Compositions for treatment of ocular diseases

Abstract

To provide a method of using certain compositions comprising Edonentan crystalline forms for the treatment of ocular diseases selected from glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), and retinopathy of prematurity (ROP).SOLUTION: The present disclosure relates to the discovery that certain ocular diseases can be treated using Edonentan crystalline forms. Specific Edonentan crystalline forms can be used alone or in combination with an intraocular pressure (IOP) reducing agent, a neuroprotective agent, or an anti-VEGF agent. Using Edonentan crystalline forms, either alone or in combination with an additional agent, provides increased perfusion to the retina or reduced IOP in certain ocular diseases and reduces damage to retinal cells.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Applications Nos. 62 / 971,002 and 63 / 010,212, filed February 6, 2020 and April 15, 2020, respectively, the entire contents of which are incorporated herein by reference for all purposes. Field

[0002] The present disclosure relates to the field of medicine and the treatment of ophthalmic diseases. More specifically, the present disclosure relates to the use of certain compositions comprising edonentan crystalline forms in the treatment or amelioration of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), and retinopathy of prematurity (ROP). [Background technology]

[0003] background Eye diseases have a significant impact on the quality of human life, yet for the most part, effective treatments remain elusive. An estimated annual economic burden of over $100 billion is attributed to vision loss, eye disease, and visual impairment in the United States. Examples of debilitating eye diseases include glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), and retinopathy of prematurity (ROP).

[0004] Glaucoma is an eye disorder characterized by visual field defects and optic disc depression.It is generally known that abnormally high intraocular pressure is harmful to the eye, and there is clear evidence that abnormally high intraocular pressure is probably the most important physical change that causes retinal degeneration in glaucoma patients.Ultimately, if not treated, vision will gradually be lost over time.However, the pathophysiological mechanism of glaucoma remains unclear.

[0005] There are three basic types of glaucoma: primary, secondary, and congenital. Primary glaucoma is the most common type and can be divided into open-angle and angle-closure glaucoma. Primary open-angle glaucoma (POAG; e.g., normal-tension glaucoma (NTG)) is the most common type of glaucoma observed in the United States. POAG is usually detected in its early stages during a routine eye examination. Primary angle-closure glaucoma, also known as acute glaucoma, usually develops suddenly and is characterized by eye pain and blurred vision. Secondary glaucoma occurs as a complication of various other conditions, such as pseudoexfoliation syndrome, injury, inflammation, systemic vascular disease, and diabetes. Congenital glaucoma results from impaired development of the eye's drainage system.

[0006] Diabetic retinopathy (DR) is the most common complication of diabetes and the leading cause of vision loss and blindness in the working-age population in developed countries. The incidence of DR increases over time as diabetes progresses. Thus, 90% of patients with type 1 diabetes and 60% of patients with type 2 diabetes have some degree of DR after 20 years of diabetes progression. The prevalence of DR in Western countries is very similar, at approximately 30%, with DR reaching a severely sight-threatening advanced stage in 10% of cases.

[0007] DR occurs when changes in blood sugar levels cause changes in retinal blood vessels. In some cases, these blood vessels swell (macular edema) and leak fluid into the back of the eye. In other cases, abnormal blood vessels grow on the surface of the retina. Unless treated, DR gradually becomes more severe, progressing from "background retinopathy" to severely affecting vision and potentially leading to blindness.

[0008] Retinal vein occlusion (RVO) is a vascular disorder of the retina and one of the most common causes of vision loss worldwide. Specifically, RVO is the second most common cause of blindness due to retinal vascular disease after diabetic retinopathy. RVO is often the result of underlying health problems (e.g., high blood pressure, 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.

[0009] Currently, there is no way to unblock retinal veins, and accepted treatments are aimed at addressing health problems associated with retinal vein occlusion. Some eyes with retinal vein occlusion may regain vision. Approximately one-third have some improvement, one-third remain the same, and one-third gradually improve, but it may take more than a year to determine final results. In some cases, blocked blood vessels lead to the accumulation of fluid within the retina. In other cases, the development of ischemia leads to the formation of new blood vessels. RVO is currently treated with intravitreal injections of anti-VEGF drugs.

[0010] Retinopathy of prematurity (ROP) can result from premature birth. Abnormal, leaky vascular growth (neovascularization) in the retina can occur secondary to other procedures in premature infants and often leads to neonatal blindness. During pregnancy, blood vessels grow from the center of the developing fetal retina by 16 weeks of the mother's pregnancy, then branch outward, reaching the retinal edges by the eighth month of pregnancy. In infants born prematurely, normal retinal vascular growth is incomplete and therefore more easily destroyed.

[0011] Endothelin ("ET") is a family of naturally occurring peptides identified as ET-1, ET-2, and ET-3. ET, which is primarily produced in endothelial cells and plays an important role in vascular homeostasis, constricts blood vessels, increases renal blood pressure, and affects glomerular hemodynamics and sodium and water homeostasis. When overexpressed, ET contributes to high blood pressure (hypertension), heart disease, kidney damage, and may contribute to other diseases such as eye disease. See, for example, Salvatore et al., J Ophthalmol. 2010, 2010:354645.

[0012] Endothelin receptor antagonists are pharmacological agents that inhibit endothelin receptors.Some agents inhibit both endothelin A receptors and endothelin B receptors, while other agents selectively inhibit only one of the two receptors (e.g., endothelin A receptor antagonists).Endothelin receptor antagonists have been shown to reduce mortality and improve hemodynamics in experimental models (e.g., heart failure).

[0013] Edonentan (BMS-207940) 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). Edonentan was tested in Phase I trials by April 2002, but its development was discontinued. There remains a need to more effectively reduce the incidence of, treat, or otherwise ameliorate glaucoma, DR, RVO, and ROP. [Prior art documents] [Non-patent literature]

[0014] [Non-Patent Document 1] Salvatore et al., J Ophthalmol.2010,2010:354645 Summary of the Invention [Means for solving the problem]

[0015] overview The present invention provides methods of using certain compositions comprising edonentan crystalline forms for treating ocular diseases selected from glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO) and retinopathy of prematurity (ROP).

[0016] In one aspect, the present invention provides a compound of formula I: [ka] a solid form of The solid form relates to a solid form that is 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 two-theta.

[0017] 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°.

[0018] 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°.

[0019] In some embodiments of the solid form, the anhydrous crystalline Form 4 has a T of about 163° C. by DSC analysis. m Shows.

[0020] In another aspect, the present invention provides a compound of formula I: [ka] A composition comprising: the compound is an anhydrous crystalline form (Form 4) having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from peaks at 5.6±0.2 degrees, 11.4±0.2 degrees, 17.7±0.2 degrees, 19.3±0.2 degrees, 21.1±0.2 degrees, and 21.9±0.2 degrees in terms of two-theta; The compound is characterized in that the composition is 90% or more by weight crystalline form 4, based on the total weight of the compound present in the composition.

[0021] In some embodiments, the compound of formula I is 95% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0022] In some embodiments, the compound of formula I is 96% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0023] In some embodiments, the compound of formula I is 97% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0024] In some embodiments, the compound of formula I is 98% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0025] In some embodiments, the compound of formula I is 99% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0026] A method of treating an ocular disease, comprising administering to a subject ocular tissue a therapeutically effective amount of a compound of formula I: [ka] contacting the composition containing Within the scope of the present invention is a method wherein the ocular disease is selected from the group consisting of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), and retinopathy of prematurity (ROP), the compound is in a crystalline form (Form 4), the crystalline Form 4 having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from 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, and the compound is 90% or more by weight of crystalline Form 4, based on the total weight of the compound present in the composition.

[0027] The present invention further provides a compound of formula I: [ka] 1. A process for preparing the anhydrous crystalline form (Form 4) of The method comprises: (a) stirring a compound of Formula I in an aqueous solution, an organic solvent, or a mixture thereof at a temperature ranging from about 40° C. to about 120° C.; (b) cooling the resulting solution to a temperature in the range of about 0° C. to room temperature (e.g., about 25° C.), and, if the compound is in a basic aqueous solution, adjusting its pH value (e.g., by adding HCl) to a range of about 3 to about 7; (c) filtering the sample thus obtained to obtain Crystalline Form 4; Including, wherein said crystalline Form 4 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 5.6±0.2 degrees, 11.4±0.2 degrees, 17.7±0.2 degrees, 19.3±0.2 degrees, 21.1±0.2 degrees, and 21.9±0.2 degrees in terms of two-theta.

[0028] In some embodiments of the above preparation methods, the aqueous solution is water.

[0029] In some embodiments of the preparation method, the aqueous solution is a basic aqueous solution. In some embodiments, the basic aqueous solution has a pH value of 8 or greater.

[0030] In some embodiments of the preparation method, the basic aqueous solution is an aqueous potassium hydroxide solution or an aqueous potassium carbonate solution.

[0031] In some embodiments of the preparation method, the organic solvent is a water-soluble organic solvent. In some embodiments, the organic solvent is tetrahydrofuran or isopropanol.

[0032] In some embodiments of the preparation method, the temperature for stirring is in the range of about 80°C to about 120°C (e.g., about 80°C to about 90°C, about 90°C to about 100°C, about 100°C to about 110°C, or about 110°C to about 120°C).

[0033] In some embodiments of the preparation method, the temperature for stirring is in the range of about 40°C to about 80°C (e.g., about 40°C to about 50°C, about 50°C to about 60°C, about 60°C to about 70°C, or about 70°C to about 80°C).

[0034] In some embodiments of the preparation method, the stirring is carried out for about 20 hours to about 50 hours (eg, about 20 hours to about 30 hours, about 30 hours to about 40 hours, or about 40 hours to about 50 hours).

[0035] In some embodiments of the above-described preparation method, a slurry of the compound of Formula I is formed in the stirring step.

[0036] In some embodiments of the preparation method, the method further comprises seeding an amount of crystalline Form 4 and holding the resulting solution for about 2 hours.

[0037] In some embodiments of the above-described preparation methods, the cooling is performed at about 5° C. for about 15 hours.

[0038] In some embodiments of the preparation method, the pH value of the basic aqueous solution is adjusted to a range of about 3 to about 4. In some embodiments of the preparation method, the pH value of the basic aqueous solution is adjusted to a range of about 3 to about 5. In some embodiments of the preparation method, the pH value of the basic aqueous solution is adjusted to a range of about 3 to about 6. In some embodiments of the preparation method, the pH value of the basic aqueous solution is adjusted to a range of about 5 to about 6. In some embodiments of the preparation method, the pH value of the basic aqueous solution is adjusted to a range of about 5.5 to about 6. In some embodiments of the preparation method, the pH value of the basic aqueous solution is adjusted to a range of about 6 to about 7.

[0039] A method of treating an ocular disease, comprising treating ocular tissue of a subject with a therapeutically effective amount of a compound of formula I, i.e., edonentan : [ka] contacting the composition containing the ocular disease is selected from the group consisting of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO) and retinopathy of prematurity (ROP), and the compound is selected from the group consisting of an anhydrous crystalline form (Form 1), a monohydrate crystalline form (Form 2) and an anhydrous crystalline form (Form 3); (i) anhydrous crystalline Form 1 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from peaks 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 two-theta, and the compound is 90% or more by weight, based on the total weight of the compound present in the composition, of crystalline Form 1; (ii) monohydrate crystalline Form 2 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from peaks 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 two-theta, wherein the compound is 90% or more by weight, based on the total weight of the compound present in the composition; Also within the scope of the invention is a method wherein (iii) anhydrous crystalline Form 3 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from peaks 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 two-theta, and the compound is 90% or more by weight, based on the total weight of the compound present in the composition, of crystalline Form 3.

[0040] In some embodiments, the contacting comprises administering a topical composition to the surface of the eye or a portion thereof, while in other embodiments, the contacting comprises injecting a composition containing an edonentan crystalline form into the eye, either broadly or in a specific area of the eye.

[0041] In some embodiments, the eye disease is glaucoma.In another embodiment, the therapeutic effectiveness of treating glaucoma is determined by detecting the amount sufficient to reduce intraocular pressure, or reduce the rate of optic nerve damage / thinning of retinal nerve fiber layer, reduce contrast sensitivity or visual field loss, alleviate or prevent optic nerve damage.In other embodiments, the therapeutic effectiveness of treating glaucoma is determined by measuring the improvement of retina, optic nerve head or tissue perfusion.

[0042] In some embodiments for the treatment of glaucoma, the regimen further comprises the addition of a therapeutically effective amount of an intraocular pressure (IOP)-lowering agent or neuroprotective agent, or any of the pharmaceutically acceptable salts thereof.In some embodiments, the IOP-lowering agent is selected from the group consisting of prostaglandins (such as latanoprost or travoprost), beta-blockers (such as timolol or betaxolol), alpha-adrenergic agonists (such as brimonidine, apraclonidine), carbonic anhydrase inhibitors (such as dorzolamide or brinzolamide), Rho kinase inhibitors (such as netarsudil) and miotics or cholinergic agents (such as pilocarpine).In some embodiments, the neuroprotective agent is selected from the group consisting of antiapoptotic agents (such as caspase-2 inhibitors) and neurotrophic factors (such as ciliary neurotrophic factor).

[0043] In some embodiments, the eye disease is diabetic retinopathy (DR). In further embodiments, the therapeutic effectiveness of treating DR is determined by reducing diabetes-induced retinal neovascularization, diabetic retinopathy severity score and neurodegeneration. In other embodiments, the therapeutic effectiveness of treating DR is determined by measuring the improvement of retinal or choroidal perfusion.

[0044] In some embodiments, the disease is retinal vein occlusion (RVO). In further embodiments, the therapeutic effectiveness of treating RVO is determined by measuring improved tissue perfusion, reduced inflammation, or a combination of the foregoing.

[0045] In some embodiments, the ocular disease is retinopathy of prematurity (ROP). In further embodiments, the therapeutic efficacy of treating ROP is determined by measuring an improvement in retinal perfusion and a reduction in abnormal neovascularization.

[0046] In some embodiments, the composition administered contains an edonentan crystalline form in a dosage of about 1 μg to about 5 mg (e.g., about 1 μg to about 10 μg, about 10 μg to about 50 μg, about 50 μg to about 100 μg, about 100 μg to about 500 μg, about 500 μg to about 1 mg, about 1 mg to about 1.5 mg, about 1.5 mg to about 2 mg, about 2 mg to 2.5 mg, about 2.5 mg to about 3 mg, about 3 mg to 3.5 mg, about 3.5 mg to 4 mg, about 4 mg to 4.5 mg, and about 4.5 mg to 5 mg). In some embodiments, the administered composition is from about 10 μg to about 3 mg (e.g., from about 10 μg to about 100 μg, from about 10 μg to about 500 μg, from about 10 μg to about 1 mg, from about 10 μg to about 2 mg, from 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 300 μg, about 350 μg, The edonentan crystalline form may be administered in a dosage of about 400 μg, about 450 μg, about 500 μg, about 550 μg, about 600 μg, about 650 μg, about 700 μg, about 750 μg, about 800 μg, about 850 μg, about 900 μg, about 950 μg, about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75 mg, about 2 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, or about 3 mg. In some embodiments, the administered composition contains an edonentan crystalline form in a dosage of about 10 μg to about 1 mg. In some embodiments, the compositions described herein are administered in a single dosage form. In some embodiments, the compositions described herein are administered in multiple dosage forms.

[0047] The present invention relates to compounds of formula I: [ka] 1. A method for preparing the monohydrate crystalline form (Form 2) of The method comprises: (a) preparing a slurry of a compound of Formula I (e.g., amorphous form) in an aqueous medium (e.g., water) at a temperature ranging from about 15°C to about 35°C for a predetermined period of time (e.g., about 15 hours) to obtain a sample; (b) filtering the sample thus obtained to obtain Crystalline Form 2; Including, Also encompassed are methods wherein said crystalline Form 2 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 9.6±0.2 degrees, 10.4±0.2 degrees, 19.6±0.2 degrees, 19.7±0.2 degrees, 22.0±0.2 degrees, 22.9±0.2 degrees, and 23.7±0.2 degrees in terms of two-theta.

[0048] In some embodiments of the above preparation methods, the aqueous medium is water.

[0049] In some embodiments of the preparation method, the predetermined period of time is about 15 hours.

[0050] Additional features and advantages of the present technology will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the present technology. The advantages of the present technology will be realized and attained by the structure particularly pointed out in the written description and aspects.

[0051] The details of one or more embodiments of the disclosure are set forth in the description below. Other features, objects, and advantages of the disclosure will become apparent from the following drawings, description, and claims. BRIEF DESCRIPTION OF THE DRAWINGS [Brief explanation of the drawings]

[0052] [Figure 1] FIG. 1 shows exemplary XRPD patterns of Forms 1-4.

[0053] [Figure 2] FIG. 2 shows an exemplary XRPD pattern of Form 1.

[0054] [Figure 3] FIG. 3 shows an exemplary XRPD pattern of Form 2.

[0055] [Figure 4] FIG. 4 shows an exemplary XRPD pattern of Form 3.

[0056] [Figure 5] FIG. 5 shows an exemplary XRPD pattern of Form 4.

[0057] [Figure 6] FIG. 6 shows an exemplary DSC curve for Form 1.

[0058] [Figure 7] FIG. 7 shows an exemplary DSC curve for Form 2.

[0059] [Figure 8] FIG. 8 shows an exemplary DSC curve for Form 3.

[0060] [Figure 9] FIG. 9 shows an exemplary DSC curve for Form 4. DETAILED DESCRIPTION OF THE INVENTION

[0061] Detailed Description The present invention arises from the discovery that certain compositions comprising edonentan crystalline forms can be used to prevent, treat, or otherwise ameliorate ocular diseases, including, but not limited to, glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), and retinopathy of prematurity (ROP). The invention is further described below.

[0062] As provided in the Summary section, the present invention encompasses methods of treating an ocular disease, comprising contacting ocular tissue of a subject with a composition containing a therapeutically effective amount of a compound of Formula I, wherein the ocular disease is selected from the group consisting of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), and retinopathy of prematurity (ROP), and wherein the compound is an anhydrous crystalline form (Form 1), the anhydrous crystalline Form 1 having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from peaks 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 two-theta, and wherein the compound is 90% or more by weight of crystalline Form 1, based on the total weight of the compound present in the composition.

[0063] In some embodiments, the anhydrous crystalline Form 1 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 6.3±0.2°, 7.5±0.2°, 11.7±0.2°, and 15.1±0.2°.

[0064] In some embodiments, the anhydrous crystalline Form 1 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 7.5±0.2°, 11.7±0.2°, and 15.1±0.2°.

[0065] In some embodiments, the anhydrous crystalline Form 1 has a T of about 151° C. by DSC analysis. m It has.

[0066] In some embodiments, the anhydrous crystalline Form 1 has a solubility of about 264 μg / mL in phosphate buffer at about pH 7.

[0067] In some embodiments, the compound of formula I is 95% or more by weight in crystalline Form 1, based on the total weight of said compound present in said composition.

[0068] In some embodiments, the compound of formula I is 96% or more by weight of crystalline Form 1, based on the total weight of said compound present in said composition.

[0069] In some embodiments, the compound of formula I is 97% or more by weight of crystalline Form 1, based on the total weight of said compound present in said composition.

[0070] In some embodiments, the compound of formula I is 98% or more by weight in crystalline Form 1, based on the total weight of said compound present in said composition.

[0071] In some embodiments, the compound of formula I is 99% or more by weight of crystalline Form 1, based on the total weight of said compound present in said composition.

[0072] The invention also encompasses a method of treating an ocular disease, comprising contacting ocular tissue of a subject with a therapeutically effective amount of a composition containing the compound of Formula I, wherein the ocular disease is selected from the group consisting of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), and retinopathy of prematurity (ROP), and wherein the compound is in a monohydrate crystalline form (Form 2), and wherein the monohydrate crystalline Form 2 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from peaks 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 two-theta, and wherein the compound of Formula I is 90% by weight or more crystalline Form 2, based on the total weight of the compound present in the composition.

[0073] In some embodiments, monohydrate crystalline Form 2 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 19.6±0.2°, 19.7±0.2°, and 9.6±0.2°.

[0074] In some embodiments, monohydrate crystalline Form 2 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 19.6±0.2°, 19.7±0.2°, 9.6±0.2°, 10.4±0.2°, 22.0±0.2°, and 22.9±0.2°.

[0075] In some embodiments, the monohydrate crystalline Form 2 has a T of about 122° C. by DSC analysis. m It has.

[0076] In some embodiments, monohydrate crystalline Form 2 has a solubility of about 35 μg / mL in phosphate buffer at about pH 7.

[0077] In some embodiments, the compound of formula I is 95% or more by weight in crystalline Form 2, based on the total weight of said compound present in said composition.

[0078] In some embodiments, the compound of formula I is 96% or more by weight in crystalline Form 2, based on the total weight of said compound present in said composition.

[0079] In some embodiments, the compound of formula I is 97% or more by weight in crystalline Form 2, based on the total weight of said compound present in said composition.

[0080] In some embodiments, the compound of formula I is 98% or more by weight in crystalline Form 2, based on the total weight of said compound present in said composition.

[0081] In some embodiments, the compound of formula I is 99% or more by weight in crystalline Form 2, based on the total weight of said compound present in said composition.

[0082] The present invention further encompasses a method of treating an ocular disease, comprising contacting ocular tissue of a subject with a therapeutically effective amount of a composition containing a compound of Formula I, wherein the ocular disease is selected from the group consisting of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), and retinopathy of prematurity (ROP), and wherein the compound is an anhydrous crystalline form (Form 3), wherein the anhydrous crystalline Form 3 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from peaks 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 two-theta, and wherein the compound of Formula I is 90% by weight or more crystalline Form 3, based on the total weight of the compound present in the composition.

[0083] In some embodiments, the anhydrous crystalline Form 3 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 9.0±0.2°, 15.8±0.2°, 7.8±0.2°, and 19.1±0.2°.

[0084] In some embodiments, the anhydrous crystalline Form 3 has the following X-ray powder diffraction pattern expressed in terms of diffraction angles (2θ): 9.0±0.2°, 15.8±0.2°, and 7.8±0.2°.

[0085] In some embodiments, the anhydrous crystalline Form 3 has a T of about 162° C. by DSC analysis. m It has.

[0086] In some embodiments, the anhydrous crystalline Form 3 has a solubility of about 251 μg / mL in phosphate buffer at about pH 7.

[0087] In some embodiments, the compound of formula I is 95% or more by weight in crystalline form 3, based on the total weight of said compound present in said composition.

[0088] In some embodiments, the compound of formula I is 96% or more by weight in crystalline form 3, based on the total weight of said compound present in said composition.

[0089] In some embodiments, the compound of formula I is 97% or more by weight in crystalline form 3, based on the total weight of said compound present in said composition.

[0090] In some embodiments, the compound of formula I is 98% or more by weight in crystalline form 3, based on the total weight of said compound present in said composition.

[0091] In some embodiments, the compound of formula I is 99% or more by weight in crystalline form 3, based on the total weight of said compound present in said composition.

[0092] Further encompassed by the present invention is a method of treating an ocular disease, comprising contacting ocular tissue of a subject with a therapeutically effective amount of a composition containing the compound of Formula I, wherein the ocular disease is selected from the group consisting of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), and retinopathy of prematurity (ROP), and wherein the compound is in a crystalline form (Form 4), and wherein the crystalline Form 4 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from 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, and wherein the compound of Formula I is 90% by weight or more crystalline Form 4, based on the total weight of the compound present in the composition.

[0093] In some embodiments, 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°.

[0094] In some embodiments, 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°.

[0095] In some embodiments, crystalline Form 4 has the following X-ray powder diffraction pattern, expressed in terms of diffraction angles (2θ): [Table 8]

[0096] In some embodiments, the anhydrous crystalline Form 4 has a T of about 163° C. by DSC analysis. m It has.

[0097] In some embodiments, the compound of formula I is 95% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0098] In some embodiments, the compound of formula I is 96% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0099] In some embodiments, the compound of formula I is 97% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0100] In some embodiments, the compound of formula I is 98% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0101] In some embodiments, the compound of formula I is 99% or more by weight in crystalline form 4, based on the total weight of said compound present in said composition.

[0102] In some embodiments, the therapeutic effectiveness of the treatment is determined by (i) detecting a reduction in intraocular pressure or a reduction in the rate of optic nerve damage by an amount sufficient to alleviate or prevent optic nerve damage; (ii) assessing the degree of improvement in visual acuity or visual field; (iii) measuring a reduction in diabetes-induced retinal neurodegeneration; (iv) measuring an improvement in tissue or retinal perfusion; or (v) measuring an improvement in tissue or retinal perfusion, a reduction in inflammation, or a combination thereof.

[0103] The methods of the present invention involve contacting ocular tissue (topically or intraocularly) with a therapeutically effective amount of a composition comprising an edonentan crystalline form, or administering a therapeutically effective amount of a composition comprising an edonentan crystalline form.

[0104] Methods for preparing edonentane (e.g., amorphous edonentane) are well known to those skilled in the art. Suitable methods are disclosed, for example, in U.S. Pat. No. 6,043,265 and WO 2002 / 32884. Edonentane 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 of 536.6 g / mol) and the structure of Formula I shown above. Edonentan crystal morphology

[0105] The crystalline forms of edonentan disclosed herein may be substantially more stable than the amorphous form of edonentan. For example, the disclosed crystalline forms (e.g., Form 4) can be stored under practical and economical storage conditions while retaining their physical properties so that they can be manufactured into dosage forms. In another series of examples, the disclosed crystalline forms (e.g., Form 1, Form 2, Form 3) are converted to the disclosed crystalline forms (e.g., Form 4). For example, heating a slurry of crystalline Form 1 in approximately 20 mL of a 1:2 mixture of isopropanol and water at 80°C for 24 hours, followed by cooling and filtering the resulting solution, yields crystalline Form 4. In another example, stirring the amorphous form of the compound of Formula I in 20 mL of water at 25°C for 15 hours yields crystalline Form 2, which is then subsequently heated at 100°C for 40 hours and filtered to yield crystalline Form 4. In yet another example, heating a slurry of anhydrous crystalline form 3 in 30 mL of water at 80° C. for 40 hours, followed by cooling and filtering a sample obtained from the solution, yields crystalline form 4. In one embodiment, crystalline form 4 is more stable (e.g., thermodynamically) than certain other crystalline forms. In one embodiment, crystalline form 4 is more stable (e.g., thermodynamically) than a crystalline form selected from the group consisting of crystalline form 1, crystalline form 2, and crystalline form 3. In one embodiment, the disclosed crystalline forms may have improved chemical and / or physical stability, e.g., when formulated in a pharmaceutical formulation, compared to, e.g., amorphous forms. In some embodiments, the crystalline form is crystalline form 4.

[0106] As used herein, the term "amorphous" refers to a solid material whose molecular positions do not have long-range order. Amorphous solids are generally supercooled liquids in which there is no clearly defined arrangement, such as molecular packing, and the molecules are randomly arranged so that there is no 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.

[0107] Hydrate forms of crystalline edonentan, e.g., edonentan·(H2O) m is contemplated, 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).

[0108] A general method for preparing crystalline Form 1 or crystalline Form 2 is provided below: Amorphous (crude) edonentan is dissolved in isopropyl alcohol (IPA). The solution is filtered and the filter is washed with a small amount of IPA. The resulting solution is heated to 60°C, and warm water is added dropwise with vigorous stirring, and the solution is stirred at the same temperature for 1-2 hours. The solution is slowly cooled to room temperature to obtain crystalline Form 1 or Form 2, depending on the ratio of IPA to water. Alternatively, in a preferred method, amorphous (crude) edonentan is slurried in 20 mL of water at 25°C for 15 hours, followed by filtration to obtain crystalline Form 2.

[0109] A general method for preparing crystalline Form 3 is provided below: Amorphous (crude) edonentane is dissolved in ethyl acetate (EA). The resulting solution is filtered and heated to 50-60°C. Hexane is added dropwise with vigorous stirring. Additional EA is added to clarify the solution, followed by stirring at the same temperature for 1-2 hours. The solution is slowly cooled to room temperature to obtain crystalline Form 3.

[0110] A general method for preparing crystalline Form 4 is provided below: Amorphous (crude) edonentane is added to a mixture of an organic solvent (e.g., tetrahydrofuran) and water. The resulting mixture is stirred at elevated temperature, cooled, and filtered to obtain Form 4. Alternatively, amorphous edonentane is dissolved in a basic aqueous solution (e.g., aqueous potassium hydroxide or aqueous potassium carbonate). The resulting solution is heated to 50-60°C, filtered while warm, and acidified with an acid (e.g., HCl). The resulting mixture is then stirred, cooled, and filtered to obtain Form 4. Alternatively, in a preferred method, amorphous (crude) edonentane is dissolved in 8 mL of isopropanol at 60°C. The resulting solution is cooled to 57°C, and small crystals of crystalline Form 4 are then added. After 2 hours, the solution is cooled to 5°C, held for 15 hours, and filtered to obtain crystalline Form 4. eye disease

[0111] The methods of the present disclosure include the use of compositions comprising the above-described edonentan crystalline forms in the treatment and amelioration of ocular diseases selected from glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO) and retinopathy of prematurity (ROP), as described below. glaucoma

[0112] In the treatment of glaucoma using compositions comprising the edonentan crystalline forms described herein, a "therapeutically effective amount" can be determined by assessing improvements in retinal blood flow (RBF) beyond that which can be achieved by standard treatment (reduction of intraocular pressure (IOP)). For glaucoma indications, blood flow improvement in a healthy rabbit eye model can be used as a predictor of pharmacodynamic responses (PDs) in humans. Due to the anatomical and functional similarities between rabbit and human eyes, rabbits are commonly used to evaluate the ocular PK / PD relationship for compounds targeting human ocular diseases. Previously, intravitreal administration of ET-1 into rabbit eyes has been shown to induce significant vasoconstriction and optic nerve damage (Sasaoka M. et al., Exp Eye Res 2006; Sugiyama T. et al., Arch Ophthalmol 2009). Pharmacodynamic responses in this model, based on the reversal of perfusion impairment induced by intravitreal ET-1 administration, can be modeled against target pharmacodynamic responses in human glaucoma patients, where ET-1 levels are observed to be elevated in plasma and aqueous humor (Li S. et al., Journal of Ophthalmology 2016).

[0113] Other examples of suitable animal glaucoma models are the Morrison rat model of chronically 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 hypertonic saline administration via the episcleral vein. In the laser-induced NHP glaucoma model, optic nerve head blood flow has been shown to decrease after sustained elevation of IOP (Wang L. et al., Invest Ophthalmol Vis Sci 2012). Furthermore, decreased optic nerve head 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). Edonentan can dose-dependently improve optic nerve head blood flow in a laser-induced NHP glaucoma model.

[0114] Efficacy in the above glaucoma models is defined as a reduction in IOP, improvement in optic nerve head or retinal blood flow from baseline, prevention or delay of progression of structural neurodegenerative changes such as retinal nerve fiber layer thickness measured by optical coherence tomography (OCT) or retinal ganglion cell count in flat mounts, and functional changes such as electroretinography (ERG) or contrast sensitivity after treatment with a composition containing edonentan crystalline form.

[0115] It is believed that the effect of compositions containing edonentan crystalline forms on retinal blood flow can be assessed by the vessel radius (r) in Poiseuille's law. An increase in (r) by an endothelin antagonist will induce a more pronounced increase in blood flow than can be achieved by increasing perfusion pressure through IOP reduction: Blood flow = (perfusion pressure × πr 4 ) / (8ηl) During the ceremony: l: Blood vessel length r: vessel radius η: Blood viscosity Perfusion pressure: Mean arterial pressure - IOP Furthermore, compositions containing edonentan crystalline forms may reduce IOP and / or prevent RGC death through mechanisms independent of improved retinal / optic nerve head tissue perfusion. Thus, by using compositions containing certain specific edonentan crystalline forms (e.g., Form 4), one (r) and / or more (IOP) of the above parameters may be altered to improve RBF and thereby achieve a therapeutic effect in treating glaucoma.

[0116] In some embodiments, glaucoma patients begin treatment as soon as they are diagnosed. In some embodiments, a composition containing an edonentan crystalline form is administered intravitreally, suprachoroidally, or locally to the back of the eye using an implant delivery platform every 3 to 12 months (e.g., every 3 to 6 months or every 4 to 6 months). In some embodiments for treating glaucoma, a composition containing an edonentan crystalline form is administered in a topical form (e.g., eye drops). Diabetic retinopathy (DR)

[0117] Diabetes can cause serious late complications, classified as microangiopathy (retinopathy, neuropathy, and diabetic nephropathy) and macroangiopathy (cardiovascular disease). Diabetic retinopathy results from damage to the small blood vessels and neurons of the retina. The earliest changes leading to diabetic retinopathy include narrowing of the retinal arteries with reduced retinal blood flow; dysfunction of inner retinal neurons with subtle changes in visual function, followed in later stages by changes in the function of the outer retina; and dysfunction of the blood-retinal barrier, which protects the retina from many substances in the blood (including toxins and immune cells), resulting in leakage of blood components into the retinal neuropil. Subsequently, the basement membrane of retinal blood vessels thickens, capillaries degenerate, and cells, particularly pericytes and vascular smooth muscle cells, are lost. This leads to loss of blood flow and progressive ischemia, as well as microscopic aneurysms, which appear as balloon-like structures protruding from the capillary walls and recruit inflammatory cells, leading to the progressive dysfunction and degeneration of retinal neurons and glial cells.

[0118] The ischemia and oxidative damage observed in DR impair blood flow and tissue ischemia, and the inventors have discovered that these can be reversed by a composition containing edonentan crystalline form. For DR indications, improving retinal perfusion is expected to have the benefit of reducing hypoxia and suppressing the upregulation of vascular endothelial growth factor (VEGF), thereby slowing vascular proliferative changes, neovascularization, and / or macular edema complications.

[0119] A preclinical mouse model of retinopathy of prematurity (ROP) can be used as a surrogate model for the ischemic retinopathy observed in DR. Oxygen-induced retinopathy in mice is a reproducible and quantifiable model of proliferative retinal neovascularization suitable for investigating the pathogenesis and therapeutic intervention of retinal neovascularization in ROP and other vascular pathologies, including DR. This model is induced by exposing 1-week-old C57BL / 6J mice to 75% oxygen for 5 days, followed by exposure to room air as previously described (Smith LEH et al., Invest Ophthalmol Vis Sci 1994). Efficacy in this preclinical model of ROP can be assessed by studying retinal hypoxia and neovascularization. Current standard treatment for DR includes anti-VEGF therapy, which only addresses advanced vascular complications of the disease. In some embodiments, patients with DR are initiated on this treatment during the non-proliferative stage of the disease. In some embodiments, the composition comprising the edonentan crystalline form is administered intravitreally, suprachoroidally, or locally to the back of the eye using an implant delivery platform every 3-12 months (e.g., every 3-6 months or every 4-6 months). In some embodiments for treating DR, the composition comprising the edonentan crystalline form is administered in a topical form (e.g., eye drops). Retinal vein occlusion (RVO)

[0120] 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 cause the edema. It would be highly desirable to use a composition containing edonentan crystalline forms to treat RVO by improving tissue perfusion and reducing inflammation, while avoiding the undesirable effects of systemic immunosuppression and / or local adverse effects of steroids.

[0121] RVO is currently treated with intravitreal steroids and anti-VEGF agents. We hypothesize that improving perfusion of existing vessels will reduce the degree of macular edema and VEGF upregulation, as well as downstream maladaptive changes manifesting as RVO. To test efficacy, a preclinical mouse model of ischemic retinopathy can be used. Oxygen-induced retinopathy in mice is a reproducible and quantifiable model of proliferative retinal neovascularization suitable for investigating the pathogenesis and therapeutic intervention of retinal neovascularization in many ischemic retinopathies, including RVO. This model is induced by exposing 1-week-old C57BL / 6J mice to 75% oxygen for 5 days, followed by exposure 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 assessed by studying retinal hypoxia and neovascularization. A "therapeutically effective amount" of a composition comprising the edonentan crystalline form described herein can add to the current standard of care by improving tissue perfusion and reducing ET-1-mediated inflammation while avoiding the undesirable effects of topical steroids. In some embodiments for treating RVO, a composition comprising an edonentan crystalline form is administered intravitreally, suprachoroidally, or locally to the back of the eye using an implanted delivery platform. The frequency of administration varies based on the patient's disease course and response to treatment. In some embodiments for treating RVO, a composition comprising an edonentan crystalline form is administered in a topical form (e.g., eye drops). Retinopathy of Prematurity (ROP)

[0122] Retinopathy of prematurity (ROP) is a retinal vascular proliferative disorder affecting premature infants. ROP remains the leading preventable cause of blindness and visual impairment worldwide. Due to improvements in perinatal care, improved survival rates for near-term preterm infants, and limited resources for oxygen delivery and monitoring, more mature preterm infants are developing severe ROP in developing countries.

[0123] The pathophysiology of ROP is characterized by two stages. Stage I ROP results from vascular occlusion beginning shortly after birth, followed by a marked decrease in vascular endothelial growth factor (VEGF) and insulin-like growth factor-1 (IGF-1). Stage II begins around 33 weeks postmenstrual age (PMA). During this stage, VEGF levels increase, especially in the presence of retinal hypoxia associated with increased retinal metabolism and oxygen demand, which leads to abnormal vascular growth. For advanced stages of ROP, laser ablation of the avascular retina, early treatment of ROP (ETROP) protocols, intravitreal injection of anti-VEGF antibodies (e.g., bevacizumab), and vitrectomy are used to protect central vision and prevent retinal detachment. Long-term complications, such as refractive error, recurrence of ROP, and risk of retinal detachment, require continued follow-up by an ophthalmologist even after adolescence.

[0124] ROP is induced by severe ischemia resulting from underdevelopment of retinal blood vessels secondary to preterm birth. Therefore, as one aspect of the present invention, improving perfusion of existing vessels with a composition containing edonentan crystalline forms is believed to reduce the degree of ischemia and VEGF upregulation, as well as downstream maladaptive changes manifesting as ROP. To test efficacy, a preclinical mouse model of ROP can be used. Oxygen-induced retinopathy in mice is a reproducible and quantifiable model of proliferative retinal neovascularization suitable for investigating the pathogenesis and therapeutic intervention of retinal neovascularization in ROP. This model is induced by exposing 1-week-old C57BL / 6J mice to 75% oxygen for 5 days, followed by exposure to room air as previously described (Smith LEH et al., Invest Ophthalmol Vis Sci 1994). Efficacy in this preclinical model of ROP can be assessed by studying retinal hypoxia and neovascularization. A "therapeutically effective amount" of a composition containing an edonentan crystalline form described herein enhances current standard of care by improving tissue perfusion and reducing VEGF-induced pathological neovascularization. In some embodiments, the medicament is administered intravitreally, suprachoroidally, or locally to the back of the eye using an implanted delivery platform every 4 to 6 weeks as needed, based on the patient's disease course and response to treatment. For example, the medicament is administered locally to the back of the eye using an intravitreal injection every 5 weeks as needed, based on the patient's disease course and response to treatment. In some embodiments for treating ROP, a composition containing an edonentan crystalline form is administered in a topical form (e.g., eye drops). Pharmaceutical Composition

[0125] Some embodiments described herein relate to pharmaceutical compositions that can include (e.g., be prepared from) a therapeutically effective amount of an edonentan crystalline form described herein and a pharmaceutically acceptable carrier, diluent, excipient, or combination thereof.

[0126] The term "pharmaceutical composition" refers to a mixture of one or both compounds disclosed herein with other chemical components, such as diluents or carriers. Pharmaceutical compositions facilitate administration of a compound to an organism. Pharmaceutical compositions are generally tailored for a specific intended route of administration.

[0127] The term "pharmaceutically 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.

[0128] As used herein, "carrier" refers to a compound that facilitates the incorporation of a compound into cells or tissues.For example, but not limited to, dimethyl sulfoxide (DMSO) is a commonly used carrier that facilitates the incorporation of many organic compounds into target cells or tissues.

[0129] As used herein, "diluent" refers to an ingredient in a pharmaceutical composition that lacks pharmacological activity but may be pharmaceutically 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. A diluent 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, such as, but not limited to, phosphate-buffered saline, which mimics the composition of human blood.

[0130] As used herein, "excipient" refers to an inert substance added to a pharmaceutical composition to impart to the composition, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegration ability, delayed dissolution, etc. A "diluent" is a type of excipient.

[0131] As used herein, "about" will be understood by one of ordinary skill in the art and can vary to some extent depending on the context in which the term is used. If there are uses of the term that are not clear to a person of ordinary skill in the art given the context in which the term is used, "about" will mean up to plus or minus 10% of the term in question.

[0132] The pharmaceutical compositions described herein can be administered to human patients by themselves or in pharmaceutical compositions mixed with other active ingredients, or carriers, diluents, excipients or combinations thereof, such as in combination therapy.Suitable formulation depends on the selected route of administration.The techniques for formulating and administering the compounds described herein are known to those skilled in the art.

[0133] 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 encapsulating processes.See, for example, Encapsulation Processes, in: Food Powders, 2005, 199-299.Furthermore, the active ingredient is contained in an amount effective to achieve its intended purpose.The compounds used in the pharmaceutical combinations disclosed herein can be provided as pharmaceutically acceptable salts.

[0134] The compound or pharmaceutical composition of the present invention is preferably administered locally as a topical ophthalmic preparation or by direct injection of the compound or pharmaceutical composition into ocular tissue, often in a depot or sustained-release preparation.The mode of local administration can be intravitreal, suprachoroidal, periocular or subconjunctival injection of the preparation, or by using implant technology or topical application.For example, the compound is administered in a liposome preparation, which slowly releases the compound to maintain the desired pharmacological effect.Alternatively, polyvinyl alcohol nanoparticles can be prepared by known methods to obtain sustained-release or sustained-release preparations for topical or intraocular application.

[0135] In some embodiments, the pharmaceutical composition is an ophthalmic preparation comprising a composition comprising a therapeutically effective amount of the edonentan crystalline form described herein. As used herein, "ophthalmic preparation" refers to a specialized dosage form designed to be instilled onto the outer surface of the eye (topical), administered inside the eye (intraocular) or adjacent to the eye (periocular), or used in combination 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, a gel-forming solution, an ocular insert, a micro / nano-particle preparation for topical or preferably intravitreal injection, or an implant.

[0136] In some embodiments, the ophthalmic preparation contains 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 esters thereof (e.g., sorbic acid, esters of p-hydroxybenzoic acid such as methyl hydroxybenzoate, propyl hydroxybenzoate, etc.), and alcohol substitutes (e.g., chlorobutanol, phenylethanol). The preservative 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 include a preservative aid. Examples of suitable preservative aids include, but are not limited to, ethylenediaminetetraacetic acid (EDTA).

[0137] In some embodiments, the ophthalmic preparation comprises one or more additional excipients or agents to impart viscosity or lubricity, stabilize the active ingredient against degradation, increase the solubility of the active ingredient or inactive ingredient, adjust tonicity, or act as a solvent. Examples of excipients or agents for imparting viscosity or lubricity include hypromellose, carbomer 974P, hydroxyethyl cellulose (HEC), polyvinyl alcohol, sodium hyaluronate, sodium carboxymethylcellulose, Carbopol 940, hydroxypropyl methylcellulose (HPMC), poloxamer, xyloglucan, alginic acid, sodium alginate, gellan gum, cellulose acetate phthalate, and xanthan gum. Examples of excipients or agents as 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 providone, glycerol, polyethylene glycol (PEG), polypropylene glycol (PPG), PEG stearate, poloxamer 407, tyloxapol, polysorbate 80, creatinine, cyclodextrin, and castor oil. Examples of excipients or agents for adjusting tonicity include 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.

[0138] In some embodiments, the ophthalmic preparation includes one or more buffers to adjust the pH. Examples of buffers for adjusting the 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 can 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).

[0139] In some embodiments, the ophthalmic preparation comprises one or more surfactants. Examples of surfactants include sorbitan ether esters of oleic acid (e.g., polysorbate or Tween® 20 and 80) and tyloxapol.

[0140] The volume that can be injected into a human eye at one time is approximately 50-90 μL via the intravitreal route, up to 450 μL via the subretinal route, and up to 200 μL via the suprachoroidal route. The needles used in these routes are typically 27-30 g in size. The dose depends on the volume, potency, and concentration that can be formulated to meet the target efficacy and pharmacokinetic profile for each indication. Generally, ocular injections are administered no more frequently than once per month per eye. In some embodiments, when the compositions of the present invention are administered topically (i.e., as eye drops), the volume administered to a human eye at one time can be approximately 50 μL (the concentration of the eye drops can be approximately 5 mg / mL).

[0141] In some embodiments, the intravitreal formulation comprises a dose of a composition comprising the edonentan crystalline form ranging from about 1 μg to about 1 mg. A first exemplary formulation comprises about 1 μg to about 1 mg of a composition comprising the edonentan crystalline form, about 10 mM histidine HCl, about 10% α,α-trehalose dihydrate, and about 0.01% polysorbate 20. A second exemplary formulation comprises about 1 μg to about 1 mg of a composition comprising the edonentan crystalline form, about 10 mM sodium phosphate, about 40 mM sodium chloride, about 0.03% polysorbate 20, and about 5% sucrose.

[0142] Without further elaboration, it is believed that one skilled in the art can, based on the foregoing, utilize the present invention to its fullest extent. The following specific examples are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever. [Example]

[0143] Example In order that the invention described herein may be more fully understood, the following examples are set forth. The examples described in this application are provided to illustrate aspects of the invention and embodiments thereof provided herein, and should not be construed in any way as limiting the scope thereof.

[0144] The compounds provided herein that are subject to crystallization can be prepared from readily available starting materials using known procedures. The compounds can be crystallized using the procedures described herein. Abbreviation: DSC Differential Scanning Calorimetry EA Ethyl acetate IPA Isopropyl Alcohol PBS Phosphate Buffered Saline XRPD X-ray powder diffraction Example 1: Exemplary method for preparing crystalline form 1

[0145] 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. 18 mL of hot water was then 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 shown in Figures 2 and 6, respectively). Example 2: Exemplary method for preparing crystalline form 2

[0146] 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, and immediately, 7.5 mL of warm water was added dropwise at 60°C with vigorous stirring, followed by stirring at 60°C for 1 hour. After slowly cooling to 25°C, the mixture was filtered to obtain crystalline Form 2 (XRPD and DSC shown in Figures 3 and 7, respectively).

[0147] 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 in Figures 3 and 7, respectively). Example 3: Exemplary method for preparing crystalline form 3

[0148] 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 at 60°C with vigorous stirring. To the resulting slightly cloudy solution, 0.1 mL of ethyl acetate was added to obtain a clear solution, which 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 shown in Figures 4 and 8, respectively). Example 4: Exemplary method for preparing crystalline form 4

[0149] Amorphous edonentane (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 give Form 4, which was confirmed to be distinct from Forms 1, 2, and 3 by XRPD ( FIG. 5 ) and DSC ( FIG. 9 ).

[0150] Alternatively, 107 mg of amorphous edonentan was added to 1 mL of water, followed by the same 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 give Form 4, which was confirmed by XRPD.

[0151] Alternatively, 150 mg of edonentane (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, demonstrating that Form 4 is thermodynamically more stable than Form 3 under these conditions.

[0152] Alternatively, 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 thus obtained was confirmed to be Form 4 by XRPD analysis, demonstrating that Form 4 is thermodynamically more stable than Form 1 under these conditions.

[0153] Alternatively, 100 mg of edonentan (amorphous) was slurried 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.

[0154] Alternatively, amorphous (crude) edonentan is dissolved in 8 volumes of isopropanol at 60° C. The resulting solution is cooled to 57° C., and then small crystals of crystalline form 4 are added. After 2 hours, the solution is cooled to 5° C., held for 15 hours, and filtered to obtain crystalline form 4. Example 5: XRPD pattern of crystalline form

[0155] XRPD patterns for crystalline forms 1-4 are shown in Figures 1-5. XRPD patterns for the crystalline forms described herein were recorded using a Polycrystalline X-ray diffractometer (Bruker, D8 ADVANCE). The 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-supported slit. The sample was placed in the center of the sample holder groove, with the surface of the sample holder flush with the sample holder surface. A lynxeye detector was used to collect data over a continuous scan at a step size of 0.02° and a rate of 8° / min.

[0156] Tables 1-4 below list certain XRPD characteristic peaks for crystalline forms 1-4, respectively. Table 1. Exemplary XRPD patterns of crystalline form 1 [Table 1] Table 2. Exemplary XRPD patterns of crystalline form 2 [Table 2] Table 3. Exemplary XRPD patterns of crystalline form 3 [Table 3] Table 4. Exemplary XRPD patterns of crystalline form 4 [Table 4]

[0157] Table 5 below lists the XRPD characteristic peaks for crystalline Form 4 shown in FIG. Table 5. XRPD characteristic peaks of crystalline form 4 [Table 5] Example 6: Physicochemical characterization of crystalline forms

[0158] Exemplary physicochemical properties of the crystalline forms are provided herein. The melting points described herein can be determined using the following procedure. Melting Point Protocol

[0159] DSC was used to identify the maximum melting point peak (T m ) was determined. 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 at the crystalline melting peak onset, peak onset, peak maximum, and peak end were collected.

[0160] The solubility described herein can be measured using the following procedure. Solubility Analysis Protocol 1. Weigh 2.0 mg or more of sample into the lower chamber of a Whatman Mini Uniprep vial (GE Healthcare). Add 450 μL of buffer into each chamber. 2. Place the filter piston of the Mini UniPrep vial and compress it to the liquid level to allow buffer and compound contact with the filter during incubation. 3. Vortex the sample for 2 minutes and then incubate at room temperature (approximately 25±2°C) with shaking at 500 rpm for 24 hours. 4. Compress the Mini Uniprep to prepare the filtrate for injection into the HPLC system. All vials are inspected for visible undissolved material before filtration and for leaks after filtration. 5. Dilute the supernatant 100-fold with buffer to produce the dilution that is analyzed by HPLC.

[0161] Table 6 below provides exemplary physicochemical properties of crystalline forms 1 to 4. The physicochemical properties can be obtained using the methods described above. Table 6. Exemplary physicochemical properties of crystalline forms 1-4 [Table 6]

[0162] Of the four crystalline forms, Form 4 has a higher melting point (T m ) Form 4 exhibited lower solubility than Form 1 and Form 3. Form 4, as shown above, exhibited higher thermodynamic stability compared to Form 1, Form 2, and Form 3. In a preferred embodiment, Form 4 is preferred for pharmaceutical development. Other Aspects

[0163] All of the features disclosed herein may be combined in any combination. Each feature disclosed herein may be replaced by an alternative feature 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.

[0164] Furthermore, from the foregoing, 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 uses and conditions without departing from the spirit and scope thereof. Accordingly, other embodiments are within the scope of the following claims. The present invention provides, for example, the following items. (Item 1) Compounds of Formula I: [ka] a solid form of The solid form is 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 two-theta. (Item 2) 2. The solid form of claim 1, wherein the anhydrous crystalline Form 4 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 5.6±0.2°, 11.4±0.2°, 17.7±0.2°, and 19.3±0.2°. (Item 3) 2. The solid form of claim 1, wherein the anhydrous crystalline Form 4 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 11.4±0.2°, 17.7±0.2°, and 19.3±0.2°. (Item 4) Item 1, wherein the crystalline form 4 has the following characteristic peaks expressed in terms of diffraction angles (2θ): [Table 7] (Item 5) The anhydrous crystalline Form 4 has a T of about 163°C by DSC analysis. m 2. The solid form according to item 1, having (Item 6) Compounds of Formula I: [ka] A composition comprising: the compound is an anhydrous crystalline form (Form 4) having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from peaks at 5.6±0.2 degrees, 11.4±0.2 degrees, 17.7±0.2 degrees, 19.3±0.2 degrees, 21.1±0.2 degrees, and 21.9±0.2 degrees in terms of two-theta; A composition, wherein the compound is 90% or more by weight in crystalline form 4, based on the total weight of the compound present in the composition. (Item 7) 7. The composition of claim 6, wherein the compound is 96% or more by weight of crystalline form 4, based on the total weight of the compound present in the composition. (Item 8) 7. The composition of claim 6, wherein the compound is 97% or more by weight of crystalline form 4, based on the total weight of the compound present in the composition. (Item 9) 7. The composition of claim 6, wherein the compound is 98% or more by weight in crystalline form 4, based on the total weight of the compound present in the composition. (Item 10) 7. The composition of claim 6, wherein the compound is 99% by weight or more crystalline form 4, based on the total weight of the compound present in the composition. (Item 11) 1. A method of treating an ocular disease, comprising: The ocular tissue of the subject is treated with a therapeutically effective amount of a compound of formula I: [ka] contacting the composition containing the ocular disease is selected from the group consisting of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO) and retinopathy of prematurity (ROP); the compound is an anhydrous crystalline form (Form 4), the crystalline Form 4 having an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from those at 5.6±0.2 degrees, 11.4±0.2 degrees, 17.7±0.2 degrees, 19.3±0.2 degrees, 21.1±0.2 degrees, and 21.9±0.2 degrees in terms of two-theta; The method, wherein the compound is 90% or more by weight of crystalline Form 4, based on the total weight of the compound present in the composition. (Item 12) 12. The method of claim 11, wherein the anhydrous crystalline Form 4 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 5.6±0.2°, 11.4±0.2°, 17.7±0.2°, and 19.3±0.2°. (Item 13) 12. The method of claim 11, wherein the anhydrous crystalline Form 4 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 11.4±0.2°, 17.7±0.2°, and 19.3±0.2°. (Item 14) The anhydrous crystalline Form 4 has a T of about 163°C by DSC analysis. m 14. The method according to any one of Items 11 to 13, comprising: (Item 15) 15. The method of any one of items 11 to 14, wherein the anhydrous crystalline Form 4 has a solubility of about 138 μg / mL in phosphate buffer at about pH 7. (Item 16) 16. The method of any one of items 11 to 15, wherein the compound is 96% by weight or more crystalline form 4, based on the total weight of the compound present in the composition. (Item 17) 16. The method of any one of items 11 to 15, wherein the compound is 97% by weight or more crystalline form 4, based on the total weight of the compound present in the composition. (Item 18) 16. The method of any one of items 11 to 15, wherein the compound is 98% by weight or more crystalline form 4, based on the total weight of the compound present in the composition. (Item 19) 16. The method of any one of items 11 to 15, wherein the compound is 99% by weight or more crystalline form 4, based on the total weight of the compound present in the composition. (Item 20) 20. The method according to any one of items 11 to 19, wherein the eye disease is glaucoma. (Item 21) 20. The method according to any one of items 11 to 19, wherein the eye disease is diabetic retinopathy (DR). (Item 22) 20. The method according to any one of items 11 to 19, wherein the eye disease is retinal vein occlusion (RVO). (Item 23) 20. The method according to any one of items 11 to 19, wherein the eye disease is retinopathy of prematurity (ROP). (Item 24) 20. The method of any one of items 11-19, wherein the therapeutic effectiveness of the treatment is determined by (i) detecting a decrease in intraocular pressure or a decrease in the rate of optic nerve damage by an amount sufficient to alleviate or prevent optic nerve damage; (ii) assessing the degree of improvement in visual acuity or visual field; (iii) measuring a reduction in diabetes-induced retinal neurodegeneration; (iv) measuring an improvement in tissue or retinal perfusion; or (v) measuring an improvement in tissue or retinal perfusion, a reduction in inflammation, or a combination thereof. (Item 25) 20. The method of any one of items 11 to 19, wherein the composition further comprises a therapeutically effective amount of an intraocular pressure (IOP)-lowering agent or a neuroprotective agent, or a pharmaceutically acceptable salt thereof. (Item 26) 20. The method of any one of items 11 to 19, wherein the composition further comprises a therapeutically effective amount of an intraocular pressure (IOP)-lowering agent or a pharmaceutically acceptable salt thereof, wherein the IOP-lowering agent is selected from the group consisting of prostaglandins (such as latanoprost or travoprost), beta-blockers (such as timolol or betaxolol), alpha-adrenergic agonists (such as brimonidine or apraclonidine), carbonic anhydrase inhibitors (such as dorzolamide or brinzolamide), Rho-kinase inhibitors (such as netarsudil), and miotics or cholinergic agents (such as pilocarpine). (Item 27) 27. The method of claim 26, wherein the composition further comprises a therapeutically effective amount of a neuroprotective agent or a pharmaceutically acceptable salt thereof, wherein the neuroprotective agent is selected from the group consisting of an anti-apoptotic agent (such as a caspase-2 inhibitor) and a neurotrophic factor (such as a ciliary neurotrophic factor). (Item 28) 12. The method of claim 11, wherein the composition is administered at a dosage of 1 μg to 5 mg. (Item 29) 12. The method of claim 11, wherein said contacting comprises topically administering said composition to the surface of the eye or a portion thereof. (Item 30) 12. The method of claim 11, wherein said contacting comprises injecting the composition into the eye or a component thereof. (Item 31) 12. The method of claim 11, wherein the composition comprises an ophthalmic preparation containing one or more preservatives, preservative aids, viscosity or lubrication adjusters, tonicity adjusters, solubilizers, buffers, surfactants, stabilizers, or combinations thereof. (Item 32) Compounds of Formula I: [ka] 1. A process for preparing the anhydrous crystalline form (Form 4) of (a) stirring a compound of Formula I in an aqueous solution, an organic solvent, or a mixture thereof at a temperature ranging from about 40° C. to about 120° C.; (b) cooling the resulting solution to a temperature in the range of about 0°C to about room temperature, and, if the compound is in a basic aqueous solution, adjusting its pH value to a range of about 3 to about 7; (c) filtering the sample thus obtained to obtain Crystalline Form 4; Including, 10. A method wherein said crystalline Form 4 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 5.6±0.2 degrees, 11.4±0.2 degrees, 17.7±0.2 degrees, 19.3±0.2 degrees, 21.1±0.2 degrees, and 21.9±0.2 degrees in terms of two-theta. (Item 33) 33. The method of claim 32, wherein the aqueous solution is water. (Item 34) 33. The method of claim 32, wherein the aqueous solution is a basic aqueous solution. (Item 35) 35. The method of claim 34, wherein the basic aqueous solution has a pH of 8 or greater. (Item 36) 35. The method of claim 34, wherein the basic aqueous solution is selected from potassium hydroxide and potassium carbonate solutions. (Item 37) Item 33. The method according to item 32, wherein the organic solvent is a water-soluble organic solvent. (Item 38) 33. The method of claim 32, wherein the organic solvent is selected from tetrahydrofuran and isopropanol. (Item 39) Item 33. The method according to item 32, wherein the temperature for stirring is in the range of about 80°C to about 120°C. (Item 40) Item 33. The method according to item 32, wherein the temperature for stirring is in the range of about 40°C to about 80°C. (Item 41) Item 33. The method according to item 32, wherein the stirring is carried out for about 20 hours to about 50 hours. (Item 42) 33. The method of claim 32, wherein a slurry of the compound of formula I is formed in the stirring step. (Item 43) 33. The method of claim 32, further comprising seeding with a quantity of crystalline Form 4 and holding the resulting solution for about 2 hours. (Item 44) 33. The method of claim 32, wherein the cooling is carried out at about 5° C. for about 15 hours. (Item 45) 1. A method of treating an ocular disease, comprising: The ocular tissue of the subject is treated with a therapeutically effective amount of a compound of formula I: [ka] contacting the composition containing the ocular disease is selected from the group consisting of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO) and retinopathy of prematurity (ROP); the compound is selected from the group consisting of an anhydrous crystalline form (Form 1), a monohydrate crystalline form (Form 2), and an anhydrous crystalline form (Form 3); the anhydrous crystalline Form 1 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 6.3±0.2 degrees, 7.5±0.2 degrees, 11.7±0.2 degrees, 15.1±0.2 degrees, and 17.3±0.2 degrees in terms of two-theta; the monohydrate crystalline Form 2 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 9.6±0.2 degrees, 10.4±0.2 degrees, 19.6±0.2 degrees, 19.7±0.2 degrees, 22.0±0.2 degrees, 22.9±0.2 degrees, and 23.7±0.2 degrees in terms of two-theta; the anhydrous crystalline Form 3 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 7.8±0.2 degrees, 9.0±0.2 degrees, 11.6±0.2 degrees, 15.8±0.2 degrees, and 19.1±0.2 degrees in terms of two-theta; when the compound is the anhydrous crystalline form 1, the compound is 90% by weight or more crystalline form 1, based on the total weight of the compound present in the composition; when the compound is the monohydrate crystalline form 2, the compound is 90% by weight or more crystalline form 2, based on the total weight of the compound present in the composition; The method, wherein when the compound is the anhydrous crystalline form 3, the compound is 90% or more by weight of crystalline form 3, based on the total weight of the compound present in the composition. (Item 46) 46. The method of claim 45, wherein the anhydrous crystalline Form 1 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 6.3±0.2°, 7.5±0.2°, 11.7±0.2°, and 15.1±0.2°. (Item 47) 46. The method of claim 45, wherein the anhydrous crystalline Form 1 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 7.5±0.2°, 11.7±0.2°, and 15.1±0.2°. (Item 48) The anhydrous crystalline Form 1 has a T of about 151°C by DSC analysis. m48. The method according to any one of Items 45 to 47, comprising: (Item 49) 49. The method of any one of items 45 to 48, wherein the anhydrous crystalline Form 1 has a solubility of about 264 μg / mL in phosphate buffer at about pH 7. (Item 50) 49. The method of any one of items 45 to 48, wherein the compound is 96% by weight or more of crystalline form 1, based on the total weight of the compound present in the composition. (Item 51) 49. The method of any one of items 45 to 48, wherein the compound is 97% by weight or more of crystalline form 1, based on the total weight of the compound present in the composition. (Item 52) 49. The method of any one of items 45 to 48, wherein the compound is 98% by weight or more crystalline form 1, based on the total weight of the compound present in the composition. (Item 53) 49. The method of any one of items 45 to 48, wherein the compound is 99% by weight or more in crystalline form 1, based on the total weight of the compound present in the composition. (Item 54) 46. The method of claim 45, wherein the monohydrate crystalline Form 2 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 19.6±0.2°, 19.7±0.2°, and 9.6±0.2°. (Item 55) 46. The method of claim 45, wherein the monohydrate crystalline Form 2 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 19.6±0.2°, 19.7±0.2°, 9.6±0.2°, 10.4±0.2°, 22.0±0.2° and 22.9±0.2°. (Item 56) The monohydrate crystalline Form 2 has a T of about 122°C by DSC analysis. m 56. The method according to any one of Items 45 and 54 to 55, comprising: (Item 57) 57. The method of any one of items 45 and 54-56, wherein the monohydrate crystalline Form 2 has a solubility of about 35 μg / mL in phosphate buffer at about pH 7. (Item 58) 58. The method of any one of items 45 and 54-57, wherein the compound is 96% by weight or more crystalline form 2, based on the total weight of the compound present in the composition. (Item 59) 58. The method of any one of items 45 and 54-57, wherein the compound is 97% by weight or more crystalline form 2, based on the total weight of the compound present in the composition. (Item 60) 58. The method of any one of items 45 and 54-57, wherein the compound is 98% by weight or more crystalline form 2, based on the total weight of the compound present in the composition. (Item 61) 58. The method of any one of items 45 and 54-57, wherein the compound is 99% by weight or more crystalline form 2, based on the total weight of the compound present in the composition. (Item 62) 46. The method of claim 45, wherein the anhydrous crystalline Form 3 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 9.0±0.2°, 15.8±0.2°, 7.8±0.2°, and 19.1±0.2°. (Item 63) 46. The method of claim 45, wherein the anhydrous crystalline Form 3 has the following characteristic peaks expressed in terms of diffraction angles (2θ): 9.0±0.2°, 15.8±0.2°, and 7.8±0.2°. (Item 64) The anhydrous crystalline Form 3 has a T of about 162°C by DSC analysis. m 64. The method according to any one of Items 45 and 62 to 63, comprising: (Item 65) 65. The method of any one of items 45 and 62-64, wherein the anhydrous crystalline Form 3 has a solubility of about 251 μg / mL in phosphate buffer at about pH 7. (Item 66) 66. The method of any one of items 45 and 62-65, wherein the compound is 96% by weight or more crystalline form 3, based on the total weight of the compound present in the composition. (Item 67) 66. The method of any one of items 45 and 62-65, wherein the compound is 97% by weight or more crystalline form 3, based on the total weight of the compound present in the composition. (Item 68) 66. The method of any one of items 45 and 62-65, wherein the compound is 98% by weight or more crystalline form 3, based on the total weight of the compound present in the composition. (Item 69) 66. The method of any one of items 45 and 62-65, wherein the compound is 99% by weight or more crystalline form 3, based on the total weight of the compound present in the composition. (Item 70) 70. The method according to any one of items 45 to 69, wherein the eye disease is glaucoma. (Item 71) 70. The method according to any one of items 45 to 69, wherein the eye disease is diabetic retinopathy (DR). (Item 72) 70. The method according to any one of items 45 to 69, wherein the eye disease is retinal vein occlusion (RVO). (Item 73) 70. The method according to any one of items 45 to 69, wherein the eye disease is retinopathy of prematurity (ROP). (Item 74) 46. The method of claim 45, wherein the therapeutic effectiveness of the treatment is determined by (i) detecting a decrease in intraocular pressure or a decrease in the rate of optic nerve damage by an amount sufficient to alleviate or prevent optic nerve damage; (ii) assessing the degree of improvement in visual acuity or visual field; (iii) measuring a reduction in diabetes-induced retinal neurodegeneration; (iv) measuring an improvement in tissue or retinal perfusion; or (v) measuring an improvement in tissue or retinal perfusion, a reduction in inflammation, or a combination thereof. (Item 75) 46. The method of claim 45, wherein the composition further comprises a therapeutically effective amount of an intraocular pressure (IOP)-lowering agent or a neuroprotective agent, or a pharmaceutically acceptable salt thereof. (Item 76) 46. The method of item 45, wherein the composition further comprises a therapeutically effective amount of an intraocular pressure (IOP)-lowering agent or a pharmaceutically acceptable salt thereof, wherein the IOP-lowering agent is selected from the group consisting of prostaglandins (such as latanoprost or travoprost), beta-blockers (such as timolol or betaxolol), alpha-adrenergic agonists (such as brimonidine, apraclonidine), carbonic anhydrase inhibitors (such as dorzolamide or brinzolamide), Rho kinase inhibitors (such as netarsudil), and miotics or cholinergic agents (such as pilocarpine). (Item 77) 46. The method of claim 45, wherein the composition is administered at a dosage of 1 μg to 5 mg. (Item 78) 46. The method of claim 45, wherein said contacting comprises topically administering said composition to the surface of the eye or a portion thereof. (Item 79) 46. The method of claim 45, wherein said contacting comprises injecting the composition into the eye or a component thereof. (Item 80) 46. The method of claim 45, wherein the composition comprises an ophthalmic preparation containing one or more preservatives, preservative aids, viscosity or lubrication adjusters, tonicity adjusters, solubilizers, buffers, surfactants, stabilizers, or combinations thereof. (Item 81) Compounds of Formula I: [ka] 1. A method for preparing the monohydrate crystalline form (Form 2) of (a) preparing a slurry of a compound of Formula I in an aqueous medium at a temperature ranging from about 15° C. to about 35° C. for a predetermined period of time to obtain a sample; (b) filtering the sample thus obtained to obtain Crystalline Form 2; wherein said crystalline Form 2 has an X-ray powder diffraction pattern comprising at least three characterizing peaks selected from the peaks at 9.6±0.2 degrees, 10.4±0.2 degrees, 19.6±0.2 degrees, 19.7±0.2 degrees, 22.0±0.2 degrees, 22.9±0.2 degrees, and 23.7±0.2 degrees in terms of two-theta.

Claims

[Claim 1] The invention described in this specification.

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