EP2648711A2 - Compositions and methods for treating ophthalmic conditions - Google Patents

Abstract

Compositions of novel fenretinide powders that reduce serum RBP, and/or serum retinol- RBP levels are used to treat ophthalmic conditions such as geographic atrophy (GA), an advanced form of dry AMD and the prevention of choroidal neovascularization (CNV, or wet AMD).

Description

COMPOSITIONS AND METHODS FOR TREATING OPHTHALMIC CONDITIONS

CROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 61/420,211, filed December 6, 2010, which is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

[0002] The methods and compositions described herein are directed to the treatment of ophthalmic conditions.

BACKGROUND OF THE INVENTION

[0003] Geographic atrophy (GA) is related to the buildup of lipofuscin and A2E in the retinal pigment epithelium (RPE) leading to the death of the RPE and to the overlying photoreceptors. End plates shed by photoreceptors require processing by the RPE to move visual chromophore material derived from retinol through the visual cycle and return it to the photoreceptors. Vitamin A (retinol) is widely available from the diet and present in many tissues in the body, but requires a complex with retinol binding protein (RBP) and transthyretin for transport into the RPE to sustain the visual cycle. One of the effects of the visual cycle is production of toxic end products lipofuscin and A2E.

SUMMARY OF THE INVENTION

[0004] Presented herein are pharmaceutical compositions comprising (a) fenretinide powder in an amount therapeutically effective to reduce the serum RBP level in a human subject by at least about 60% or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients, wherein the diameter of at least 80% of the particles in the fenretinide powder in the composition is about or less than 10 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the composition is in the range of about 2 micron to about 10 micron. In some embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the composition is in the range of about 2 micron to about 6 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the composition is less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the composition is about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In some embodiments, the fenretinide powder in the composition is substantially free of particles having a diameter of 10 micron or more. In certain embodiments, the diameter of at least 90% of the particles in the fenretinide powder in the composition is less than about 10 micron.

[0005] In one aspect of the invention, provided herein are pharmaceutical compositions comprising (a) fenretinide powder that has been produced from a suspension comprising corn oil and/or polysorbate in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients.

[0006] In another aspect of the invention, provided herein pharmaceutical compositions comprising (a) a non-micronized fenretinide powder in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients.

[0007] In one aspect of the invention, provided herein are pharmaceutical compositions comprising (a) fenretinide powder, characterized by an X-ray powder diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1, in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients.

[0008] In another aspect of the invention, there are provided pharmaceutical compositions comprising (a) fenretinide powder, characterized by a DSC phase transition at a temperature of about 164°C, in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients.

[0009] In one aspect of the invention, provided herein are suspension pharmaceutical compositions prepared by (a) premixing corn oil, polysorbate 80, and fenretinide powder, and (b) high shear mixing the premixed corn oil, polysorbate 80, and fenretinide powder. In certain embodiments, the fenretinide powder does not aggregate in the suspension. In some

embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the composition is in the range of about 2 micron to about 10 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the composition is less than about 10 micron. [0010] In some embodiments of the pharmaceutical compositions provided herein, the one or more pharmaceutically acceptable excipients are selected from the group consisting of corn oil, polysorbate, micro crystalline cellulose, starch, hydro xypropylcellulose, sodium starch glycolate, isopropyl alcohol, magnesium stearate, and combinations thereof. In certain embodiments, the composition comprises corn oil. In some embodiments, the composition is administered orally. In certain embodiments, the composition is in a solid dosage form. In some embodiments, the solid dosage form is a tablet or a capsule. In certain embodiments, the composition is in suspension form. In some embodiments ,the diameter of at least 80% of the particles in the fenretinide powder in the composition is in the range of about 2 micron to about 6 micron. In some embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the composition is about 6 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the composition is greater than about 2 micron. In some embodiments, the fenretinide powder is characterized by a predominant endotherm at about 164°C as measured by DSC and a powder x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1. In certain embodiments, the therapeutically effective amount of fenretinide powder is at least about 100 mg. In some embodiments, the

therapeutically effective amount of fenretinide powder is at least about 300 mg.

[0011] In another aspect of the present invention, provided herein are methods for reducing the serum RBP level by at least about 60% in a human subject or reducing the rate of geographic atrophy lesion size increase in an eye of the human subject, comprising administering the invention pharmaceutical composition to a human subject in need thereof. Also provided herein are methods for reducing the serum RBP level of a human subject by at least about 2 mg/dL to about 2.5 mg/dL, comprising administering the invention pharmaceutical composition to a human subject in need thereof. Further provided herein are methods for treating Stargardt disease, comprising administering the invention pharmaceutical composition to a human subject in need thereof. In some embodiments, the effective amount of the pharmaceutical composition is administered orally to the human. In certain embodiments, the methods comprise multiple administrations of the effective amount of the pharmaceutical composition. In some

embodiments, the methods induce or cause delayed dark adaptation after administration of the composition. In certain embodiments, the methods reduce at least about 40% the rate of geographic atrophy lesion size increase after administration of the composition for 12 months. In certain embodiments, the methods reduce about 60% to about 80% the rate of geographic atrophy lesion size. In certain embodiments, the methods reduce the rate of geographic atrophy lesion size increase to less than about 1.0 mm² per year after administration of the composition for 12 months. In certain embodiments, the methods reduce the rate of geographic atrophy lesion size increase in less than about 0.5 mm² per year after administration of the composition for 12 months. In certain embodiments, the methods reduce the rate of geographic atrophy lesion size increase to about 0.5 mm² to about 1.0 mm² per year after administration of the composition for 12 months.

[0012] In another aspect are serum RBP monitoring kits for determining the therapeutically effective amount of fenretinide to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject comprising an RBP assay unit, and a means for obtaining a fluid sample from a patient who has been administered fenretinide. In some embodiments, the means for obtaining a fluid sample comprises a needle and the fluid is blood or serum.

[0013] In another aspect are compositions or methods for treating geographic atrophy in a mammal comprising reducing the serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP level in the mammal by a desired percentage. In certain embodiments, the desired percentage of serum retinol, serum retinol binding protein (RBP), and/or serum retinol- RBP reduction is relative to pre-therapeutic levels; in alternative embodiments, the desired percentage of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP reduction is relative to a pre-determined threshold level. In certain embodiments, the desired percentage of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP reduction is at least about 60%, at least about 70%, or at least about 80%. In certain

embodiments, the desired percentage of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP reduction is maintained for at least 1 week, for at least 1 month, for at least 6 months, for at least 12 months, for at least 18 months, for at least 24 months, for the lifetime of the mammal.

[0014] In some embodiments, the compositions or methods are also used to treat a lipofuscin-based retinal disease by reducing the serum level of retinol, RBP, and/or retinol-RBP in the body of a mammal, including embodiments wherein (a) the lipofuscin-based retinal disease is juvenile macular degeneration, including Stargardt Disease; (b) the lipofuscin-based retinal disease is dry form age-related macular degeneration; (c) the lipofuscin-based retinal disease is cone-rod dystrophy; (d) the lipofuscin-based retinal disease is retinitis pigmentosa; (e) the lipofuscin-based retinal disease is wet-form age-related macular degeneration; (f) the lipofuscin-based retinal disease is or presents geographic atrophy and/or photoreceptor degeneration; or (g) the lipofuscin-based retinal disease is a lipofuscin-based retinal

degeneration. [0015] In further embodiments, are compositions or methods for treating a lipofusin-based retinal disease in a mammal comprising reducing the serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP level in the mammal by a desired percentage. In certain embodiments, the desired percentage of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP reduction is relative to pre-therapeutic levels; in alternative embodiments, the desired percentage of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP reduction is relative to a pre-determined threshold level. In certain embodiments, the desired percentage of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP reduction is at least about 60%, at least about 70%, or at least about 80%. In certain embodiments, the desired percentage of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP reduction is maintained for at least 1 week, for at least 1 month, for at least 6 months, for at least 1 year, for the lifetime of the mammal.

[0016] In another aspect are compositions or methods for treating a lipofuscin-based retinal disease in a mammal comprising maintaining the serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP level in the mammal within a desired range. In certain embodiments, the desired range of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP is greater than a level that leads to diseases or conditions associated with Vitamin A deficiency and less than a level that increases the accumulation of A2E in at least one eye of the mammal. In certain embodiments, the level of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP that increases the accumulation of A2E in at least one eye of the mammal is at least about 60%, at least about 70%, or at least about 80% of the pre- therapy serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP level. In certain embodiments, the desired percentage of serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP reduction is maintained for at least 1 week, for at least 1 month, for at least 6 months, for at least 1 year, for the lifetime of the mammal. In certain embodiments, the serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP level in the mammal is measured at periodic levels to make sure that the serum retinol, serum retinol binding protein (RBP), and/or serum retinol-RBP level is maintained within a desired range.

[0017] In another aspect are compositions or methods for treating a lipofuscin-based retinal disease in a mammal comprising reducing the retinol level in at least one RPE of the mammal by a desired percentage. In certain embodiments, the desired percentage of retinol reduction is relative to pre-therapeutic levels; in alternative embodiments, the desired percentage of retinol reduction is relative to a pre-determined threshold level. In certain embodiments, the desired percentage of retinol reduction is at least about 60%, at least about 70%, or at least about 80%. In certain embodiments, the desired percentage of retinol reduction is maintained for at least 1 week, for at least 1 month, for at least 6 months, for at least 1 year, for the lifetime of the mammal.

[0018] The level of serum retinol, serum RBP, and serum retinol-RBP are inter-related. Reduction of the level of any one of these biological materials will lead to a reduction in the levels of the other two biological materials. Thus, hereinafter, the term "serum retinol" refers to any one or all of serum retinol, serum RBP, and serum retinol-RBP.

[0019] In another aspect are methods for reducing the formation of lipofuscin in an eye of a mammal comprising modulating the serum retinol level in the mammal by administering to the mammal at least once an effective amount of invention compositions.

[0020] In another aspect are methods for reducing the formation of drusen in an eye of a mammal comprising modulating the serum retinol level in the mammal by administering to the mammal at least once an effective amount of invention compositions.

[0021] In another aspect are methods for reducing and/or inhibiting choroidal

neovascularization in the eye of a mammal comprising modulating the serum retinol levels in the mammal by administering to the mammal at least once an effective amount of invention compositions.

[0022] In another aspect are methods for reducing the formation of abnormal blood vessel growth beneath the macula in an eye of a mammal comprising modulating the serum RBP level in the mammal by administering to the mammal at least once an effective amount of invention compositions.

[0023] In another aspect are methods for protecting the photoreceptors in any eye of a mammal comprising modulating the serum RBP level in the mammal by administering to the mammal at least once an effective amount of invention compositions.

[0024] In another aspect are methods for protecting an eye of a mammal from light comprising modulating the serum RBP level in the mammal by administering to the mammal at least once an effective amount of invention compositions.

[0025] In any of the aforementioned aspects are further embodiments comprising multiple administrations of the effective amount of the compound, including further embodiments in which (i) the time between multiple administrations is at least one week; (ii) the time between multiple administrations is at least one day; and (iii) the compound is administered to the mammal on a daily basis; or (iv) the compound is administered to the mammal every 12 hours. [0026] In any of the aforementioned aspects are further embodiments comprising (a) monitoring serum RBP levels; (b) monitoring formation of drusen in the eye of the mammal; (c) measuring levels of lipofuscin in the eye of the mammal by auto fluorescence; (d) measuring visual acuity in the eye of the mammal; (e) conducting a visual field examination on the eye of the mammal, including embodiments in which the visual field examination is a Humphrey visual field exam and/or microperimetry; (f) measuring the auto fluorescence or absorption spectra of N-retinylidene-phosphatidylethanolamine, dihydro-N-retinylidene-N-retinyl- phosphatidylethanolamine, N-retinylidene-N-retinyl-phosphatidylethanolamine, dihydro-N- retinylidene-N-retinyl-ethanolamine, and/or N-retinylidene-phosphatidylethanolamine in the eye of the mammal; (g) conducting a reading speed and/or reading acuity examination; (h) measuring scotoma size; or (i) measuring the size and number of the geographic atrophy lesions.

[0027] In any of the aforementioned aspects are further embodiments comprising an additional treatment for retinal degeneration.

[0028] In further embodiment of the pharmaceutical composition aspect, (a) the

pharmaceutically acceptable carrier comprises corn oil and a non-ionic surfactant; (b) the pharmaceutically acceptable carrier further comprises flour, a sweetener, and a humectant; (c) the pharmaceutically acceptable carrier comprises lysophosphatidylcholine, monoglyceride and a fatty acid; (d) the pharmaceutically acceptable carrier comprises dimyristoyl

phosphatidylcholine, soybean oil, t-butyl alcohol and water; (e) the pharmaceutically acceptable carrier comprises ethanol, alkoxylated caster oil, and a non- ionic surfactant; (f) the

pharmaceutically acceptable carrier comprises an extended release formulation; or (g) the pharmaceutically acceptable carrier comprises a rapid release formulation.

[0029] In another aspect is a polymorphic form of fenretinide characterized by a powder x- ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1.

[0030] In another aspect is a polymorphic form of fenretinide characterized by a

predominant endotherm at about 164°C as measured by DSC.

[0031] In another aspect is a polymorphic form of fenretinide characterized by: (i) a powder x-ray diffraction pattern wherein said x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1 and (ii) a predominant endotherm at about 164°C as measured by DSC. In another aspect is a polymorphic form of fenretinide prepared by precipitation of ethanol water solution at suitable temperature.

[0032] In a further aspect provided herein is a solid dosage form wherein the invention polymorphic form of fenretinide is present in at least about 50% to about 99.99% of the total weight of the fenretinide. BRIEF DESCRIPTION OF THE FIGURES

[0033] FIG. 1A-1C show particle size difference between lot No. 1 and lot No. 2 of fenretinide powders before and after formulation in a suspension comprising corn oil and polysorbate. FIG 1 A shows the appearance of both lots after retrieval from the formulation. FIG. IB illustrate the particle size distribution of both lots after formulation and FIG. 1C illustrates the particle size distribution before formulation.

[0034] FIG. 2 illustrates XRPD diagrams of lot No. 1 of fenretinide powders before and after formulation (suspension in corn oil and polysorbate).

[0035] FIG. 3 illustrates DSC diagrams of lot No. 1 of fenretinide powders before (3B) and after (3A) formulation (suspension in corn oil and polysorbate).

[0036] FIGS. 4A-4D illustrate pharmacokinetic analysis of three lots of fenretinide in rat. Concentrations of serum fenretinide (μΜ) for lot No. 1, lot No. 2 and lot No. 3 (2.4 micron particle size) were recorded for 24 hours following a single oral dose of the indicated lot (10 mg/kg) (FIG. 4A and 4C). Similarly, concentrations of serum retinol (μΜ) for lot No. 1, lot No. 2 and lot No. 3 were recorded (FIG. 4B and 4D).

[0037] FIG. 5 illustrates temporal analysis of serum RBP levels of patients with lot No. 1 (300 mg "Early EnroUers") vs. patients with lot No. 2 of fenretinide (300 mg "Late EnroUers") for 24 months.

[0038] FIG. 6A illustrates results of median GA lesion size growth over 24 months among all patients. FIG. 6B illustrates results of median GA lesion size growth among patients who received only lot No 1 for the entire 24 months.

[0039] FIG. 7A-7F show changes in lesion size (mm²) or as a percentage change from baseline vs. RBP reduction among patients in the placebo and the 300 mg cohorts ("Early EnroUers" received only lot No. 1, "Late EnroUers" received a mixture of lot No. 1 and lot No. 2) at 24 months. The y-axis of FIG. 7A-7C is lesion size in mm² while the y-axis of 7D-7F is lesion size increase in percentage.

[0040] FIGS. 8A-8D show correlation between RBP reduction (% from baseline) and lesion size growth. FIG. 8A and 8B show results from the placebo group and FIG. 8C and 8D show results from the 300 mg cohorts.

[0041] FIG. 9 shows mean visual acuity by ETDRS letters lost between patients who took 300 mg lot No. 1 ("Early EnroUers") of fenretinide and lot No. 2 ("Late EnroUers").

[0042] FIG. 10 shows mean visual acuity by ETDRS letters read between patients who took 300mg lot No. 1 ("Early EnroUers") of fenretinide and lot No. 2 ("Late EnroUers"). [0043] FIG. 11 shows results of fenretinide decreasing the emergence of choroidal neovascularization (CNV) by 50%. The incidence of CNV at 24 months in placebo patients was 22% (15/68), in patients receiving 100 mg fenretinide 13.5% (7/52), and in patients receiving 300 mg fenretinide 13.8% (8/58).

DETAILED DESCRIPTION OF THE INVENTION

[0044] Fenretinide can be used to provide benefit to patients suffering from or susceptible to various macular degenerations and dystrophies, including but not limited to dry- form age-related macular degeneration and Stargardt Disease. Specifically, fenretinide provide at least some of the following benefits to such human patients: reduction in the amount of all-trans-retinal (atRAL), reduction in the formation of A2E, reduction in the formation of lipofuscin, reduction in the formation of drusen, and reduction in light sensitivity. There is a reduced tendency to form A2E in ophthalmic and ocular tissues caused, in part, by a reduction in the over- accumulation of all-trans-retinal in these tissues. Because A2E itself is cytotoxic to the retinal pigment epithelium (RPE) (which can lead to retina cell death), administration of the invention fenretinide powder reduces the rate of accumulation of A2E, a cytotoxic agent, thus providing patient benefit. In addition, because A2E is the major fluorophore of lipofuscin, reduced quantities of A2E in ophthalmic and ocular tissues also results in a reduced tendency to accumulate lipofuscin in such tissues. Thus, in some respects the methods and compositions described herein can be considered to be lipofuscin-based treatments because administration of the invention fenretinide powder (alone, or in combination with other agents, as described herein) reduces, lowers or otherwise impacts the accumulation of lipofuscin in ophthalmic and/or ocular tissues. A reduction in the rate of accumulation of lipofuscin in ophthalmic and/or ocular tissues benefits patients that have diseases or conditions such as macular degenerations and/or dystrophies.

[0045] In addition, because dry- form age-related macular degeneration is often a precursor to wet-form age-related macular degeneration, the use of fenretinide can also be used as a preventative therapy for this latter ophthalmic condition. In addition, fenretinide may provide further therapeutic effect for wet-form age-related macular degeneration because such compounds additionally have anti-angiogenic activity.

[0046] Macular or Retinal Degenerations and Dystrophies. Macular degeneration (also referred to as retinal degeneration) is a disease of the eye that involves deterioration of the macula, the central portion of the retina. Approximately 85% to 90% of the cases of macular degeneration are the "dry" (atrophic or non-neovascular) type. In dry macular degeneration, the deterioration of the retina is associated with the formation of small yellow deposits, known as drusen, under the macula; in addition, the accumulation of lipofuscin in the RPE leads to photoreceptor degeneration and geographic atrophy. This phenomena leads to a thinning and drying out of the macula. The location and amount of thinning in the retina caused by the drusen directly correlates to the amount of central vision loss. Degeneration of the pigmented layer of the retina and photoreceptors overlying drusen become atrophic and can cause a slow loss of central vision. Ultimately, loss of retinal pigment epithelium and underlying photoreceptor cells results in geographic atrophy. Administration of fenretinide to a mammal can reduce the formation of, or limit the spread of, photoreceptor degeneration and/or geographic atrophy in the eye of the mammal. By way of example only, administration of hydro xypinaco lone retinoate (HPR) and/or N-(4-methoxyphenyl) retinamide (MPR) to a mammal, can be used to treat photoreceptor degeneration and/or geographic atrophy in the eye of the mammal.

[0047] In "wet" macular degeneration new blood vessels form (i.e., neovascularization) to improve the blood supply to retinal tissue, specifically beneath the macula, a portion of the retina that is responsible for our sharp central vision. The new vessels are easily damaged and sometimes rupture, causing bleeding and injury to the surrounding tissue. Although wet macular degeneration only occurs in about 10 percent of all macular degeneration cases, it accounts for approximately 90% of macular degeneration-related blindness. Neovascularization can lead to rapid loss of vision and eventual scarring of the retinal tissues and bleeding in the eye. This scar tissue and blood produces a dark, distorted area in the vision, often rendering the eye legally blind. Wet macular degeneration usually starts with distortion in the central field of vision. Straight lines become wavy. Many people with macular degeneration also report having blurred vision and blank spots (scotoma) in their visual field. Growth promoting proteins called vascular endothelial growth factor, or VEGF, have been targeted for triggering this abnormal vessel growth in the eye. This discovery has lead to aggressive research of experimental drugs that inhibit or block VEGF. Studies have shown that anti-VEGF agents can be used to block and prevent abnormal blood vessel growth. Such anti-VEGF agents stop or inhibit VEGF stimulation, so there is less growth of blood vessels. Such anti-VEGF agents may also be successful in anti-angiogenesis or blocking VEGF's ability to induce blood vessel growth beneath the retina, as well as blood vessel leakiness. Administration of fenretinide and the like to a mammal can reduce the formation of, or limit the spread of, wet-form age-related macular degeneration in the eye of the mammal. By way of example only, administration of HPR and/or MPR to a mammal, can be used to treat wet-form age-related macular degeneration in the eye of the mammal. Similarly, fenretinide and the like can be used to treat choroidal neovascularization and the formation of abnormal blood vessels beneath the macula of the eye of a mammal. Such therapeutic effect can result from a number of effects: lowering of serum retinol and thus ocular retinol levels; anti-angiogenic activity, and/or the quelling of geographic atrophy.

[0048] Geographic atrophy. Geographic atrophy the "dry" form of advanced AMD, results from atrophy to the retinal pigment epithelial layer below the retina, which causes vision loss through loss of photoreceptors (rods and cones) in the central part of the eye. Atrophy refers to the degeneration of the deepest cells, the retinal pigment epithelium (RPE), of the retina.

Degeneration of RPE cells leads to the death of rods and cones. GA tends to progress slowly. Progression is studied by auto fluorescence (AF) imaging to define the areas of GA or by high density optical coherence tomography (OCT) to determine when cells are becoming thinned or destroyed. By measuring enlargement of the area of atrophy, a doctor can estimate the loss in visual function. No medical or surgical treatment is available for this condition, however vitamin supplements with high doses of antioxidants, lutein and zeaxanthin, have been suggested by the National Eye Institute and others to slow the progression of dry macular degeneration and, in some patients, improve visual acuity.

[0049] In some embodiments, provided herein are methods for reducing the serum RBP level by at least about 60% in a human subject or reducing the rate of geographic atrophy lesion size increase in an eye of the human subject, comprising administering the invention

pharmaceutical composition to a human subject in need thereof. In some embodiments, the effective amount of the pharmaceutical composition is administered orally to the human. In certain embodiments, the methods comprise multiple administrations of the effective amount of the pharmaceutical composition. In some embodiments, the methods reduce visual acuity after administration of the composition for 18 months. In certain embodiments, the methods reduce at least about 40% the rate of geographic atrophy lesion size increase after administration of the composition for 12 months. In certain embodiments, the methods reduce about 60% to about 80% the rate of geographic atrophy lesion size increase.

[0050] The term "fenretinide powder" as used herein refers to the fenretinide solid in a particulate form, in which the diameter of at least 80% of the particles in the solid is about or less than 10 micron. In some embodiments, the diameter is less than 9 micron, less than 8 micron, less than 7 micron, less than 6 micron, less than 5 micron, less than 4 micron, or less than 2 micron. In some embodiments, the diameter of at least 80% of the particles in the solid is about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 4 micron, about 2 micron, or about 1 micron. The fenretinide powder may be prepared by precipitation, by crystallization or by micronization. Micronization is the reduction in particle size. In some embodiments, the invention fenretinide powder has been produced from a suspension

comprising corn oil and/or polysorbate.

[0051] Stargardt Disease is a macular dystrophy that manifests as a recessive form of macular degeneration with an onset during childhood. See e.g., Allikmets et al., Science, 277: 1805-07 (1997); Lewis et al, Am. J. Hum. Genet., 64:422-34 (1999); Stone et al, Nature Genetics, 20:328-29 (1998); Allikmets, Am. J. Hum. Gen., 67:793-799 (2000); Klevering, et al, Ophthalmology, 111 :546-553 (2004). Stargardt Disease is characterized clinically by

progressive loss of central vision and progressive atrophy of the RPE overlying the macula. Mutations in the human ABCA4 gene for Rim Protein (RmP) are responsible for Stargardt Disease. Early in the disease course, patients show delayed dark adaptation but otherwise normal rod function. Histologically, Stargardt Disease is associated with deposition of lipofuscin pigment granules in RPE cells.

[0052] In addition, there are several types of macular degenerations that affect children, teenagers or adults that are commonly known as early onset or juvenile macular degeneration. Many of these types are hereditary and are looked upon as macular dystrophies instead of degeneration. Some examples of macular dystrophies include: Cone-Rod Dystrophy, Corneal Dystrophy, Fuch's Dystrophy, Sorsby's Macular Dystrophy, Best Disease, and Juvenile

Retinoschisis, as well as Stargardt Disease.

[0053] The methods and compositions described herein include the use of N-oxides, crystalline forms (also known as polymorphs), or pharmaceutically acceptable salts of the fenretinide powder, as well as active metabolites of these compounds having the same type of activity. By way of example only, a known metabolite of fenretinide is N-(4-methoxyphenyl)retinamide, also known as 4-MPR or MPR. Another known metabolite of fenretinide is 4-oxo fenretinide. In some situations, compounds may exist as tautomers. All tautomers are included within the scope of the compounds presented herein. In addition, the compounds described herein can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. In some instances, the compounds described herein can exist in solvated forms with solvents such as pyridine, MEK, and the like. The solvated forms of the compounds presented herein are also considered to be disclosed herein.

PHARMACEUTICAL COMPOSITIONS

[0054] Another aspect are pharmaceutical compositions comprising (a) fenretinide powder in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients, wherein the diameter of at least 80% of the particles in the fenretinide powder is about or less than 10 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In certain embodiments, the fenretinide powder is substantially free of particles having the diameter of 10 micron or more. In some embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is less than about 10 micron, less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is about 10 micron, about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In certain embodiments, the fenretinide powder in the pharmaceutical composition is substantially free of particles having the diameter of 10 micron or more. In certain embodiments, the fenretinide powder of the composition is characterized by a predominant endotherm at about 164°C as measured by DSC and a powder x-ray diffraction pattern wherein said x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1.

[0055] In some embodiments provide herein pharmaceutical compositions comprising (a) fenretinide powder that has been produced from a suspension comprising corn oil and/or polysorbate in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is about or less than 10 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In certain embodiments, the fenretinide powder is substantially free of particles having the diameter of 10 micron or more. In some embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is less than about 10 micron, less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is about 10 micron, about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In certain embodiments, the fenretinide powder in the pharmaceutical composition is substantially free of particles having the diameter of 10 micron or more. In certain embodiments, the fenretinide powder of the composition is characterized by a predominant endotherm at about 164°C as measured by DSC and a powder x-ray diffraction pattern wherein said x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1.

[0056] Polysorbates are oily liquids derived from PEG-ylated sorbitan (a derivative of sorbitol) esterified with fatty acids. Exemplary polysorbates include, but not limited to, Polysorbate 20 (Tween 20 or polyoxy ethylene (20) sorbitan monolaurate), Polysorbate 40 (Tween 40 or polyoxy ethylene (20) sorbitan monopalmitate), Polysorbate 60 (Tween 60 or polyoxy ethylene (20) sorbitan monostearate), Polysorbate 80 (Tween 80 or polyoxy ethylene (20) sorbitan monooleate). The number 20 following the polyoxyethylene part refers to the total number of oxyethylene -(CH₂CH₂O)- groups found in the molecule. The number following the polysorbate part is related to the type of fatty acid associated with the polyoxyethylene sorbitan part of the molecule. Monolaurate is indicated by 20, monopalmitate is indicated by 40, monostearate by 60 and monooleate by 80. In some embodiments, the invention fenretinide powder is produced from any portion of PEG-ylated sorbitan esterified with fatty acids. For example, the invention fenretinide powder is produced from a composition comprising

Polysorbate 20, Polysorbate 40, Polysorbate 60, Polysorbate 80, or the like and/or combinations thereof.

[0057] Refined corn oil is 99%> triglyceride, with proportions of approximately 55%> polyunsaturated fatty acid, 30% monounsaturated fatty acid, and 15% saturated fatty acid. Of the saturated fatty acids, 80% are palmitic acid (lipid number of C16:0), 14% stearic acid (C18:0), and 3% arachidic acid (C20:0). Over 99% of the monounsaturated fatty acids are oleic acid (CI 8: 1 c). 98% of the polyunsaturated fatty acids are the omega-6 linoleic acid (CI 8:2 n-6 c,c) with the 2% remainder being the omega-3 alpha- lino lenic acid (CI 8:3 n-3 c,c,c). In some embodiments, the invention fenretinide powder is produced from any composition of corn oil or component thereof. [0058] In some embodiments, there are provided pharmaceutical compositions comprising (a) a non-micronized fenretinide powder in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more

pharmaceutically acceptable excipients In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is about or less than 10 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In certain embodiments, the fenretinide powder is substantially free of particles having the diameter of 10 micron or more. In some embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is less than about 10 micron, less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is about 10 micron, about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In certain embodiments, the fenretinide powder in the pharmaceutical composition is substantially free of particles having the diameter of 10 micron or more. In certain embodiments, the fenretinide powder of the composition is characterized by a predominant endotherm at about 164°C as measured by DSC and a powder x-ray diffraction pattern wherein said x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1.

[0059] In some instances, during the micronization processing, the particle gains charges due to constant grinding and/or interacting between particles. The charge is magnified in a very non- polar environment such as corn oil and/or polysorbate. As a result, in some instances, the particle aggregates and agglomerates into spheres and creates a larger particle. In one embodiment, the process of micronization provides a build-up of charges (e.g. ionizes) on the surface of the fenretinide powder, as such, in some embodiments, such surface charges promote agglomeration in nonpolar liquids, such as corn oil.

[0060] In some embodiments, there are provided pharmaceutical compositions comprising (a) fenretinide powder, characterized by an X-ray powder diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1, in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more

pharmaceutically acceptable excipients. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is about or less than 10 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In some embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is less than about 10 micron, less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is about 10 micron, about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In certain embodiments, the fenretinide powder in the pharmaceutical composition is substantially free of particles having the diameter of 10 micron or more. In certain embodiments, the fenretinide powder of the composition is characterized by a predominant endotherm at about 164°C as measured by DSC.

[0061] In some embodiments provide herein pharmaceutical compositions comprising (a) fenretinide powder, characterized by a DSC phase transition at a temperature of about 164°C, in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is about or less than 10 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder is less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 90% of the particles in the fenretinide powder is about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In some embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is less than about 10 micron, less than about 9 micron, less than about 8 micron, less than about 7 micron, less than about 6 micron, less than about 5 micron, less than about 4 micron, less than about 2 micron. In certain embodiments, the diameter of at least 80% of the particles in the fenretinide powder in the pharmaceutical composition is about 10 micron, about 9 micron, about 8 micron, about 7 micron, about 6 micron, about 5 micron, about 4 micron, about 2 micron, or about 1 micron. In certain embodiments, the fenretinide powder in the pharmaceutical composition is substantially free of particles having the diameter of 10 micron or more. In certain embodiments, the fenretinide powder of the composition is characterized by a powder x- ray diffraction pattern wherein said x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1.

[0062] In certain embodiments, the one or more pharmaceutically acceptable excipients of the invention compositions are selected from the group consisting of corn oil, polysorbate, micro crystalline cellulose, starch, hydroxypropylcellulose, sodium starch glycolate, isopropyl alcohol, magnesium stearate, and combinations thereof. In certain embodiments, the one or more pharmaceutically acceptable excipients comprise corn oil.

[0063] In some embodiments, provided herein are solid dosage forms of the invention fenretinide powder, wherein the polymorphic form of fenretinide is present in at least about 50% to about 99.99% of the total weight of fenretinide.

[0064] The term "diluent" refers to chemical compounds that are used to dilute the compound of interest prior to delivery. Diluents can also be used to stabilize compounds because they can provide a more stable environment. Salts dissolved in buffered solutions (which also can provide pH control or maintenance) are utilized as diluents in the art, including, but not limited to a phosphate buffered saline solution.

[0065] The term "physiologically acceptable" refers to a material, such as a carrier or diluent, that does not abrogate the biological activity or properties of the compound, and is nontoxic.

[0066] A "metabolite" of a compound disclosed herein is a derivative of that compound that is formed when the compound is metabolized. The term "active metabolite" refers to a biologically active derivative of a compound that is formed when the compound is metabolized. The term "metabolized" refers to the sum of the processes (including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes) by which a particular substance is changed by an organism. Thus, enzymes may produce specific structural alterations to a compound. For example, cytochrome P450 catalyzes a variety of oxidative and reductive reactions while uridine diphosphate glucuronyltransferases catalyze the transfer of an activated glucuronic-acid molecule to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines and free sulphydryl groups. Further information on metabolism may be obtained from The

Pharmacological Basis of Therapeutics, 9th Edition, McGraw-Hill (1996).

[0067] Metabolites of the compounds disclosed herein can be identified either by administration of compounds to a host and analysis of tissue samples from the host, or by incubation of compounds with hepatic cells in vitro and analysis of the resulting compounds. Both methods are well known in the art.

[0068] By way of example only, MPR is a known metabolite of HPR. MPR accumulates systemically in patients that have been chronically treated with HPR. One of the reasons that MPR accumulates systemically is that MPR is only (if at all) slowly metabolized, whereas HPR is metabolized to MPR. In addition, MPR may undergo relatively slow clearance. Thus, (a) the pharmacokinetics and pharmacodynamics of MPR must be taken into consideration when administering and determining the bioavailability of HPR, (b) MPR is more stable to

metabolism than HPR, and (c) MPR can be more immediately bio available than HPR following absorption. Another known metabolite of fenretinide is 4-oxo fenretinide.

[0069] MPR may also be considered an active metabolite. MPR (like HPR) can bind to Retinol Binding Protein (RBP) and prevent the binding of RBP to transthyretin (TTR). As a result, when either HPR or MPR is administered to a patient, one of the resulting expected features is that MPR will accumulate and bind to RBP and inhibit binding of retinol to RBP, as well as the binding of RBP to TTR. Accordingly, MPR can (a) serve as an inhibitor of retinol binding to RBP, (b) serve as an inhibitor of RBP to TTR, (c) limit the transport of retinol to certain tissues, including ophthalmic tissues, and (d) be transported by RBP to certain tissues, including ophthalmic tissues. MPR appears to bind more weakly to RBP than HPR, and is thus a less strong inhibitor of retinol binding to RBP. Nevertheless, both MPR and HPR are expected to inhibit, approximately equivalently, the binding of RBP to TTR. MPR has, in these respects, the same mode of action as HPR and can serve as a therapeutic agent in the methods and compositions described herein.

[0070] The invention fenretinide powder described herein can be administered to a human patient per se, or in pharmaceutical compositions where they are mixed with other active ingredients, as in combination therapy, or suitable carrier(s) or excipient(s). Techniques for formulation and administration of the compounds of the instant application may be found in "Remington: The Science and Practice of Pharmacy," 20th ed. (2000).

Routes of Administration

[0071] In some embodiments, invention compositions are administered orally. In certain embodiments, invention compositions are in a solid dosage form. In certain embodiments, the solid dosage form is a tablet or a capsule. In other embodiments, invention compositions are in suspension form. In certain embodiments, the suspension comprises corn oil or a component thereof and a non-ionic surfactant.

Composition/Formulation

[0072] Invention compositions may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or compression processes.

[0073] Invention compositions may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically.

Proper formulation is dependent upon the route of administration chosen. Any of the well- known techniques, carriers, and excipients may be used as suitable and as understood in the art; e.g., in Remington's Pharmaceutical Sciences, above.

[0074] The invention compositions can be administered in a variety of ways, including systemically, such as orally.

[0075] The invention compositions can illustratively take the form of a liquid where the agents are present in solution, in suspension or both. Typically when the composition is administered as a solution or suspension a first portion of the agent is present in solution and a second portion of the agent is present in particulate form, in suspension in a liquid matrix. In some embodiments, a liquid composition may include a gel formulation. In other embodiments, the liquid composition is aqueous. Alternatively, the composition can take the form of an ointment.

[0076] Useful aqueous suspension can also contain one or more polymers as suspending agents. Useful polymers include water-soluble polymers such as cellulosic polymers, e.g., hydroxypropyl methylcellulose, and water-insoluble polymers such as cross-linked carboxyl- containing polymers. Useful compositions can also comprise an acceptable mucoadhesive polymer, selected for example from carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methylmethacrylate), polyacrylamide, polycarbophil, acrylic acid/butyl acrylate copolymer, sodium alginate and dextran.

[0077] Useful compositions may also include solubilizing agents to aid in the solubility of invention fenretinide power. The term "solubilizing agent" generally includes agents that result in formation of a micellar solution or a true solution of the agent. Certain acceptable nonionic surfactants, for example polysorbate 80, can be useful as solubilizing agents, as can acceptable glycols, polyglycols, e.g., polyethylene glycol 400, and glycol ethers. [0078] Useful compositions may also include one or more pH adjusting agents or buffering agents, including acids such as acetic, boric, citric, lactic, phosphoric and hydrochloric acids; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate and tris-hydroxymethylaminomethane; and buffers such as

citrate/dextrose, sodium bicarbonate and ammonium chloride. Such acids, bases and buffers are included in an amount required to maintain pH of the composition in an acceptable range.

[0079] Useful compositions may also include one or more acceptable salts in an amount required to bring osmolality of the composition into an acceptable range. Such salts include those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions; suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite and ammonium sulfate.

[0080] Other useful compositions may also include one or more acceptable preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as merfen and thiomersal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide and cetylpyridinium chloride.

[0081] Still other useful compositions may include one or more acceptable surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxy ethylene fatty acid glycerides and vegetable oils, e.g., polyoxy ethylene (60)

hydrogenated castor oil; and polyoxy ethylene alkylethers and alkylphenyl ethers, e.g., octoxynol 10, octoxynol 40.

[0082] Still other useful compositions may include one or more antioxidants to enhance chemical stability where required. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.

[0083] Aqueous suspension compositions can be packaged in single-dose non-reclosable containers. Alternatively, multiple-dose reclosable containers can be used, in which case it is typical to include a preservative in the composition.

[0084] One useful formulation for solubilizing higher quantities of the invention

compositions are, by way of example only, positively, negatively or neutrally charged phospholipids, or bile salt/phosphatidylcholine mixed lipid aggregate systems, such as those described in Li, C.Y., et al., Pharm. Res. 13:907-913 (1996). An additional formulation that can be used for the same purpose with the invention compositions involves use of a solvent comprising an alcohol, such as ethanol, in combination with an alkoxylated caster oil. See, e.g., U.S. Patent Publication Number 2002/0183394. Or, alternatively, a formulation comprising the invention compositions is an emulsion composed of a lipoid dispersed in an aqueous phase, a stabilizing amount of a non-ionic surfactant, optionally a solvent, and optionally an isotonic agent. See id. Yet another formulation comprising the invention compositions includes corn oil and a non-ionic surfactant. See U.S. Patent No. 4,665,098. Still another formulation comprising the invention compositions includes lysophosphatidylcholine, monoglyceride and a fatty acid. See U.S. Patent No. 4,874,795. Still another formulation comprising the invention compositions includes flour, a sweetener, and a humectant. See International Publication No. WO

2004/069203. And still another formulation comprising the invention compositions includes dimyristoyl phosphatidylcholine, soybean oil, t-butyl alcohol and water. See U.S. Patent Application Publication No. US 2002/0143062.

[0085] For oral administration, the invention compositions can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers or excipients well known in the art. Such carriers enable the compounds described herein to be formulated as tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained by mixing one or more solid excipient with one or more of the compounds described herein, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as: for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, micro crystalline cellulose,

hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or others such as:

polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. If desired, disintegrating agents may be added, such as the cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0086] Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc,

polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different

combinations of active compound doses.

[0087] Pharmaceutical preparations which can be used orally include push- fit capsules made of gelatin, including by way of example only, soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol; or hard-gel capsules or tablets. The push- fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration.

[0088] In some embodiments, a pharmaceutical carrier for the invention compositions is a cosolvent system comprising benzyl alcohol, a nonpolar surfactant (e.g. a polysorbate), a water- miscible organic polymer, and an aqueous phase. The cosolvent system may be a 10% ethanol, 10%) polyethylene glycol 300, 10%> polyethylene glycol 40 castor oil (PEG-40 castor oil) with 70%o aqueous solution. This cosolvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. Naturally, the proportions of a cosolvent system may be varied considerably without destroying its solubility and toxicity characteristics. Furthermore, the identity of the cosolvent components may be varied: for example, other low-toxicity nonpolar surfactants may be used instead of PEG-40 castor oil, the fraction size of polyethylene glycol 300 may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides maybe included in the aqueous solution.

[0089] Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are well known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as N-methylpyrrolidone also may be employed, although usually at the cost of greater toxicity. Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.

[0090] One formulation for the administration of the invention compositions has been used with fenretinide powder in accordance with the present invention in the treatment of

neuroblastoma, prostate and ovarian cancers, and is marketed by Avanti Polar Lipids, Inc.

(Alabaster, Alabama) under the name Lym-X-Sorb™. This formulation, which comprises an organized lipid matrix that includes lysophosphatidylcholine, monoglyceride and fatty acid, is designed to improve the oral availability of fenretinide. Such a formulation, i.e., an oral formulation that includes lysophosphatidylcholine, monoglyceride and fatty acid, is proposed to also provide improved bioavailability of the invention compositions for the treatment of ophthalmic and ocular diseases and conditions, including but not limited to the macular degenerations and dystrophies. This formulation can be used in a range of orally- administered compositions, including by way of example only, a capsule and a powder that can be suspended in water to form a drinkable composition.

[0091] All of the formulations described herein may benefit from antioxidants, metal chelating agents, thiol containing compounds and other general stabilizing agents. Examples of such stabilizing agents, include, but are not limited to: (a) about 0.5% to about 2% w/v glycerol, (b) about 0.1%) to about 1% w/v methionine, (c) about 0.1%> to about 2% w/v monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% w/v ascorbic acid, (f) 0.003% to about 0.02% w/v polysorbate 80, (g) 0.001% to about 0.05% w/v. polysorbate 20, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrins, (1) pentosan polysulfate and other heparinoids, (m) divalent cations such as magnesium and zinc; or (n) combinations thereof.

TREATMENT METHODS AND DOSAGES

[0092] The term "subject" or "mammal" means all mammals including humans. Mammals include, by way of example only, humans, non-human primates, cows, dogs, cats, goats, sheep, pigs, rats, mice and rabbits.

[0093] The term "effective amount" as used herein refers to that amount of the compound being administered which will relieve to some extent one or more of the symptoms of the disease, condition or disorder being treated.

[0094] The invention compositions containing the compound(s) described herein can be administered for prophylactic and/or therapeutic treatments. The term "treating" is used to refer to either prophylactic and/or therapeutic treatments. In therapeutic applications, the

compositions are administered to a patient already suffering from a disease, condition or disorder, in an amount sufficient to cure or at least partially arrest the symptoms of the disease, disorder or condition. Amounts effective for this use will depend on the severity and course of the disease, disorder or condition, previous therapy, the patient's health status and response to the drugs, and the judgment of the treating physician. It is considered well within the skill of the art for one to determine such therapeutically effective amounts by routine experimentation (e.g., a dose escalation clinical trial).

[0095] In prophylactic applications, compositions containing the compounds described herein are administered to a patient susceptible to or otherwise at risk of a particular disease, disorder or condition. Such an amount is defined to be a "prophylactically effective amount or dose." In this use, the precise amounts also depend on the patient's state of health, weight, and the like. It is considered well within the skill of the art for one to determine such prophylactically effective amounts by routine experimentation (e.g., a dose escalation clinical trial).

[0096] The terms "enhance" or "enhancing" means to increase or prolong either in potency or duration a desired effect. Thus, in regard to enhancing the effect of therapeutic agents, the term "enhancing" refers to the ability to increase or prolong, either in potency or duration, the effect of other therapeutic agents on a system. An "enhancing-effective amount," as used herein, refers to an amount adequate to enhance the effect of another therapeutic agent in a desired system. When used in a patient, amounts effective for this use will depend on the severity and course of the disease, disorder or condition, previous therapy, the patient's health status and response to the drugs, and the judgment of the treating physician.

[0097] In the case wherein the patient's condition does not improve, upon the doctor's discretion the administration of the compounds may be administered chronically, that is, for an extended period of time, including throughout the duration of the patient's life in order to ameliorate or otherwise control or limit the symptoms of the patient's disease or condition.

[0098] In the case wherein the patient's status does improve, upon the doctor's discretion the administration of the compounds may be given continuously or temporarily suspended for a certain length of time (i.e., a "drug holiday").

[0099] Once improvement of the patient's conditions has occurred, a maintenance dose is administered if necessary. Subsequently, the dosage or the frequency of administration, or both, can be reduced, as a function of the symptoms, to a level at which the improved disease, disorder or condition is retained. Patients can, however, require intermittent treatment on a long-term basis upon any recurrence of symptoms.

[00100] The amount of a given agent that will correspond to such an amount will vary depending upon factors such as the particular compound, disease condition and its severity, the identity (e.g., weight) of the subject or host in need of treatment, but can nevertheless be routinely determined in a manner known in the art according to the particular circumstances surrounding the case, including, e.g., the specific agent being administered, the route of administration, the condition being treated, and the subject or host being treated. In general, however, doses employed for adult human treatment will typically be in the range of 0.02-5000 mg per day, preferably 1-1500 mg per day. The desired dose may conveniently be presented in a single dose or as divided doses administered simultaneously (or over a short period of time) or at appropriate intervals, for example as two, three, four or more sub-doses per day.

[00101] In some embodiments, the pharmaceutical compositions provided herein are used for reducing serum RBP levels by at least about 60% in a human subject. In some embodiments, serum RBP levels are reduced by at least 30%, at least 40%, at least 50%>, at least 60%>, at least 70%), or at least 80%>. In certain embodiments, serum RBP levels are reduced by about 30%>, about 40%, about 50%, about 60%, about 70%, or about 80%.

[00102] In certain embodiments, the pharmaceutical compositions provided herein are used for reducing the rate of geographic atrophy lesion size increase in an eye by at least about 60% in a human subject. In some embodiments, the rate of geography atrophy lesion size increase in an eye is reduced by at least 30%>, at least 40%>, at least 50%>, at least 60%>, at least 70%>, or at least 80%. In certain embodiments, the rate of geography atrophy lesion size increase in an eye is reduced by about 30%, about 40%, about 50%, about 60%>, about 70%, or about 80%.

[00103] In some embodiments, the pharmaceutical compositions provided herein are used for reducing the serum RBP level of a human by about 2 mg/dL to about 2.5 mg/dL. In certain embodiments, the serum RBP levels of a human are reduced by about 1.0 mg/dL, about 1.2 mg/dL, about 1.4 mg/dL, about 1.5 mg/dL, about 1.6 mg/dL, about 1.8 mg/dL, about 2.0 mg/dL, about 2.2 mg/dL, about 2.4 mg/dL, about 2.5 mg/dL, about 2.6 mg/dL, about 2.8 mg/dL, or about 3.0 mg/dL.

[00104] In certain embodiments, the pharmaceutical compositions provided herein are used for treating Stargardt Disease.

[00105] In some embodiments, the therapeutically effective amount of fenretinide powder in the invention compositions is at least about 100 mg. In other embodiments, the therapeutically effective amount of fenretinide powder is at least about 300 mg.

[00106] In certain instances, it may be appropriate to administer the invention fenretinide powder (or a pharmaceutically acceptable salt, ester, amide, or solvate) in combination with another therapeutic agent. By way of example only, if one of the side effects experienced by a patient upon receiving one of the compounds herein is inflammation, then it may be appropriate to administer an anti-inflammatory agent in combination with the initial therapeutic agent. Or, by way of example only, the therapeutic effectiveness of one of the compounds described herein may be enhanced by administration of an adjuvant (i.e., by itself the adjuvant may only have minimal therapeutic benefit, but in combination with another therapeutic agent, the overall therapeutic benefit to the patient is enhanced). Or, by way of example only, the benefit of experienced by a patient may be increased by administering one of the compounds described herein with another therapeutic agent (which also includes a therapeutic regimen) that also has therapeutic benefit. By way of example only, in a treatment for macular degeneration involving administration of one of the compounds described herein, increased therapeutic benefit may result by also providing the patient with other therapeutic agents or therapies for macular degeneration. In any case, regardless of the disease, disorder or condition being treated, the overall benefit experienced by the patient may simply be additive of the two therapeutic agents or the patient may experience a synergistic benefit.

[00107] In addition, the invention fenretinide powder or other agents that result in the reduction of serum retinol levels can be administered with (meaning before, during or after) agents that treat or alleviate side effects arising from serum retinol reduction. Such side effects include dry skin and dry eye. Accordingly, agents that alleviate or treat either dry skin or dry eye may be administered with the invention fenretinide powder or other agents that reduce serum retinol levels.

Detection of modulator activity

[00108] The compound and compositions disclosed herein can also be used in assays for detecting perturbations in RBP or TTR availability through conventional means. For example, a subject may be treated with invention fenretinide powder or compositions disclosed herein, and RBP or TTR levels quantified using conventional assay techniques. See Sundaram, M., et al, Biochem. J. 362:265-271 (2002). For example, a typical non-competitive sandwich assay is an assay disclosed in U.S. Pat. No. 4,486,530, incorporated herein by reference. In this method, a sandwich complex, for example an immune complex, is formed in an assay medium. The complex comprises the analyte, a first antibody, or binding member, that binds to the analyte and a second antibody, or binding member that binds to the analyte or a complex of the analyte and the first antibody, or binding member. Subsequently, the sandwich complex is detected and is related to the presence and/or amount of analyte in the sample. The sandwich complex is detected by virtue of the presence in the complex of a label wherein either or both the first antibody and the second antibody, or binding members, contain labels or substituents capable of combining with labels. The sample may be plasma, blood, feces, tissue, mucus, tears, saliva, or urine, for example for detecting modulation of clearance rates for RBP or TTR. For a more detailed discussion of this approach see U.S. Pat. Nos. Re 29,169 and 4,474,878, the relevant disclosures of which are incorporated herein by reference.

[00109] In a variation of the above sandwich assay, the sample in a suitable medium is contacted with labeled antibody or binding member for the analyte and incubated for a period of time. Then, the medium is contacted with a support to which is bound a second antibody, or binding member, for the analyte. After an incubation period, the support is separated from the medium and washed to remove unbound reagents. The support or the medium is examined for the presence of the label, which is related to the presence or amount of analyte. For a more detailed discussion of this approach see U.S. Pat. No. 4,098,876, the relevant disclosure of which is incorporated herein by reference.

[00110] The modulators disclosed herein may also be used in in vitro assays for detecting perturbations in RBP or TTR activity. For example, the modulator may be added to a sample comprising RBP, TTR and retinol to detect complex disruption. A component, for example, RBP, TTR, retinol or the modulator, may be labeled to determine if disruption of complex formation occurs. Complex formation and subsequent disruption may be detected and/or measured through conventional means, such as the sandwich assays disclosed above. Other detection systems may also be used to detect modulation of RBP or TTR binding, for example, FRET detection of RBP-TTR-retinol complex formation. See U.S. Provisional Patent

Application No. 60/625,532 "Fluorescence Assay for Modulators of Retinol Binding," herein incorporated by reference in its entirety.

[00111] In vitro gene expression assays may also be used to detect modulation of

transcription or translation of RBP or TTR by the modulators disclosed herein. For example, as described in Wodicka et al, Nature Biotechnology 15 (1997), (hereby incorporated by reference in its entirety), because mRNA hybridization correlates to gene expression level, hybridization patterns can be compared to determine differential gene expression. As a non-limiting example, hybridization patterns from samples treated with the modulators may be compared to hybridization patterns from samples which have not been treated or which have been treated with a different compound or with different amounts of the same compound. The samples may be analyzed using DNA array technology, see U.S. Patent No. 6,040,138, herein incorporated by reference in its entirety. Gene expression analysis of RBP or TTR activity may also be analyzed using recombinant DNA technology by analyzing the expression of reporter proteins driven by RBP or TTR promoter regions in an in vitro assay. See, e.g., Rapley and Walker, Molecular Biomethods Handbook (1998); Wilson and Walker, Principals and Techniques of Practical Biochemistry (2000), hereby incorporated by reference in its entirety.

[00112] In vitro translation assays may also be used to detect modulation or translation of RBP or TTR by the modulators disclosed herein. By way of example only, modulation of translation by the modulators may be detected through the use of cell- free protein translation systems, such as E. coli extract, rabbit reticulocyte lysate and wheat germ extract, see Spirin, A.S., Cell-free protein synthesis bioreactor (1991), herein incorporated by reference in its entirety, by comparing translation of proteins in the presence and absence of the modulators disclosed herein. Modulator effects on protein translation may also be monitored using protein gel electrophoretic or immune complex analysis to determine qualitative and quantitative differences after addition of the modulators.

[00113] In addition, other potential modulators which include, but are not limited to, small molecules, polypeptides, nucleic acids and antibodies, may also be screened using the in vitro detection methods described above. For example, the methods and compositions described herein may be used to screen small molecule libraries, nucleic acid libraries, peptide libraries or antibody libraries in conjunction with the teachings disclosed herein. Methods for screening libraries, such as combinatorial libraries and other libraries disclosed above, can be found in U.S. Pat. Nos. 5,591,646; 5,866,341; and 6,343,257, which are hereby incorporated by reference in its entirety.

In vivo detection of modulator activity

[00114] In addition to the in vitro methods disclosed above, the methods and compositions disclosed herein may also be used in conjunction with in vivo detection and/or quantitation of modulator activity on TTR or RBP availability. For example, labeled TTR or RBP may be injected into a subject, wherein a candidate modulator added before, during or after the injection of the labeled TTR or RBP. The subject may be a mammal, for example a human; however other mammals, such as primates, horse, dog, sheep, goat, rabbit, mice or rats may also be used. A biological sample is then removed from the subject and the label detected to determine TTR or RBP availability. A biological sample may comprise, but is not limited to, plasma, blood, urine, feces, mucus, tissue, tears or saliva. Detection of the labeled reagents disclosed herein may take place using any of the conventional means known to those of ordinary skill in the art, depending upon the nature of the label. Examples of monitoring devices for

chemiluminescence, radio labels and other labeling compounds can be found in U.S. Pats. No. 4,618,485; 5,981,202, the relevant disclosures of which are herein incorporated by reference.

On-site monitoring of RBP levels for dosage

[00115] The present invention also provide a serum RBP monitoring kit for determining a real time and accurate dosage for a patient who takes the invention fenretinide powder in a suitable formulation. The kit determines the therapeutically effective amount of fenretinide to reduce the serum RBP level by at least about 60% or reduce the rate of geographic atrophy lesion size increase in an eye comprise a RBP assay unit. The RBP assay unit may be any assay units known in the art. The RBP assay unit in some instances is an electronic basis assay unit that comprises reagents useful to detect serum RBP levels. PREPARATION OF THE FENRETINIDE POWDER

[00116] Optionally, fenretinide is synthesized using standard synthetic techniques known to those of skill in the art or using methods known in the art in combination with methods described herein. See, e.g., U.S. Patent Application Publication 2004/0102650; Um, S. J., et al., Chem. Pharm. Bull., 52:501-506 (2004). In addition, fenretinide is optionally purchased from various commercial suppliers. Optionally, the invention fenretinide powder is produced from a suspension comprising corn oil and/or polysorbate at suitable conditions, or by milled down (e.g., micronized) fenretinide solid (said solid can be produced by any means, including precipitation and crystallization) to a desired size that is about or less than 10 micron (e.g., 6 micron).

[00117] In some embodiments, the invention fenretinide powder is prepared by precipitation, e.g., from ethanol water solution, in which the solubility of the fenretinide powder is limited. In certain embodiment, the diameter of at least 80% of the particles in the fenretinide powder is about or less than 10 micron (e.g. about 6 micron).

[00118] In other embodiments, the invention fenretinide powder is prepared by particle size reduction (e.g. milling or micronization) of a fenretinide solid (prepared by any known methods including, but not limited to, a synthetic method before crystallization procedure; a

crystallization process from any solvent systems such as isopropanol water, ethanol water or other solvent systems; a precipitation procedure from any solvent systems that produce a supernatant comprising fenretinide) where the micronization reduces the diameter of most of (e.g. at least 50% , at least 60% or at least 70% or at least 80%) the particles in the fenretinide solid to about or less than 10 micron, or to about or less than 9 micron, or to about or less than 8 micron, or to about or less than 7 micron, or to about or less than 6 micron, or to about or less than 5 micron, or to about or less than 4 micron, or to about or less than 2 micron, or to about or less than 1 micron. An ordinary skill in the art would readily recognize other suitable methods to reduce the particle size of fenretinide solid to the desired diameter in accordance with the practice of the present invention.

[00119] In some embodiments, the present invention provides a polymorphic form of fenretinide characterized by a powder x-ray diffraction pattern wherein said x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1.

[00120] In other embodiments, the present invention provides a polymorphic form of fenretinide characterized by a predominant endotherm at 164°C as measured by DSC. [00121] In certain embodiments, the present invention provides a polymorphic form of fenretinide characterized by: (i) a powder x-ray diffraction pattern wherein said x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1 and (ii) a predominant endotherm at 164°C as measured by DSC.

[00122] The invention fenretinide powder is further formulated in accordance with the practice of the present invention. For example, the fenretinide powder is formulated to be a solid dosage form or a suspension comprising corn oil and a surfactant.

[00123] In some embodiments, provided herein is a solid dosage form of the invention fenretinide polymorphic form, wherein the polymorphic form of fenretinide is present in at least about 50% to about 99.99% of the total weight.

ILLUSTRATIVE EXAMPLES

[00124] The following examples provide illustrative methods for testing the effectiveness and safety of the invention compositions. These examples are provided for illustrative purposes only and not to limit the scope of the claims provided herein.

Example 1: Preparation of fenretinide powder lot No. 1 in capsule

[00125] Fenretinide was prepared in accordance with the known methods. The solid was then re-dissolved in ethanol-water solution either at room temperature or at other suitable temperature to produce a precipitate. The precipitate was filtered and dried under high vacuum to afford fenretinide powder. The powder was formulated in corn oil and polysorbate in capsule for clinical trial.

[00126] Upon measurement, the diameter of the fenretinide powder is about 5.4 micron (before formulation, FIG 1C). The powder was formulated in a suspension comprising corn oil and polysorbate in a capsule. The diameter of the fenretinide powder after retrieved from the suspension was about 5.5 micron (FIG. IB and 1 A). The after retrieved powder was tested by x- ray diffraction and differential scanning calorimetry (DSC) (See FIGS 2 and 3A). The x-ray diffraction of the solid shows peaks at angles 2 theta of 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1. The DSC plot shows a predominant endothermic peak at about 164°C. Lot No. 1 in capsule was used for subjects ("Early Enroller") in the clinical trial as in Example 4.

Example 2: Preparation of fenretinide powder lot No. 2 and lot No. 3.

[00127] Fenretinide was prepared in accordance with the known methods (e.g. crystallization from isopropyl alcohol water). The solid was further milled down to about 7.2 micron (D50) as lot No. 2 and about 2.4 micron (D50) as lot No. 3. Example 3: Preparation of fenretinide lot No. 2 in capsule

[00128] Lot No. 2 was further formulated to a suspension comprising corn oil and

polysorbate in a capsule. FIG 1 A and IB show the diameter of the particle in lot No. 2 retrieved from capsule formation is about 12 micron (D50) due to agglomeration. The distribution diagram of particle size in FIG IB shows that more particles in lot No. 2 have more than 10 micron in diameter (about 50%). On the other hand, particle size distribution of lot No. 1 retrieved from capsule formulation has similar distribution as before.

Example 4: Standard preparation of suspension versus pre-mixing

[00129] In the standard process for preparing a fenretinide suspension comprising corn oil and polysorbate fenretinide and corn oil are high shear mixed followed by the addition of polysorbate. In some instances (see for instance Example 3), fenretinide particles agglomerate after 4-6 days in suspension, increasing the particle size of the fenretinide in the pharmaceutical composition. Several modified processes were evaluated and the results are summarized below.

Experiment Particle Size t = 0 Particle Size t = 4-6 days

Standard Process: D(0.5) = = 4.6 μιη D(0 5) = = 21.4 μιη

Add corn oil, API then high shear mix; D(0.9) = = 9.5 μιη D(0 9) = = 35.6 μιη add polysorbate 80

Pre-mix Process: D(0.5) = = 9.8 μιη D(0 5) = = 8.5 μιη

Pre-mix corn oil, polysorbate 80, API; D(0.9) = = 18. 9 μιη D(0 9) = = 19.2 μιη high shear mix

Modified Standard Process with 50% D(0.5) = = 4.1 μιη* D(0 5) = = 4.4 μιη

less polysorbate 80 D(0.9) = = 10. 9 μιη D(0 9) = = 10.5 μιη

Modified Standard Process with 50%> D(0.5) = = 4.1 μιη D(0 5) = = 10.7 μιη more polysorbate 80 D(0.9) = = 8.7 μιη D(0 9) = = 21.9 μιη

Modified Standard Process without D(0.5) = = 2.5 μιη D(0 5) = = 2.5 μιη

polysorbate D(0.9) = = 5.2 μιη D(0 9) = = 4.8 μιη

* Initially formed a sticky mass, but continued slow stirring formed a suspension.

[00130] In a process that used less polysorbate, the particles did not appear to agglomerate after 4-6 days, but processing was difficult because the mixture initially formed a sticky mass. In a process that was using more polysorbate, agglomeration still occurred. But in a process that did not use polysorbate, no agglomeration was observed. In a pre-mixing process, wherein the corn oil and polysorbate excipients were pre-mixed with the fenretinide API before the high- shear mixing, no agglomeration was observed after 4-6 days. Example 5: PK/PD animal study of Lot No. 1 and Lot No. 2 fenretinide

[00131] Pharmacokinetic analysis of fenretinide powder prepared by Example 1 and 3 was conducted in rats. Fenretinide concentrations were measured against time (FIG. 4A). The results indicate Lot No. 1 has a better bioavailability as it has higher concentration initially (up to 8 hours) compared with Lot No. 2. Retinol concentrations were measured against time (FIG. 4B). The results show that fenretinide Lot No. 1 reduced retinol better compared to fenretinide Lot No. 2.

[00132] Interestingly, when lot No. 3 was used against lot No. 1 the results indicate Lot No. 3 (particle size of 2.4 micron) has about the same (or comparable) bioavailability as lot No. 1. See FIG. 4C and 4D.

Example 6: Analysis of fenretinide clinical trial data

[00133] Phase II proof-of concept, multicenter, double-masked, placebo-controlled study was conducted and analyzed.

[00134] Inclusion criteria:

Male or female, 50-89 yrs.

GA within 500 μιη of fovea

Total atrophic area 1 - 8 DA

(2.54 - 20.32 mm2)

FAF, other than none, focal or patchy

BCVA 20/20 - 20/100

Methods:

[00135] 246 patients were enrolled initially. Among them, 178 Subjects completed the study (24 months); of which 68 were in placebo group, 52 subjects received 100 mg of fenretinide, and 58 subjects received 300 mg of fenretinide. In particular, data of 137 subjects with RBP and lesion measurement at baseline and 24 months was collected. Among them, 52 patients received placebo; 41 patients received 100 mg fenretinide; and 44 patients received 300 mg fenretinide. Arms Assigned Interventions

100 mg fenretinide softgel capsules: Drug: Fenretinide

Active Comparator Once daily 30 minutes after the evening meal

Three (3) 100-mg fenretinide softgel for 24 months

capsules

Intervention: Drug: Fenretinide

Fenretinide and placebo softgel Drug: Fenretinide

capsules: Active Comparator Once daily 30 minutes after the evening meal

One (1) 100-mg fenretinide softgel for 24 months

capsule and two (2) placebo softgel

capsules

Intervention: Drug: Fenretinide

Placebo softgel capsules: Placebo Drug: Fenretinide

Comparator Once daily 30 minutes after the evening meal

Three (3) placebo softgel capsules for 24 months

Intervention: Drug: Fenretinide

Results:

[00136] Temporal analysis of serum RBP levels of the 300 mg cohorts shows that patients who took 300 mg fenretinide reduced RBP levels significantly in comparison with patients in the placebo group. In addition, the patients who took fenretinide lot No. 1 ("Early Enrollers") reduced more RBP than the patients who took lot No. 2 ("Late Enrollers") (FIG. 5).

[00137] An interim analysis was planned to take place 1 year after completion of patient enrollment (246 patients). Due to the "staggered" nature of enrollment, the 1-year time point included a number of patients who had reached 18-months of treatment (-30% from each arm). Lesion growth was measured in these patients using color fundus photography. The 18-month measurement from each patient was compared to their baseline lesion measurement. The results are shown below as the percent change from baseline over time. This analysis showed a time- and dose-dependent effect of fenretinide on slowing lesion growth, compared to placebo. The therapeutic effect of fenretinide was evident as early as 12 months and was most profound at 24 months in the 300 mg dosage group (-50% treatment effect, FIG. 6). Surprisingly, those patients who took lot No. 1 fenretinide powder ("Early Enrollers") (300mg dosage) have less change of lesion size growth from baseline compared to those who took lot No. 2 fenretinide powder ("Late Enrollers"). Following those patients who took lot No. 1 fenretinide powder from 18 to 24 months shows a therapeutic effect in 300 mg dosage group (FIG. 6A). Analysis of lesion growth in these same patients shows that the treatment effect was evident as early as 12 months (FIG 6B).

[00138] Lesion data from the remainder of patients initially analyzed at 24 months did not show the same therapeutic trend as observed for the 18-month cohort. However, when stratified by RBP levels, the data becomes quite strong (FIG. 7A-7F). In an effort to understand the absence of treatment efficacy at 24 months, an analysis of RBP levels in all patients at the 24- month time point was performed. The rationale for this analysis was based upon the hypothesis that slowing of lesion growth with fenretinide is dependent upon reductions in serum RBP. This analysis revealed that levels of RBP (the key bio marker) were highly variable among patients in the fenretinide arms. This outcome was unexpected when considered in the context of 30 years of clinical data to the contrary. In studies of hundreds of patients at doses of fenretinide ranging from 100 mg - 300 mg, RBP levels were consistently reduced 40% - 80% (FIG. 7B, 7C, 7E and 7F). Furthermore, the patients ("Early Enrollers") who took lot No. 1 fenretinide powder (formulated in corn oil and polysorbate) showed a surprising consistent 60%> to 80%> RBP level reduction compared to those who took lot No. 2 ("Late Enrollers"). The "Early Enrollers" also showed much less increased of GA lesion size (FIG. 7B v. 7C).

[00139] A secondary analysis of lesion growth as a function of reduction in RBP was also performed. In these correlation analyses, only patients with RBP and lesion data at baseline and 24 months were considered. Accordingly, only patients with reductions in RBP at 24 months could be considered. The y-axis in each plot shows the change (increase) in lesion size (FIG. 7A-7C) or in percentage (FIG. 7D-F), relative to baseline and the x-axis shows reduction in RBP relative to baseline. Symbols in each plot represent individual patients. The horizontal dashed line in each plot shows the median growth of lesions in the placebo group (-50% increase from baseline, ~2mm²/year). It is clear from the analysis that as RBP levels are more profoundly reduced, there is trend towards reducing lesion growth below the median lesion growth line of placebo. This trend was particularly evident in patients within the 300 mg dosage group. Data from the 300 mg group shows a marked reduction in median lesion growth (FIG. 7B, 7C, 7E and 7F) when RBP levels are reduced greater than 60% for early enrollers (i.e., patients who took lot No. 1 fenretinide, 7B and 7E) and greater than 40% for late enrollers (patients who took lot No. 2 fenretinide, 7C and 7F). These data indicate a "therapeutic window" of RBP reduction which must be achieved for slowing of lesion growth, approximately 60% RBP reduction when invention fenretinide powers are applied. Collectively, these findings support the proof of concept that reduction of circulating RBP reduces lesion growth in patients with GA. [00140] A total of 27 patients in the combined fenretinide treatment arms (100 mg and 300 mg) achieved an RBP reduction of 60% or greater for the duration of the trial. Of these 27 patients, 23 had a lesion growth which was below the median lesion growth of placebo (i.e., less than 2 mmVyear, less than 50% increase in lesion size growth relative to baseline). This indicates that 85% of patients with a sustained RBP reduction of 60% or greater will experience a treatment benefit against lesion growth. A similar observation is shown in FIG. 8 for the 300 mg dosage group.

Visual Acuity

[00141] Visual acuity (VA) is acuteness or clearness of vision, which is dependent on the sharpness of the retinal focus within the eye and the sensitivity of the interpretative faculty of the brain. The duration and extent of dark adaptation varies with the criterion of threshold. Luminance threshold can be measured for the resolution of a visual-acuity object.

[00142] Visual acuity data was collected on all patients at all time points. Consistent with published data, the placebo group showed a 9-11 letter loss over the 24 months of the study. In contrast, appropriately matched patients who took lot No. 1 fenretinide in the 300 mg cohort that showed a decrease in lesion growth only had a 6-letter loss (FIG. 9) or 61 > of letters read (FIG. 10) as measured by ETDRS letters.

Incidence of CNV

[00143] Although the study was not powered for prevention of the incidence of CNV (i.e., conversion from dry to wet AMD), the incidence of CNV was monitored. Fenretinide demonstrated a strong effect on slowing the conversion from dry to wet AMD. During the course of the fenretinide trial, more than 20% of patients in the placebo arm experienced CNV. Meanwhile, less than 15% of patients receiving fenretinide experienced CNV (FIG. 11).

Calculated odds ratios indicated a ~2.2-fold lower chance of developing CNV in patients receiving either dose of fenretinide, relative to placebo. At 24-months the combined therapeutic effect of fenretinide (100 mg and 300 mg patients) reached statistical significance (p = 0.039), compared to placebo. These data indicate that fenretinide is useful to slow, or prevent, the conversion from dry to wet AMD.

[00144] All of the methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. It will be apparent to those of skill in the art that variations may be applied to the methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents that are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

Claims

CLAIMS WHAT IS CLAIMED IS:

1. A pharmaceutical composition comprising (a) fenretinide powder in an amount

therapeutically effective to reduce the serum retinol binding protein (RBP) level in a human subject by at least about 60% or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients, wherein the diameter of at least 80% of the particles in the fenretinide powder in the composition is in the range of about 2 micron to about 10 micron.

2. The composition of claim 1, wherein the diameter of at least 80% of the particles in the fenretinide powder in the composition is in the range of about 2 micron to about 6 micron.

3. The composition of claim 1, wherein the fenretinide powder in the composition is

substantially free of particles having a diameter of 10 micron or more.

4. A pharmaceutical composition comprising (a) fenretinide powder that has been produced from a suspension comprising corn oil and/or polysorbate in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients.

5. A pharmaceutical composition comprising (a) a non-micronized fenretinide powder in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients.

6. A pharmaceutical composition comprising (a) fenretinide powder, characterized by an X-ray powder diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1, in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients.

7. A pharmaceutical composition comprising (a) fenretinide powder, characterized by a DSC phase transition at a temperature of about 164°C, in an amount therapeutically effective to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject, and (b) one or more pharmaceutically acceptable excipients.

8. A suspension pharmaceutical composition prepared by (a) premixing corn oil, polysorbate 80, and fenretinide powder and (b) high shear mixing the premixed corn oil, polysorbate 80 and fenretinide powder.

9. The composition of claim 8, wherein the fenretinide powder does not aggregate in the

suspension.

10. The composition of any of claims 1-7, wherein the one or more pharmaceutically acceptable excipients are selected from the group consisting of corn oil, polysorbate, micro crystalline cellulose, starch, hydro xypropylcellulose, sodium starch glycolate, isopropyl alcohol, magnesium stearate, and combinations thereof.

11. The composition of claim 10, wherein the composition comprises corn oil.

12. The composition of any of claims 1-9, wherein the composition is administered orally.

13. The composition of any of claims 1-9, wherein the composition is in a solid dosage form.

14. The composition of claim 13, wherein the solid dosage form is a tablet or a capsule.

15. The composition of any of claims 1-9, wherein the composition is in suspension form.

16. The composition of any of the preceding claims, wherein the diameter of at least 80% of the particles in the fenretinide powder in the composition is in the range of about 2 micron to about 6 micron.

17. The composition of any of the preceding claims, wherein the diameter of at least 80% of the particles in the fenretinide powder in the composition is greater than about 2 micron.

18. The composition of any of the preceding claims, wherein the fenretinide powder is

characterized by a predominant endotherm at about 164°C as measured by DSC and a powder x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1.

19. The composition of any of the preceding claims, wherein the therapeutically effective

amount of fenretinide powder is at least about 100 mg.

20. The composition of any of the preceding claims, wherein the therapeutically effective

amount of fenretinide powder is at least about 300 mg.

21. A method for reducing the serum RBP level by at least about 60% in a human subject or reducing the rate of geographic atrophy lesion size increase in an eye of the human subject, comprising administering the composition of any of claims 1-9 to a human in need thereof.

22. A method for reducing the serum RBP level of a human by at least about 2 mg/dL to about 2.5 mg/dL, comprising administering the composition of any of claims 1-9 to a human in need thereof.

23. A method for treating Stargardt disease, comprising administering the composition of any of claims 1-9 to a human in need thereof.

24. The method of any of claims 21-23, wherein the effective amount of the composition is administered orally to the human.

25. The method of any of claims 21-23, comprising multiple administrations of the effective amount of the composition.

26. The method of any of claims 21-23, wherein the method induces or causes delayed dark adaptation after administration of the composition.

27. The method of any of claims 21-23, wherein the method reduces by at least about 40% the rate of geographic atrophy lesion size increase after administration of the composition for 12 months.

28. The method claim 27, wherein the method reduces about 60% to about 80% of the rate of geographic atrophy lesion size increase.

29. The method of any of claims 21-23, wherein the method reduces the rate of geographic atrophy lesion size increase to less than about 1.0 mm² per year after administration of the composition for 12 months.

30. The method of claim 29, wherein the method reduces the rate of geographic atrophy lesion size increase to less than about 0.5 mm² per year after administration of the composition for 12 months.

31. The method of claim 29, wherein the method reduces the rate of geographic atrophy lesion size increase to about 0.5 mm² to about 1.0 mm² per year after administration of the composition for 12 months.

32. A serum RBP monitoring kit for determination the therapeutically effective amount of

fenretinide to reduce the serum RBP level by at least about 60% in a human subject or reduce the rate of geographic atrophy lesion size increase in an eye of the human subject comprising: (a) a case, said case comprising a layer of cushioned material, and (b) a RBP assay unit.

33. A polymorphic form of fenretinide characterized by a powder x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1.

34. A polymorphic form of fenretinide characterized by a predominant endotherm at about 164°C as measured by DSC.

35. A polymorphic form of fenretinide characterized by: (i) a powder x-ray diffraction pattern having 2 theta values at 7.0 ± 0.1, 17.2 ± 0.1, and 20.2 ± 0.1 and (ii) a predominant endotherm at about 164°C as measured by DSC.

36. A polymorphic form of fenretinide prepared by precipitation or crystallization from a

suspension comprising corn oil and/or polysorbate at suitable conditions.

37. A solid dosage form, wherein the polymorphic form of fenretinide of any of claims 33-36 is present in at least about 50% to about 99.99% of the total weight.