Treatment of ocular diseases using endothelin receptor antagonists

Abstract

To provide methods for treating ocular diseases.SOLUTION: The present disclosure relates to the discovery that certain diseases of the eye that profoundly affect the human visual system and, as a result, quality of life, may be treated using Edonentan or A-182086. Edonentan or A-182086 can be used alone or, for example, in combination with an intra-ocular pressure (IOP) reducing agent, a neuroprotective agent, an anti-VEGF agent, or all. Using Edonentan or A-182086, alone or in combination with an additional agent, provides increased perfusion to the retina in certain 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 Application Nos. 62 / 928,092 and 63 / 068,215, filed on October 30, 2019 and August 20, 2020, respectively, the entire contents of which are incorporated herein by reference for all purposes.

[0002] Field The present disclosure relates to the field of medicine and the treatment of eye diseases. More specifically, the present disclosure relates to the use of edonentan and A-182086 endothelin receptor antagonists in the treatment or improvement of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), non-arteritic anterior ischemic optic neuropathy (NAION), arteritic anterior ischemic optic neuropathy (AION), and retinopathy of prematurity (ROP).

Background Art

[0003] Background Eye diseases have a very significant impact on human quality of life, but effective treatments remain very elusive. In the United States, the annual economic burden resulting from vision loss, eye diseases, and visual impairments is estimated to exceed $100 billion. Examples of degenerative eye diseases include glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), non-arteritic anterior ischemic optic neuropathy (NAION), arteritic anterior ischemic optic neuropathy (AION), and retinopathy of prematurity (ROP).

[0004] Glaucoma is an eye disorder characterized by visual field defects and optic nerve head cupping. Abnormally high intraocular pressure is generally known to be harmful to the eye, and in glaucoma patients, there is clear evidence that this is probably the most important physical change that causes retinal degeneration. Ultimately, untreated, vision is gradually lost over time. However, the pathophysiological mechanism of glaucoma remains unknown.

[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 glaucoma and closed-angle glaucoma. Primary open-angle glaucoma ("POAG") is the most frequent type of glaucoma observed in the United States. POAG is usually detected at its early stage during routine eye examinations. Primary closed-angle glaucoma, also known as acute glaucoma, usually has a sudden onset and is characterized by eye pain and blurred vision. Secondary glaucoma occurs as a complication of various other conditions (e.g., injury, inflammation, general vascular diseases, and diabetes). Congenital glaucoma is due to a developmental defect in the eye's drainage mechanism.

[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 of developed countries. The incidence of DR increases with the duration of diabetes progression. 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, approximately 30%, and in 10% of those cases, DR is at an advanced stage that severely threatens vision.

[0007] DR occurs when changes in blood glucose levels cause changes in the retinal blood vessels. In some cases, these vessels swell up (macular edema) and leak fluid into the back of the eye. In other cases, abnormal blood vessels grow on the surface of the retina. If left untreated, DR gradually worsens, progressing from "background retinopathy" and can severely affect vision and lead to blindness.

[0008] Retinal vein occlusion (RVO) is a retinal vascular disorder and one of the most common causes of vision loss worldwide. Specifically, it is the second most common cause of blindness resulting from retinal vascular diseases 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 the occlusion of the main retinal vein, and branch retinal vein occlusion (BRVO) is the occlusion of one of the smaller branch veins.

[0009] Currently, there is no way to unblock retinal vein occlusions, and the approved treatments are aimed at addressing the health problems associated with RVO. Vision may return in eyes affected by RVO. Approximately one-third have some improvement, one-third remain the same, and one-third improve gradually, but it can take one year or longer to determine the final outcome. In some cases, the blocked vessels cause fluid accumulation in the retina. In other cases, the occurrence of ischemia triggers the formation of new blood vessels. RVO is currently treated with intravitreal injections of anti-vascular endothelial growth factor (VEGF) drugs.

[0010] Anterior ischemic optic neuropathy (AION) results from ischemic damage to the anterior part of the optic nerve (the area mainly supplied by the posterior ciliary artery circulation). Anterior ischemic optic neuropathy is divided into two types: arteritic AION (AAION) secondary to vasculitis (especially giant cell arteritis) and non-arteritic AION (NAION) secondary to non-inflammatory small vessel disease. NAION constitutes 95% of all AION and is the most common cause of acute optic neuropathy in people over 50 years old, affecting approximately 2 to 10 individuals per 100,000 (approximately 1500 to 6000 new cases per year in the United States). Currently, there is no generally accepted treatment or secondary prevention for NAION, although steroids have been used in some patients for a long time.

[0011] Retinopathy of prematurity (ROP) can occur due to premature birth. Abnormal leaky blood vessel growth (neovascularization) in the retina occurs as a sequel to other treatments related to premature birth and can often lead to blindness in newborns. During pregnancy, blood vessels grow from the center of the developing child's retina at the 16th week of the mother's pregnancy and then branch out laterally to reach the edge of the retina at the 8th month of pregnancy. In premature children, normal retinal blood vessel growth is incomplete and thus more prone to rupture.

[0012] Edonerpic is a highly selective and very potent endothelin A receptor antagonist. Edonerpic was developed as a second-generation analog after the discontinuation of BMS-193884, which was the first clinical candidate and was being developed for the treatment of congestive heart failure (CHF). Edonerpic was in Phase I clinical trials until April 2002, but its development was discontinued.

[0013] A-182086 is a potent dual ET A / ET B receptor antagonist with 4-fold ET A / ET B selectivity. A-182086 has not been studied in the clinical setting to date.

[0014] There remains a need to more effectively reduce the incidence of glaucoma, DR, RVO, NAION, AION, and ROP, treat these, or otherwise improve them.

Summary of the Invention

Means for Solving the Problems

[0015] Abstract The present invention provides a method of using an endothelin receptor antagonist for treating an eye disease selected from glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), non-arteritic anterior ischemic optic neuropathy (NAION), arteritic anterior ischemic optic neuropathy (AION), and retinopathy of prematurity (ROP).

[0016] The method includes contacting the optical tissue in a subject with a composition comprising a therapeutically effective amount of edonentan or A-182086, or a pharmaceutically acceptable salt thereof, wherein the endothelin receptor antagonist is edonentan or A-182086. Such antagonist or its pharmaceutically acceptable salt can be in crystalline or amorphous form. Each of these can be for pharmacologically acceptable use.

[0017] In some embodiments, the contacting step includes administering a topical composition to the surface of the eye or a part thereof. In other embodiments, the contacting step includes injecting edonentan or A-182086 into the eye, either globally or to a specific region thereof.

[0018] In yet other embodiments, the contacting step includes administering the composition via an eye implant, such as a port delivery system. Examples of eye implant technologies include, but are not limited to, ApidCOR, BioSeizer-ProDex, Vitrasert, Retisert, Iluvien, I-Vation, Nanoporous Silicon, Ozurdex / Novadur, OcuLief, Port Delivery System (PDS), PEA Implant, PEG-PLA Microspheres, PRINT Technology, Q-Sphera, SKS Microparticles, Verisome, Capsule Ring Device, MicroPump, Microneedle Injector, Microneedle / Needle-less Injectors, EyeCET, Gemini Refractive Capsule, IVMED, Ciliary Sulcus Ring, Episcleral Exoplant, Eye-D Implant, and Nanoliposomes.

[0019] In some embodiments, the ocular implant technologies that can be used in the methods of the present disclosure are selected from ApidCOR, BioSeizer-ProDex, Vitrasert, Retisert, Iluvien, I-Vation, Nanoporous Silicon, Ozurdex / Novadur, OcuLief, Port Delivery System (PDS), PEA Implant, PEG-PLA Microspheres, PRINT Technology, Q-Sphera, SKS Microparticles, Verisome, Capsule Ring Device, MicroPump, Microneedle Injector, Microneedle / Needle-less Injectors, EyeCET, Gemini Refractive Capsule, and IVMED. Preferably, the ocular implant technology is ApidCOR, BioSeizer-ProDex, Vitrasert, Retisert, Iluvien, I-Vation, Nanoporous Silicon, Ozurdex / Novadur, OcuLief, Port Delivery System (PDS), PEA Implant, PEG-PLA Microspheres, PRINT Technology, Q-Sphera, SKS Microparticles, Verisome, Capsule Ring Device, or MicroPump.

[0020] In some embodiments, the ocular disease is glaucoma. In some embodiments, the therapeutic efficacy in treating glaucoma is determined by detecting a reduction in intraocular pressure or a reduction in the rate of optic nerve damage / retinal nerve fiber layer thinning in an amount sufficient to reduce or prevent optic nerve damage. In further embodiments, the therapeutic efficacy of the treatment is determined by improvement of optic nerve head blood flow. In other embodiments, the therapeutic efficacy of treating glaucoma is determined by measuring improvement in the retina, optic nerve head, or tissue perfusion.

[0021] In some embodiments related to the treatment of glaucoma, the regimen further includes an addition of a therapeutically effective amount of an intraocular pressure (IOP) lowering agent or a neuroprotective agent, or a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the IOP lowering agent is selected from the group consisting of prostaglandins (e.g., latanoprost or travoprost), beta blockers (e.g., timolol or betaxolol), alpha adrenergic agonists (e.g., brimonidine, apraclonidine), carbonic anhydrase inhibitors (e.g., dorzolamide or brinzolamide), Rho kinase inhibitors (e.g., netarsudil), and miotics or cholinergic agents (e.g., pilocarpine). In some embodiments, the neuroprotective agent is selected from the group consisting of anti-apoptotic agents (e.g., caspase-2 inhibitors) and neurotrophic factors (e.g., ciliary neurotrophic factor).

[0022] In some embodiments, the eye disease is diabetic retinopathy (DR). In further embodiments, the therapeutic efficacy of treating DR is determined by a reduction in retinal neovascularization, the diabetic retinopathy severity score, and diabetes-induced neurodegeneration. In other embodiments, the therapeutic efficacy of treating DR is determined by measuring an improvement in retinal or choroidal perfusion.

[0023] In some embodiments, the disease is retinal vein occlusion (RVO). In further embodiments, the therapeutic efficacy of treating RVO is determined by measuring an improvement in tissue perfusion, a reduction in inflammation, or a combination of the foregoing.

[0024] In some embodiments, the disease is NAION. In further embodiments, the therapeutic efficacy of treating NAION is determined by measuring an improvement in tissue perfusion, a reduction in inflammation, or a combination of the foregoing.

[0025] In some embodiments, the disease is AION. In further embodiments, the therapeutic efficacy of treating AION is determined by measuring improvement in tissue perfusion, reduction of inflammation, or a combination of the foregoing.

[0026] In some embodiments, the eye disease is retinopathy of prematurity (ROP). In further embodiments, the therapeutic efficacy of treating ROP is determined by measuring improvement in retinal perfusion and reduction of abnormal neovascularization.

[0027] In some embodiments, the endothelin receptor antagonist or a pharmaceutically acceptable salt thereof is edonentan. In other embodiments, the endothelin receptor antagonist or a pharmaceutically acceptable salt thereof is A-182086. In any case, the antagonist or a pharmaceutically acceptable salt thereof can be in crystalline or amorphous form. Each of them can be for pharmacologically acceptable use.

[0028] In some embodiments, the endothelin receptor antagonist is administered at a dosage between about 1 μg and about 4 mg (e.g., between about 1 μg and about 10 μg, between about 10 μg and about 100 μg, between about 100 μg and about 500 μg, and between about 500 μg and about 4 mg). In some embodiments, the endothelin receptor antagonist is administered at a dosage of about 0.1 μg to about 10 μg. In some embodiments, the endothelin receptor antagonist is administered at a dosage of about 1 μg to about 10 μg. In further embodiments, the endothelin receptor antagonist is administered at a dosage of about 10 μg to about 100 μg.

[0029] In further embodiments, the endothelin receptor antagonist is administered at dosages between about 100 μg and about 500 μg, and between about 500 μg and about 4 mg.

[0030] Further features and advantages of the technology of the present invention are shown in the following description, are partly apparent from the description, or can be learned by the practice of the technology of the present invention. The advantages of the technology of the present invention are realized and obtained by the structures and their embodiments specifically pointed out in the written description.

[0031] Details of one or more embodiments of the present disclosure are set forth in the following description. Other features, objects, and advantages of the present disclosure will be apparent from the following drawings, description, and claims.

Brief Description of the Drawings

[0032]

Figure 1

[0033]

Figure 2

[0034]

Figure 3

[0035]

Figure 4

[0036]

Figure 5-1

[0037]

Figure 5-2

[0038]

Figure 6-1

[0039]

Figure 6-2

[0040]

Figure 7-1

Figure 7-2

Figure 7-3

Figure 7-4

Figure 7-5

Figure 7-6

Figure 7-7

Figure 7-8

Figure 7-9

Figure 7-10

Figure 7-11

Figure 7-12

Figure 7-13

Figure 7-14

Figure 7-15

[0041]

Figure 8

[0042]

Figure 9-1

Figure 9-2

Figure 9-3

Figure 9-4

[0043]

Figure 10

Mode for Carrying Out the Invention

[0044] Detailed Description The present invention results from the discovery that endothelin and A-182086 can be used to prevent, treat, or otherwise improve eye diseases (including, but not limited to, glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), non-arteritic anterior ischemic optic neuropathy (NAION), arteritic anterior ischemic optic neuropathy (AION), and retinopathy of prematurity (ROP)). The present invention is further described below.

[0045] Endothelin Receptor Antagonist The methods of the present invention involve contacting or administering an eye tissue with a therapeutically effective amount of endothelin and A-182086, or a pharmaceutically acceptable salt thereof, either locally or intravitreally. The antagonists are specifically endothelin and A-182086, as described below.

[0046] Methods for preparing endothelin are well known to those skilled in the art. Suitable methods are disclosed, for example, in U.S. Patent No. 6,043,265. Endothelin 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) and the following structure:

Chemical formula

[0047] The method for preparing A-182086 is well-known to those skilled in the art. Suitable methods are disclosed, for example, in U.S. Patent No. 6,162,927. 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) and the following structure: [Chemical formula] and has the following structure:

[0048] Eye diseases The methods of the present disclosure include the use of edonentan and A-182086 described above in the treatment and improvement of eye diseases selected from glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), non-arteritic anterior ischemic optic neuropathy (NAION), arteritic anterior ischemic optic neuropathy (AION), and retinopathy of prematurity (ROP), which are described below.

[0049] Glaucoma In the treatment of glaucoma using edonentan or A-182086 described herein, a "therapeutically effective amount" can be determined by evaluating an improvement in retinal blood flow (RBF) that exceeds what can be achieved by standard care (a decrease in intraocular pressure (IOP)). With regard to glaucoma symptoms, improvement in blood flow in a healthy rabbit eye model can be used as a predictor of the pharmacodynamic response (PD) in humans. Rabbits are commonly used to evaluate the eye's PK / PD relationship for compounds targeting human eye diseases due to the anatomical and functional similarities between rabbit and human eyes. Previously, it has been shown that intravitreal administration of ET-1 to the rabbit eye induces significant vasoconstriction and optic nerve damage (Sasaoka M. et al., Exp Eye Res 2006; Sugiyama T. et al., (Arch Ophthalmol 2009). The efficacy in this model is benchmarked against the reversal of perfusion impairment induced by intravitreal ET-1 administration at a specific concentration. For example, the above efficacy can be achieved at a concentration equal to the levels observed in the plasma and aqueous humor of human glaucoma patients (Li S. et al., Journal of Ophthalmology 2016).

[0050] Other examples of relevant animal glaucoma models are Morrison's rat model of IOP elevation and the laser-induced non-human primate (NHP) glaucoma model. Glaucoma in Morrison's rat model is induced by a sustained increase in IOP via administration of hypertonic saline through the episcleral vein. In the above laser-induced NHP glaucoma model, it has been shown that after a sustained increase in IOP, the optic nerve head blood flow is reduced (Wang L. et al., Invest Ophthalmol Vis Sci 2012). Furthermore, it has been shown that the reduction in optic nerve head blood flow correlates with long-term structural changes in the optic nerve (Cull G. et al., Invest Ophthalmol Vis Sci 2013).

[0051] The 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 slowing of the progression of structural neurodegenerative changes on flat mounts (e.g., thickness of the retinal nerve fiber layer measured by optical coherence tomography (OCT) or number of retinal ganglion cells), and functional changes such as electroretinogram (ERG) or contrast sensitivity after treatment with edonentan or A-182086.

[0052] It is thought that the effect of edonentan or A-182086 on retinal blood flow can be evaluated by the vascular radius (r) in Poiseuille's law. An increase in (r) by an endothelin antagonist induces a more significant increase in blood flow than can be achieved by an increase in perfusion pressure via IOP reduction: Blood flow = (Perfusion pressure × πr 4) / (8ηl) Here l: length of the blood vessel r: radius of the blood vessel η: viscosity of the blood 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 a specific specific endothelin receptor antagonist, one (r) or more (IOP) of the above parameters can be changed to improve RBF, thus achieving therapeutic efficacy in treating glaucoma.

[0053] In some embodiments, the glaucoma patients are treated as soon as they are diagnosed. In some embodiments, edonentan or A-182086 is locally administered to the posterior side of the eye at a frequency of every 3 to 12 months (e.g., every 3 to 6 months or every 4 to 6 months) using an intravitreal, topical, suprachoroidal, or implant delivery platform.

[0054] Diabetic retinopathy (DR) Diabetes can cause severe late complications classified as microangiopathic (retinopathy, neuropathy, and diabetic nephropathy) and macroangiopathic (cardiovascular disease). Diabetic retinopathy is a result of damage to the small blood vessels and neurons of the retina. The earliest changes leading to diabetic retinopathy include narrowing of the retinal arteries associated with reduced retinal blood flow; dysfunction of neurons in the inner retina, followed at later stages by changes in the function of the outer retina associated with subtle changes in visual function; dysfunction of the blood-retinal barrier that protects the retina from many substances in the blood (including toxins and immune cells), leading to leakage of blood components into the retinal neuropile. Later, the basement membrane of the retinal blood vessels thickens, and the capillaries degenerate and lose cells, particularly pericytes and vascular smooth muscle cells. This leads to loss of blood flow and progressive ischemia, as well as microscopic aneurysms that appear as balloon-like structures protruding from the capillary walls (which recruit inflammatory cells); and progressive dysfunction and degeneration of retinal neurons and glial cells.

[0055] The ischemia and oxidant injury observed in DR impair blood flow, and the tissue ischemia discovered by the inventors can be reversed by edonentan and A-182086. With regard to DR symptoms, it is understood that improvement of retinal perfusion suppresses vascular endothelial growth factor (VEGF) upregulation, with the resultant benefit of reducing hypoxia and delaying angiogenic changes, neovascularization, and / or macular edema complications.

[0056] As a surrogate model for ischemic retinal lesions observed in DR, a preclinical mouse model of retinopathy of prematurity (ROP) can be used. Oxygen-induced retinopathy in the above mice is a reproducible and quantifiable model of proliferative retinal neovascularization suitable for testing the etiology and therapeutic interventions related to retinal neovascularization in ROP and other vascular pathologies (including DR). As previously described, the above model is induced by exposing 1-week-old C57BL / 6J mice to 75% oxygen for 5 days and then to room air (Smith LEH et al., Invest Ophthalmol Vis Sci 1994). The efficacy in this preclinical model of ROP can be evaluated by testing retinal hypoxia and neovascularization. Current standard care in DR includes anti-VEGF therapy that only addresses the advanced vascular complications of the disease. In some embodiments, this treatment is initiated in patients with DR during the non-proliferative stage of the disease. In some embodiments, either edonentan or A-182086 is locally administered to the posterior side of the eye at a frequency of every 3 to 12 months (e.g., every 3 to 6 months or every 4 to 6 months) using an intravitreal, topical, suprachoroidal, or implant delivery platform.

[0057] 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 cause edema. It is highly desirable to use edonentan and A-182086 treatments to treat RVO by reducing inflammation while improving tissue perfusion and avoiding the unwanted effects of systemic immunosuppression and / or the local harmful effects of steroids.

[0058] RVO is currently being treated with intravitreal steroids and anti-VEGF agents. Improving the perfusion of existing blood vessels reduces the degree of macular edema as well as changes in VEGF upregulation and downstream maladaptive changes manifesting as RVO. To test efficacy, a preclinical mouse model of ischemic retinopathy can be used. Oxygen-induced retinopathy in the said mice is a reproducible and quantifiable proliferative retinovascularization model suitable for testing the etiology and therapeutic interventions regarding retinal neovascularization in many ischemic retinopathies (including RVO). The said model is induced, as previously described, by exposing 1-week-old C57BL / 6J mice to 75% oxygen for 5 days and then to room air (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 current standard care by improving tissue perfusion and reducing ET-1-mediated inflammation while avoiding the unwanted effects of topical steroids. In some embodiments of the treatment of RVO, edonentan or A-182086 is administered locally to the posterior side of the eye using an intravitreal, topical, suprachoroidal, or implant delivery platform. The dosing frequency varies based on the patient's disease course and response to treatment.

[0059] Non-arteritic anterior ischemic optic neuropathy (NAION) In nonarteritic anterior ischemic optic neuropathy (NAION), there is an obstruction of blood flow to the small vessels that supply the anterior part of the optic nerve. Visual loss in NAION is painless, rapid, and usually permanent. Risk factors for NAION include atherosclerosis (which impairs blood flow through the vessels that supply the optic nerve) and a "tight" optic nerve. The optic nerve with a small cup or no cup, also known as the "disc at risk," when entering the eye, forms a "tight" pathway through the sclera. This tight pathway through the sclera is thought to impose additional pressure on the small vessels that supply the optic nerve. Atherosclerosis causes an increase in the outer diameter (and a decrease in the inner diameter) of these small vessels, leaving no room for the vessels to expand because they are constricted by the "tight" optic nerve. This process ultimately results in a loss of adequate blood flow to the optic nerve, leading to subsequent ischemic optic neuropathy. Attempts to treat NAION have included radial neurotomy to relieve mechanical pressure on the optic nerve and its supporting vasculature. This procedure has all the risks of intraocular surgery and is difficult to perform. The area to be penetrated is extremely delicate, as are the surrounding structures. Collateral damage to these structures is not uncommon.

[0060] Regarding NAION, there are no relevant preclinical models to test efficacy along with endothelin antagonism. Furthermore, there are no approved treatments for non-arteritic anterior ischemic optic neuropathy. Regarding the treatment of NAION, an amount of endothelin antagonist that is “therapeutically effective” is one that causes a clinically significant improvement in visual acuity and / or visual field by increasing perfusion of the optic nerve. As predictors of this preclinical endpoint, improvement in optic nerve head perfusion evaluated by laser flowmeter or optical coherence tomography angiography-A (OCT-A), and anatomical changes in retinal nerve fiber layer (RNFL) thickness determined by (OCT) are used. In some embodiments, the drug treatment is locally administered to the posterior side of the eye using an intravitreal, topical, suprachoroidal, or implant delivery platform at a frequency of every 4 to 6 weeks as needed, based on the patient's disease course and response to treatment. For example, the above drug treatment is locally administered to the posterior side of the eye using intravitreal injection of a suspension at a frequency of every 5 weeks as needed, based on the patient's disease course and response to treatment.

[0061] Arteritic anterior ischemic optic neuropathy (AION) Arteritic anterior ischemic optic neuropathy (AION) mainly occurs in elderly patients over 50 years old and is an acute, often painful optic neuropathy with an increasing incidence every 10 years thereafter, which can lead to permanent loss of vision. Ischemia occurs at the head of the optic nerve in relation to the structural density of nerve fibers, impairs perfusion, and results in optic disc edema. There is thought to be a genetic component to the disease as evidence shows a higher proportion of white people are affected, but it has also been reported in many different races and ethnic groups.

[0062] Symptom onset includes the sudden occurrence of decreased vision (typically severe: <20 / 200 in over 60% of patients), visual field defects (horizontal hemianopsia being the most common), or both, associated with unilateral visual loss. For AION, there are no relevant preclinical models to test efficacy along with endothelin antagonism. Furthermore, there are no approved treatments for non-arteritic anterior ischemic optic neuropathy; however, steroids have been traditionally used in most patients. Without treatment, visual loss occurs in 54 - 95% of GCA patients (16), typically within 4 months (10). With corticosteroid treatment, such rates of decline are reduced to approximately 13% (16). Visual recovery in treated affected eyes is insufficient with improvement rates of 15 - 34%, which is higher with intravenous treatment. Visual worsening has been reported in 9 - 17% despite treatment. Bilateral visual loss can occur; however, most cases are shown to be frequent when a patient does not notice the visual loss in the first eye. This bilateral incidence is highly dependent on timing, as well as how aggressively corticosteroid treatment is utilized. However, if left untreated, bilateral visual loss can rapidly progress in up to 50% of cases from either optic nerve ischemia, retinal ischemia, or choroidal ischemia. For the treatment of AION, a "therapeutically effective" amount of an endothelin antagonist is one that causes an additional improvement in vision achieved by steroids by improving perfusion of the optic nerve and retina. As predictive factors for improvement in vision and / or visual field, improvement in optic nerve head perfusion evaluated by laser flowmeter or OCT-A, and anatomical changes in retinal nerve fiber layer (RNFL) thickness determined by optical coherence tomography (OCT) are used. In some embodiments, drug treatment is locally administered posterior to the eye using intravitreal, topical, suprachoroidal, or implant delivery platforms at a frequency of every 4 - 6 weeks as needed, based on the patient's disease course and response to treatment.For example, the above drug treatment is topically administered posterior to the eye using intravitreal injection of the suspension at a frequency of every five weeks, if needed, based on the patient's disease course and response to treatment.

[0063] Retinopathy of prematurity (ROP) Retinopathy of prematurity (ROP) is a retinal vascular proliferative disease that affects premature infants. ROP continues to be a major preventable cause of blindness and visual impairment worldwide. With the improvement of perinatal care, the survival of moderately preterm infants, and the limitation of resources related to oxygen delivery and monitoring, more mature preterm infants are developing severe ROP in developing countries.

[0064] The pathophysiology of ROP is characterized by two stages. Stage I ROP begins immediately after birth due to vaso-obliteration following 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, particularly in the presence of retinal hypoxia and oxygen demand associated with increased retinal metabolism, leading to abnormal blood vessel growth. For advanced stage ROP, laser ablation of the avascular retina, the Early Treatment for Retinopathy of Prematurity (ETROP) protocol, intravitreal injection of anti-VEGF antibodies (e.g., bevacizumab), and vitrectomy are used to protect central vision and prevent retinal detachment. Long-term complications (e.g., refractive error, recurrence of ROP, and risk of retinal detachment) require continued ophthalmologist follow-up during puberty and thereafter. ROP is induced by severe ischemia resulting from incomplete development of retinal blood vessels following preterm birth. Thus, as one aspect of the present invention, the inventors believe that improvement of perfusion of existing blood vessels with edonentan or A-182086 reduces the degree of ischemia and VEGF upregulation, and the downstream maladaptive changes manifesting as ROP. To test efficacy, a preclinical mouse model of ROP can be used. Oxygen-induced retinopathy in the above mice is a reproducible and quantifiable proliferative retinopathy model suitable for testing the etiology and therapeutic interventions regarding retinal neovascularization in ROP. The above model is induced, as previously described, by exposing 1-week-old C57BL / 6J mice to 75% oxygen for 5 days and then to room air (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. The "therapeutically effective amount" of edonentan or A-182086 is an addition to current standard care by improving tissue perfusion and reducing pathological neovascularization induced by VEGF. In some embodiments, the above drug treatment is topically administered to the posterior side of the eye using an intravitreal, topical, suprachoroidal, or implant delivery platform at a frequency of every 4 to 6 weeks as needed, based on the patient's disease course and response to treatment. For example, the above drug treatment is topically administered to the posterior side of the eye using intravitreal injection at a frequency of every 5 weeks as needed, based on the patient's disease course and response to treatment.

[0065] Pharmaceutical composition 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 pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, excipient, or combinations thereof. Such antagonists or pharmaceutically acceptable salts thereof may be in crystalline or amorphous form. Each of these may be for pharmacologically acceptable use.

[0066] The term "pharmaceutical composition" refers to a mixture of one or both of the compounds disclosed herein with other chemical components (e.g., diluents or carriers). The pharmaceutical composition facilitates the administration of the compound to a living organism. Pharmaceutical compositions are generally made in accordance with a particular intended route of administration.

[0067] 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. Examples of 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, glucaronate, 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 salts, calcium salts, lithium salts, magnesium salts, potassium salts, sodium salts, zinc salts, and diethanolamine salts. For a review of pharmaceutically acceptable salts, see Berge et al., 66 J. PHARM. SCI, 1-19 (1977).

[0068] The term "pharmaceutically acceptable" defines a carrier, diluent, excipient, salt, or composition that is safe and effective for its intended use and has the desired biological and pharmacological activity.

[0069] As used herein, "carrier" refers to a compound that facilitates the incorporation of a compound into cells or tissues. For example, without limitation, dimethyl sulfoxide (DMSO) is a commonly used carrier that facilitates the uptake of many organic compounds into the cells or tissues of a subject.

[0070] As used herein, "diluent" refers to a component in a pharmaceutical composition that lacks pharmacological activity but is pharmaceutically necessary or desirable. For example, a diluent can be used to increase the bulk of a potent drug that is too small in mass for manufacture and / or administration. It can also be a liquid for dissolving a drug to be administered by injection, ingestion, or inhalation. Common forms of diluents in the art are buffered aqueous solutions (such as, but not limited to, phosphate buffered saline that mimics the composition of human blood).

[0071] As used herein, "excipient" refers to an inert substance added to a pharmaceutical composition to provide, without limitation, bulk, viscosity, stability, binding ability, lubrication, disintegration ability, etc. to the composition. "Diluent" is a type of excipient.

[0072] The pharmaceutical compositions described herein can be administered to a human patient, either by themselves or in pharmaceutical compositions that are mixed with other active ingredients (as in combination therapy) or with carriers, diluents, excipients, or combinations thereof. Appropriate formulations depend on the route of administration selected. Techniques for formulating and administering the compounds described herein are known to those of skill in the art.

[0073] The pharmaceutical compositions disclosed herein can be manufactured 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. Further, 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 pharmaceutically acceptable salts.

[0074] It is often preferred to administer the compounds or pharmaceutical compositions of the present invention in a topical manner as a topical ophthalmic formulation or directly into the ocular tissues, either via injection of the compound or pharmaceutical composition, often in a depot or sustained release formulation. The modes of topical administration can be intravitreal, suprachoroidal, periocular, or subconjunctival injection of the formulation, or the use of implant technology or topical application. For example, the compounds are administered in a liposomal preparation that slowly releases a compound having the desired pharmacological effect. Alternatively, polyvinyl alcohol nanoparticles can be prepared by well-known methods to provide a sustained release or long-term release formulation for topical or intravitreal application.

[0075] Furthermore, the compounds can be administered in a targeted drug delivery system. Examples of targeted drug delivery systems include ApidCOR, BioSeizer-ProDex, Vitrasert, Retisert, Iluvien, I-Vation, Nanoporous Silicon, Ozurdex / Novadur, OcuLief, Port Delivery System (PDS), PEA Implant, PEG-PLA Microspheres, PRINT Technology, Q-Sphera, SKS Microparticles, Verisome, Capsule Ring Device, MicroPump, Microneedle Injector, Microneedle / Needle-less Injectors, EyeCET, Gemini Refractive Capsule, IVMED, Ciliary Sulcus Ring, Episcleral Exoplant, Eye-D Implant, and Nanoliposomes, but are not limited thereto.

[0076] In some embodiments, the targeted drug delivery systems that can be used in the methods of the present disclosure are selected from ApidCOR, BioSeizer-ProDex, Vitrasert, Retisert, Iluvien, I-Vation, Nanoporous Silicon, Ozurdex / Novadur, OcuLief, Port Delivery System (PDS), PEA Implant, PEG-PLA Microspheres, PRINT Technology, Q-Sphera, SKS Microparticles, Verisome, Capsule Ring Device, MicroPump, Microneedle Injector, Microneedle / Needle-less Injectors, EyeCET, Gemini Refractive Capsule, and IVMED. Preferably, the targeted drug delivery system is ApidCOR, BioSeizer-ProDex, Vitrasert, Retisert, Iluvien, I-Vation, Nanoporous Silicon, Ozurdex / Novadur, OcuLief, Port Delivery System (PDS), PEA Implant, PEG-PLA Microspheres, PRINT Technology, Q-Sphera, SKS Microparticles, Verisome, Capsule Ring Device, or MicroPump.

[0077] In some embodiments, the pharmaceutical composition is an ophthalmic preparation comprising a therapeutically effective amount of one or more endothelin receptor antagonists, or pharmaceutically acceptable salts thereof, as described herein. As used herein, "ophthalmic preparation" refers to a specialized dosage form designed to be instilled onto the outer surface (topically) of the eye, administered internally (intraocularly) or adjacent to the eye (periocularly), 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 / nanoparticle preparation for topical, or preferably, intravitreal injection, or implant.

[0078] 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 mercuries (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). The preservative may be present in the ophthalmic preparation in an amount in the range of about 0.002% w / v to about 0.5% w / v (e.g., 0.01 - 0.25% w / v). The ophthalmic preparation may further contain a preservative aid. Examples of suitable preservative aids include, but are not limited to, ethylenediaminetetraacetic acid (EDTA).

[0079] In some embodiments, the ophthalmic preparation contains one or more additional excipients or agents to impart viscosity or lubrication, to stabilize the active ingredient against degradation, to increase the solubility of the active or inactive ingredient, to adjust the tonicity, or to act as a solvent. Examples of excipients or agents for imparting viscosity or lubrication include hypromellose, carbomer 974P, hydroxyethyl cellulose (HEC), polyvinyl alcohol, sodium hyaluronate, sodium carboxymethyl cellulose, Carbopol 940, hydroxypropyl methyl cellulose (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 pyrosulfite, sodium thiosulfate, and sodium sulfate / sulfuric acid (which can act as antioxidants). Examples of excipients or agents as solubilizers include providone, creatinine, castor oil, and cyclodextrin (e.g., γ-cyclodextrin), but are not limited thereto. Examples of excipients or agents for adjusting the 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, but are not limited thereto.

[0080] In some embodiments, the ophthalmic preparation contains one or more buffering substances for adjusting the pH. Examples of buffering substances 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 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).

[0081] In some embodiments, the ophthalmic preparation contains one or more surfactants. Examples of surfactants include sorbitan ether esters of oleic acid (e.g., polysorbate or Tween® 20 and 80) and tyloxapol.

[0082] 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 of sizes 27 - 30G. The dose depends on the concentration that can be formulated to be compatible with this volume, potency, target efficacy, and the pharmacokinetic profile for each indication. Generally, eye injections are not administered at a frequency of more than once per 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 more than twice a day.

[0083] In some embodiments, the intravitreal formulation comprises an endothelin receptor antagonist in an amount ranging from about 1 μg to about 1 mg. A first exemplary formulation comprises about 1 μg to about 1 mg of the endothelin receptor antagonist described above, 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 the endothelin receptor antagonist, about 10 mM sodium phosphate, about 40 mM sodium chloride, about 0.03% polysorbate 20, and about 5% sucrose.

[0084] In some embodiments, the intravitreal formulation comprises an endothelin receptor antagonist in an amount ranging from about 10 μg to about 100 μg. A first exemplary formulation comprises about 10 μg to about 100 μg of the endothelin receptor antagonist described above, about 10 mM histidine HCl, about 10% α,α-trehalose dihydrate, and about 0.01% polysorbate 20. A second exemplary formulation comprises about 10 μg to about 100 μg of the endothelin receptor antagonist, about 10 mM sodium phosphate, about 40 mM sodium chloride, about 0.03% polysorbate 20, and about 5% sucrose.

[0085] In further embodiments, the intravitreal formulation comprises an endothelin receptor antagonist in an amount ranging from about 500 μg to about 4 mg. A first exemplary formulation comprises about 500 μg to about 1 mg of the endothelin receptor antagonist described above, 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 the endothelin receptor antagonist described above, 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.

[0086] Without further elaboration, one of ordinary skill in the art will appreciate that the present invention can be utilized to its fullest extent based on the foregoing description. Accordingly, the following specific examples, namely Examples 1-8, are illustrative only and should in no way be construed as limiting the remainder of the disclosure.

Example

[0087] Example 1: Physicochemical and Biochemical Characterization of Compounds The following Table 1 provides the physicochemical and biochemical data for the above edonentan and A-182086. As shown in Table 1, at pH 2, A-182086 has better solubility than that of edonentan. On the other hand, at pH 7, edonentan has better solubility than that of A-182086. Table 1. Physicochemical and Biochemical Characterization of Compounds

Table 1

[0088] In the above table, the physicochemical data (e.g., solubility) was obtained according to standard protocols known in the art (see, for example, Reis et al., Mini Rev Med Chem., 2010, 10(11):1071-6; Avdeef et al., Expert Opin Drug Metab Toxicol., 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); the biochemical data (i.e., ET A / ET BThe efficacy) was obtained according to protocols known in the art (see, for example, Kirkby et al., Br J Pharmacol., 2008, 153(6):1105-19; and Maguire et al., Br J Pharmacol., 2014, 171(24):5555-72).

[0089] Example 2: Formulation of edonentan for intravitreal use in rabbits An appropriate amount of edonentan is dissolved in neat PEG400, and subsequently a 15% CD (HP-β-cyclodextrin) solution is added. The final concentration of PEG400 is measured to be 20%. The target concentrations are 5 mg / ml and 0.5 mg / ml based on the amount of edonentan. The resulting solution is filtered using a 0.25 micron filter.

[0090] 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 of ET-1, followed 30 minutes after ET-1 administration by a 20 μl intravitreal injection of 10 - 100 μg of edonentan. Intraocular pressure (IOP), optical coherence tomography-angiography (OCT-A), and fluorescein angiogram (FA) were performed at predetermined time points (30 minutes, 45 minutes, 60 minutes, and 75 minutes) 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 a marked vasoconstriction in the retinal vascular bed within 45 minutes. Figure 2 shows that the effect of ET-1 was then reversed by administration of 10 μg of edonentan within 90 minutes (60 minutes after edonentan administration).

[0091] Example 4: Preparation of a long-term release formulation containing edonentan A concentrated edentan dispersion is prepared by combining edentan with water, vitamin E-TPGS, and γ-cyclodextrin. These components are mixed to disperse the edentan, and then autoclaved. Sodium hyaluronate can be purchased as a sterile powder or a dilute solution can be sterilized by filtration and subsequently lyophilized to obtain a sterile powder. The sterile sodium hyaluronate is dissolved in water to form an aqueous concentrate. The concentrated edentan dispersion is mixed and added to the sodium hyaluronate concentrate as a slurry. Water is added in a sufficient amount (sufficient to the extent required in this case 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. Compositions of Sustained Release Formulations Containing Edentan

Table 2

[0092] 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 sodium hyaluronate used is less than 2 million, and more preferably, the average molecular weight of the sodium hyaluronate used is between about 1.3 million and 1.6 million. The edentan particles are actually trapped or retained within this viscous plug of sodium hyaluronate, such that undesirable pluming does not occur upon intravitreal injection of the above formulation. Thus, the risk that drug particles will inconveniently precipitate directly onto the retinal tissue is substantially reduced compared to using a composition having viscosity such as water (e.g., Kenalog® 40). Since the sodium hyaluronate solution undergoes dramatic shear thinning, these formulations can be easily injected via a 25-gauge, 27-gauge, or even 30-gauge needle.

[0093] Example 5: Preparation of a Topical Edentan Formulation The local edonentan preparation can be prepared according to a known method (for example, 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 stirred until completely dissolved. Then, 1.5 g of tromethamine is added to the resulting solution and stirred for 15 minutes to achieve complete dissolution. 0.5 g of edonentan is added and stirred for 15 minutes to ensure complete dissolution. Then, 2 g of glycine and 1 g of boric acid are added and stirred until completely dissolved. A sufficient amount of the resulting solution is added to 100 mL of deionized water. When the final solution is filtered through filter paper, a colorless transparent solution having a pH of 8.06 is obtained. The above solution is filled into an eye drop in an eye drop bottle having a volume of 5 mL.

[0094] Example 6: Local ophthalmic solution nanoparticles containing edonentan Nanoparticles were prepared by solvent evaporation technique. A solution of 120 mg of 50:50 PLGA in 60 mL of ethyl acetate was prepared. An aqueous solution of 50 ml of water, 12 mg of edonentan and 0.5 mg of polyvinyl alcohol was incorporated into this solution with vigorous stirring. The resulting mixture was left under continuous stirring and under vacuum for 2 hours. Then, the resulting preparation was ultracentrifuged and washed three times with water to remove the nanoparticles from the medium. The nanoparticles thus obtained were dried in a vacuum oven and, after evaluation, dispersed in an isotonic aqueous solution sufficient for a concentration of 5 mg / 1 mL of edonentan.

[0095] Example 7: Preclinical glaucoma test The healthy rabbit model is used to (in vivo) evaluate the pharmacodynamic effects of edonentan and / or A-182086 or their pharmaceutically acceptable salts. These tests are conducted at various doses of the selected endothelin antagonists. Further animal tests are conducted by combining the endothelin antagonist with current standard care. The Morrison rat model of glaucoma, the rat model of acutely elevated IOP, and the laser-induced glaucoma model in non-human primates are used to evaluate the optic nerve head blood flow and the rate of retinal ganglion cell loss associated with various doses of the selected endothelin antagonists with and without standard care.

[0096] Improvement in blood flow in the healthy rabbit model is measured at various doses for the indicated endothelin receptor antagonists after induction of perfusion impairment by locally administered ET-1. Changes in optic nerve head blood flow and retinal nerve fiber layer (RNFL) thickness in the non-human primate glaucoma model are measured at various doses for the indicated endothelin receptor antagonists. The results show improvement in RGC survival, retinal and optic nerve head blood flow, and slowing of RNFL thinning resulting from the use of the selected endothelin receptor antagonists. The dosing regimen for humans is inferred from the results of the healthy rabbit and non-human primate glaucoma models.

[0097] Pharmacodynamic tests to evaluate changes in retinal blood flow in rabbits To evaluate the effect of intravitreally administered endothelin-1 (ET-1) followed by administration of the antagonist edonentan on retinal blood flow in rabbits, 20 μL of intravitreal injection of ET-1 was given to the left eyes of rabbits (Oryctolagus cuniculus), followed by 20 μL of intravitreal injection of 2 (or 3) different doses (e.g., 0.1 μg, 0.5 μg, 2.5 μg) of edonentan. 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 FIGS. 8A and 8B.

[0098] Pharmacokinetics and Tolerance Analysis of Endothelin Administered Intravitreally in Rabbits To determine the pharmacokinetics and safety profile of endothelin following 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 by 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 collected from the marginal ear vein or by cardiac puncture (for terminal blood collection only) into K2EDTA tubes for plasma collection and processed for analysis.

[0099] Immediately following euthanasia, the eyes were enucleated. The aqueous humor of both eyes was removed via syringe and snap frozen for analysis. The eyes were dissected when frozen to isolate various eye tissues and minimize drug diffusion to adjacent tissues. The left and right eye tissues were collected in separate vials. The list of tissues collected included plasma and aqueous humor, iris / ciliary body (ICB), retina, vitreous humor, and RPE / choroid. The pharmacokinetic characteristics of endothelin administered intravitreally in rabbits are shown in FIGS. 9A, 9B, 9C, and 9D.

[0100] Pharmacokinetic Analysis of Endothelin Administered Topically in Rabbits To determine the pharmacokinetic properties of edonentan after topical administration in rabbits, both eyes of rabbits (Dutch-belted rabbits) were instilled with eye drops (100 μg of edonentan, 35 μL dose volume / eye). After the administration, the animals (N = 2) were euthanized at various time points (e.g., 10 minutes (immediately after pot-dose), 2 hours, and 7 hours), and tissues were collected for analysis. The list of collected tissues included plasma, retina, vitreous humor, and conjunctiva. The pharmacokinetic properties of topically delivered edonentan in rabbits are shown in Figure 10. This indicates that edonentan was detected in all tissues tested at all time points after a single topical application.

[0101] Efficacy test in Morrison's rat model of glaucoma Adult male and female Brown Norway rats that had finished breeding (approximate age group of 8 - 11 months) were obtained from Envigo (Indianapolis, IN). Baseline IOP measurements and pattern electroretinogram (PERG) amplitudes were collected before surgery for the elevation of IOP (to ensure that IOP and PERG amplitudes were within the predicted range). While IOP was elevated in one eye (left eye) of the rat, the corresponding right eye was used as a contralateral control. The Morrison method for elevating IOP in rats was performed by injecting 50 μL of hypertonic saline via the episcleral vein to sclerose the trabecular meshwork. IOP was measured twice a week throughout the duration of the experiment. Seven to ten days after surgery, an elevation in IOP was observed in the surgically treated eye of the rat. After detecting an elevation in IOP for two consecutive days, topical administration of eye drops (20 μL (100 μg) / dose of the test compound in the eye with elevated IOP) was initiated and carried out for 5 days a week for a total of 4 weeks. At the fourth week of treatment, PERG analysis was performed, and the rats were sacrificed by an overdose of pentobarbital (Fatal-Plus). Aqueous humor was collected from the eyes of the rats, frozen, and sent for analysis. Flat mounts of the retina were prepared, immunostained with the RGC marker, Brn3a antibody, and the surviving RGCs were counted at two eccentricities (central and peripheral).

[0102] For this study, the Morrison model was used to induce ocular hypertension in adult male Brown Norway rats that had completed breeding, 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).

[0103] Immunostained retinal flat mounts were obtained and the number of retinal ganglion cells (RGCs) was measured. To obtain immunostained retinal flat mounts, the animals were euthanized after treatment and then their eyes were enucleated. The eyecups were fixed overnight in 4% paraformaldehyde (PFA) at 4 °C and retinal flat mounts were prepared for image collection. Retinal ganglion cell (RGC) counts were performed using 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 (central and peripheral). Figure 5A shows a comparison of the number of RGCs in the peripheral retina between vehicle and edonerpic, and Figure 6A shows a comparison between vehicle and A-182086.

[0104] The retinal ganglion cell (RGC) function was evaluated using pattern electroretinogram (PERG). To obtain the PERG recordings, a 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 tool to monitor progressive retinal ganglion cell dysfunction in the DBA / 2J mouse model of glaucoma. Invest Ophthalmol Vis Sci. 2007;48(2):745-751). Briefly, the PERG signals were acquired from a DTL-plus electrode placed on the inferior part of the corneal surface, and the PERG waves were analyzed using EMWIN software (LKC). The difference between the amplitudes of the major positive (P1) and negative (N2) waves was calculated to decipher the PERG amplitude. Figure 5B shows the IOP-mediated PERG changes between vehicle and edonerpic, and Figure 6B shows the changes between vehicle and A-182086.

[0105] As shown in Figures 5A, 5B, 6A, and 6B, the RGC counts and PERG changes demonstrate that both edonerpic and A-182086 prevented RGC loss and maintained RGC function in the Morrison rat model of glaucoma.

[0106] Pharmacokinetic analysis of edonerpic or A-182086 delivered locally or orally in rats To determine the pharmacokinetic properties of edonentan or A-182086 after local administration in rats, rats (Brown Norway rats) were given eye drops (100 μg of edonentan, 20 μL dose volume / eye; or 100 μg of 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 an oral dose of 10 mg / kg or 50 mg / kg of edonentan, or 1.7 mg / kg or 17 mg / kg of A-182086. After administration, the 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 properties of edonentan administered locally or orally in rats are shown in Figure 5C. The pharmacokinetic properties of A-182086 administered locally or orally in rats are shown in Figure 6C. Figures 5C and 6C show that both edonentan and A-182086 were detected in the retina / RPE / choroid, aqueous humor, and vitreous humor 4 hours and 8 hours after local administration. These data also revealed that edonentan was detectable in aqueous humor at 17 mg / kg after oral administration of edonentan, and in the retina / RPE / choroid and vitreous humor at 1.7 and 17 mg / kg, and that A-182086 was detectable in the retina / RPE / choroid at 50 mg / kg after oral administration of A-182086.

[0107] Test in mice with oxygen-induced ischemic retinopathy The relevant mouse models were used to obtain retinal hypoxic regions in mice with oxygen-induced ischemic retinopathy (OIR) at various time points as shown in Figure 4. The improvement of retinal hypoxia in the mouse oxygen-induced ischemic retinopathy (OIR) model by edonentan was revealed. Briefly, at P17, mice with OIR (n = 60) were given an injection (1 μL of 2 μg / μL edonentan formulation, single dose) of an ET-1 antagonist in one eye and PBS in the other eye. At 24 hours, 48 hours, and 96 hours after injection, the mice (n = 10 for each time point) were euthanized, the retinas were dissected, and stained with GSA lectin and hypoxyprobe. The areas of NV, retinal hypoxia, and retinal non-perfusion were determined for each retina.

[0108] Example 8: Laser-Induced Non-Human Primate Study - Pharmacodynamic Study Non-human primates (rhesus monkeys, Macaca Mulatta) were obtained for this study. In one eye of each animal, an increase in intraocular pressure (IOP) was induced by repeated laser photocoagulation of the fiber column network. Imaging sessions were repeated to monitor changes in the optic nerve head (ONH) and retinal structure.

[0109] Effect of Edonentan on Optic Nerve Head Blood Flow after IVT Administration As an index of ONH blood flow in a laser-induced glaucoma model, a study was conducted to compare the overall mean blur rate (MBR) and the change in MBR over time from baseline in the experimental glaucomatous eyes and the contralateral healthy eyes (controls) of three non-human primates. More specifically, vehicle control, 0.02 mg / mL edonerpic, 0.2 mg / mL edonerpic, or 2.0 mg / mL edonerpic was intravitreally administered (50 μL) to each glaucomatous eye of three non-human primates (rhesus monkeys, Macaca Mulatta). Subsequently, ONH blood flow was measured over 6 hours using laser speckle flowgraphy (LSFG) as shown in FIGS. 7A-7L. These graphs show ONH blood flow in three non-human primates after intravitreal administration of vehicle alone (FIGS. 7A, 7E, and 7I), 0.02 mg / mL edonerpic (FIGS. 7B, 7F, and 7J), 0.2 mg / mL edonerpic (FIGS. 7C, 7G, and 7K), or 2.0 mg / mL edonerpic (FIGS. 7D, 7H, and 7L). FIGS. 7A-7L reveal an improvement in ONH blood flow in a dose-dependent manner after treatment with edonerpic. The pooled results of the three non-human primates are shown in FIG. 7M. This indicates that edonerpic clearly shows a dose-related increase in ONH blood flow resulting from the dilation of retinal arteries, veins, and capillaries in experimental glaucomatous eyes compared to control eyes.

[0110] In one of the above three non-human primates, LSFG scans were performed at various selected time points when edonerpic was administered at 2.0 mg / mL. The results are shown in FIG. 7N.

[0111] Effect of edonerpic on intraocular pressure after topical administration A single dose of 0.5% timolol or a single dose of 2 mg / mL edonerpic was topically administered to three non-human primates with laser-induced glaucoma in the right eye (OD) in a randomized order with a 1-week washout.

[0112] Test results: Control 1: A single dose of 50 μL of topical timolol 0.5% in each eye showed an IOP reduction of approximately 20% from before administration to after administration (for 120 minutes).

[0113] Control 2: A single dose of 50 μL of topical timolol 0.5% in each eye showed an IOP reduction of approximately 30% from before administration to after administration (for 120 minutes).

[0114] Non - human primate 1: 50 μL of edonerpic eye drops (2 mg / mL) in the experimentally glaucomatous eyes showed an IOP reduction of approximately 60% from before administration to after administration (for 120 minutes), and in the contralateral healthy eyes, it showed an IOP reduction of approximately 10% from before administration to after administration (for 120 minutes).

[0115] Non - human primate 2: 50 μL of edonerpic eye drops (2 mg / mL) in the experimentally glaucomatous eyes showed an IOP reduction of approximately 50% from before administration to after administration (for 15 minutes) and approximately 30% from before administration to after administration (for 120 minutes). 50 μL of edonerpic eye drops (2 mg / mL) in the contralateral healthy eyes showed an IOP reduction of approximately 20% from before administration to after administration (for 15 minutes) and approximately 0% from before administration to after administration (for 120 minutes).

[0116] Non - human primate 3: 50 μL of edonerpic eye drops (2 mg / mL) in the experimentally glaucomatous eyes showed an IOP reduction of approximately 40% from before administration to after administration (for 15 minutes) and approximately 40% from before administration to after administration (for 120 minutes). 50 μL of edonerpic eye drops (2 mg / mL) in the contralateral healthy eyes showed an IOP reduction of approximately 10% from before administration to after administration (for 15 minutes) and approximately 40% from before administration to after administration (for 120 minutes).

[0117] Other embodiments All features disclosed in this specification may be combined in any combination. Each feature disclosed in this specification may be replaced by alternative features that serve the same, equivalent, or similar purpose. Accordingly, unless expressly stated otherwise, each feature disclosed is only an example of a broad series of equivalent or similar features.

[0118] Furthermore, from the above description, those skilled in the art can easily identify the essential features of the present invention and make various changes and modifications to the present invention without departing from its spirit and scope, so as to adapt to various usage methods and conditions. Accordingly, other embodiments are also within the scope of the claims. The present invention provides, for example, the following items. (Item 1) A method for treating an eye disease, the method comprising: contacting the visual tissue of a subject with a composition comprising a therapeutically effective amount of either edonentan or A-182086, or a pharmaceutically acceptable salt thereof, or a crystalline or amorphous form thereof; wherein the eye disease is selected from the group consisting of glaucoma, diabetic retinopathy (DR), retinal vein occlusion (RVO), non-arteritic anterior ischemic optic neuropathy (NAION), arteritic anterior ischemic optic neuropathy (AION), and retinopathy of prematurity (ROP). (Item 2) The method according to item 1, wherein the therapeutic effectiveness of the treatment is determined by evaluating the degree of improvement in visual acuity or visual field. (Item 3) The method according to item 1, wherein the eye disease is glaucoma. (Item 4) The method according to item 3, wherein the therapeutic effectiveness of the treatment is determined by detecting a sufficient amount of a decrease in intraocular pressure or a reduction in the rate of optic nerve damage to reduce or prevent optic nerve damage. (Item 5) The method according to item 3, wherein the therapeutic effectiveness of the treatment is determined by improvement in optic nerve head blood flow. (Item 6) The method according to item 1, wherein the eye disease is DR, RVO, NAION, AION, or ROP. (Item 7) The therapeutic effectiveness of the treatment is determined by a decrease in retinal neurodegeneration induced by diabetes. The method according to item 6. (Item 8) The method according to item 6, wherein the therapeutic effectiveness of the treatment is indicated by an improvement in tissue or retinal perfusion. (Item 9) The method according to item 1, wherein the therapeutic effectiveness of the treatment is determined by measuring an improvement in tissue or retinal perfusion, a reduction in inflammation, or a combination thereof. (Item 10) The method according to item 1, 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 11) The method according to item 10, 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 (e.g., latanoprost or travoprost), β-blockers (e.g., timolol or betaxolol), α-adrenergic agonists (e.g., brimonidine, apraclonidine), carbonic anhydrase inhibitors (e.g., dorzolamide or brinzolamide), Rho kinase inhibitors (e.g., netarsudil), and miotics or cholinergic agents (e.g., pilocarpine). (Item 12) The method according to item 10, 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 anti-apoptotic agents (e.g., caspase-2 inhibitor) and neurotrophic factors (e.g., ciliary neurotrophic factor). (Item 13) The method according to item 1, wherein the composition comprises edonentan. (Item 14) The method according to item 13, wherein the eye disease is glaucoma. (Item 15) The therapeutic effectiveness of the treatment is determined by detecting a decrease in intraocular pressure or a reduction in the rate of optic nerve damage in an amount sufficient to reduce or prevent optic nerve damage, according to the method of item 14. (Item 16) The composition contains A-182086, according to the method of item 1. (Item 17) The eye disease is glaucoma, according to the method of item 16. (Item 18) The therapeutic effectiveness of the treatment is determined by detecting a decrease in intraocular pressure, or a reduction in the rate of optic nerve damage, or a combination thereof, in an amount sufficient to reduce or prevent optic nerve damage, according to the method of item 17. (Item 19) The composition is administered at a dosage between 1 μg and 4 mg, according to the method of item 1. (Item 20) The composition is administered at a dosage between 10 μg and 100 μg, according to the method of item 1. (Item 21) The step of contacting includes the step of locally administering the composition to the surface of the eye or a part thereof, according to the method of item 1. (Item 22) The step of contacting includes the step of injecting the composition into the eye or its components, according to the method of item 1. (Item 23) The composition includes an ophthalmic preparation containing one or more preservatives, preservative aids, viscosity or lubrication modifiers, tonicity modifiers, solubilizing agents, buffering substances, surfactants, stabilizers, or combinations thereof, according to the method of item 1. (Item 24) The step of contacting includes the step of administering the composition via a targeted drug delivery system, where the targeted drug delivery system is selected from the group consisting of ApidCOR, BioSeizer-ProDex, Vitrasert, Retisert, Iluvien, I-Vation, Nanoporous Silicon, Ozurdex / Novadur, OcuLief, Port Delivery System (PDS), PEA Implant, PEG-PLA Microspheres, PRINT Technology, Q-Sphera, SKS Microparticles, Verisome, Capsule Ring Device, MicroPump, Microneedle Injector, Microneedle / Needle-less Injectors, EyeCET, Gemini Refractive Capsule, IVMED, Ciliary Sulcus Ring, Episcleral Exoplant, Eye-D Implant, and Nanoliposomes, the method according to item 1.

Claims

1. A composition for the treatment of diabetic retinopathy, comprising a therapeutically effective amount of edonentan, a pharmaceutically acceptable salt thereof, or a crystalline form thereof.

2. The composition according to claim 1, wherein the therapeutic effectiveness of the treatment is determined by a reduction in the diabetic retinopathy severity score compared to baseline.

3. The composition according to claim 1, wherein the therapeutic effectiveness of the treatment is determined by an improvement in visual acuity or visual field.

4. The composition according to claim 1, wherein the therapeutic effectiveness of the treatment is determined by a reduction in retinal neurodegeneration induced by diabetes.

5. The composition according to claim 1, wherein the therapeutic effectiveness of the treatment is determined by the improvement of retinal perfusion in the patient.

6. The composition according to claim 1, wherein the therapeutic effectiveness of the treatment is determined by delaying macular edema complications in the patient.

7. The composition according to claim 1, characterized in that the composition is administered in a dose between 100 μg and 500 μg.

8. The composition according to claim 1, wherein the treatment comprises the step of bringing the visual tissue of a patient into contact with the composition.

9. The composition according to claim 8, wherein the step of contacting the eye or a component thereof comprises the step of injecting the composition into the eye or a component thereof.

10. The composition according to claim 9, wherein the step of contacting the eye comprises the step of injecting the composition behind the eye.

11. The composition according to claim 10, wherein the step of injecting the composition into the posterior side of the eye is performed intravitreously, on the choroid, subretinally, or using an implantable delivery platform.

12. The composition according to claim 1, comprising an ophthalmic preparation containing one or more preservatives, preservative aids, viscosity or lubrication modifiers, tension modifiers, solubilizers, buffering substances, surfactants, stabilizers, or combinations thereof.

13. The composition according to claim 1, characterized in that it is administered via a targeted drug delivery system.

14. The composition according to claim 13, wherein the targeted drug delivery system is selected from the group consisting of nanoporous silicon, a port delivery system (PDS), a PEA implant, poly(ethylene glycol)-poly(lactic acid) (PEG-PLA) microspheres, a micropump, a microneedle injector, a needleless injector, and nanoliposomes.

15. An intravitreal implant composition for the treatment of diabetic retinopathy in a patient requiring treatment for diabetic retinopathy, wherein the intravitreal implant composition comprises about 100 μg to about 500 μg of edonentan, and the treatment comprises the step of injecting the intravitreal implant composition behind the eye of the patient every 3 to 6 months.

16. The intravitreous implant composition according to claim 15, wherein the edonentan is in crystalline form.

17. The intravitreous implant composition according to claim 15, wherein the patient is in the non-proliferative stage of diabetic retinopathy.

18. An ophthalmic preparation for the treatment of diabetic retinopathy in a patient requiring treatment for diabetic retinopathy, wherein the ophthalmic preparation comprises a therapeutically effective amount of edonentan, and the treatment comprises the step of injecting the ophthalmic preparation into the posterior side of the patient's eye every three to six months.

19. The ophthalmic preparation according to claim 18, wherein the edonentane is in crystalline form.

20. The ophthalmic preparation according to claim 18, wherein the ophthalmic preparation delivers approximately 100 μg to approximately 500 μg of edonentan to the posterior side of the eye when injected.

21. The ophthalmic preparation according to claim 18, wherein, upon injection of the ophthalmic preparation, the patient shows a reduction in the diabetic retinopathy severity score compared to baseline.

22. The ophthalmic preparation according to claim 18, wherein, upon injection of the ophthalmic preparation, the patient exhibits an improvement in visual acuity or visual field.

23. The ophthalmic preparation according to claim 18, wherein, upon injection of the ophthalmic preparation, the patient exhibits a reduction in diabetes-induced retinal neurodegeneration.

24. The ophthalmic preparation according to claim 18, wherein, upon injection of the ophthalmic preparation, the patient exhibits improved retinal perfusion.

25. The ophthalmic preparation according to claim 18, wherein, upon injection of the ophthalmic preparation, the patient exhibits delayed macular edema complications.

Images