed in the SAM9 mice and the R4 mice treated with genistein. Approximately 0-1 layer of photoreceptor nuclei was observed in control groups. At week 3, an average of 0-1 layers of photoreceptor nuclei was observed in both treated and untreated animals of both strains.

iw a w ~m WO 00/67738 PCT/USOO/1 2339 An additional strain of senile accelerated mice, strain 8 (SAM8), along with resistant strain 1 were exposed to a light source in order to determine the effect of genistein on light-induced photoreceptor cell loss. All subjects were exposed to constant, white fluorescent light of 800-lux, after dark adaptation for 24 hours. Each of the SAM8 and R1 strains of mice was divided into two groups: the treated group, which received genistein (50 mg/kg body weight/day) orally by the lavage method, and the control group, which received no treatment. After one week of light exposure, the thickness and cell count in the outer nuclear layer (ONL) was greater in the genistein treated groups of both SAM8 and R1, than in the control groups. After two weeks of light exposure, the ONL thickness was greater in the genistein treated R1 than in the corresponding control group. After three weeks of light exposure, the ONL thickness was greater in the genistein treated R1 than in the corresponding control group. There was no difference by treatment in the SAM8 group after two and three weeks of light exposure.

The example illustrates the ability of genistein to inhibit photoreceptor loss, such as that associated with degeneration of the retina, such as that which is due to aging, and age-related macular degeneration.

EXAMPLE 3 This example demonstrates the ability of genistein to inhibit the progression of age-related macular degeneration after disruption of Bruch's membrane.

Bruch's membrane in three cynomolgus monkeys was disrupted using a laser beam and subsequent choroidal neovascularization was evaluated. Genistein was I riii- i AIjMA' 4*ML~1nAt?2 WO 00/67738 PCT/US00/12339 26 administered to two monkeys via a single intraocular injection of 100 mg before laser-induced disruption of Bruch's membrane. Thirteen of seventeen lesions developed neovascularization. One monkey was administered genistein orally (300 mg/kg body weight), in addition to a single intraocular injection (100 mg), every other day starting two days prior to laser treatment, for a total of three doses. Of eighteen lesions resulting from laser treatment, only two developed choroidal neovascularization.

These results indicate that genistein, when given prior to disruption of Bruch's membrane, can inhibit or prevent secondary development of neovascularization, thereby inhibiting the progression of age-related macular degeneration.

EXAMPLE 4 This example describes a method of determining the usefulness of a PTK pathway inhibitor as described herein to inhibit or ameliorate degeneration of the retina, such as that which is due to aging.

Two strains of mice, a senile accelerated prone strain (SAMP) and a resistant strain (SAMR) are obtained from Takeda Chemical Industries Ltd., Osaka, Japan. As discussed above, SAMP mice are prone to accelerated ageassociated changes in the eye, while SAMR mice are resistant to physical changes associated with accelerated aging.

Subjects of each strain are divided into control and treatment groups. All mice are bred under specific pathogen-free conditions with a high efficiency particulate device with a temperature- and a lightcontrolled regimen (68-72° F and a light/dark cycle with aF~?nn~:~a ~uw~intpr~or r m~ilo~~r~p' i~~Om4YEiq'n, n g~y~l ~L WO 00/67738 PCT/US00/12339 27 lights on at 7:00 a.m. to 9:00 and free access to food (PMI Nutrition International, Inc, 1401, St. Louis, MO) ad libitum and water from an automated filtered system. The treatment groups receive genistein mg/kg/day) by oral lavage once daily or, alternatively, genistein is incorporated into food (150 mg/kg food).

For analysis, eyes are enucleated and, optionally, cut serially for histological study. For light or electron microscopy, eye cups are fixed for 2 hours in 2% osmium tetroxide in phosphate buffer, alcohol dehydrated and embedded in epoxy resin. Two micron thick sections are stained with toluidine blue and are used for light microscopy. For electron microscopy, thin sections are stained with lead citrate and uranyl acetate and examined in a JOEL JEM-100 CX 2 electron microscope (Hitachi, Tokyo, Japan). Ganglion cell loss and retinal pigment epithelium changes in control and treated subjects are then examined.

Specifically, if desired, the ganglion cell nuclei are counted to determine the degree of ganglion cell loss.

Example This example describes a method of determining the usefulness of a PTK pathway inhibitor as described herein to inhibit or ameliorate degeneration of the choroid, such as that which is due to aging.

Two strains of mice, a senile accelerated prone strain (SAMP) and a resistant strain (SAMR), obtained from Takeda Chemical Industries Ltd., Osaka, Japan, are employed to analyze the effect of genistein on degeneration of the choroid. The animals are kept as described in Example 4. Changes in the choriocapillaries, including atrophy of the choriocapillaries, are examined FWA-9. -11 "ThI--i zn-wa:X&jix4mt AAL-" WO 00/67738 PCT/US00/12339 28 via light and electron microscopy, as described in Example 4.

Choroidal vascular casts are also useful in analyzing the extent of choriocapillary atrophy. Subjects are anaesthetized with an intramuscular injection of 50 mg/kg body weight of Ketamine Xylazine. The left ventricle is perfused with 50 ml of heparinized lactated Ringer's solution. The mice are then sacrificed prior to injecting mercox solution (Ladd Research Industries, Burlington, VT) through the left ventricle. The eyes are enucleated and the anterior segment is separated by micro-dissection.

Corrosion casts of the posterior segment are made in 0.1 mol KOH. After complete bleaching of the tissues, the retinal vessels are separated from the choroidal vasculature by careful micro-dissection under water.

Choroidal vascular casts are mounted on aluminum tubs and sputter-coated with gold palladium prior to scanning electron microscopy (JOEL JSM-840A scanning electron microscope, Hitachi, Tokyo, Japan). Quantitative analysis of the choroidal vascular bed is performed by recording random areas in the posterior pole of each eye at 400 X magnification. Microplan II image analysis program (Laboratory Computer Systems Inc., MA) is used for measuring the area of choroidal vascular bed in each 400x photograph by tracing the area of choriocapillaries. The resultant values are tabulated and analyzed by paired student's t test. P values less than 0.05 are typically regarded as significant.

ki. S 'tL.uy~. A. AA.r. WO 00/67738 PCTIUSOO/1 2339 29 Example 6 This example describes a method of determining the usefulness of a PTK pathway inhibitor as described herein to inhibit or ameliorate thickening of Bruch's membrane, which is often associated with aging.

Two strains of mice, a senile accelerated prone strain (SAMP) and a resistant strain (SAMR), are obtained from Takeda Chemical Industries Ltd., Osaka, Japan, and employed to analyze the effect of genistein on thickening of Bruch's membrane. The animals are kept as described in Example 4. Changes in Bruch's membrane, including thickening of Bruch's membrane, are examined via light and electron microscopy, as described in Example 4.

All of the references cited herein, including patents, patent applications, literature publications, and the like, are hereby incorporated in their entireties by reference.

While this invention has been described with an emphasis upon preferred embodiments, it will be obvious to those of ordinary skill in the art that variations of the preferred compounds and methods may be used and that it is intended that the invention may be practiced otherwise than as specifically described herein.

Accordingly, this invention includes all modifications encompassed within the spirit and scope of the invention as defined by the following claims.

U-11 PZ M 1Ix

Claims (65)

1. A method of prophylactically or therapeutically treating an animal for age-related macular degeneration, which method comprises topically, subconjunctivally, retrobulbarly, subretinally, suprachoroidally, intraocularly, or periocularly administering to said animal an inhibitor of the protein tyrosine kinase pathway in an amount sufficient to treat said animal for age-related macular degeneration prophylactically or therapeutically.

2. The method of claim 1, wherein said inhibitor of the protein tyrosine kinase pathway is a compound of formula: x 7 w o v wherein V, W and X are selected from the group consisting of hydro, alkoxy, hydroxyl, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, and Y is selected from the group consisting of oxygen, sulfur, C(OH), and C=O, and Z is selected from the group consisting of hydro and C(O)ORi, wherein RI is an alkyl, or a protein tyrosine kinase-inhibiting prodrug or pharmaceutically acceptable salt thereof. 11111111 LWARMIM1-"1-~ WO 00/67738 PCT/US00/12339 31

3. A method of prophylactically or therapeutically treating an animal for age-related macular degeneration, which method comprises administering to said animal an inhibitor of the protein tyrosine kinase pathway, wherein said inhibitor is a compound of formula: I I w 0 \V wherein V and W are selected from the group consisting of hydro, alkoxy, hydroxyl, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, X is selected from the group consisting of hydro, alkoxy, halo, an ester, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, Y is selected from the group consisting of oxygen, sulfur, C(OH), and C=O, and Z is selected from the group consisting of hydro and C(O)OR 1 wherein R 1 is an alkyl, or a protein tyrosine kinase-inhibiting prodrug or pharmaceutically acceptable salt thereof, in an amount sufficient to treat said animal for age-related macular degeneration prophylactically or therapeutically.

4. A method of prophylactically or therapeutically treating an animal for age-related macular degeneration, which method comprises administering to said animal an -tn r y -8m li~n r U~ u ir~i~ i wuamontx..W' A W.Cnl~l'" W''rl_-L V f. WO 00/67738 PCT/US00/12339 32 inhibitor of the protein tyrosine kinase pathway, wherein said inhibitor is a compound of formula: x z W 0 1 V wherein W and X are selected from the group consisting of hydro, alkoxy, hydroxyl, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, V is selected from the group consisting of hydro, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, Y is selected from the group consisting of oxygen, sulfur, C(OH), and C=O, and Z is selected from the group consisting of hydro and C(O)OR 1 wherein R 1 is an alkyl, or a protein tyrosine kinase- inhibiting prodrug or pharmaceutically acceptable salt thereof, in an amount sufficient to treat said animal for age-related macular degeneration prophylactically or therapeutically. A method of prophylactically or therapeutically treating an animal for age-related macular degeneration, which method comprises administering to said animal an inhibitor of the protein tyrosine kinase pathway of formula: R MA' itjK WO 00/67738 PCT/US00/12339 33 X Z YV v wherein V and X are selected from the group consisting of hydro, alkoxy, hydroxyl, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, W is selected from the group consisting of alkoxy, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, Y is selected from the group consisting of oxygen, sulfur, C(OH), and C=O, and Z is selected from the group consisting of hydro and C(O)OR 1 wherein R 1 is an alkyl, or a protein tyrosine kinase- inhibiting prodrug or pharmaceutically acceptable salt thereof, in an amount sufficient to treat said animal for age-related macular degeneration prophylactically or therapeutically.

6. A method of prophylactically or therapeutically treating an animal for age-related macular degeneration, which method comprises administering to said animal an inhibitor of the protein tyrosine kinase pathway, wherein said inhibitor is a compound of formula: WO 00/67738 PCT/USOO/1 2339 34 X 7 w o v wherein V, W and X are selected from the group consisting of hydro, alkoxy, hydroxyl, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, Y is selected from the group consisting of sulfur, C(OH), and C=O, and Z is selected from the group consisting of hydro and C(0)ORI, wherein Ri is an alkyl, or a protein tyrosine kinase-inhibiting prodrug or pharmaceutically acceptable salt thereof, in an amount sufficient to treat said animal for age-related macular degeneration prophylactically or therapeutically.

7. A method of prophylactically or therapeutically treating an animal for age-related macular degeneration, which method comprises administering to said animal an inhibitor of the protein tyrosine kinase pathway, wherein said inhibitor is a compound of formula: x 7 WO 00/67738 PCT/US00/12339 wherein V, W and X are selected from the group consisting of hydro, alkoxy, hydroxyl, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, and Y is selected from the group consisting of oxygen, sulfur, C(OH), and C=O, and Z is C(O)ORI, wherein RI is an alkyl, or a protein tyrosine kinase-inhibiting prodrug or pharmaceutically acceptable salt thereof, in an amount sufficient to treat said animal for age-related macular degeneration prophylactically or therapeutically.

8. The method of any of claims 2-7, wherein the halo group is selected from the group consisting of fluorine, chlorine and bromine.

9. The method of any of claims 2-7, wherein the ester is a C 1 -C6 ester.

10. The method of any of claims 2-7, wherein the ether is a Ci-C 6 ether.

11. The method of any of claims 2-7, wherein said pharmaceutically acceptable salt of a carboxylic acid group is a sodium salt or a potassium salt.

12. The method of any of claims 2-7, wherein the alkyl groups are Ci-C 6 alkyl groups and the alkoxy group is a Ci-C alkoxy group.

13. The method of claim 2, wherein said inhibitor of the protein tyrosine kinase pathway is genistein. -i"~cnr n~l~mmi~i ml WM~ai% i m'Wni' I!rn~W'lil PCT/US00/12339 WO 00/67738 36

14. The method of any of claims 1-7, wherein age- related macular degeneration is that which results from age, hyperopia, systemic disease, smoking, light exposure, or nutritional deficiency.

15. The method of claim 14, wherein said systemic disease is cardiovascular disease or hypertension.

16. The method any of claims 1-7, wherein age- related macular degeneration is associated with an exudative complication.

17. The method of claim 16, wherein said exudative complication is choroidal neovascularization.

18. The method of claim 16, wherein said exudative complication is a retinal pigment epithelial detachment, a retinal pigment epithelial tear, disciform scarring, a vitreous hemorrhage or a subretinal hemorrhage.

19. The method of any of claims 1-7, wherein age- related macular degeneration is associated with an atrophic complication.

20. The method of claim 19, wherein said atrophic complication is drusen, a basal laminar deposit, a thickening of Bruch's membrane or retinal pigment epithelium atrophy.

21. The method of any of claims 3-7, wherein said inhibitor of the protein tyrosine kinase pathway is administered systemically. -I:R~rrMM V WO 00/67738 PCT/US00/12339 37

22. The method of claim 21, wherein said inhibitor of the protein tyrosine kinase pathway is administered in an amount from about 1 mg/kg/day to about 100 mg/kg/day.

23. The method of claim 22, wherein said inhibitor of the protein tyrosine kinase pathway is administered in an amount from about 15 mg/kg/day to about 50 mg/kg/day.

24. The method of any of claims 21-23, wherein said inhibitor of the protein tyrosine kinase pathway is administered orally or by injection. The method of any of claims 21-23, wherein said inhibitor of the protein tyrosine kinase pathway is administered upon appearance of retinal pigment epithelial hyperpigmentation and/or drusen.

26. The method of any of claims 3-7, wherein said inhibitor of the protein tyrosine kinase pathway is administered topically, subconjunctivally, retrobulbarly, periocularly, subretinally, suprachoroidally, or intraocularly.

27. The method of claim 1 or 26, wherein said inhibitor of the protein tyrosine kinase pathway is administered intraocularly in an amount from about 0.1 mg total to about 5 mg total.

28. The method of claim 27, wherein said inhibitor of the protein tyrosine kinase pathway is administered intraocularly in an amount from about 0.5 mg total to about 1 mg total. rwnrar~iY csc~rrio~i~iKnrunEnun~ri~iK~i~~L~z~n~~m WO 00/67738 PCT/US00/12339 38

29. The method of claim 13, wherein genistein is administered topically, subconjunctivally, retrobulbarly, periocularly, subretinally, suprachoroidally, or intraocularly. The method of claim 29, wherein genistein is administered intraocularly in an amount from about 0.1 mg total to about 5 mg total.

31. The method of claim 30, wherein genistein is administered intraocularly in an amount from about 0.5 mg total to about 1 mg total.

32. A method of prophylactically or therapeutically treating an animal for an exudative complication of age- related macular degeneration, which method comprises administering to said animal an inhibitor of the protein tyrosine kinase pathway in an amount sufficient to treat said animal for said exudative complication prophylactically or therapeutically.

33. The method of claim 32, wherein said exudative complication is a retinal pigment epithelial detachment, a retinal pigment epithelial tear, disciform scarring, a vitreous hemorrhage or a subretinal hemorrhage.

34. The method of claim 32 or 33, wherein said inhibitor of the protein tyrosine kinase pathway is a compound of formula: -;-~u~.~L~lrum~~~u.i~Rl~~nurrri~r ~lrumu~w~a~iviioum~~I11~.4U~~R~NIU~ WO 00/67738 PCT/US00/12339 39 x I w o v wherein V, W and X are selected from the group consisting of hydro, alkoxy, hydroxyl, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, and Y is selected from the group consisting of oxygen, sulfur, C(OH), and C=O, and Z is selected from the group consisting of hydro and C(O)ORI, wherein Ri is an alkyl, or a protein tyrosine kinase-inhibiting prodrug or pharmaceutically acceptable salt thereof. The method of claim 34, wherein the halo group is selected from the group consisting of fluorine, chlorine and bromine.

36. The method of claim 34, wherein the ester is a Cz-C 6 ester.

37. The method of claim 34, wherein the ether is a Cz-C 6 ether.

38. The method of claim 34, wherein said pharmaceutically acceptable salt of a carboxylic acid group is a sodium salt or a potassium salt. ~~~I~S~r3Y1*hlRli~ ~orrr WO 00/67738 PCT/US00/12339

39. The method of claim 34, wherein the alkyl groups are Ci-C6 alkyl groups and the alkoxy group is a C 1 -C6 alkoxy group.

40. The method of claim 34, wherein said inhibitor of the protein tyrosine kinase pathway is genistein.

41. The method of any of claims 32-39, wherein said inhibitor of the protein tyrosine kinase pathway is administered systemically.

42. The method of claim 41, wherein said inhibitor of the protein tyrosine kinase pathway is administered in an amount from about 1 mg/kg/day to about 100 mg/kg/day.

43. The method of claim 42, wherein said inhibitor of the protein tyrosine kinase pathway is administered in an amount from about 15 mg/kg/day to about 50 mg/kg/day.

44. The method of any of claims 41-43, wherein said inhibitor of the protein tyrosine kinase pathway is administered orally or by injection. The method of any of claims 32-39, wherein said inhibitor of the protein tyrosine kinase pathway is administered topically, subconjunctivally, retrobulbarly, periocularly, subretinally, suprachoroidally, or intraocularly.

46. The method of claim 45, wherein said inhibitor of the protein tyrosine kinase pathway is administered intraocularly in an amount from about 0.1 mg total to about 5 mg total. e- C' YW.A r rnuJ4~j L A~M' f.'H!n a WO 00/67738 PCT/USOO/12339 41

47. The method of claim 46, wherein said inhibitor of the protein tyrosine kinase pathway is administered intraocularly in an amount from about 0.5 mg total to about 1 mg total.

48. The method of claim 40, wherein genistein is administered systemically.

49. The method of claim 48, wherein genistein is administered in an amount from about 1 mg/kg/day to about 100 mg/kg/day. The method of claim 49, wherein genistein is administered in an amount from about 15 mg/kg/day to about 50 mg/kg/day.

51. The method of claim 48-50, wherein genistein is administered orally by injection.

52. The method of claim 40, wherein genistein is administered topically, subconjunctivally, retrobulbarly, periocularly, subretinally, suprachoroidally, or intraocularly.

53. The method of claim 52, wherein genistein is administered intraocularly in an amount from about 0.1 mg total to about 5 mg total.

54. The method of claim 53, wherein genistein is administered intraocularly in an amount from about 0.5 mg total to about 1 mg total. "L n fl'!C A method of prophylactically or therapeutically treating an animal for degeneration of the choroid, which method comprises administering to said animal an inhibitor of the protein tyrosine kinase pathway in an amount sufficient to treat said animal for degeneration of the choroid prophylactically or therapeutically.

56. A method of prophylactically or therapeutically treating an animal for thickening of Bruch's membrane, which method comprises administering to said animal an inhibitor of the protein tyrosine kinase pathway in an amount sufficient to treat said animal for thickening of Bruch's membrane prophylactically or therapeutically.

57. The method of claim 55 or claim 56, wherein said inhibitor of the protein tyrosine kinase pathway is a compound of formula: Y z w o oo *o 11 AMIM..,. .K .Iv2A i 2L jum r'y*i 43 wherein V, W and X are selected from the group consisting of hydro, alkoxy, hydroxyl, halo, an ester, an ether, a carboxylic acid group, a pharmaceutically acceptable salt of a carboxylic acid group, and -SR, in which R is hydrogen or an alkyl group, and Y is selected from the group consisting of oxygen, sulfur, C(OH), and C=O, and Z is selected from the group consisting of hydro and C(O)ORI, wherein R, is an alkyl, or a protein tyrosine kinase-inhibiting prodrug or pharmaceutically acceptable salt thereof.

58. The method of claim 57, wherein the halo group is selected from the group consisting of fluorine, chlorine and bromine.

59. The method of claim 57, wherein the ester is a Ci-C6 ester. The method of claim 57, wherein the ether is a C 1 -C 6 ether.

61. The method of claim 57, wherein said pharmaceutically acceptable salt of a carboxylic acid group is a sodium salt or a potassium salt. C 1 -C 6 alkyl groups and the alkoxy group is a C 1 -C 6 alkoxy group.

63. The method of claim 57, wherein said inhibitor of the protein tyrosine kinase pathway is genistein. **oo L uhii~mni.llruinnn.urm------ iL;~~min n 44

64. The method of any one of claims 55 to 62, wherein said inhibitor of the protein tyrosine kinase pathway is administered systemically.

65. The method of claim 64, wherein said inhibitor of the protein tyrosine kinase pathway is administered in an amount from 1 mg/kg/day to 100 mg/kg/day.

66. The method of claim 65, wherein said inhibitor of the protein tyrosine kinase pathway is administered in an amount from 15 mg/kg/day to 50 mg/kg/day.

67. The method of claims 64, wherein said inhibitor of the protein tyrosine kinase pathway is administered orally or by injection.

68. The method of claim 63, wherein genistein is administered systemically. 20 69. The method of claim 68, wherein genistein is administered in an amount from 1 mg/kg/day to 100 mg/kg/day.

70. The method of claim 69, wherein genistein is ~administered in an amount from 15 mg/kg/day to 50 mg/kg/day.

71. The method of claim 68, wherein said inhibitor of the protein tyrosine kinase pathway is administered orally or by injection. L* 30 72. The method of any one of claims 55 to 62, wherein said inhibitor of the protein tyrosine kinase pathway is I n~ iU,1,AOi il~~Uin~kAM m lIYUi1li~/l~i.i jli ii-1 administered topically, subconjunctivally, retrobulbarly, periocularly, subretinally, suprachoroidally, or intraocularly.

73. The method of claim 72, wherein said inhibitor of the protein tyrosine kinase pathway is administered intraocularly in an amount from 0.1 mg total to 5 mg total.

74. The method of claim 73, wherein said inhibitor of the protein tyrosine kinase pathway is administered intraocularly in an amount from 0.5 mg total to 1 mg total. The method of claim 63, wherein genistein is administered topically, subconjunctivally, retrobulbarly, periocularly, subretinally, suprachoroidally, or intraocularly.

76. The method of claim 75, wherein genistein is administered intraocularly in an amount from 0.1 mg total to •20 mg total. *.25

77. The method of claim 76, wherein genistein is administered intraocularly in an amount from 0.5 mg total to 1 mg total. DATED THIS FIFTEENTH DAY OF APRIL 2004 SJOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE BY PIZZEYS PATENT AND TRADE MARK ATTORNEYS V i w l a F i