Treatment of ocular diseases using endothelin receptor antagonists - Patents.com

Abstract

The present disclosure relates to the discovery that certain ocular diseases that significantly affect the human visual system and, consequently, quality of life, can be treated using edonentan or A-182086. Examples of diseases include, but are not limited to, ocular neovascularization, neovascular glaucoma, vascular leakage, macular edema, and neovascular age-related macular degeneration. The present disclosure provides a method of preventing, treating, or ameliorating an ocular disease in a subject in need thereof, comprising contacting the subject's ocular tissue with a composition comprising a therapeutically effective amount of an endothelin receptor antagonist or a pharma- ceutically acceptable salt thereof.

Description

[Technical field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 182,750, filed April 30, 2021, the entire contents of which are incorporated by reference herein for all purposes. [Background technology]

[0002] background Examples of debilitating eye diseases include neovascular glaucoma, ocular neovascularization, vascular leak, macular edema, neovascular age-related macular degeneration, retinal vein occlusion (RVO), and retinopathy of prematurity (ROP). These eye diseases can cause various long-term damage to the eye and can ultimately cause blindness. They affect newborns, juveniles, adults of all ages, and the elderly, but only a handful of treatments exist. These treatments are for only a subset of eye diseases and slow but do not prevent blindness. The annual economic burden in the United States alone is over $100 billion.

[0003] Ocular neovascularization (the formation of new blood vessels from the existing vascular tree) is a leading cause of severe vision loss and significant visual impairment worldwide. It can affect various structures in the eye, including the retina, choroid, and cornea. It occurs when new abnormal blood vessels grow and spread throughout the retina and / or other parts of the eye (e.g., the tissue lining the back of the eye and the anterior chamber). The new abnormal blood vessels, in contrast to normal blood vessels, tend to leak, allowing fluid from the blood to enter the retina. The fluid can quickly distort vision and damage the retina.

[0004] Neovascular glaucoma (NVG) is a potentially blinding secondary glaucoma characterized by neovascularization of the iris, the development of elevated intraocular pressure (IOP), and often poor visual prognosis. NVG is a severe form of glaucoma that is attributed to the obstruction of aqueous humor outflow by new blood vessels secondary to posterior segment ischemia. It is associated with the development of a fibrovascular tissue membrane on the anterior surface of the iris and the iridocorneal angle of the anterior chamber.

[0005] Retinal vein occlusion (RVO) is a vascular disorder of the retina and is one of the most common causes of vision loss worldwide. Specifically, it is the second most common cause of blindness resulting from retinal vascular disease after diabetic retinopathy. RVO is often the result of underlying health problems (e.g., hypertension, high cholesterol levels, diabetes, and other health problems). There are two types of retinal vein occlusion: central retinal vein occlusion (CRVO) is a blockage of the main retinal vein, and branch retinal vein occlusion (BRVO) is a blockage of one of the smaller branch veins.

[0006] Currently, there is no way to unblock the retinal vein blockage, and accepted treatments are directed at addressing the health problems associated with retinal vein occlusion. Vision may return in eyes that have been affected by retinal vein occlusion. About one-third have some improvement, about one-third remain the same, and about one-third improve gradually, but it may take a year or longer to determine the final outcome. In some cases, the blocked blood vessels lead to fluid accumulation in the retina. In other cases, the development of ischemia causes the formation of new blood vessels. RVO is currently treated with intravitreal injections of anti-vascular endothelial growth factor (VEGF) drugs.

[0007] Retinopathy of prematurity (ROP) can occur due to premature birth. Abnormal leaky blood vessel growth (neovascularization) in the retina can occur secondary to other procedures related to premature birth and often results in newborn blindness. During pregnancy, blood vessels grow from the center of the developing child's retina at the mother's 16th week of pregnancy, then branch outward to reach the edge of the retina at the 8th month of pregnancy. In premature children, normal retinal blood vessel growth is incomplete and therefore can rupture more easily.

[0008] Thus, there is an unmet need to more effectively reduce the incidence of, treat or otherwise ameliorate neovascular glaucoma, ocular neovascularization, vascular leakage, macular edema, neovascular age-related macular degeneration, retinal vein occlusion (RVO), and retinopathy of prematurity (ROP). Summary of the Invention [Means for solving the problem]

[0009] Abstract The present disclosure provides a method for preventing, treating, or ameliorating ocular diseases in a subject in need thereof, comprising contacting the subject's ocular tissue with a composition comprising a therapeutically effective amount of an endothelin receptor antagonist or its pharma- ceutically acceptable salt.The ocular diseases that can be treated using the methods described herein include, but are not limited to, neovascular glaucoma, retinal vein occlusion (RVO), retinopathy of prematurity (ROP), ocular neovascularization, vascular leakage, neovascular age-related macular degeneration, and macular edema.

[0010] The method includes contacting the subject's ocular tissue with a composition comprising a therapeutically effective amount of an endothelin receptor antagonist or a pharma- ceutically acceptable salt thereof. In various embodiments, the endothelin receptor antagonist is selected from the group consisting of edonentan, tezosentan, A-182086, clazosentan, S1255, ACT-132577, enrasentan, and sparsentan. Preferably, the endothelin receptor antagonist is edonentan or A-182086.

[0011] The present disclosure also provides a method of preventing, treating, or ameliorating ocular neovascularization, vascular leakage, macular edema, or neovascular age-related macular degeneration in a subject in need thereof, comprising administering to the subject's ocular tissue a therapeutically effective amount of a compound of formula I: [ka] or a pharma- ceutically acceptable salt thereof. [Brief description of the drawings]

[0012] [Figure 1] Figure 1 shows optical coherence tomography-angiography (OCT-A) images from a representative experiment, revealing severe vasospasm of focal rabbit retinal vasculature 45 min after administration of 0.5 μg of endothelin-1 (ET-1) via intravitreal (IVT) injection.

[0013] [Diagram 2] FIG. 2 shows fluorescein angiography (FA) images revealing reversal of ET-1-induced vasospasm following IVT administration of 10 μg edonentan.

[0014] [Diagram 3]FIG. 3 shows a comparison of fluorescein dye velocity as an index of retinal blood flow in healthy rabbits (n=5 / group) following IVT administration of vehicle alone (control group), or 0.5 μg ET-1 alone, or 0.5 μg ET-1 and 10 μg edonentan, or 0.5 μg ET-1 and 10 μg A-182086 - revealing prolongation of dye velocity / reduced flow in ET-1 treated rabbits that is improved relative to control levels following treatment with edonentan or A-182086.

[0015] [Figure 4] FIG. 4 shows a comparison of neovascular area (NV) in 7-day-old neonatal C57BL / 6 mice with oxygen-induced ischemic retinopathy (OIR) following topical eye drops of edonentan, vehicle control, or intraperitoneal injection of 1 mg / kg aflibercept.

[0016] [Figure 5-1] FIG. 5A shows a comparison of retinal ganglion cell (RGC) numbers in the peripheral retina of rats with elevated intraocular pressure (IOP) after topical administration of vehicle alone (control group) or edonentan (n=4 rats / group for control, n=6 rats / group for edonentan). FIG. 5B shows a comparison of pattern electroretinogram (PERG) changes in rats with elevated intraocular pressure (IOP) after topical administration of vehicle alone (control group) or edonentan (n=4 rats / group for control, n=5 rats / group for edonentan). FIG. 5A and FIG. 5B demonstrate the prevention of RGC loss and the maintenance of RGC function after treatment with edonentan. [Figure 5-2] Figure 5C shows the pharmacokinetic profiles of topically or orally administered edonentan in rat plasma, retina / retinal pigment epithelium (RPE) / choroid, vitreous humor and aqueous humor. Figure 5C demonstrates the ability of edonentan to penetrate through the cornea / sclera and achieve retinal exposure after topical administration.

[0017] [Figure 6-1] FIG. 6A shows a comparison of retinal ganglion cell (RGC) numbers in the peripheral retina in rats with elevated intraocular pressure (IOP) following topical administration of vehicle alone (control group) or A-182086 (n=4 rats / group for control, n=6 rats / group for A-182086). FIG. 6B shows a comparison of pattern electroretinogram (PERG) changes in rats with elevated intraocular pressure (IOP) following topical administration of vehicle alone (control group) or A-182086 (n=4 rats / group for control, n=5 rats / group for A-182086). FIG. 6A and FIG. 6B demonstrate prevention of RGC loss and maintenance of RGC function following treatment with A-182086.

[0018] [Figure 6-2] Figure 6C shows the pharmacokinetic profiles of topically or orally administered A-182086 in rat plasma, retina / retinal pigment epithelium (RPE) / choroid, vitreous humor, and aqueous humor. Figure 6C demonstrates the ability of A-182086 to penetrate through the cornea / sclera and achieve retinal exposure after topical administration.

[0019] [Figure 7-1] Figures 7A-7L show laser speckle flow graphs (LSFG) of experimental glaucoma eyes and contralateral healthy eyes (controls) of three non-human primates compared in global average mean blur rate (MBR) or MBR change from baseline over time as an index of optic nerve head (ONH) blood flow in a laser-induced glaucoma model. Figure 7M shows the aggregate results from the three non-human primates. Figure 7N shows LSFG scans of one of the non-human primates at various selected time points. [Figure 7-2] Same as above. [Figure 7-3] Same as above. [Figure 7-4] Same as above. [Figure 7-5] Same as above. [Figure 7-6] Same as above. [Figure 7-7] Same as above. [Figure 7-8] Same as above. [Figure 7-9] Same as above. [Figure 7-10] Same as above. [Figure 7-11] Same as above. [Figure 7-12] Same as above. [Figure 7-13] Same as above. [Figure 7-14] Same as above. [Figure 7-15] Same as above.

[0020] [Figure 8] Figures 8A and 8B show a comparison of fluorescein dye velocity as an index of retinal blood flow in ET-1-induced rabbits (n=5 rabbits / group) following IVT administration of vehicle alone (control), 0.1 μg ET-1 and 10 μg edonentan, 0.1 μg ET-1 and 2.5 μg edonentan, 0.1 μg ET-1 and 0.5 μg edonentan, 0.1 μg ET-1 and 0.1 μg edonentan, or 0.1 μg ET-1 alone - revealing a dose response in the rabbit ET-1-induced vasospasm model.

[0021] [Figure 9-1] Figures 9A, 9B, 9C and 9D show the pharmacokinetic profiles of intravitreally delivered edonentan in rabbit plasma, retina, iris-ciliary body (ICB), retinal pigment epithelium (RPE) / choroid, vitreous humor or aqueous humor (Figures 9A, 9B, 9C and 9D) - revealing a longer t1 / 2 for edonentan. [Figure 9-2] Same as above. [Figure 9-3] Same as above. [Figure 9-4] Same as above.

[0022] [Figure 10]FIG. 10 shows the pharmacokinetic profile of topically administered edonentan in rabbit plasma, retina, vitreous humor and bulbar conjunctiva - demonstrating the ability of edonentan to penetrate through the ocular layers following a single topical administration to the eye.

[0023] [Figure 11] Figures 11A and 11B show the pharmacokinetic profiles of intravitreally delivered edonentan in the retina and retinal pigment epithelium (RPE) / choroid of rabbits administered two implants of the injection molded and ram extruded products (Figures 11A, 11B).

[0024] [Figure 12] FIG. 12 shows an exemplary overlay of the XRPD patterns of Forms 1-4.

[0025] [Figure 13] FIG. 13 shows an exemplary XRPD pattern of Form 1.

[0026] [Figure 14] FIG. 14 shows an exemplary XRPD pattern of Form 2.

[0027] [Figure 15] FIG. 15 shows an exemplary XRPD pattern of Form 3.

[0028] [Figure 16] FIG. 16 shows an exemplary XRPD pattern of Form 4.

[0029] [Figure 17] FIG. 17 shows an exemplary DSC curve for Form 1.

[0030] [Figure 18] FIG. 18 shows an exemplary DSC curve for Form 2.

[0031] [Figure 19] FIG. 19 shows an exemplary DSC curve for Form 3.

[0032] [Figure 20] FIG. 20 shows an exemplary DSC curve for Form 4.

[0033] [Figure 21] FIG. 21 shows the XRPD characteristic peaks of crystalline Form 4 shown in FIG.

[0034] [Figure 22] FIG. 22 shows the time course of edonentan retinal levels during a 12-week, single-dose intravitreal ocular pharmacokinetic study in pigmented rabbits receiving two implants of the injection molded product.

[0035] [Diagram 23] FIG. 23 shows the time course of edonentan RPE / choroid levels during a 12-week, single-dose intravitreal ocular pharmacokinetic study in pigmented rabbits receiving two implants of the injection molded product. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0036] Detailed Description The present disclosure provides a method for preventing, treating, or improving ocular neovascularization in a subject in need of such prevention, treatment, or improvement.Also provided herein is a method for preventing, treating, or improving vascular leakage or neovascular age-related macular degeneration in a subject in need of such prevention, treatment, or improvement.The present disclosure arises from the discovery that edonentan and A-182086 can be used to prevent, treat, or otherwise improve ocular diseases, including but not limited to neovascular glaucoma, retinal vein occlusion (RVO), and retinopathy of prematurity (ROP).

[0037] compound The methods of the present invention involve contacting or administering to ocular tissues (e.g., topically, intraocularly, intravitreally) a therapeutically effective amount of a compound described herein or a pharma- ceutically acceptable salt thereof. Compounds contemplated herein are endothelin receptor antagonists (e.g., edonentan, tezosentan, A-182086, clazosentan, S1255, ACT-132577, enrasentan, and sparsentan).

[0038] In certain embodiments, the compound is a compound of formula I: [ka] or a pharma- ceutically acceptable salt thereof.

[0039] The compound of formula I is also known as edonentan. Edonentan has the chemical name N-[[2'-[[(4,5-dimethyl-3-isoxazolyl)amino]sulfonyl]-4-(2-oxazolyl)[1,1'-biphenyl]-2-yl]methyl]-N,3,3-trimethylbutanamide (molecular weight 536.6 g / mol). Methods for preparing edonentan are well known to those skilled in the art. A suitable method is disclosed, for example, in U.S. Pat. No. 6,043,265. Edonentan is a highly selective and highly potent endothelin A receptor antagonist. Edonentan was developed as a second generation analogue after the discontinuation of the first clinical candidate, BMS-193884, which was being developed for the treatment of congestive heart failure (CHF). Edentan was in Phase I clinical trials until April 2002, when its development was discontinued.

[0040] In some embodiments, the compositions described herein have the following structure: [ka] or a pharma- ceutically acceptable salt thereof.

[0041] A-182086 has the chemical name (2R,3R,4S)-4-(2H-1,3-benzodioxol-5-yl)-2-(3-fluoro-4-methoxyphenyl)-1-[2-(N-propylpentane-1-sulfonamido)ethyl]pyrrolidine-3-carboxylic acid (molecular weight 578.7 g / mol). Methods for preparing A-182086 are well known to those skilled in the art. Suitable methods are disclosed, for example, in U.S. Pat. No. 6,162,927. A-182086 has a 4-fold increase in ET A / ET B Potent dual ET with selectivity A / ET B A-182086 is a receptor antagonist. To date, A-182086 has not been tested in a clinical setting.

[0042] As described herein, the disclosure provides a method for preventing, treating, or ameliorating ocular neovascularization in a subject in need thereof, the method comprising contacting an ocular tissue of the subject with a therapeutically effective amount of a compound of formula I or a composition comprising A-182086.

[0043] Also provided herein is a method of preventing, treating, or ameliorating vascular leakage in a subject in need thereof, the method comprising contacting ocular tissue of the subject with a therapeutically effective amount of a compound of formula I or a composition comprising A-182086.

[0044] The disclosure also provides a method of preventing, treating, or ameliorating neovascular age-related macular degeneration in a subject in need thereof, comprising contacting an ocular tissue of the subject with a therapeutically effective amount of a compound of formula I or a composition comprising A-182086.

[0045] Also provided herein is a method of preventing, treating, or ameliorating macular edema in a subject in need thereof, the method comprising contacting the subject's ocular tissue with a therapeutically effective amount of a compound of formula I or a composition comprising A-182086.

[0046] Crystal morphology The methods of the present invention involve contacting ocular tissue with or administering (e.g., topically, intraocularly, intravitreally) a solid form of a compound of formula I. In certain embodiments, a compound of formula I: [ka] is an anhydrous crystalline form (Form 4) having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from those at 5.6±0.2°, 11.4±0.2°, 17.7±0.2°, 19.3±0.2°, 21.1±0.2°, and 21.9±0.2° in terms of 2θ.

[0047] In some embodiments of the solid form, the anhydrous crystalline Form 4 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 5.6±0.2°, 11.4±0.2°, 17.7±0.2°, 19.3±0.2°, and 21.9±0.2°. In some embodiments of the solid form, the anhydrous crystalline Form 4 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 11.4±0.2°, 17.7±0.2°, and 19.3±0.2°. In some embodiments of the solid form, the anhydrous crystalline Form 4 has a T of about 163° C. by DSC analysis. m In some embodiments of the solid form, the anhydrous crystalline Form 4 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 5.6±0.2°, 11.4±0.2°, 17.7±0.2°, 19.3±0.2°, and 21.9±0.2°. In some embodiments of the solid form, the anhydrous crystalline Form 4 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 11.4±0.2°, 17.7±0.2°, and 19.3±0.2°. In some embodiments of the solid form, the anhydrous crystalline Form 4 has a T of about 163° C. by DSC analysis. m Shows.

[0048] In some embodiments, the compound is 90% or more by weight in crystalline form 4 based on the total weight of the compounds present in the composition. In some embodiments, the compound is 95% or more by weight in crystalline form 4 based on the total weight of the compounds present in the composition. In some embodiments, the compound is 96% or more by weight in crystalline form 4 based on the total weight of the compounds present in the composition. In some embodiments, the compound is 97% or more by weight in crystalline form 4 based on the total weight of the compounds present in the composition. In some embodiments, the compound is 98% or more by weight in crystalline form 4 based on the total weight of the compounds present in the composition. In some embodiments, the compound is 99% or more by weight in crystalline form 4 based on the total weight of the compounds present in the composition.

[0049] In certain embodiments, the compound of Formula I is in an anhydrous crystalline form (Form 1), wherein said anhydrous crystalline Form 1 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from those at 6.3±0.2°, 7.5±0.2°, 11.7±0.2°, 15.1±0.2°, and 17.3±0.2° in terms of 2θ; said compound is 90% by weight or more in crystalline Form 1, based on the total weight of compound present in the composition.

[0050] In certain embodiments, the compound of Formula I is a monohydrate crystalline form (Form 2), wherein the monohydrate crystalline Form 2 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from those at 9.6±0.2°, 10.4±0.2°, 19.6±0.2°, 19.7±0.2°, 22.0±0.2°, 22.9±0.2°, and 23.7±0.2° in terms of 2θ; the compound is 90% by weight or more in crystalline Form 2, based on the total weight of compound present in the composition.

[0051] In certain embodiments, the compound of Formula I is anhydrous crystalline (Form 3), wherein the anhydrous crystalline Form 3 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from those at 7.8±0.2°, 9.0±0.2°, 11.6±0.2°, 15.8±0.2°, and 19.1±0.2° in terms of 2θ; the compound is 90% by weight or more in crystalline Form 3, based on the total weight of compound present in the composition.

[0052] As used herein, the term "amorphous" refers to a solid material that does not have long-range order in the position of its molecules. Amorphous solids are generally supercooled liquids in which the molecules are arranged in a random manner, resulting in neither a well-defined arrangement (e.g., molecular packing) nor long-range order. Amorphous solids are generally isotropic, i.e., they exhibit similar properties in all directions, and do not have a distinct melting point. For example, an amorphous material is a solid material that does not have a sharp characteristic crystalline peak in its X-ray powder diffraction (XRPD) pattern (i.e., it is not crystalline as determined by XRPD). Instead, one or several broad peaks (e.g., halos) appear in its XRPD pattern.

[0053] Hydrate forms of crystalline edonentan are contemplated (e.g., edonentan·(HO) m (where m is a fraction or integer between about 0 and about 4, inclusive). For example, anhydrous or monohydrate forms of crystalline edonentan are contemplated herein. In one embodiment, the disclosed crystalline forms of edonentan may have a water level of about 1-10% by weight (e.g., 3-9% by weight or 5-8% by weight).

[0054] Biodegradable eye implants The methods of the present invention involve contacting ocular tissue with or administering (e.g., topically, intraocularly, intravitreally) a biodegradable ocular implant comprising a compound of formula I (also referred to herein as edonentan).

[0055] Biodegradable ocular implants containing edonentan as described herein may be used to prevent, treat, or ameliorate ocular neovascularization, vascular leakage, neovascular age-related macular degeneration, neovascular age-related macular degeneration, or macular edema in a subject in need thereof.

[0056] The biodegradable ocular implants described herein comprise a biodegradable polymer that includes a compound incorporated therein. In a preferred embodiment, the compound is a compound of Formula I.

[0057] In various embodiments, the implant has a diameter of about 300 μm to about 400 μm (e.g., about 300 μm, about 325 μm, about 350 μm, about 375 μm, and about 400 μm) and a length of about 4 mm to about 5 mm (e.g., about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, and about 5 mm). In certain embodiments, the implant has a diameter of about 300 μm and a length of about 4 mm. In certain embodiments, the implant has a diameter of about 340 μm and a length of about 4 mm.

[0058] In various embodiments, the implant has a total weight of about 250 μg to about 450 μg (e.g., about 250 μg, about 270 μg, about 290 μg, about 310 μg, about 330 μg, about 350 μg, about 370 μg, about 390 μg, about 410 μg, about 430 μg, and about 450 μg). In various embodiments, the implant has a total weight of about 300 μg to about 450 μg. In various embodiments, the implant has a total weight of about 350 μg to about 450 μg. In some embodiments, the implant has a total weight of about 380 μg.

[0059] In various embodiments, the concentration of the compound (e.g., the compound of Formula I) present in the biodegradable polymer in the biodegradable ocular implant is from about 5% w / w to about 95% w / w. (For example, about 10% w / w to about 95% w / w, about 15% w / w to about 95% w / w, about 20% w / w to about 95% w / w, about 25% w / w to about 95% w / w, about 30% w / w to about 95% w / w, about 35% w / w to about 95% w / w, about 40% w / w to about 95% w / w, about 45% w / w to about 95% w / w, about 50% w / w to about 95% w / w, about 55% w / w to about 95% w / w, about 60% w / w to about 95% w / w, about 65% w / w to about 95% w / w, about 70% w / w to about 95% w / w, about 75% w / w to about 95% w / w, about 80% w / w to about 95% w / w, about 85% w / w, about 95% w / w, about 90% w / w to about 95% w / w About 95% w / w, about 5% w / w to about 10% w / w, about 5% w / w to about 15% w / w, about 5% w / w to about 20% w / w, about 5% w / w to about 25% w / w, about 5% w / w to about 30% w / w, about 5% w / w to about 35% w / w, about 5% w / w to about 40% w / w, about 5% w / w to about 45% w / w, about 5% w / w In certain embodiments, the concentration of the compound present in the biodegradable polymer in the biodegradable ocular implant is about 20% w / w to about 60% w / w, about 5% w / w to about 55% w / w, about 5% w / w to about 60% w / w, about 5% w / w to about 65% w / w, about 5% w / w to about 70% w / w, about 5% w / w to about 75% w / w, about 5% w / w to about 80% w / w, about 5% w / w to about 85% w / w, and about 5% w / w to about 90% w / w. w / w (e.g., about 20% w / w to about 55% w / w, about 20% w / w to about 50% w / w, about 20% w / w to about 45% w / w, about 20% w / w to about 40% w / w, about 20% w / w to about 35% w / w, about 20% w / w to about 30% w / w, about 20% w / w to about 25% w / w, about 25% w / w to about 60% w / w, about 30% w / w to about 60% w / w, about 35% w / w to about 60% w / w, about 40% w / w to about 60% w / w, about 45% w / w to about 60% w / w, about 50% w / w to about 60% w / w, about 55% w / w to about 60% w / w). In certain embodiments, the concentration of compound present in the biodegradable polymer in the biodegradable ocular implant is from about 25% w / w to about 45% w / w.In certain embodiments, the concentration of the compound present in the biodegradable polymer in the biodegradable ocular implant is about 40% w / w to about 50% w / w (e.g., about 40% w / w to about 45% w / w, about 45% w / w to about 50% w / w). In various embodiments, the concentration of the compound is about 5% w / w, about 10% w / w, about 15% w / w, about 20% w / w, about 25% w / w, about 30% w / w, about 35% w / w, about 40% w / w, about 45% w / w, or about 50% w / w. In various embodiments, the concentration of the compound is about 30% w / w. In various embodiments, the concentration of the compound is about 40% w / w. In various embodiments, the concentration of the compound is about 45% w / w. In various embodiments, the concentration of the compound is about 50% w / w.

[0060] In an embodiment, the amount of the compound (e.g., compound of formula I, A-182086) present in the biodegradable polymer in the biodegradable ocular implant is from about 1 μg to about 500 μg (e.g., from about 10 μg to about 500 μg, from about 20 μg to about 500 μg, from about 30 μg to about 500 μg, from about 40 μg to about 500 μg, from about 50 μg to about 500 μg, from about 60 μg to about 500 μg, from about 70 μg to about 500 μg, from about 80 μg to about 500 μg, from about 90 μg to about 500 μg, from about 10 ... about 125 μg to about 500 μg, about 150 μg to about 500 μg, about 175 μg to about 500 μg, about 200 μg to about 500 μg, about 225 μg to about 500 μg, about 250 μg to about 500 μg, about 275 μg to about 500 μg, about 300 μg to about 500 μg, about 325 μg to about 500 μg, about 350 μg to about 500 μg, about 375 μg to about 500 μg, about 400 μg to about 500 μg, about 425 μg to about 500 μg, about 450 μg to about 500 μg, and about 475 μg to about 500 μg). In various embodiments, the amount of compound (e.g., compound of Formula I, A-182086) present in the biodegradable polymer in the biodegradable ocular implant is from about 70 μg to about 230 μg (e.g., about 70 μg, about 75 μg, about 80 μg, about 85 μg, about 90 μg, about 95 μg, about 100 μg, about 105 μg, about 110 μg, about 115 μg, about 120 μg, about 125 μg, about 130 μg, about 135 μg, about 140 μg, about 145 μg, about 150 μg, about 155 μg, about 160 μg, about 165 μg, about 170 μg, about 175 μg, about 180 μg, about 185 μg, about 190 μg, about 195 μg, about 200 μg, about 205 μg, about 210 μg, about 215 μg, about 220 μg, about 225 μg, and about 230 μg). In various embodiments, the amount of compound (e.g., compound of Formula I, A-182086) present in the biodegradable polymer in the biodegradable ocular implant is about 165 μg to about 220 μg (e.g., about 165 μg, about 170 μg, about 175 μg, about 180 μg, about 185 μg, about 190 μg, about 195 μg, about 200 μg, about 205 μg, about 210 μg, about 215 μg, and about 220 μg).In various embodiments, the amount of the compound (e.g., a compound of formula I, A-182086) present in the biodegradable polymer in the biodegradable ocular implant is about 150 μg to about 250 μg, about 300 μg to about 550 μg, or about 300 μg to about 600 μg. In various embodiments, the amount of the compound (e.g., a compound of formula I, A-182086) present in the biodegradable polymer in the biodegradable ocular implant is about 330 μg to about 500 μg (e.g., about 330 μg, about 335 μg, about 340 μg, about 345 μg, about 350 μg, about 355 μg, about 360 μg, about 365 μg, about 370 μg, about 375 μg, about 380 μg, about 385 μg, about 390 μg, about 400 μg, about 400 μg, about 45 ... 5 μg, about 390 μg, about 395 μg, about 400 μg, about 405 μg, about 410 μg, about 415 μg, about 420 μg, about 425 μg, about 430 μg, about 435 μg, about 440 μg, about 445 μg, about 450 μg, about 455 μg, about 460 μg, about 465 μg, about 470 μg, about 475 μg, about 480 μg, about 485 μg, about 490 μg, about 495 μg, and about 500 μg).

[0061] In some embodiments, the biodegradable ocular implant initially comprises at least about 95% to about 99% (e.g., about 95%, about 96%, about 97%, about 98%, and about 99%) of the matrix of the biodegradable polymer and the compound. In some embodiments, the biodegradable ocular implant initially comprises at least about 95% of the matrix of the biodegradable polymer and the compound. In some embodiments, the biodegradable ocular implant initially comprises at least about 80% to about 95% (e.g., about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, and about 95%) of the matrix of the biodegradable polymer and the compound.

[0062] The rate of therapeutic agent (e.g., a compound of Formula I) release from an intravitreal implant or particle suspension (e.g., a biodegradable ocular implant of the present disclosure) can depend on several factors, including, but not limited to, the surface area of ​​the implant, the therapeutic agent content, and the aqueous solubility of the therapeutic agent, as well as the rate of polymer degradation.

[0063] In some embodiments, less than 40% (e.g., about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, about 10%, and about 5%) of the compound is released from the biodegradable ocular implant when placed in phosphate buffered saline (PBS) for about one month. In some embodiments, less than 90% (e.g., about 90%, about 85%, about 80%, about 75%, about 70%, about 65%, about 60%, about 55%, about 50%, about 45%, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, about 10%, and about 5%) of the compound is released from the biodegradable ocular implant when placed in phosphate buffered saline (PBS) for about 1 month to about 12 months (e.g., about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months).

[0064] In various embodiments, the implant is administered as an intravitreal administration. Intravitreal administration refers to drug administration into the vitreous humor of the eye. In some embodiments, the implant is administered locally to the back of the eye. In some embodiments, the implant is injected into the intravitreal space using a needle and applicator. In some embodiments, the biodegradable ocular implant is administered in a dose range of about 1 μg to about 1 mg (e.g., about 1 μg, about 10 μg, about 25 μg, about 50 μg, about 75 μg, about 100 μg, about 125 μg, about 150 μg, about 175 μg, about 200 μg, about 225 μg, about 250 μg, about 275 μg, about 300 μg, about 325 μg, about 350 μg, about 375 μg, about 400 μg, about 425 μg, about 450 μg, about 475 μg, , about 500 μg, about 525 μg, about 550 μg, about 575 μg, about 600 μg, about 625 μg, about 650 μg, about 675 μg, about 700 μg, about 725 μg, about 750 μg, about 775 μg, about 800 μg, about 825 μg, about 850 μg, about 875 μg, about 900 μg, about 925 μg, about 950 μg, and about 975 μg). In some embodiments, the biodegradable ocular implant comprises a dose of the compound (e.g., a compound of formula I or a crystalline form thereof) in the range of about 10 μg to about 100 μg. In some embodiments, the biodegradable ocular implant comprises a dose of the compound (e.g., a compound of Formula I or a crystalline form thereof) in the range of about 500 μg to about 4 mg (e.g., about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, and about 3.5 mg). In some embodiments, the dose is about 150 μg to about 250 μg. In certain embodiments, the dose is about 165 μg to about 220 μg (e.g., about 165 μg, about 170 μg, about 175 μg, about 180 μg, about 185 μg, about 190 μg, about 195 μg, about 200 μg, about 205 μg, about 210 μg, about 215 μg, and about 220 μg). In some embodiments, the dose is about 300 μg to about 500 μg. In some embodiments, the dose is from about 300 μg to about 550 μg.In some embodiments, the dose is about 300 μg to about 600 μg. In certain embodiments, the dose is about 330 μg to about 500 μg (e.g., about 330 μg, about 335 μg, about 340 μg, about 345 μg, about 350 μg, about 355 μg, about 360 μg, about 365 μg, about 370 μg, about 375 μg, about 380 μg, about 385 μg, about 390 μg, about 395 μg, about 400 μg, about 4 05 μg, about 410 μg, about 415 μg, about 420 μg, about 425 μg, about 430 μg, about 435 μg, about 440 μg, about 445 μg, about 450 μg, about 455 μg, about 460 μg, about 465 μg, about 470 μg, about 475 μg, about 480 μg, about 485 μg, about 490 μg, about 495 μg, and about 500 μg). In some embodiments, the dose is about 200 μg to about 400 μg (e.g., about 200 μg, about 210 μg, about 220 μg, about 230 μg, about 240 μg, about 250 μg, about 260 μg, about 270 μg, about 280 μg, about 290 μg, about 300 μg, about 310 μg, about 320 μg, about 330 μg, about 340 μg, about 350 μg, about 360 μg, about 370 μg, about 380 μg, about 390 μg, about 400 μg). In some embodiments, the dose is about 175 μg.

[0065] In some embodiments, the biodegradable ocular implant may be a sterile biodegradable ocular implant. As used herein, "sterile" refers to a composition that meets the requirements of sterility enforced by drug regulatory authorities (e.g., MCA in the UK or FDA in the US). Tests are included within the current versions of compendia (e.g., British Pharmacopoeia and US Pharmacopoeia). In some embodiments, the biodegradable ocular implant is a substantially pure biodegradable ocular implant. In some embodiments, the biodegradable ocular implant is a medical grade biodegradable ocular implant. In some embodiments, the biodegradable ocular implant is administered into the intravitreal space every 3-12 months.

[0066] Biodegradable Polymers Suitable polymeric materials or compositions for use in the implants described herein include materials that are compatible with the eye, i.e., biocompatible, so as not to cause substantial interference with the function or physiology of the eye. Such polymeric materials may be biodegradable, bioerodible, or both biodegradable and bioerodible.

[0067] The terms "biodegrade" or "biodegradable," as used herein, generally refer to the biologically assisted degradation process that the polymers that make up the implant undergo in a biological environment (e.g., within a subject's body). Biodegradation is understood to include within its scope the processes of absorption, dissolution, breaking down, degradation, assimilation, or otherwise removal of the implant from the body (biological environment).

[0068] The term "polymer", as used herein, includes both homopolymers (polymers with only one type of repeat unit) and copolymers (polymers with more than one type of repeat unit).

[0069] The term "biodegradable polymer" as used herein refers to a polymer that degrades under physiological conditions in vivo. Release of the therapeutic agent occurs simultaneously with or subsequent to the degradation of the biodegradable polymer over time.

[0070] In a preferred embodiment, the biodegradable polymer is PLGA (poly(lactic-co-glycolic acid)). PLGA polymers are known to degrade via backbone hydrolysis (bulk erosion), with the end degradation products being lactic acid and glycolic acid, which are non-toxic and considered natural metabolic compounds. Lactic acid and glycolic acid are safely eliminated via the Krebs cycle by conversion to carbon dioxide and water.

[0071] PLGA is synthesized via random ring-opening copolymerization of cyclic dimers of glycolic acid and lactic acid. Successive monomer units of glycolic acid or lactic acid are linked together by ester linkages. The ratio of lactide to glycolide can be varied to change the biodegradation characteristics of the product. By varying the ratio, it is possible to tune the polymer degradation time. Importantly, drug release characteristics are influenced by the rate of biodegradation, molecular weight, and degree of crystallinity of the drug release system. By modifying and customizing the biodegradable polymer matrix, drug delivery profiles can be altered.

[0072] PLGA is primarily cleaved by non-enzymatic hydrolysis of its ester linkages throughout the polymer matrix in the presence of water in the surrounding tissue. PLGA polymers are biocompatible because they undergo hydrolysis in the body to produce the original monomers, lactic acid and / or glycolic acid. Lactic acid and glycolic acid are non-toxic and are safely eliminated by conversion to carbon dioxide and water via the Krebs cycle. The biocompatibility of PLGA polymers has been further tested in both non-ocular and ocular tissues in animals and humans. This finding indicates that the polymer is well tolerated.

[0073] Examples of PLGA polymers that may be utilized in embodiments of the present disclosure include, but are not limited to, Evonik Industries' RESOMER® product line (identified as RG502, RG502H, RG503, RG503H, RG504, RG504H, RG505, RG653H, RG750S, RG752H, RG752S, RG753H, RG753S, RG755S, RG756S, RG757S, and RG858S).

[0074] Such PLGA polymers include both acid and ester terminated polymers with intrinsic viscosities ranging from approximately 0.14 to approximately 1.7 dL / g as measured in an Ubbelhode size 0c glass capillary viscometer at 0.1% w / v in CHCl3 at 25° C. Exemplary polymers for use in various embodiments of the present disclosure may include variations in the molar ratio of D,L-lactide to glycolide from approximately 50:50 to approximately 85:15, including but not limited to 50:50, 65:35, 75:25, and 85:15.

[0075] Other examples of PLGA polymers that may be utilized in embodiments of the present disclosure include those produced by Lakeshore Biomaterials (identified, but not limited to, DLG 1A, DLG 3A, or DLG 4A). Such DLG polymers include both acid (A) and ester (E) terminated polymers with intrinsic viscosities ranging from approximately 0.0.5 to approximately 1.0 dL / g as measured in an Ubbelhode size 0c glass capillary viscometer at 0.1% w / v in CHCl3 at 25° C. Exemplary polymers used in various embodiments of the present disclosure may include variations in the molar ratio of D,L-lactide to glycolide from approximately 1:99 to approximately 99:1, including, but not limited to, 50:50, 65:35, 75:25, and 85:15.

[0076] RESOMERS® identified by “RG” or “DLG” in the product name (e.g., RG752S) have the general structure (V): [ka] The compound is poly(D,L-lactide-co-glycolide) or PLGA having the formula:

[0077] Synthesis of DLG of various molecular weights with various D,L-lactide-glycolide ratios is possible. In one embodiment, DLG with an intrinsic viscosity of about 0.05 to about 0.15 dL / g (e.g., 1A) can be used. In another embodiment, DLG with an intrinsic viscosity of about 0.15 to about 0.25 dL / g (e.g., 2A) can be used.

[0168] Poly(D,L-lactide-co-glycolide) or PLGA copolymers can be synthesized in various ratios of lactide to glycolide (e.g., lactide:glycolide ratio 75:25). These copolymers can be ester-terminated PLGA copolymers, as identified by the suffix "S" in the product name, or acid-terminated PLGA copolymers, as identified by the suffix "H" in the product name.

[0078] In some embodiments, the biodegradable ocular implants of the present disclosure comprise at least one PLGA, where each PLGA is independently selected from the group consisting of RG502, RG502S, RG502H, RG503, RG503H, RG504, RG504H, RG505, RG506, RG653H, RG752H, RG752S, RG753H, RG753S, RG755, RG755S, RG756, RG756S, RG757S, RG750S, RG858, and RG858S. In some embodiments, the biodegradable polymer comprises poly(lactic-co-glycolic acid) (PLGA), where the PLGA is selected from the group consisting of RG502, RG503H, RG503, RG752S, RG753S, RG755S, RG756S, and RG858S. In some embodiments, the biodegradable polymer comprises poly(lactic-co-glycolic acid) (PLGA), wherein the PLGA is selected from the group consisting of RG502, RG503, RG752S, RG753S, RG755S, RG756S, and RG858S. In some embodiments, the biodegradable ocular implant of the present disclosure comprises one PLGA. In some embodiments, the PLGA has a ratio of PLA and PLG of about 65:35.

[0079] In some embodiments, the biodegradable ocular implants of the present disclosure include at least two types of PLGA. In some embodiments, the biodegradable polymer includes at least three types of PLGA (e.g., 3-6 types of PLGA, 3 types of PLGA, 4 types of PLGA, 5 types of PLGA).

[0080] In some embodiments, the biodegradable ocular implants of the present disclosure comprise at least two types of PLGA, where each PLGA is independently selected from the group consisting of RG502, RG502H, RG503, RG503H, RG504, RG504H, RG505, RG653H, RG750S, RG752H, RG752S, RG753H, RG753S, RG755S, RG756S, RG757S, and RG858S. In some embodiments, the biodegradable ocular implants of the present disclosure comprise at least two types of PLGA in a ratio of about 99%:about 1% (e.g., about 98%:about 2%, about 97%:about 3%, about 96%:about 4%, about 95%:about 5%, about 94%:about 6%, about 95%:about 5%, about 94%:about 6%, about 93%:about 7%, about 92%:about 8%, about 91%:about 9%, about 90%:about 10%, about 90%:about 10%, about 89%:about 11%, about 88%:about 12%, about 87%:about 13%, about 87%:about 14%, about 88%:about 15%, about 88%:about 16%, about 88%:about 17%, about 88%:about 18%, about 88%:about 19%, about 98%:about 20%, about 98%:about 21%, about 98%:about 22%, about 98%:about 23%, about 98%:about 24%, about 98%:about 25%, about 98%:about 26%, about 98%:about 27%, about 98%:about 28%, about 98%:about 29%, about 98%:about 30%, about 98%:about 31%, about 98%:about 32%, about 98%:about 33%, about 98%:about 34%, about 98%:about 35%, about 98%:about 36%, about 98%:about 37%, about 98%:about 38%, about 98%:about 39%, about 98%:about 40%, about 98%:about 40%, about %: approx. 13%, approx. 86%: approx. 14%, approx. 85%: approx. 15%, approx. 84%: approx. 16%, approx. 83%: approx. 17%, approx. 82%: approx. 18%, approx. 81%: approx. 19%, approx. 80%: approx. 20%, approx. 79%: approx. 21%, approx. 78%: approx. 22%, approx. 77%: approx. 23%, approx. 76%: approx. 24%, approx. 75%: approx. 25%, approx. 74%: approx. 26%, approx. 73%: approx. 27%, approx. 72%: approx. 28%, approx. 71%: approx. 29%, approx. 70%: approx. 30%, approx. 69%: approx. 31%, approx. 68%: approx. 32%, approx. 67%: 33%, 66%: 34%, 65%: 35%, 64%: 36%, 63%: 37%, 62%: 38%, 61%: 39%, 60%: 40%, 59%: 41%, 58%: 42%, 57%: 43%, 56%: 44%, 55%: 45%, 54%: 46%, 53%: 47%, 52%: 48%, 51%: 49%, 50%: 50%, 49%: 51%, 48%: 52%, 47%: 5 3%, about 46%: about 54%, about 45%: about 55%, about 44%: about 56%, about 43%: about 57%, about 42%: about 58%, about 41%: about 59%, about 40%: about 60%, about 39%: about 61%, about 38%: about 62%, about 37%: about 63%, about 36%: about 64%, about 35%: about 65%, about 34%: about 66%, about 33%: about 67%, about 32%: about 68%, about 31%: about 69%, about 30%: about 70%, about 29%: about 71%, about 28%: about 72%, about 27%: about 73%,26%: 74%, 25%: 75%, 24%: 76%, 23%: 77%, 22%: 78%, 21%: 79%, 20%: 80%, 19%: 81%, 18%: 82%, 17%: 83%, 16%: 84%, 15%: 85%, 14%: 86%, The ratios of PLGA are 13%: about 87%, 12%: about 88%, 11%: about 89%, 10%, 90%, 9%: about 91%, 8%: about 92%, 7%: about 93%, 6%: about 94%, 5%: about 95%, 4%: about 96%, 3%: about 97%, 2%: about 98%, and 1%: about 99%. In some embodiments, the biodegradable ocular implant of the present disclosure comprises at least two types of PLGA in a ratio of about 50% to about 75%: about 25% to about 50% (e.g., about 50% to about 70%: about 30% to about 50%, about 50% to about 65%: about 35% to about 50%, about 50% to about 60%: about 40% to about 50%, and about 55%: about 45%). In certain embodiments, the biodegradable ocular implant of the present disclosure comprises at least two types of PLGA in a ratio of about 50%:about 50%. In an embodiment, the two types of PLGA are RG503 and RG503H. In an embodiment, the two types of PLGA are RG502 and RG502H. In an embodiment, the two types of PLGA are RG504 and RG504H.

[0081] In some embodiments, the biodegradable polymer comprises at least three different biodegradable polymers. In some embodiments, the biodegradable polymer comprises at least three PLGAs, where each PLGA is independently selected from the group consisting of RG502, RG502H, RG503, RG503H, RG504, RG504H, RG505, RG653H, RG750S, RG752H, RG752S, RG753H, RG753S, RG755S, RG756S, RG757S, and RG858S. In some embodiments, the biodegradable polymer comprises at least three types of PLGA, the at least three types being about 1% to about 95% (e.g., about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, and about 95%): about 1% to about 95% (e.g., about 1%, about 5%, about 10%, about 15%, about 20%, about 25% , about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, and about 95%): about 1% to about 95% (e.g., about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, and about 95%).

[0082] In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 40%:about 40%:about 20%. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 50%:about 10%:about 40%. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 10%:about 50%:about 40%. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 40%:about 40%:about 20%. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 10%:about 50%:about 40%. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 20%:about 60%:about 20%. In some embodiments, the biodegradable polymer comprises at least three PLGAs in a ratio of about 20%:about 50%:about 30%. In some embodiments, the biodegradable polymer comprises at least three PLGAs in a ratio of about 15%:about 50%:about 35%. In some embodiments, the biodegradable polymer comprises at least three PLGAs in a ratio of about 15%:about 45%:about 40%. In embodiments, each PLGA is independently selected from the group consisting of RG503, RG503H, and RG753S. In embodiments, each PLGA is independently selected from the group consisting of RG502, RG503, and RG753S. In embodiments, each PLGA is independently selected from the group consisting of RG502, RG503, and RG752S. In certain embodiments, each PLGA is independently selected from the group consisting of RG502, RG503, and RG755S. In certain embodiments, each PLGA is independently selected from the group consisting of RG502, RG503, and RG756S.

[0083] In some embodiments, the biodegradable polymer comprises at least four different biodegradable polymers. In some embodiments, the biodegradable polymer comprises at least four PLGAs, where each PLGA is independently selected from the group consisting of RG502, RG502H, RG503, RG503H, RG504, RG504H, RG505, RG653H, RG750S, RG752H, RG752S, RG753H, RG753S, RG755S, RG756S, RG757S, and RG858S. In certain embodiments, the biodegradable polymer comprises at least four PLGAs, where each PLGA is independently selected from the group consisting of RG502, RG503, RG753S, RG755S, RG756S, and RG858S. In certain embodiments, the biodegradable polymer comprises at least four PLGAs, where each PLGA is independently selected from the group consisting of RG502, RG503, RG753S, and RG858S.

[0084] In some embodiments, the biodegradable polymer comprises at least four types of PLGA, the at least four types of PLGA being about 1% to about 95% (e.g., about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, and about 95%): about 1% to about 95% (e.g., about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, and about 95%): about 1% to about 95% (e.g., about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, and about 95%): about 1% to about 95% (e.g., about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, and about 95%). In some embodiments, the biodegradable polymer comprises at least four types of PLGA in a ratio of about 10% to about 30% (e.g., about 10%, about 15%, about 20%, about 25%, and about 30%): about 20% to about 40% (e.g., about 20%, about 25%, about 30%, about 35%, about 40%): about 20% to about 40% (e.g., about 20%, about 25%, about 30%, about 35%, about 40%): about 10% to about 30% (e.g., about 10%, about 15%, about 20%, about 25%, and about 30%). In some embodiments, the biodegradable polymer contains at least four types of PLGA in a ratio of about 1% to about 20% (e.g., about 1%, about 5%, about 10%, about 15%, about 20%): about 40% to about 60% (e.g., about 40%, about 45%, about 50%, about 55%, about 60%): about 20% to about 40% (e.g., about 20%, about 25%, about 30%, about 35%, about 40%): about 1% to about 20% (e.g., about 1%, about 5%, about 10%, about 15%, about 20%).

[0085] In certain embodiments, the biodegradable polymer comprises at least four kinds of PLGA in a ratio of about 20%:about 30%:about 30%:about 20%. In certain embodiments, the biodegradable polymer comprises at least four kinds of PLGA in a ratio of about 10%:about 50%:about 30%:about 10%. Each of the four kinds of PLGA in the biodegradable polymer can be independently selected from the group consisting of RG502, RG503, RG753S, RG755S, RG756S, and RG858S. In some embodiments, each PLGA is independently RG502, RG503, RG753S, or RG858S.

[0086] In some embodiments, the biodegradable polymer (e.g., PLGA) is substantially biodegraded in about 1 month to about 24 months (e.g., about 2 months to about 24 months, about 5 months to about 24 months, about 7 months to about 10 months, about 10 months to about 24 months, about 12 months to about 24 months, about 15 months to about 24 months, about 17 months to about 24 months, about 20 months to about 24 months, and about 22 months to about 24 months). In some embodiments, the biodegradable polymer (e.g., PLGA) is substantially biodegraded in about 3 months to about 12 months (e.g., about 4 months to about 12 months, about 5 months to about 12 months, about 5 months to about 12 months, about 6 months to about 12 months, about 7 months to about 12 months, about 8 months to about 12 months, about 9 months to about 12 months, about 10 months to about 12 months, and about 11 months to about 12 months). In some embodiments, the biodegradable polymer (e.g., PLGA) is substantially biodegraded in about 12 months to about 18 months (e.g., about 13 months to about 18 months, about 14 months to about 18 months, about 15 months to about 18 months, about 16 months to about 18 months, and about 17 months to about 18 months). In some embodiments, the biodegradable polymer (e.g., PLGA) is substantially biodegraded in about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 12 months.

[0087] Also provided herein is a method for preventing, treating, or improving ocular neovascularization in a subject in need thereof, comprising contacting a biodegradable ocular implant comprising a biodegradable polymer, the biodegradable polymer comprising a compound incorporated therein; wherein the compound is a compound of formula I or a pharma- ceutically acceptable salt thereof. In certain embodiments, the biodegradable polymer comprises at least three types of PLGA. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 50%:about 10%:about 40%. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 20%:about 20%:about 60%. In certain embodiments, the three types of PLGA are selected from the group consisting of RG503, RG502, and RG753S. In some embodiments, the concentration of the compound of formula I in the biodegradable polymer is about 45% w / w, and the biodegradable polymer comprises RG503, RG502 and RG753S in a ratio of about 20%:about 20%:about 60%. In some embodiments, the concentration of the compound of formula I in the biodegradable polymer is about 45% w / w, and the biodegradable polymer comprises RG503, RG502 and RG753S in a ratio of about 50%:about 10%:about 40%. In some embodiments, the compound of formula I is an anhydrous crystalline form (e.g., form 1, 3, or 4) or a monohydrate crystalline form (e.g., form 2). In some embodiments, the compound of formula I is an anhydrous crystalline form (e.g., form 4).

[0088] Also provided herein is a method for preventing, treating, or ameliorating vascular leakage in a subject in need thereof, comprising contacting a biodegradable ocular implant comprising a biodegradable polymer, the biodegradable polymer comprising a compound incorporated therein; wherein the compound is a compound of formula I or a pharma- ceutically acceptable salt thereof. In certain embodiments, the biodegradable polymer comprises at least three types of PLGA. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 50%:about 10%:about 40%. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 20%:about 20%:about 60%. In certain embodiments, the three types of PLGA are selected from the group consisting of RG503, RG502, and RG753S. In some embodiments, the concentration of the compound of formula I in the biodegradable polymer is about 45% w / w, and the biodegradable polymer comprises RG503, RG502 and RG753S in a ratio of about 20%:about 20%:about 60%. In some embodiments, the concentration of the compound of formula I in the biodegradable polymer is about 45% w / w, and the biodegradable polymer comprises RG503, RG502 and RG753S in a ratio of about 50%:about 10%:about 40%. In some embodiments, the compound of formula I is an anhydrous crystalline form (e.g., form 1, 3, or 4) or a monohydrate crystalline form (e.g., form 2). In some embodiments, the compound of formula I is an anhydrous crystalline form (e.g., form 4).

[0089] Also provided is a method for preventing, treating, or ameliorating neovascular age-related macular degeneration in a subject in need thereof, comprising contacting a biodegradable ocular implant comprising a biodegradable polymer, the biodegradable polymer comprising a compound incorporated therein; wherein the compound is a compound of formula I or a pharma-ceutically acceptable salt thereof. In certain embodiments, the biodegradable polymer comprises at least three types of PLGA. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 50%:about 10%:about 40%. In some embodiments, the biodegradable polymer comprises at least three types of PLGA in a ratio of about 20%:about 20%:about 60%. In certain embodiments, the three types of PLGA are selected from the group consisting of RG503, RG502, and RG753S. In some embodiments, the concentration of the compound of formula I in the biodegradable polymer is about 45% w / w, and the biodegradable polymer comprises RG503, RG502 and RG753S in a ratio of about 20%:about 20%:about 60%. In some embodiments, the concentration of the compound of formula I in the biodegradable polymer is about 45% w / w, and the biodegradable polymer comprises RG503, RG502 and RG753S in a ratio of about 50%:about 10%:about 40%. In some embodiments, the compound of formula I is an anhydrous crystalline form (e.g., form 1, 3, or 4) or a monohydrate crystalline form (e.g., form 2). In some embodiments, the compound of formula I is an anhydrous crystalline form (e.g., form 4).

[0090] Also provided is a method for preventing, treating, or ameliorating macular edema in a subject in need thereof, comprising contacting a biodegradable ocular implant comprising a biodegradable polymer, the biodegradable polymer comprising a compound incorporated therein; wherein the compound is a compound of formula I or a pharma-ceutically acceptable salt thereof. In some embodiments, the biodegradable polymer comprises at least three PLGAs. In certain embodiments, the three PLGAs are selected from the group consisting of RG503, RG502, and RG753S. In some embodiments, the biodegradable polymer comprises at least three PLGAs in a ratio of about 20%:about 20%:about 60%. In some embodiments, the biodegradable polymer comprises at least three PLGAs in a ratio of about 50%:about 10%:about 40%. In some embodiments, the concentration of the compound of formula I in the biodegradable polymer is about 45% w / w, and the biodegradable polymer comprises RG503, RG502 and RG753S in a ratio of about 20%:about 20%:about 60%. In some embodiments, the concentration of the compound of formula I in the biodegradable polymer is about 45% w / w, and the biodegradable polymer comprises RG503, RG502 and RG753S in a ratio of about 50%:about 10%:about 40%. In some embodiments, the compound of formula I is an anhydrous crystalline form (e.g., form 1, 3, or 4) or a monohydrate crystalline form (e.g., form 2). In some embodiments, the compound of formula I is an anhydrous crystalline form (e.g., form 4).

[0091] How to Make an Implant The method of making the biodegradable ocular implants described herein includes subjecting a biodegradable polymer containing a compound to solvent casting, injection molding, or extrusion, wherein the compound is a compound of Formula I: [ka] or a pharma- ceutically acceptable salt thereof.

[0092] Prior to implant fabrication, the blend of the polymer matrix and therapeutic agent can be dissolved and mixed with a solvent to produce a therapeutic agent that is homogeneously dispersed throughout the body of the implant. The blends prepared can each contain multiple, for example, three, different PLGA polymers in various ratios. The PLGA polymers used to produce the pharmaceutical compositions of the present invention can include, but are not limited to, RESOMER® RG502, RG503, RG752S, RG753S, and 65 / 35 PLA / PLG, all of which are commercially available.

[0093] The following is an exemplary procedure used to prepare the compositions of the present invention: For example, the polymer is dissolved in an organic solvent (e.g., methylene chloride) in a specific ratio. The therapeutic agent (e.g., edonentan) is then added to the polymer solution and dissolved. The methylene chloride is then evaporated at room temperature in a polytetrafluoroethylene (PTFE) dish. After the methylene chloride is evaporated, a thin film of homogeneous material remains. In one embodiment, the thin film ranges in thickness from 200 μm to 300 μm.

[0094] The remaining homogenous film is then ground into a powder using a cryogenic mill. A small portion of the film is added to a stainless steel cryogenic mill vessel containing 2-3 appropriately sized grinding balls and pre-cooled using liquid nitrogen at 5 Hz for 2-3 minutes. The material is then milled for 1 minute at 20 Hz-25 Hz and rested for 1 minute at 5 Hz. This mill / rest cycle is repeated 2-5 times. The resulting material is a coarse to fine powder of homogenous material.

[0095] In one embodiment, the implant of the present invention can be prepared using the homogenous material described above. In one embodiment, the implant is formed by injection molding. Injection molding can be performed, for example, by a suitable injection molding machine, such as a modified Haake MiniJet (ThermoFisher Scientific). The following is an exemplary procedure used to prepare the implant of the present invention.

[0096] The homogenous powder is loaded and injected into a mould consisting of a suitable sized channel (e.g. 300 μm×12 mm). The powder is loaded into a barrel leading to the mould and the mould is placed under vacuum. The temperature of the mould is held at 15° C. to 75° C. The cylinder around the barrel loaded with the powder is held at 145° C. to 220° C. for 10 to 15 minutes to melt the powder blend. Injection is carried out using an injection pressure of 220 bar to 330 bar, held for 2 to 10 minutes. The post-injection pressure is held at 50 bar for 2 to 10 minutes. The mould is then cooled to 15 to 23° C., after which the mould is removed from the injection moulding machine. The moulded fibre is then removed from the mould and then cut into implants of the target weight and length. In some embodiments, the implant is 4 mm in length and contains about 165 μg to about 220 μg of active ingredient (eg, edonentan).

[0097] In one embodiment, the implant of the present invention can be prepared using the homogenous material. In one embodiment, the implant is formed by extrusion, for example, hot melt extrusion. Hot melt extrusion can be performed using ThermoFisher Pharma mini HME Micro Compounder, ThermoFisher FP-Pharma-11-Twin-230x100, ThermoFisher Pharma 11 Twin-Screw Extruder, ThermoFisher FP-Pharma-16-230x100, ThermoFisher Pharma 16 Twin-Screw Extruder, or Barrell Engineering Micro Syringe Type Extruder.

[0098] eye disease The methods of the present disclosure include the use of edonentan and A-182086 as described above in the prevention, treatment and amelioration of ocular diseases selected from the group consisting of ocular neovascularization, vascular leakage, neovascular age-related macular degeneration, neovascular glaucoma, retinal vein occlusion (RVO), and retinopathy of prematurity (ROP), which are described below.

[0099] As clearly demonstrated herein, the therapeutic efficacy of the method is determined by assessment of the reduction in new blood vessel formation or by the reduction in the rate of ocular neovascularization. In further embodiments, the therapeutic efficacy of the method or treatment is demonstrated by an improvement in tissue perfusion, retinal perfusion, visual acuity, visual field, contrast sensitivity, or color vision.

[0100] Neovascularization and Vascular Leakage Ocular neovascularization (also called angiogenesis) occurs when abnormal blood vessels grow and spread throughout the retina and tissues lining the back of the eye and / or other structures of the eye (e.g., the anterior chamber). These abnormal blood vessels are fragile and often leak, scarring the retina and pulling it out of position or causing blockage of aqueous humor drainage, resulting in increased intraocular pressure (i.e., neovascular glaucoma). An eye disorder in which neovascularization plays a role is age-related macular degeneration (AMD), which is the leading cause of severe vision loss in the elderly. Vision loss in AMD results from choroidal neovascularization (CNV). Neovascularization begins in choroidal vessels and grows through Bruch's membrane into the subretinal pigment epithelium space and / or the retina, usually at multiple sites. Leakage and bleeding from these new vessels results in vision loss.

[0101] Ocular neovascularization (also called angiogenesis) occurs when abnormal blood vessels grow and spread throughout the retina, tissues lining the back of the eye, and / or other structures of the eye (e.g., the anterior chamber). These abnormal blood vessels are fragile and often leak, scarring the retina, pulling it out of place, or causing blockage of aqueous humor drainage, resulting in increased intraocular pressure (i.e., neovascular glaucoma).

[0102] Types of ocular neovascularization (e.g., choroidal neovascularization) include histoplasmosis and pathologic myopia, angioidal streaks, anterior ischemic optic neuropathy, bacterial endocarditis, Best's disease, birdshot retinochoroidopathy, choroidal hemangioma, choroidal nevus, choroidal nonperfusion, choroidal osteoma, choroidal rupture, choroideremia, chronic retinal detachment, retinal coloboma, drusen, endogenous Candida endophthalmitis, extrapapillary hamartomas of the retinal pigmented epithelium, fundus flaviformis, idiopathic macular hole, malignant melanoma, membranoproliferative glomerulonephritis (type II), intraocular metallic foreign body, morning glory disc syndrome, and chronic retinal detachment. syndrome, multiple evanescent white dot syndrome (MEWDS), neovascularization at the ora serrata, surgical microscope burn, optic disc pits, photocoagulation, punctate inner choroidopathy, rubella, sarcoidosis, creeping or geographic choroiditis, subretinal drainage, tilted disc syndrome, toxoplasmic retinopathy, tuberculosis, Vogt-Koyanagi-Harada syndrome, diabetic retinopathy, nondiabetic retinopathy, branch vein occlusion, central retinal vein occlusion, retinopathy in premature infants, rubeosis iridis, neovascular glaucoma, perifoveal telangiectasis, sickle cell retinopathy, Eales' disease, retinal vasculitis, von Hippel-Lindau disease, radiation retinopathy, retinal cryopathies These conditions include, but are not limited to, corneal neovascularization due to cryoinjury, retinitis pigmentosa, retinal choroidal coloboma, herpes simplex virus keratitis, corneal ulcer, keratoplasty, pterygium, and neovascularization due to trauma.

[0103] In embodiments, the ocular neovascularization or vascular leakage disorder is rubeosis iridis, neovascular glaucoma, pterygium, vascularized glaucoma filtering bleb, or the like. blebs), conjunctival papilloma); choroidal neovascularization (e.g., neovascular age-related macular degeneration (AMD)), myopia, previous uveitis, trauma, or idiopathic; macular edema (e.g., postoperative macular edema, macular edema secondary to uveitis including inflammation of the retina and / or choroid, macular edema secondary to diabetes, and macular edema secondary to retinal vascular occlusive disease (i.e., branch retinal vein occlusion and central retinal vein occlusion); retinal neovascularization due to diabetes (e.g., retinal vein occlusion), uveitis, ocular ischemic syndrome from carotid artery disease, ophthalmic or retinal artery occlusion, sickle cell retinopathy, other ischemic or occlusive neovascular retinopathies, retinopathy of prematurity, or Eales' disease; and edema or neovascularization related to any occlusive or inflammatory retinal vascular disease, such as genetic disorders (e.g., von Hippel-Lindau syndrome).

[0104] In some embodiments, the ocular neovascularization is associated with a condition selected from the group consisting of retinopathy of prematurity, retinal vein occlusion, macular edema, sickle cell retinopathy, choroidal neovascularization, radiation retinopathy, neovascular glaucoma, microangiopathy, retinal hypoxia, diabetic retinopathy, diabetic macular edema, ablation-induced neovascularization, age-related macular degeneration, and vascular leakage.

[0105] In one embodiment, the neovascular age-related macular degeneration is wet age-related macular degeneration. In another embodiment, the neovascular age-related macular degeneration is dry age-related macular degeneration, and the patient is characterized as being at increased risk of developing wet age-related macular degeneration.

[0106] In embodiments, the ocular neovascularization is associated with a condition selected from the group consisting of retinopathy of prematurity, retinal vein occlusion, macular edema, sickle cell retinopathy, choroidal neovascularization, radiation retinopathy, neovascular glaucoma, microangiopathy, retinal hypoxia, diabetic retinopathy, diabetic macular edema, ablation-induced neovascularization, age-related macular degeneration, and vascular leakage.

[0107] Neovascular glaucoma In the treatment of glaucoma (e.g., neovascular glaucoma) using edonentan or A-182086 as described herein, a "therapeutically effective amount" can be determined by evaluating the improvement in retinal blood flow (RBF) beyond that which can be achieved by standard care (reduction in intraocular pressure (IOP)). For glaucoma (e.g., neovascular glaucoma) indications, the improvement in blood flow in a healthy rabbit eye model can be used as a predictor of pharmacodynamic response (PD) in humans. Rabbits are commonly used to evaluate the ocular PK / PD relationship for compounds targeting human ocular diseases due to the anatomical and functional similarities between rabbit and human eyes. Previously, intravitreal administration of ET-1 in rabbit eyes was shown to induce significant vasoconstriction and optic nerve damage (Sasaoka M. et al., Exp Eye Res 2006; Sugiyama T. et al., Arch Ophthalmol 2009). Efficacy in this model is benchmarked against the reversal of perfusion damage induced by intravitreal edonentan-1 (ET-1) administration at a certain concentration. For example, the efficacy can be achieved at concentrations equivalent to those observed in the plasma and aqueous humor of human glaucoma patients (Li S. et al., Journal of Ophthalmology 2016).

[0108] Other examples of relevant animal glaucoma models are the Morrison rat model of elevated IOP and the laser-induced non-human primate (NHP) glaucoma model. Glaucoma in the Morrison rat model is induced by sustained elevation of IOP via administration of hypertonic saline via the episcleral vein. In the laser-induced NHP glaucoma model, optic nerve head blood flow was shown to be reduced after sustained elevation of IOP (Wang L. et al., 2011). Invest Ophthalmol Vis Sci 2012). Furthermore, reduced optic nerve blood flow has been shown to correlate with long-term structural changes in the optic nerve (Cull G. et al., Invest Ophthalmol Vis Sci 2013).

[0109] Efficacy in the above glaucoma models is defined as lowering of IOP, improvement in optic nerve head or retinal blood flow from baseline, prevention or slowing of progression of structural neurodegenerative changes on flat mounts (e.g., retinal nerve fiber layer thickness measured by optical coherence tomography (OCT) or retinal ganglion cell count), and functional changes such as electroretinogram (ERG) or contrast sensitivity following treatment with edonentan or A-182086.

[0110] It is believed that the effect of edonentan or A-182086 on retinal blood flow can be evaluated by the vessel radius (r) in Poiseuille's law. An increase in (r) by an endothelin antagonist induces a more pronounced increase in blood flow than can be achieved by increasing the perfusion pressure via a drop in IOP: Blood flow = (perfusion pressure × πr 4 ) / (8ηl) where l: length of blood vessel r: radius of the vessel η: Blood viscosity Perfusion pressure: Mean arterial pressure - IOP Furthermore, edonentan or A-182086 can lower IOP and / or prevent RGC death through a mechanism independent of the improvement in retinal / optic nerve head tissue perfusion.Thus, by using certain specific endothelin receptor antagonists, one (r) or more (IOP) of the above parameters can be altered to improve RBF, thus achieving therapeutic efficacy in treating glaucoma.

[0111] In some embodiments, the glaucoma patients are started on treatment as soon as they are diagnosed. In some embodiments, edonentan or A-182086 is administered intravitreally, topically, suprachoroidally, or locally to the back of the eye using an implant delivery platform (e.g., a biodegradable ocular implant) at a frequency of every 3-12 months (e.g., every 3-6 months or every 4-6 months).

[0112] Retinal vein occlusion (RVO) Retinal vein occlusion (RVO), a vascular disorder of the retina, is currently treated with intravitreal injections of anti-VEGF drugs to inhibit growth factors that cause macular edema and corticosteroids to combat the inflammatory components that lead to edema. The use of edonentan and A-182086 therapy to treat RVO by improving tissue perfusion and reducing inflammation while avoiding the unwanted effects of systemic immunosuppression and / or the local deleterious effects of steroids would be highly desirable.

[0113] RVO is currently treated with intravitreal steroids and anti-VEGF agents. Improving perfusion of existing vessels reduces the degree of macular edema and downstream maladaptive changes manifested as VEGF upregulation and RVO. To test efficacy, a preclinical mouse model of ischemic retinopathy can be used. Oxygen-induced retinopathy in the mouse is a reproducible and quantifiable proliferative retinal neovascularization model suitable for testing the pathogenesis and therapeutic interventions of retinal neovascularization in many ischemic retinopathies, including RVO. The model is induced by exposing 1-week-old C57BL / 6J mice to 75% oxygen for 5 days and then to room air, as previously described (Smith LEH et al., Invest Ophthalmol Vis Sci 1994). Efficacy in this preclinical model of ischemic retinopathy can be evaluated by testing retinal hypoxia and neovascularization. The "therapeutically effective amount" of edonentan or A-182086 described herein can be an addition to the current standard of care by improving tissue perfusion and reducing ET-1-mediated inflammation while avoiding the unnecessary effects of topical steroids. In some embodiments of the treatment of RVO, edonentan or A-182086 is administered intravitreally, topically, suprachoroidally, or locally to the back of the eye using an implant delivery platform (e.g., a biodegradable ocular implant). The frequency of administration varies based on the patient's disease course and response to treatment.

[0114] Retinopathy of Prematurity (ROP) Retinopathy of prematurity (ROP) is a retinal vascular proliferative disorder that affects preterm infants. ROP remains the leading preventable cause of blindness and visual impairment worldwide. With improvements in perinatal care, improved survival of moderately preterm infants, and limited resources for oxygen delivery and monitoring, more mature preterm infants are developing severe ROP in developing countries.

[0115] The pathophysiology of ROP is characterized by two stages. Stage I ROP results from vaso-obliteration beginning shortly after birth secondary to a marked decrease in VEGF and insulin-like growth factor-1 (IGF-1). Stage II begins at approximately 33 weeks postmenstrual age (PMA). During this stage, VEGF levels increase, especially in the presence of retinal hypoxia and oxygen demand associated with increased retinal metabolism, resulting in abnormal vascular proliferation. For advanced stages of ROP, laser ablation of the avascular retina, early treatment of ROP (ETROP) protocols, intravitreal injections of anti-VEGF antibodies (e.g., bevacizumab) and vitrectomy are used to preserve central vision and prevent retinal detachment. Long-term complications (e.g., refractory error, risk of recurrence of ROP and retinal detachment) require continued follow-up by an ophthalmologist during adolescence and beyond.

[0116] ROP is induced by severe ischemia due to impaired development of retinal blood vessels secondary to premature birth. Thus, as an aspect of the present invention, we believe that improving perfusion of existing vessels with edonentan or A-182086 will reduce the extent of ischemia and VEGF upregulation, and downstream maladaptive changes that manifest as ROP. To test efficacy, a preclinical mouse model of ROP can be used. Oxygen-induced retinopathy in the mouse is a reproducible and quantifiable proliferative retinal neovascularization model suitable for testing pathogenesis and therapeutic interventions related to retinal neovascularization in ROP. The model is induced by exposing 1-week-old C57BL / 6J mice to 75% oxygen for 5 days, followed by room air, as previously described (Smith LEH et al., Invest Ophthalmol Vis Sci 1994). Efficacy in this preclinical model of ROP can be evaluated by testing retinal hypoxia and neovascularization. A "therapeutically effective amount" of edonentan or A-182086 is an addition to the current standard of care by improving tissue perfusion and reducing pathological neovascularization induced by VEGF, as described herein. In some embodiments, the drug therapy is administered intravitreally, topically, suprachoroidally, or locally to the back of the eye using an implant delivery platform (e.g., a biodegradable ocular implant) as frequently as every 4-6 weeks, as needed, based on the patient's disease history and response to treatment. For example, the drug therapy is administered locally to the back of the eye using an intravitreal injection as frequently as every 5 weeks, as needed, based on the patient's disease history and response to treatment.

[0117] Pharmaceutical Compositions Some embodiments described herein relate to pharmaceutical compositions that may include a therapeutically effective amount of one of edonentan and A-182086 (described herein), or a pharma- ceutically acceptable salt thereof, and a pharma-ceutically acceptable carrier, diluent, excipient, or combination thereof. Such antagonists or their pharma-ceutically acceptable salts may be in crystalline or amorphous form. Each of these may be for pharmacologically acceptable use.

[0118] The term "pharmaceutical composition" refers to a mixture of one or both compounds disclosed herein with other chemical components (e.g., diluents or carriers). The pharmaceutical composition facilitates administration of the compound to an organism. Pharmaceutical compositions are generally tailored for a particular intended route of administration.

[0119] Some pharmaceutical compositions require the preparation of pharmaceutically acceptable salts. Pharmaceutically acceptable salts include salts of acidic or basic groups present in the compounds of the present invention. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). Certain compounds of the present invention can form pharmaceutically acceptable salts with various amino acids.Suitable base salts include, but are not limited to, aluminum salt, calcium salt, lithium salt, magnesium salt, potassium salt, sodium salt, zinc salt and diethanolamine salt.For a review of pharmaceutically acceptable salts, see Berge et al., 66 J. PHARM. SCI, 1-19 (1977).

[0120] The term "pharmacologically acceptable" defines a carrier, diluent, excipient, salt or composition that is safe and effective for its intended use and possesses the desired biological and pharmacological activity.

[0121] As used herein, "carrier" refers to a compound that facilitates the incorporation of a compound into cells or tissues. For example, and without limitation, dimethylsulfoxide (DMSO) is a commonly used carrier that facilitates the incorporation of many organic compounds into cells or tissues of a subject.

[0122] As used herein, "diluent" refers to an ingredient in a pharmaceutical composition that lacks pharmacological activity but may be pharmacologic necessary or desirable. For example, a diluent may be used to increase the bulk of a potent drug whose mass is too small for manufacture and / or administration. It may also be a liquid for dissolving a drug to be administered by injection, ingestion, or inhalation. A common form of diluent in the art is a buffered aqueous solution, including but not limited to phosphate buffered saline, which mimics the composition of human blood.

[0123] As used herein, "excipient" refers to an inert substance added to a pharmaceutical composition to provide the composition with, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegration ability, etc. A "diluent" is a type of excipient.

[0124] The pharmaceutical compositions described herein can be administered to human patients by themselves or in pharmaceutical compositions in which they are mixed with other active ingredients (as in combination therapy) or with carriers, diluents, excipients or combinations thereof.The appropriate formulation depends on the route of administration selected.The techniques for formulating and administering the compounds described herein are known to those skilled in the art.

[0125] The pharmaceutical compositions disclosed herein can be prepared in a manner known per se, for example, by conventional mixing, dissolving, granulating, levigating, emulsifying, encapsulating or entrapping processes.See, for example, Encapsulation Processes, in: Food Powders, 2005, 199-299.Furthermore, the active ingredient is included in an amount effective to achieve its intended purpose.The compounds used in the pharmaceutical combinations disclosed herein can be provided as pharma-ceutically acceptable salts.

[0126] It is preferred to administer the compound or pharmaceutical composition of the present invention in a localized manner, either as a topical ophthalmic preparation or via injection of the compound or pharmaceutical composition directly into ocular tissue, often in a depot or sustained release preparation.The local administration mode can be intravitreal, suprachoroidal, periocular, or subconjunctival injection of the preparation, or use of implantation techniques or topical application.For example, the compound is administered in a liposome preparation, which slowly releases the compound to prolong the desired pharmacological effect.Alternatively, polyvinyl alcohol nanoparticles can be prepared by well-known methods to provide sustained or extended release preparations for topical or intraocular application.

[0127] Furthermore, the compound can be administered in targeted drug delivery system.The example of targeted drug delivery system includes but is not limited to the biodegradable eye implant that is made of edonentan homogeneously dispersed in PLGA polymer.In some embodiments, the biodegradable eye implant is a sustained release biodegradable eye implant.

[0128] In some embodiments, the pharmaceutical composition is an ophthalmic preparation comprising a therapeutically effective amount of one or more endothelin receptor antagonists described herein, or pharma- ceutically acceptable salts thereof. As used herein, "ophthalmic preparation" refers to a specialized dosage form designed to be instilled on the outer surface of the eye (topical), administered inside (intracorcular) or adjacent to the eye (periocular), or used with an ophthalmic device. In some embodiments, the ophthalmic preparation is in the form of a solution, suspension, or ointment. In other embodiments, the ophthalmic preparation is in the form of a gel, gel-forming solution, ophthalmic insert, micro / nano-particle preparation for topical or, preferably, intravitreal injection, or implant.

[0129] In some embodiments, the ophthalmic preparation comprises a preservative. Examples of suitable preservatives include, but are not limited to, cationic wetting agents (e.g., benzalkonium chloride), organic mercurials (e.g., phenylmercuric nitrate, phenylmercuric acetate), organic acids or their esters (e.g., sorbic acid, esters of p-hydroxybenzoic acid (e.g., methyl hydroxybenzoate, propyl hydroxybenzoate), and alcohol substitutes (e.g., chlorobutanol, phenylethanol). Preservatives may be present in the ophthalmic preparation in an amount ranging from about 0.002% w / v to about 0.5% w / v (e.g., 0.01-0.25% w / v). The ophthalmic preparation may further comprise a preservative aid. Examples of suitable preservatives include, but are not limited to, ethylenediaminetetraacetic acid (EDTA).

[0130] In some embodiments, the ophthalmic preparation comprises one or more additional excipients or agents to provide viscosity or lubrication, stabilize active ingredients against degradation, increase the solubility of active or inactive ingredients, adjust tonicity, or act as a solvent. Examples of excipients or agents to provide viscosity or lubrication include hypromellose, carbomer 974P, hydroxyethyl cellulose (HEC), polyvinyl alcohol, sodium hyaluronate, sodium carboxymethyl cellulose, Carbopol 940, hydroxypropyl methylcellulose (HPMC), poloxamer, xyloglucan, alginic acid, sodium alginate, gellan gum, cellulose acetate phthalate, and xanthan gum. Examples of agents as excipients or stabilizers include sodium bisulfite, sodium metabisulfite, sodium thiosulfate, and sodium sulfate / sulfuric acid (which can act as antioxidants). Examples of excipients or agents as solubilizers include, but are not limited to, povidone, creatinine, castor oil, and cyclodextrin (e.g., gamma-cyclodextrin). Examples of excipients or agents for adjusting tonicity include, but are not limited to, sodium chloride, potassium chloride, calcium chloride dehydrate, magnesium chloride hexahydrate, sugars (e.g., sucrose, maltose, dextrose, etc.), glycerin, propylene glycol, mannitol, ascorbic acid, and acetylcysteine.

[0131] In some embodiments, the ophthalmic preparation comprises one or more buffer substances for adjusting pH. Examples of buffer substances for adjusting pH include, but are not limited to, sodium citrate, monobasic sodium phosphate, dibasic sodium phosphate, boric acid, hepatahydrate, sodium acetate trihydrate, sodium citrate dihydrate, histidine, and phosphate buffered saline (PBS). The resulting composition may have a pH value of 5.0 to 8.5 (e.g., 5.0 to 6.0, 5.2 to 5.8, 6.0 to 8.0, 6.6 to 7.8, 6.2 to 8.2, and 6.2 to 7.5).

[0132] In some embodiments, the ophthalmic preparation comprises one or more surfactants. Examples of surfactants include sorbitan ether esters of oleic acid (e.g., polysorbates or Tween® 20 and 80) and tyloxapol.

[0133] The volume that can be injected into the human eye at one time is approximately 50-90 μL via the intravitreal route, up to 450 μL via the subretinal route, up to 200 μL via the suprachoroidal route, and about 40-50 μL via the topical route (e.g., topical administration as eye drops). The needles used in these routes are typically 27-30G in size. The dose depends on the volume, the potency, the target efficacy, and the concentration that can be formulated to match the pharmacokinetic profile for each indication. In general, ocular injections are not administered more frequently than once a month per eye. For topical administration (e.g., eye drops), in most cases, the frequency of administration to the eye does not exceed more than once or twice a day.

[0134] In some embodiments, the intravitreal formulation comprises a dose of the compound (e.g., an endothelin receptor antagonist, e.g., a compound of formula I) in the range of about 1 μg to about 1 mg (e.g., about 1 μg, about 5 μg, about 10 μg, about 25 μg, about 50 μg, about 75 μg, about 100 μg, about 125 μg, about 150 μg, about 175 μg, about 200 μg, about 250 μg, about 500 μg, about 700 μg, and about 1 mg). A first exemplary formulation comprises about 1 μg to about 1 mg of the compound (e.g., an endothelin receptor antagonist, e.g., a compound of formula I), about 10 mM histidine HCl, about 10% α,α-trehalose dihydrate, and about 0.01% polysorbate 20. A second exemplary formulation includes about 1 μg to about 1 mg of a compound (e.g., an endothelin receptor antagonist, e.g., a compound of Formula I), about 10 mM sodium phosphate, about 40 mM sodium chloride, about 0.03% polysorbate 20, and about 5% sucrose.

[0135] In some embodiments, the intravitreal preparation contains about 1 μg to about 500 μg (e.g., about 10 μg to about 500 μg, about 20 μg to about 500 μg, about 30 μg to about 500 μg, about 40 μg to about 500 μg, about 50 μg to about 500 μg, about 60 μg to about 500 μg, about 70 μg to about 500 μg, about 80 μg to about 500 μg, about 90 μg to about 500 μg, about 100 μg to about 500 μg, about 100 μg to about 500 μg, about 125 μg to about 500 μg, about 150 μg to about 500 μg, about 175 μg to about 500 μg , about 200 μg to about 500 μg, about 225 μg to about 500 μg, about 250 μg to about 500 μg, about 275 μg to about 500 μg, about 300 μg to about 500 μg, about 325 μg to about 500 μg, about 350 μg to about 500 μg, about 375 μg to about 500 μg, about 400 μg to about 500 μg, about 425 μg to about 500 μg, about 450 μg to about 500 μg, and about 475 μg to about 500 μg).

[0136] In some embodiments, the intravitreal formulation comprises a dose of the compound (e.g., an endothelin receptor antagonist, e.g., a compound of Formula I) in the range of about 10 μg to about 500 μg. In some embodiments, the intravitreal formulation comprises a dose of the compound (e.g., an endothelin receptor antagonist, e.g., a compound of Formula I) in the range of about 10 μg to about 300 μg. In some embodiments, the intravitreal formulation contains about 1 μg, about 5 μg, about 10 μg, about 15 μg, about 20 μg, about 25 μg, about 30 μg, about 35 μg, about 40 μg, about 45 μg, about 50 μg, about 55 μg, about 60 μg, about 65 μg, about 70 μg, about 75 μg, about 80 μg, about 85 μg, about 90 μg, about 95 μg, about 100 μg, about 110 μg, about 120 μg, about 130 μg, about 140 μg, about 150 μg, about 160 μg, about 170 μg, about 180 μg, about 190 μg, about 200 μg, about 210 μg, about 220 μg, about 230 μg, about These include doses of the compound (e.g., an endothelin receptor antagonist, e.g., a compound of Formula I) at about 240 μg, about 250 μg, about 260 μg, about 270 μg, about 280 μg, about 290 μg, about 300 μg, about 310 μg, about 320 μg, about 330 μg, about 340 μg, about 350 μg, about 360 μg, about 370 μg, about 380 μg, about 390 μg, about 400 μg, about 410 μg, about 420 μg, about 430 μg, about 440 μg, about 450 μg, about 460 μg, about 470 μg, about 480 μg, about 490 μg, and about 500 μg. A first exemplary formulation includes about 10 μg to about 500 μg (e.g., 300 μg) of the compound (e.g., an endothelin receptor antagonist), about 10 mM histidine HCl, about 10% α,α-trehalose dihydrate, and about 0.01% polysorbate 20. A second exemplary formulation includes about 10 μg to about 500 μg (e.g., 300 μg) of the compound (e.g., an endothelin receptor antagonist), about 10 mM sodium phosphate, about 40 mM sodium chloride, about 0.03% polysorbate 20, and about 5% sucrose.

[0137] In some embodiments, the intravitreal formulation comprises a dose of the compound (e.g., an endothelin receptor antagonist, e.g., a compound of Formula I) in the range of about 150 μg to about 300 μg. In some embodiments, the intravitreal formulation comprises a dose of the compound (e.g., an endothelin receptor antagonist, e.g., a compound of Formula I) in the range of about 165 μg to about 220 μg (e.g., about 165 μg, about 170 μg, about 175 μg, about 180 μg, about 185 μg, about 190 μg, about 195 μg, about 200 μg, about 205 μg, about 210 μg, about 215 μg, and about 220 μg).

[0138] In some embodiments, the intravitreal formulation comprises a dose of the compound (e.g., an endothelin receptor antagonist, e.g., a compound of Formula I) in the range of about 300 μg to about 600 μg. In some embodiments, the intravitreal formulation comprises a dose of about 330 μg to about 500 μg (e.g., about 330 μg, about 335 μg, about 340 μg, about 345 μg, about 350 μg, about 355 μg, about 360 μg, about 365 μg, about 370 μg, about 375 μg, about 380 μg, about 385 μg, about 390 μg, about 395 μg, about 400 μg, about 405 μg, about 410 μg, about 415 μg, about 420 μg, about 430 μg, about 440 μg, about 450 μg, about 460 μg, about 470 μg, about 480 μg, about 490 μg, about 500 μg, about 510 μg, about 520 μg, about 530 μg, about 540 μg, about 550 μg, about 560 μg, about 570 μg, about 580 μg, about 590 μg, about 600 μg, about 610 μg, about 620 μg, about 630 μg, about 640 μg, about 650 μg, about 660 μg, about 670 μg, about 680 μg, about 690 μg, about 700 μg, about 710 μg, about 720 μg, about 730 μg, about 740 μg, about 750 μg, about and about 500 μg).

[0139] In further embodiments, the intravitreal formulation comprises a dose of the compound (e.g., an endothelin receptor antagonist) in the range of about 500 μg to about 4 mg (e.g., about 500 μg, about 725 μg, about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, and about 3.5 mg). A first exemplary formulation comprises about 500 μg to about 1 mg of the compound (e.g., an endothelin receptor antagonist), about 0.014% monobasic potassium phosphate, 0.08% dibasic sodium phosphate, 0.7% sodium chloride, 0.02% polysorbate, and 0.5% sodium carboxymethylcellulose. A second exemplary formulation comprises about 500 μg to about 1 mg of a compound described above (e.g., an endothelin receptor antagonist), about 0.04% monobasic sodium phosphate monohydrate, about 0.3% dibasic sodium phosphate heptahydrate, 0.63% sodium chloride, and about 1% to about 2.3% sodium hyaluronate.

[0140] Without further elaboration, it is believed that one skilled in the art can utilize the present invention to its full extent based on the above description. Accordingly, the following specific examples, i.e., Examples 1-15, are to be construed as merely illustrative, and not limiting in any way to the remainder of the disclosure. EXAMPLES

[0141] Example 1: Physicochemical and biochemical characterization of the compounds Provided below in Table 1 are the physicochemical and biochemical data for the above edonentan and A-182086. As shown in Table 1, at pH 2, A-182086 has a better solubility than that of edonentan. On the other hand, at pH 7, edonentan has a better solubility than that of A-182086. Table 1. Physicochemical and biochemical characterization of compounds. [Table 1-1] [Table 1-2] a The data is for the amorphous form. b Appropriate calculations were made taking into account the surface charge distribution (mainly O and N). Compounds with a PSA of approximately 90 or less are predicted to cross the blood-brain barrier.

[0142] In the above tables, physicochemical data (e.g., solubility) were obtained according to standard protocols known in the art (e.g., Reis et al., Mini Rev Med Chem., 2010, 10(11):1071-6; Avdeef et al., Expert Opin Drug Metab ~xicol. 2005, 1(2):325-42; Bharate et al., Comb Chem High Throughput Screen. 2016, 19(6):461-9; and Jain et al., J Pharm Biomed Anal., 2013, 86:11-35); biochemical data (i.e., ET A / ET B Potency of the antibody was obtained according to protocols known in the art (see, e.g., Kirkby et al., Br J Pharmacol., 2008, 153(6):1105-19; and Maguire et al., Br J Pharmacol., 2014, 171(24):5555-72).

[0143] Example 2: Formulation of edonentan for intravitreal use in rabbits An appropriate amount of edonentan is dissolved in neat PEG400, followed by the addition of 15% CD (HP-β-cyclodextrin) solution. The final concentration of PEG400 is determined to be 20%. The target concentrations are 5mg / ml and 0.5mg / ml, based on the amount of edonentan. The resulting solution is filtered using a 0.25 micron filter.

[0144] Example 3: Effects of edonentan and ET-1 in a rabbit model Adult male Dutch-belted rabbits were given a 20 μl intravitreal injection (IVT) of 0.5 μg ET-1, followed by a 20 μl intravitreal injection of 10–100 μg edonentan 30 min after ET-1 administration. IOP, optical coherence tomography-angiograms (OCT-A), and fluorescein angiograms (FA) were performed at predefined time points (30, 45, 60, and 75 min) after ET-1 and edonentan administration to evaluate retinal blood flow changes induced by ET-1 ± edonentan. As shown in Figure 1, ET-1 administration effectively induced obvious vasoconstriction in the retinal vascular bed within 45 min. Figure 2 shows that the effect of ET-1 was then reversed with 10 μg edonentan administration within 90 min (60 min after edonentan administration).

[0145] Example 4: Preparation of an extended release formulation containing edonentan Concentrated edonentan dispersion is made by combining edonentan with water, vitamin E-TPGS and gamma-cyclodextrin. These components are mixed to disperse edonentan, then autoclaved. Sodium hyaluronate can be purchased as a sterile powder, or a dilute solution can be sterilized by filtration, followed by lyophilization to obtain a sterile powder. The sterile sodium hyaluronate is dissolved in water to form an aqueous concentrate. The concentrated edonentan dispersion is mixed and added to the sodium hyaluronate concentrate as a slurry. A sufficient amount of water is added (enough, in this case, enough to prepare a homogeneous mixture, dispersion, gel or suspension), and the mixture is mixed until homogeneous. Examples of these compositions are provided in Table 2 below: Table 2. Composition of extended release formulations containing edonentan [Table 2]

[0146] These exemplary compositions contain a sufficient concentration of high molecular weight (i.e., polymeric) sodium hyaluronate to form a gelatinous plug or drug depot upon intravitreal injection into the human eye. Preferably, the average molecular weight of the hyaluronate used is less than 2 million, and more preferably, the average molecular weight of the hyaluronate used is between about 1.3 million and 1.6 million. The edentan particles are actually trapped or held within this viscous plug of hyaluronate, so that undesirable pluming does not occur upon intravitreal injection of the formulation. Thus, the risk of drug particles undesirably precipitating directly onto retinal tissue is substantially reduced, for example, when compared to using compositions with a water-like viscosity (e.g., Kenalog® 40). Because sodium hyaluronate solutions undergo dramatic shear thinning, these formulations are easily injected through 25-gauge, 27-gauge, or even 30-gauge needles.

[0147] Example 5: Preparation of a topical edonentan formulation Topical edonentan formulations can be prepared according to known methods (e.g., WO 2016156639 A1). More specifically, 20 g of Cremophor (登録商標) RH40 is dissolved in 75 mL of deionized water by magnetic stirring and allowed to stir until completely dissolved. Then, 1.5 g of trometamol is added to the resulting solution and allowed to stir for 15 minutes to achieve complete dissolution. 0.5 g of edonentan is added and allowed to stir for 15 minutes to ensure complete dissolution. Then, 2 g of glycine and 1 g of boric acid are added and allowed to stir until completely dissolved. The resulting solution is added in sufficient quantity to 100 mL of deionized water. The final solution is filtered through filter paper to obtain a colorless, clear solution having a pH of 8.06. The above solution is filled into an eye dropper bottle with a volume of 5 mL.

[0148] Example 6: Topical Ophthalmic Solution Nanoparticles Containing Edonentan Nanoparticles were prepared by solvent evaporation technique. A solution of 120 mg 50:50 PLGA in 60 mL ethyl acetate was prepared. To this solution, 50 ml water and an aqueous solution of 12 mg edonentan and 0.5 mg polyvinyl alcohol were incorporated under vigorous stirring. The resulting mixture was left under continuous stirring and vacuum for 2 hours. The resulting preparation was then ultracentrifuged and washed three times with water to remove the nanoparticles from the medium. The nanoparticles so obtained were dried in a vacuum oven and dispersed in an aqueous solution sufficient for a concentration of 5 mg / 1 mL edonentan after evaluation.

[0149] Example 7: Glaucoma Preclinical Trials The healthy rabbit model is used to evaluate (in vivo) the pharmacodynamic effects of edonentan and / or A-182086 or their pharmaceutically acceptable salts. These studies are performed with different doses of the selected endothelin antagonist. Further animal studies are performed by combining the endothelin antagonist with the current standard of care. The Morrison rat model of glaucoma, the rat model of acutely elevated IOP elevation and the non-human primate laser-induced glaucoma model are used to evaluate the optic nerve head blood flow and the rate of retinal ganglion cell loss associated with different doses of the selected endothelin antagonist with and without standard of care.

[0150] The improvement of blood flow in a healthy rabbit model is measured for the indicated endothelin receptor antagonists at various doses after induction of perfusion impairment by topically administered ET-1. The change in optic nerve head blood flow and retinal nerve fiber layer (RNFL) thickness in a non-human primate glaucoma model is measured for the indicated endothelin receptor antagonists at various doses. The results show improvement of RGC survival, retinal and optic nerve head blood flow, and slowing of RNFL thinning due to the use of selected endothelin receptor antagonists. The dosing regimen for humans is extrapolated from the results of healthy rabbit and non-human primate glaucoma models.

[0151] Pharmacodynamic studies to assess retinal blood flow changes in rabbits To evaluate the effect of intravitreally administered endothelin-1 (ET-1) followed by the antagonist edonentan on retinal blood flow in rabbits, rabbits (Oryctolagus cuniculus) were given a 20 μL intravitreal injection of ET-1 followed by a 20 μL intravitreal injection of edonentan at two (or three) different doses (e.g., 0.1 μg, 0.5 μg, 2.5 μg). Pulse oximetry, intraocular pressure measurement, optical coherence tomography-angiography (OCTA), fluorescein angiography (FA) and retinal leakage scoring were performed for evaluation. The dose response in rabbits is shown in Figure 8A and Figure 8B.

[0152] Pharmacokinetic and tolerability analysis of intravitreally delivered edonentan in rabbits To determine the pharmacokinetics and safety profile of edentan after intravitreal administration in rabbits, rabbits (Oryctolagus cuniculus) were given bilateral intravitreal injections (20 μL injection volume / eye). After the injection, the animals were sedated with a ketamine / xylazine cocktail and then euthanized with an overdose of sodium pentobarbital (Euthasol). Animals designated for pharmacokinetic analysis were euthanized at various time points (e.g., 12 hours, 16 hours, 24 hours, 36 hours, and 48 hours). At least 1.0 mL of whole blood was drawn from the marginal ear vein or cardiac puncture (terminal blood draws only) into K2EDTA tubes for plasma collection and processed for analysis.

[0153] Immediately after euthanasia, the eyes were enucleated. Aqueous humor from both eyes was removed via syringe and flash frozen for analysis. When frozen, the eyes were dissected to isolate various ocular tissues and minimize drug diffusion to adjacent tissues. Left and right eye tissues were collected into separate vials for analysis. The list of tissues collected included plasma and aqueous humor, iris / ciliary body (ICB), retina, vitreous humor and RPE / choroid. The pharmacokinetic profile of edonentan delivered intravitreally in rabbits is shown in Figure 9A, Figure 9B, Figure 9C and Figure 9D.

[0154] Pharmacokinetic analysis of topically administered edonentan in rabbits To determine the pharmacokinetic properties of edonentan after topical administration in rabbits, rabbits (Dutch-belted rabbits) were given eye drops (100 μg edonentan, 35 μL dose volume / eye) in both eyes. After administration, animals (N=2) were euthanized at various time points (e.g., 10 min (immediately after pot-dose), 2 h and 7 h) and tissues were collected for analysis. The list of tissues collected includes plasma, retina, vitreous humor and bulbar conjunctiva. The pharmacokinetic properties of edonentan delivered topically in rabbits are shown in FIG. 10. It shows that edonentan was detected in all tissues tested at all time points after a single topical application.

[0155] Efficacy studies in the Morrison rat model of glaucoma Adult male and female post-breed Brown Norway rats (age group approximately 8-11 months) were obtained from Envigo (Indianapolis, IN). Baseline IOP measurements and pattern electroretinogram (PERG) amplitudes were collected prior to surgery for IOP elevation (to ensure that IOP and PERG amplitudes were within the expected range of values). IOP was elevated in one eye (left eye) of the rats, while the corresponding right eye served as a contralateral control. The Morrison method of IOP elevation in rats was performed by injecting 50 μL of hypertonic saline via the episcleral vein to stiffen the trabecular meshwork. IOP was measured twice weekly for the entire duration of the experiment. Seven to 10 days after surgery, IOP elevation was observed in the operated eye of the rats. After detecting elevated IOP for two consecutive days, topical administration of eye drops (20 μL (100 μg) / dose of test compound in eyes with elevated IOP) was initiated, 5 days per week for a total of 4 weeks. During the fourth week of treatment, PERG analysis was performed and rats were sacrificed by an overdose of pentobarbital (Fatal-Plus). Aqueous humor was collected from rat eyes, frozen, and sent for analysis. Retinal flat mounts were prepared and immunostained with RGC marker, Brn3a antibody, and surviving RGCs were counted in two eccentricities (central and peripheral).

[0156] For this study, Morrison's model was used to induce ocular hypertension in adult post-breeding male Brown Norway rats as previously described by Morrison et al. (Morrison JC, Moore CG, Deppmeier LM, Gold BG, Meshul CK, Johnson EC. A rat model of chronic pressure-induced optic nerve damage. Exp Eye Res. 1997;64(1):85-96).

[0157] Immunostained retinal flat mounts were obtained to measure retinal ganglion cell (RGC) numbers. To obtain immunostained retinal flat mounts, the animals were euthanized after treatment and then their eyes were enucleated. The eye cups were fixed overnight in 4% paraformaldehyde (PFA) at 4°C, and retinal flat mounts were prepared for image collection. Retinal ganglion cell (RGC) counting was performed using the images of immunostained retinal flat mounts. The images were uploaded to ImageJ (a photo editor designed for biological research (Rasband, 1997-2018)), and the labeled retinal ganglion cells were manually counted at two eccentricities (center and periphery). Figure 5A shows the comparison of RGC numbers in the peripheral retina between vehicle and edentan, and Figure 6A shows the comparison between vehicle and A-182086.

[0158] Pattern ERG (PERG) was used to assess RGC function. To obtain the pattern ERG recordings, the UTAS Visual Electrodiagnostic System (LKC, Gaithersburgh, MD, USA) was used according to the method described by Porciatti et al. (Porciatti V, Saleh M, Nagaraju M. The pattern electroretinogram as a ~ol ~ monitor progressive retinal ganglion cell dysfunction in the DBA / 2J mouse model of glaucoma. Inves~phthalmol Vis Sci. 2007;48(2):745-751). Briefly, PERG signals were acquired from a DTL-plus electrode placed on the inferior part of the corneal surface, and PERG waves were analyzed using EMWIN software (LKC). The difference between the amplitudes of the main positive (P1) and negative (N2) waves was calculated to deciper the PERG amplitude. FIG. 5B shows the IOP-mediated PERG changes between vehicle and edonentan, and FIG. 6B shows the changes between vehicle and A-182086.

[0159] RGC number and PERG changes, as shown in Figures 5A, 5B, 6A, and 6B, reveal that both edonentan and A-182086 prevented RGC loss and maintained RGC function in the Morrison's rat model of glaucoma.

[0160] Pharmacokinetic analysis of edonentan or A-182086 delivered topically or orally in rats To determine the pharmacokinetic properties of edonentan or A-182086 after topical administration in rats, rats (Brown Norway rats) were given eye drops (100 μg edonentan, 20 μL dose volume / eye; or 100 μg A-182086, 20 μL dose volume / eye). To determine the pharmacokinetic properties of edonentan or A-182086 after oral administration in rats, rats (Brown Norway rats) were given oral doses of 10 mg / kg or 50 mg / kg edonentan, or oral doses of 1.7 mg / kg or 17 mg / kg A-182086. After administration, animals (N=2) were euthanized at various time points (e.g., 4 hours and 8 hours) and tissues were collected for analysis. The list of tissues collected included plasma, retina / retinal pigment epithelium (RPE) / choroid, vitreous humor, and aqueous humor. The pharmacokinetic profile of edonentan administered topically or orally in rats is shown in FIG. 5C. The pharmacokinetic profile of A-182086 administered topically or orally in rats is shown in FIG. 6C. FIG. 5C and FIG. 6C show that both edonentan and A-182086 are detected in the retina / RPE / choroid, aqueous humor and vitreous humor 4 and 8 hours after topical administration. These data also revealed that edonentan is detectable in the aqueous humor at 17 mg / kg and in the retina / RPE / choroid and vitreous humor at 1.7 and 17 mg / kg after oral administration of edonentan, and A-182086 is detectable in the retina / RPE / choroid at 50 mg / kg after oral administration of A-182086.

[0161] Example 8: Laser-induced glaucoma, non-human primate study - Pharmacodynamics study Non-human primates (rhesus monkeys, Macaca Mulatta) were obtained for this study. One eye of each animal underwent induction of elevated intraocular pressure (IOP) by repeated laser photocoagulation of the trabecular meshwork. Repeated imaging sessions were performed to monitor changes in the optic nerve head (ONH) and retinal structure.

[0162] Effect of edonentan on optic nerve head blood flow after IVT administration A study was conducted to compare experimentally glaucomatous eyes and contralateral healthy eyes (controls) of three non-human primates in overall mean blur ratio (MBR) and MBR change over time from baseline as an index of ONH blood flow in a laser-induced glaucoma model. More specifically, vehicle control, 0.02 mg / mL edonentan, 0.2 mg / mL edonentan, or 2.0 mg / mL edonentan were administered intravitreally (50 μL) to each glaucomatous eye of three non-human primates (rhesus monkeys, Macaca Mulatta). ONH blood flow was then measured over a 6-hour period using laser speckle flowgraphy (LSFG), as shown in Figures 7A-7L. These graphs show ONH blood flow in three non-human primates after IVT administration of vehicle alone (Figures 7A, 7E, and 7I), 0.02 mg / mL edonentan (Figures 7B, 7F, and 7J), 0.2 mg / mL edonentan (Figures 7C, 7G, and 7K), or 2.0 mg / mL edonentan (Figures 7D, 7H, and 7L). Figures 7A-7L reveal improvement in ONH blood flow in a dose-dependent manner after treatment with edonentan. The collective results of the three non-human primates are shown in Figure 7M. This shows that edonentan clearly shows a dose-related increase in ONH blood flow resulting from dilation of retinal arteries, veins, and capillaries in experimental glaucoma eyes compared to control eyes.

[0163] In one of the three non-human primates, LSFG scans were performed at various selected time points when edonentan was administered at 2.0 mg / mL, and the results are shown in Figure 7N.

[0164] Effect of edonentan on intraocular pressure after topical administration A single dose of 0.5% timolol or a single dose of 2 mg / mL edonentan was administered topically with a washout in randomized order for 1 week to three non-human primates with laser-induced glaucoma in the right eye (OD).

[0165] Test results: Control 1: A single dose of 50 μL topical timolol 0.5% in each eye demonstrated an IOP reduction of approximately 20% from pre-dose to post-dose (120 minutes).

[0166] Control 2: A single dose of 50 μL topical timolol 0.5% in each eye demonstrated an approximately 30% IOP reduction from pre-dose to post-dose (120 minutes).

[0167] Non-human primate 1: In experimental glaucoma eyes, 50 μL of edonentan eye drops (2 mg / mL) reduced IOP by approximately 60% from pre- to post-treatment (120 min), and in the contralateral healthy eye, reduced IOP by approximately 10% from pre- to post-treatment (120 min).

[0168] Non-human primate 2: In experimental glaucoma eyes, 50 μL of edonentan eye drops (2 mg / mL) reduced IOP by approximately 50% from pre-to-post (15 min) and approximately 30% from pre-to-post (120 min). In the contralateral healthy eye, 50 μL of edonentan eye drops (2 mg / mL) reduced IOP by approximately 20% from pre-to-post (15 min) and approximately 0% from pre-to-post (120 min).

[0169] Non-human primate 3: In experimental glaucoma eyes, 50 μL of edonentan eye drops (2 mg / mL) reduced IOP by approximately 40% from pre-to-post (15 min) and approximately 40% from pre-to-post (120 min). In the contralateral healthy eye, 50 μL of edonentan eye drops (2 mg / mL) reduced IOP by approximately 10% from pre-to-post (15 min) and approximately 40% from pre-to-post (120 min).

[0170] Example 9: Formulation of edonentan for testing in mice with oxygen-induced ischemic retinopathy Suitable topical formulations of edonentan were prepared in a physiologically compatible system containing hydroxypropyl beta cyclodextrin (HPβCD) and sodium carboxymethylcellulose (CMC) (both available from Sigma-Aldrich) at concentrations of 0.05% w / w and 0.2% w / w active. The HPβCD was dissolved in PBS (pH 7.4) at a concentration of 15% w / w. To this solution, CMC (low molecular weight) was added at a concentration of 0.3% w / w. The solution was mixed until the polymer was completely dissolved and wetted. The active ingredient was then dissolved in an appropriate volume of 15% HPβCD with 0.3% w / w CMC. The active solution was placed in an autoclave and heated to 120°C for 15 minutes and allowed to cool to room temperature. The solution was then filtered through a 0.22 μm PVDF filter.

[0171] Example 10: Study in mice with oxygen-induced ischemic retinopathy A mouse model was used to obtain retinal hypoxic regions in mice with oxygen-induced ischemic retinopathy (OIR) at various time points, as shown in Figure 4. 7-day-old neonatal C57BL / 6 mice were exposed to 75% oxygen from postnatal day (P) 7 to P12. Upon return to normoxia on P12, mice were treated with topical eye drops (5 μL) twice daily of edonentan (0.05% and 0.2% solutions, Example 9) or vehicle control, and intraperitoneal injections once daily of 1 mg / kg aflibercept. Tissues were harvested after 5 days of treatment and stained for isolectin-IB4 for visualization and analysis of NV. Separate studies were performed to determine the drug levels achieved by the 0.2% solution in the retina and RPE / choroid as the target therapeutic level.

[0172] Example 11:Edonentan biodegradable ocular implants - materials and preparation methods Biodegradable implants were prepared using various grades of PLGA polymer. The polymer was dissolved in methylene chloride in a specific ratio. Then, a therapeutic agent (e.g., edentan) was added to the polymer solution and dissolved. Then, the methylene chloride was evaporated at room temperature in a polytetrafluoroethylene (PTFE) dish. After removing the methylene chloride, a thin film of homogeneous material remained.

[0173] Exemplary polymers were dissolved in methylene chloride at a particular ratio (e.g., 50% RG503 and 50% RG503H (50 / 50 RG503 / RG503H). Edonentane was then added to the polymer solution at 30% w / w and dissolved. The methylene chloride was then evaporated in a polytetrafluoroethylene (PTFE) dish at room temperature for 72-120 hours. After the methylene chloride was removed, a thin film of a homogenous mixture of polymer and edonentane remained. The thin film could be 200-300 μm thick. The thin film was then cut into 3.5 mm long implants that could be loaded into a 22 gauge needle. Implants were cut to range in weight from approximately 200 μg to 380 μg, resulting in drug loadings of 60 μg to 114 μg.

[0174] Example 12: Edonentan biodegradable ocular implants – Pharmacokinetic and tolerability analysis The biodegradable ocular implants of Example 11 were designed for intravitreal delivery of edonentan over a period of 3 months. For in vitro drug release testing, the three implants were incubated in 3 mL of PBS (pH 7.4) in a shaking incubator set at 37°C and 50 rpm. Drug release was sampled at the designated time points and the drug content was analyzed by HPLC assay. The release medium was completely replaced with fresh medium during each sampling time point. The pharmacokinetics and tolerability of edonentan biodegradable implants were evaluated in rabbits up to 21 days after administration. Macroscopic ophthalmic examination was performed and the ocular matrix containing the residual content in the implant was processed and analyzed by LC-MS / MS at 14 and 21 days after administration.

[0175] Example 13: Edonentan biodegradable ocular implants - materials and preparation methods Using the procedure for preparing homogenous films of Example 11, additional formulations were prepared using injection molding and ram extrusion.

[0176] Exemplary polymers were dissolved in methylene chloride at specific ratios (e.g., 50% RG503, 10% RG502, and 40% RG753S). Exemplary formulations with various polymer and drug ratios are shown in Table 3. Edentan was then added to the polymer solution at 45% w / w and dissolved. The methylene chloride was then evaporated in a polytetrafluoroethylene (PTFE) dish at room temperature for 24 hours, and then dried under vacuum at 25°C and 20 mbar for 24 hours. The film was then ground into powder using a cryo-mill. A small portion of the film was added to a stainless steel cryo-mill vessel containing 2-3 appropriately sized grinding balls and pre-cooled using liquid nitrogen at 5 Hz for 2-3 minutes. The material was then ground at 20 Hz-25 Hz for 1 minute, with a 1 minute pause at 5 Hz. This grind / rest cycle was repeated 2-5 times. The resulting material was a coarse to fine powder of homogeneous material.

[0177] Implants were formed by injection molding in a modified Haake MiniJet (ThermoFisher Scientific). The homogenous powder was loaded and injected into a mold consisting of channels of appropriate size (e.g., 300 μm×12 mm or 325 μm×12 mm). The powder was loaded into a barrel leading to the mold, and the mold was placed under vacuum. The temperature of the mold was held at 15° C. to 25° C. The cylinder around the barrel loaded with the powder was held at 145° C. to 165° C. for 12 to 15 minutes to melt the powder blend. Injection was performed using an injection pressure of 230 bar to 320 bar, held for 2 to 5 minutes. Post-injection pressure was held at 50 bar for 2 to 5 minutes. The mold was then cooled to 15 to 23° C., after which the mold was removed from the injection machine. The molded fibers were then removed from the mold and they were then cut into 4 mm implants containing 165 μg to 220 μg of edonentan per implant.

[0178] Implants of selected formulations were also formed by ram extrusion using a modified Barrell Micro Extruder (Barrell Engineering). The homogenous powder was loaded into a 3 mm barrel and extruded through a 0.30 μm die maintaining a temperature of 68° C. to 80° C. at a flow rate of 5 μL / min to 6 μL / min. The extruded filaments were then cut into 4 mm implants containing 165 μg to 220 μg of edonentan per implant. The resulting implants have similar performance characteristics to those produced by injection molding. Table 3. Exemplary Formulations [Table 3]

[0179] Example 14: Edonentan biodegradable ocular implants – Pharmacokinetic and tolerability analysis The biodegradable ocular implant of Example 13 was designed for intravitreal delivery of edonentan over a period of 3 months. For in vitro drug release testing, the three implants were incubated in 3 mL of PBS (pH 7.4) in a shaking incubator set at 37°C and 50 rpm. Drug release was sampled at designated time points and its drug content was analyzed by HPLC assay. The release medium was completely replaced with fresh medium during each sampling time point.

[0180] In a non-GLP 12-week ocular and systemic pharmacokinetic study in DB rabbits, two Edentan intravitreal implants (total implant weight IM 423 μg / implant; 380 μg Edentan / 2 implants, RE 461 μg / implant, 415 μg Edentan / 2 implants) from either an injection molding (IM) or ram extrusion (RE) manufacturing process were administered as one bilateral IVT injection (2 animals and 4 eyes per time point) in DB rabbits. The implants contained 45% Edentan in a blend of Resomer® containing 50% RG503, 10% RG502, and 40% RG753S. Rabbits were euthanized at weeks 4, 8, 10, 11, and 12 and drug concentrations were determined in the aqueous humor, lens, vitreous humor, retina, RPE / choroid, and plasma.

[0181] Ocular tissues and plasma were analyzed for edonentane content using an analytical method based on protein precipitation and liquid-liquid extraction followed by reversed-phase LC-MS / MS analysis. An Agilent 1290 UPLC coupled to an Agilent 6430 triple quadrupole mass spectrometer was used for the analysis. The quantification range for edonentane was 1-250 ng / mL. Tissue and plasma samples were homogenized and extracted with 0.1% formic acid in acetonitrile spiked with deuterated edonentane at approximately 10 ng / mL. The extracts were analyzed using reversed-phase liquid chromatography separation and tandem mass spectrometry detection in positive ion mode following quantitative transitions m / z 537.2-439.1 for edonentane and m / z 540.2-442.1 for deuterated edonentane.

[0182] The IVT sustained delivery of 45% edonentan in the PLGA implants clearly demonstrated the achievement of sustained therapeutic target tissue levels of edonentan over the duration of the study (Figure 11A, Figure 11B). The cumulative edonentan released from the implants was 100% at 8 weeks, as seen in Table 4 below. Table 4. Cumulative edonentan released from the implant during a 12-week ocular and systemic pharmacokinetics of edonentan intravitreal implant in a rabbit study. [Table 4-1] [Table 4-2]

[0183] Example 15. Crystalline form of edonentan Exemplary Preparations of Crystalline Form 1 Amorphous edonentan (840 mg) was dissolved in 12 mL of IPA. The resulting solution was filtered and the filter was washed with an additional 2.5 mL of IPA. The filtrate was concentrated to dryness, dissolved in 11.8 mL of IPA and heated to 60° C. with stirring. Then, 18 mL of hot water was added dropwise at 60° C. with vigorous stirring and the solution was stirred at 60° C. for 1 hour. The solution was slowly cooled to 25° C., filtered and dried under vacuum at 25° C. to obtain 660 mg of crystalline Form 1 (XRPD and DSC are in FIG. 13 and FIG. 17, respectively).

[0184] Exemplary Preparation of Crystalline Form 2 Amorphous edonentan (250 mg) was dissolved in 3.5 mL of IPA. The resulting solution was filtered and the filter was washed with an additional 0.25 mL of IPA. The solution was then heated to 60° C., during which 7.5 mL of hot water was added dropwise at 60° C. with vigorous stirring, and then stirred at 60° C. for 1 hour. After slowly cooling to 25° C., the mixture was filtered to obtain crystalline form 2 (XRPD and DSC are in Figures 3 and 7, respectively). Alternatively, a preferred method for preparing crystalline form 2 is as follows: Amorphous edonentan (1 g) was slurried in 20 mL of water at 25° C. for 15 hours. The solution was then filtered to obtain crystalline form 2 (XRPD and DSC are in Figures 14 and 18, respectively).

[0185] Exemplary Preparations of Crystalline Form 3 Amorphous edonentan (250 mg) was dissolved in 0.5 mL of ethyl acetate. The resulting solution was filtered and heated to 60° C., and 1.5 mL of hexane was added dropwise with vigorous stirring at 60° C. To the resulting slightly cloudy solution, 0.1 mL of ethyl acetate was added, resulting in a clear solution. This was then stirred at 60° C. for 1 hour. The solution was slowly cooled to 25° C., and the resulting precipitate was filtered to obtain crystalline Form 3 (XRPD and DSC are in FIG. 15 and FIG. 19, respectively).

[0186] Exemplary Preparations of Crystalline Form 4 Amorphous edonentan (100 mg) was added to 2 mL of water containing 0.2 mL of tetrahydrofuran (THF). The resulting mixture was stirred at 50° C. for 24 hours, cooled, and filtered to obtain Form 4, which was confirmed to be distinct from Forms 1, 2, and 3 by XRPD (FIG. 16) and DSC (FIG. 20).

[0187] In an alternative method, 107 mg of amorphous edonentan was added to 1 mL of water, followed by the equivalent amount of KOH in 1 mL of water. The resulting solution was heated to 60° C. for 20 minutes, filtered warm, and acidified with 1 mL of 0.2 N HCl. The resulting mixture was stirred at 60° C. for 5 hours, cooled, and filtered to obtain Form 4, which was confirmed by XRPD.

[0188] In an alternative method, 150 mg of edonentan (form 3) was added to a mixture of isopropanol and water (1 mL and 2 mL, respectively). The resulting slurry was stirred at 15° C. for 48 hours and then filtered. The sample was confirmed to be form 4 by XRPD analysis. This clearly indicates that form 4 is more thermodynamically stable than form 3 under these conditions.

[0189] In an alternative method, 200 mg of edonentan (form 1) was added to a mixture of isopropanol and water (1.3 mL and 2.6 mL, respectively). The resulting solution was heated to 80° C. and stirred for 24 hours, then cooled and filtered. The sample so obtained was confirmed to be form 4 by XRPD analysis. This clearly indicates that under these conditions, form 4 is more thermodynamically stable than form 1.

[0190] In an alternative method, 100 mg of edonentan (amorphous) was stirred in 10 mL of water and heated to 100° C. for 40 hours. The resulting solution was cooled to ambient temperature and filtered to obtain Form 4. In an alternative method, amorphous (crude) edonentan was dissolved in 8 volumes of isopropanol at 60° C. The resulting solution was cooled to 57° C. and then small crystals of crystalline Form 4 were added. After 2 hours, the solution was cooled to 5° C., held for 15 hours and filtered to obtain crystalline Form 4.

[0191] XRPD pattern of the crystalline form XRPD patterns of crystalline forms 1-4 are shown in Figures 12-16. XRPD patterns of the crystalline forms described herein were recorded using a Polycrystalline X-ray diffractometer (Bruker, D8 ADVANCE). CuKa radiation was operated at a voltage of 40 kv and a current of 40 mA with a 1.0 mm transmission slit and a 0.4° cable-stayed slit. The sample was placed in the center of the sample holder groove, with the surface of the sample holder flush with the surface of the sample holder. Data were collected using a lynxeye detector with a step size of 0.02° and a continuous scan at a rate of 8° / min.

[0192] Tables 5-8 below list certain XRPD characteristic peaks for crystalline forms 1-4, respectively. Table 5. Exemplary XRPD patterns of crystalline form 1 [Table 5] Table 6. Exemplary XRPD patterns of crystalline form 2 [Table 6] Table 7. Exemplary XRPD patterns of crystalline form 3 [Table 7] Table 8. Exemplary XRPD patterns of crystalline form 4 [Table 8-1] [Table 8-2]

[0193] Physicochemical properties of crystalline forms Exemplary physicochemical properties of the crystalline forms are provided herein. The melting points described herein can be measured using the following procedure:

[0194] i. Melting Point Protocol The maximum melting point peak of each crystalline form (T m ) was determined using DSC. DSC of the crystalline forms described herein was measured using a TA Instruments DSC Q2000. Samples (1.3010 mg) were weighed into aluminum crucibles and heated from 30° C. to 300° C. at a heating rate of 10° C. / min. The temperatures of the crystalline melting peak start, peak onset, peak maximum, and peak end were collected.

[0195] The solubility described herein may be measured using the following procedure: ii. Solubility Analysis Protocol 1. 2.0 mg or more of sample was weighed into the lower chamber of a whatman mini uniprep vial (GE Healthcare). 450 μL of buffer was added to each chamber. 2. Place the filter piston of the Mini Uniprep vial and push it down to liquid level to allow buffer and compound contact with the filter during incubation. 3. The samples are vortexed for 2 minutes and then incubated at room temperature (approximately 25±2° C.) for 24 hours with shaking at 500 rpm. 4. Press the Mini Uniprep to prepare the filtrate for injection into the HPLC system. Inspect all vials for visible undissolved material before filtration and for leakage after filtration. 5. Dilute the supernatant 100-fold with buffer to prepare a dilution for analysis by HPLC.

[0196] Exemplary physicochemical properties of crystalline forms 1-4 are provided below in Table 9. The physicochemical properties may be obtained using the methods described above. Table 9. Exemplary physicochemical properties of crystalline forms 1-4 [Table 9]

[0197] Other embodiments All features disclosed herein may be combined in any combination. Each feature disclosed herein may be replaced by alternative features serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

[0198] Furthermore, from the above description, those skilled in the art can easily ascertain the essential features of the present invention, and can make various changes and modifications to the present invention to adapt it to various usages and conditions without departing from the spirit and scope thereof. Accordingly, other embodiments are also within the scope of the claims.

Claims

1. 1. A composition for use in a method for preventing, treating, or ameliorating ocular neovascularization in a subject in need thereof, comprising a compound of formula I: 【Chemistry 7】 or a pharma- ceutically acceptable salt thereof, wherein the method comprises contacting an ocular tissue of the subject with the composition.

2. 2. The composition for use of claim 1, wherein the ocular neovascularization is associated with a condition selected from the group consisting of retinopathy of prematurity, retinal vein occlusion, macular edema, sickle cell retinopathy, choroidal neovascularization, radiation retinopathy, neovascular glaucoma, microangiopathy, retinal hypoxia, diabetic retinopathy, diabetic macular edema, ablation-induced neovascularization, age-related macular degeneration, and vascular leakage.

3. The composition for use according to any one of claims 1 to 2, characterized in that the composition is administered in a dosage of between about 1 μg and about 4 mg.

4. The composition for use according to claim 1 , wherein the contacting step comprises topically administering the composition to a surface of the eye or a portion thereof.

5. The composition for use according to claim 1 , wherein the contacting step comprises injecting the composition into the eye or a component thereof.

6. The composition for use according to claim 1 , wherein the contacting step comprises administering the composition intravitreally in a biodegradable ocular implant.

7. The composition for use according to claim 1, wherein edonentan or the compound of formula I is in an anhydrous crystalline form (Form 4) having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 5.6±0.2°, 11.4±0.2°, 17.7±0.2°, 19.3±0.2°, 21.1±0.2°, and 21.9±0.2° in terms of 2θ.

8. The biodegradable ocular implant comprises a biodegradable polymer having incorporated therein a compound; wherein said compound is a compound of Formula I or a pharma- ceutically acceptable salt thereof, wherein the concentration of said compound in said biodegradable polymer is about 45% w / w; and said biodegradable polymer comprises (i) RG503, RG502 and RG753S in a ratio of about 50%:about 10%:about 40%, or (ii) RG503, RG502 and RG753S in a ratio of about 20%:about 20%:about 60% 7. A composition for use according to claim 6 comprising:

9. 1. A composition for use in a method for preventing, treating, or ameliorating vascular leakage in a subject in need thereof, comprising a compound of formula I: 【Chemistry 8】 or a pharma- ceutically acceptable salt thereof, The method includes the step of contacting an ocular tissue of the subject with the composition.

10. 10. The composition for use of claim 9, wherein the vascular leakage is associated with a condition selected from the group consisting of retinopathy of prematurity, retinal vein occlusion, macular edema, sickle cell retinopathy, choroidal neovascularization, radiation retinopathy, neovascular glaucoma, microangiopathy, retinal hypoxia, diabetic retinopathy, diabetic macular edema, ablation-induced neovascularization, age-related macular degeneration, and vascular leakage.

11. The composition for use according to any one of claims 9 to 10, characterized in that the composition is administered in a dosage of between about 1 μg and about 4 mg.

12. The composition for use according to claim 9 , wherein the contacting step comprises topically administering the composition to a surface of the eye or a portion thereof.

13. The composition for use according to claim 9 , wherein the contacting step comprises injecting the composition into the eye or a component thereof.

14. The composition for use according to claim 9 , wherein the contacting step comprises administering the composition intravitreally in a biodegradable ocular implant.

15. The composition for use according to claim 9, wherein edonentan or the compound of formula I is in an anhydrous crystalline form (form 4) having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 5.6±0.2°, 11.4±0.2°, 17.7±0.2°, 19.3±0.2°, 21.1±0.2°, and 21.9±0.2° in terms of 2θ.

16. The biodegradable ocular implant comprises a biodegradable polymer having incorporated therein a compound; wherein said compound is a compound of Formula I or a pharma- ceutically acceptable salt thereof, wherein the concentration of said compound in said biodegradable polymer is about 45% w / w; and said biodegradable polymer comprises (i) RG503, RG502 and RG753S in a ratio of about 50%:about 10%:about 40%, or (ii) RG503, RG502 and RG753S in a ratio of about 20%:about 20%:about 60% 15. A composition for use according to claim 14 comprising:

17. 1. A composition for use in a method for preventing, treating, or ameliorating neovascular age-related macular degeneration in a subject in need thereof, comprising a compound of formula I: 【Chemistry 9】 or a pharma- ceutically acceptable salt thereof, The method includes the step of contacting an ocular tissue of the subject with the composition.

18. The composition for use according to claim 17, characterized in that the composition is administered in a dosage of between about 1 μg and about 4 mg.

19. The composition for use according to any one of claims 17 to 18, wherein the contacting step comprises topically administering the composition to a surface of the eye or a portion thereof.

20. The composition for use according to any one of claims 17 to 18, wherein the step of contacting comprises injecting the composition into the eye or a component thereof.

21. The composition for use according to any one of claims 17 to 18, wherein the contacting step comprises intravitreally administering the composition in a biodegradable ocular implant.

22. The composition for use according to claim 17, wherein edonentan or the compound of formula I is in an anhydrous crystalline form (form 4) having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 5.6±0.2°, 11.4±0.2°, 17.7±0.2°, 19.3±0.2°, 21.1±0.2°, and 21.9±0.2° in terms of 2θ.

23. The biodegradable ocular implant comprises a biodegradable polymer having incorporated therein a compound; wherein said compound is a compound of Formula I or a pharma- ceutically acceptable salt thereof, wherein the concentration of said compound in said biodegradable polymer is about 45% w / w; and said biodegradable polymer comprises (i) RG503, RG502 and RG753S in a ratio of about 50%:about 10%:about 40%, or (ii) RG503, RG502 and RG753S in a ratio of about 20%:about 20%:about 60% 22. A composition for use according to claim 21 comprising:

24. 1. A composition for use in a method for preventing, treating, or ameliorating macular edema in a subject in need thereof, comprising a compound of formula I: 【Chemistry 10】 or a pharma- ceutically acceptable salt thereof, A composition, wherein the method comprises the step of contacting an ocular tissue of the subject with the composition.

25. The composition for use according to claim 24, characterized in that the composition is administered in a dosage of between about 1 μg and about 4 mg.

26. The composition for use according to any one of claims 24 to 25, wherein the contacting step comprises topically administering the composition to a surface of the eye or a portion thereof.

27. The composition for use according to any one of claims 24 to 25, wherein the step of contacting comprises injecting the composition into the eye or a component thereof.

28. The composition for use according to any one of claims 24 to 25, wherein the contacting step comprises intravitreally administering the composition in a biodegradable ocular implant.

29. The composition for use according to claim 24, wherein edonentan or the compound of formula I is in an anhydrous crystalline form (form 4) having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 5.6±0.2°, 11.4±0.2°, 17.7±0.2°, 19.3±0.2°, 21.1±0.2°, and 21.9±0.2° in terms of 2θ.

30. The biodegradable ocular implant comprises a biodegradable polymer having incorporated therein a compound; wherein said compound is a compound of Formula I or a pharma- ceutically acceptable salt thereof, wherein the concentration of said compound in said biodegradable polymer is about 45% w / w; and said biodegradable polymer comprises (i) RG503, RG502 and RG753S in a ratio of about 50%:about 10%:about 40%, or (ii) RG503, RG502 and RG753S in a ratio of about 20%:about 20%:about 60% 29. A composition for use according to claim 28 comprising:

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