Local intraocular delivery of cromakalim

Abstract

Aqueous topical ophthalmic pharmaceutical formulations are provided that deliver an effective amount of (lev)cromakalim or a pharmaceutically acceptable salt thereof to treat anterior or posterior ocular disorders that respond to (lev)cromakalim, including glaucoma.

Description

[Technical Field]

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 422,805, filed November 4, 2022, U.S. Provisional Patent Application No. 63 / 424,434, filed November 10, 2022, and U.S. Provisional Patent Application No. 63 / 523,616, filed June 27, 2023, the entire contents of each of which are incorporated herein by reference for all purposes.

[0002] [STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT] This invention was made with government support under Grant No. EY021727 awarded by the National Institutes of Health. The United States Government has certain rights in this invention.

[0003] The present application provides pharmaceutical formulations for local intraocular delivery of cromakalim, including levcromakalim, for use in therapies including the reduction of intraocular pressure and the general treatment of glaucoma, including normal tension glaucoma, as further described herein. [Background technology]

[0004] Cromakalim and its use as an antihypertensive agent were first described in U.S. Patent No. 5,629,499, which claims priority to an application filed in 1983 and issued in 1990, and is assigned to Beecham Group, Inc. Disclosures of the effects of cromakalim on intraocular pressure and glaucoma were reported in U.S. Patent No. 5,629,499, U.S. Patent No. 5,629,499, and U.S. Patent No. 5,629,499.

[0005] Cromakalim has been reported to lower blood pressure in Non-Patent Document 3. Cromakalim deposited in membrane patches from rabbit mesenteric artery smooth muscle cells lowers the blood pressure by a single K ATP The open state probability of the channel (P open) by more than 9-fold (Non-Patent Document 4). Cromakalim exists as a mixture of stereoisomers in the trans configuration (a mixture of the (3R,4S) stereoisomer and the (3S,4R) stereoisomer).

[0006] [ka]

[0007] The (3S,4R)-stereoisomer is also called (-)-cromakalim or levcromakalim, and the (3R,4S)-stereoisomer is also called (+)-cromakalim or dexcromakalim.

[0008] [ka]

[0009] Most of the reported activity of cromakalim comes from the (3S,4R)-stereoisomer, levcromakalim (Non-Patent Document 5 and Non-Patent Document 6).

[0010] Cromakalim is poorly soluble in water and aqueous buffer formulations and tends to crystallize or co-crystallize out of solution. Cromakalim is also poorly soluble in oils or nonpolar hydrophobic solvents. Cromakalim is soluble in highly polar "general purpose" organic solvents such as DMSO (dimethyl sulfoxide), DMF (dimethylformamide), or NMP (1-methylpyrrolidone), which allow for hydrogen-bonding and hydrophobic interactions, but these are not preferred solvents for in vivo drug delivery. Cromakalim is also somewhat soluble in alcohols such as ethanol, but these low molecular weight monoalcohols are not suitable for topical intraocular administration in humans.

[0011] Despite its potent biological activity, cromakalim has never been approved as a drug. Cromakalim was tested in human clinical trials for systemic hypertension, but the program was discontinued. Levcromakalim has never been considered for local administration for ophthalmic use, such as via the topical or intraocular route, due to its insolubility in water.

[0012] Cromakalim has been solubilized with DMSO and cremophor (Cremophor EL, now called Kolliphor EL by BASF), which is also used for the water-insoluble anticancer drug Taxol. However, cromakalim showed poor solubility even in DMSO co-solvent mixtures, which required the use of a high percentage of DMSO, as demonstrated in Non-Patent Document 7, or in combination with Cremophor, as demonstrated in Non-Patent Document 2 (Roy Chowdhury, U et al. PLOS ONE. 2015, 10 (11), e0141783).

[0013] Chiang and Lin investigated the effects of cromakalim in rabbit eyes using three levels of intraocular pressure (Chiang and Lin, "Effects of Cromakalim and Nicorandil on Intraocular Pressure After Topical Administration in Rabbit Eyes", J. Ocular Pharmacology; 1995 Vol 11(3), 195-202). The cromakalim test solution used in the study was reported to be 0.5% (estimated weight / weight) cromakalim in 0.02 M phosphate buffer. The authors reported that cromakalim tends to increase IOP after topical administration and may be contraindicated in patients with glaucoma. They reported that cromakalim reduced IOP in the α-chymotrypsin model after a delay that included an increase in IOP for the first few hours. That being said, Chiang and Lin's report must be taken with a pinch of salt. This is because, in 1993, SmithKline Beecham Pharmaceuticals, Harlow, UK, had previously reported that levcromakalim had a solubility limit in water of 0.48 mg / ml (see Non-Patent Document 8), which is 0.048% (wt / wt), or one-tenth the value used by Chiang and Lin at the National Defence Medical Centre in Taiwan in 1995. Therefore, the materials tested by Chiang and Lin undoubtedly contained insoluble substances that would have adverse effects on the eye and invalidated their results. Hamilton's paper also states that cromakalim is soluble in ethanol, DMSO, and PEG, but that none of these are acceptable topical ocular carriers for administration to humans.

[0014] In response to the need to create a cromakalim formulation with suitable solubility for administration to an aqueous environment in vivo, the Mayo Foundation for Medical Education and Research and the Regents of The University of Minnesota created the phosphate prodrug CKLP1 (also known as QLS-101), also reported as the sodium salt.

[0015] [ka]

[0016] CKLP1, combined with in vivo hydrolysis to the parent levcromakalim, provides improved aqueous solubility, facilitating administration. See U.S. Patent No. 5,629,997, filed by the Mayo Foundation for Medical Education and Research and Reagents of The University of Minnesota. See also U.S. Patent No. 5,629,997 and U.S. Patent No. 5,629,997, filed by the Mayo Foundation for Medical Education and Research and Qlaris Bio, Inc. However, as a prodrug, it suffers from many of the typical prodrug problems, such as increased manufacturing cost and complexity, as well as metabolic complications.

[0017] (2003) reported that a phosphate prodrug is more water-soluble than cromakalim and reduces intraocular pressure (IOP) in a normotensive (i.e., normal IOP) mouse model, but the drug was only administered for 7 days. This paper also reported the efficacy of increasing doses of certain cromakalim derivatives over 8 days in rabbit eyes.

[0018] The effects of CKLP1 on episcleral venous pressure and distal outflow resistance were described in Non-Patent Document 10. Pharmacokinetic parameters in rabbits after topical and intravenous administration were described in Non-Patent Document 11. The synthesis of CKLP1 and the corresponding (3R,4S)-enantiomer was described in Non-Patent Document 9 (Roy Chowdhury et al. (J. Med. Chem. 2016, 59, 6221)).

[0019] Qlaris Bio, Inc. reported results from a Phase 2 clinical trial of QLS-101, demonstrating a promising safety and tolerability profile and positive efficacy signals for the cromakalim prodrug. The clinical trial was conducted to investigate its ability to reduce intraocular pressure in the treatment of glaucoma. The study showed no evidence of hyperemia (redness of the eye) and demonstrated positive efficacy signals in patients with primary open-angle glaucoma.

[0020] Delivering ocular therapeutic drugs through the ocular mucosal barrier is a particularly challenging task. The mucus barrier layer, among other functions, traps and excludes foreign particles such as allergens, pathogens, and debris, which can also trap drugs. One solution has been to use mucus-penetrating particles, such as nanoparticles, with particle sizes smaller than the mesh size of mucus. See Non-Patent Document 12.

[0021] Qlaris Bio and the Mayo Foundation for Medical Education and Research have filed a patent application describing polymeric controlled-release formulations of levcromakalim for medical applications, including intraocular delivery. See U.S. Patent Application Publication No. 2007 / 0122994.

[0022] In addition to these stated challenges, although solutions and suspensions remain the preferred dosage form for intraocular drug delivery (if the drug is sufficiently soluble), only a small portion of the drug administered as conventional eye drops (sometimes as little as 0.1% to 5% of the dispensed dose) reaches the anterior segment tissues required to treat glaucoma due to poor corneal permeability, reflex blinking, lacrimation, dose extravasation, and nasolacrimal drainage.

[0023] Given the potential but unrealized therapeutic efficacy of cromakalim, it would be beneficial to provide new routes for this drug to lower intraocular pressure and unlock its activity to treat a variety of disorders affected by elevated IOP, or normal intraocular pressure disorders such as normal tension glaucoma, or ocular disorders that respond differently to treatment with cromakalim. [Prior art documents] [Patent documents]

[0024] [Patent Document 1] European Patent No. 0120428 [Patent Document 2] International Publication No. 89 / 10757 [Patent Document 3] International Publication No. 2015 / 117024 [Patent Document 4] International Publication No. 2021 / 1119503 [Patent Document 5] International Publication No. 2021 / 158992 [Patent Document 6] No. PCT / US2022 / 040197 [Non-patent literature]

[0025] [Non-Patent Document 1] Lin et al., "Effects of Cromakalim and Nicorandil on Intraocular Pressure after Topical Administration in Rabbit Eyes" Journal of Ocular Pharmacology and Therapeutics, 1995, 11, 195

Non-licensed Document 2

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

[0026] Surprisingly, it has been discovered that by using a specially discovered combination of excipient components, parent cromakalim drugs, such as levcromakalim (collectively referred to as (lev)cromakalim) or their pharmaceutically acceptable salts, can be efficiently delivered locally in the form of drops in therapeutic amounts effective to treat the anterior segment of the human eye, without the use of covalent prodrug approaches, polymeric delivery systems, or high levels of toxic components, and with a shelf-life stability of at least 4 or 5 months. The topical formulations of the present invention can be used to treat anterior or posterior ocular disorders that respond to cromakalim. For example, the cromakalim formulations described herein can be used to reduce intraocular pressure and generally treat glaucoma, including normal-tension glaucoma, open-angle glaucoma, and angle-closure glaucoma, among other therapeutic uses further described herein.

[0027] Nearly 40 years after the discovery of (rev)cromakalim, the availability of a stable, pharmaceutically acceptable topical formulation of this drug for human use is a major advance.

[0028] The formulations of the present invention containing levcromakalim are well tolerated and can be used as effective topical intraocular pressure (IOP)-lowering agents in generally healthy, normotensive hosts, such as humans. A study in Dutch-belted rabbits (Example 11) demonstrated that the formulations of the present invention exhibit a benign toxicity profile without significant adverse events up to the maximum formulateable concentration (5 mM). No serious ocular or systemic AEs, such as hyperemia (redness of the eye), were observed in this study.

[0029] Human clinical data have established that the levcromakalim formulation of the present invention has a safe and well-tolerated profile for daily ophthalmic administration, and that the formulation readily demonstrates IOP reduction. In certain embodiments, IOP reduction occurs within hours, e.g., within 4 hours, after the initial topical administration of two different concentrations of (lev)cromakalim (0.5 mM and 5 mM). Furthermore, the formulation maintains IOP-lowering efficacy throughout an exemplary, non-limiting treatment period of 14 days (see Examples 8 and 9). The formulation is also effective not only in hypertensive hosts, but also in normotensive hosts.

[0030] The formulations of the present invention containing levcromakalim relax blood vessels in vascular and vascular-like tissue distal to the trabecular meshwork (TM), thereby decreasing distal outflow resistance and lowering episcleral venous pressure (EVP), thereby lowering IOP. Current therapies target only three of the four components of IOP: aqueous humor inflow rate, uveoscleral outflow rate, and conventional outflow capacity. The (lev)cromakalim formulations of the present invention target the fourth component, lowering EVP. Because EVP may be the greatest determinant of overall IOP, this addresses a deficiency in the ability to maximally lower IOP while maintaining the normal vascular integrity of the venous system without causing congestion.

[0031] In certain embodiments, the (rev)cromakalim formulation is a clear aqueous solution, including but not limited to a micellar aqueous solution or a nanomicellar aqueous solution. In another embodiment, the topical formulation is not an emulsion. Emulsions, for example, may be less clear or even milky, which may interfere with vision. In another embodiment, the topical formulation does not include oil, as that term is specifically defined below.

[0032] In certain embodiments, the topical formulations provided herein may represent a paradigm shift in the use of (lev)cromakalim for intraocular delivery to reduce intraocular pressure, thereby reducing the risk of damage to the optic nerve and therefore can be used to treat glaucoma and diseases caused by or exacerbated by elevated intraocular pressure, as well as diseases in which further reduction of intraocular pressure is beneficial to slow disease progression, such as normal-tension glaucoma (NTG). The present invention does not require the production of microparticle or nanoparticle formulations or covalently linked prodrugs. The (lev)cromakalim formulations may advantageously be used as topical drops or via other routes for intraocular delivery.

[0033] In certain aspects of the embodiment, the topical ophthalmic pharmaceutical formulation, which can be used as an eye drop, comprises at least 0.01 mM or at least 0.05 mM, and typically not more than about 5 mM, of cromakalim, typically levcromakalim, or a pharmaceutically acceptable salt thereof, in an aqueous liquid, which is stable for at least 5 months under ambient conditions without significant crystallization or excessive separation of the cromakalim from the liquid, as described in more detail herein, and which does not contain DMSO, DMF, or NMP, or other topical carriers that are not acceptable for human use, and which contains an optional buffer in addition to at least two excipients.

[0034] In certain aspects of this embodiment, the topical ophthalmic pharmaceutical formulation, which can be used as an eye drop, comprises at least 0.1 mM, and typically no more than about 5 mM, cromakalim, typically levcromakalim, or a pharmaceutically acceptable salt thereof, in an aqueous liquid, which is stable for at least 5 months under ambient conditions without significant crystallization or excessive separation of the cromakalim from the liquid, as described in more detail herein, and which does not contain DMSO, DMF, or NMP, or other topical carriers that are not acceptable for human use, and which contains an optional buffer in addition to at least two excipients.

[0035] In certain aspects of this embodiment, the topical ophthalmic pharmaceutical formulation, which can be used as an eye drop, comprises at least 0.25 mM or even at least 0.5 mM (which can range to 1 mM, 1.5 mM, 2.0 mM, or 2.5 mM) (lev)cromakalim, typically levcromakalim, and typically no more than about 5 mM (lev)cromakalim, or a pharmaceutically acceptable salt thereof, in an aqueous liquid, which is stable for at least 5 months under ambient conditions without significant crystallization or excessive separation of the cromakalim from the liquid, as described in more detail herein, and which does not contain DMSO, DMF, or NMP, or other topical carriers that are not acceptable for human use, and which contains an optional buffer in addition to at least two excipients.

[0036] In additional aspects of these embodiments, the topical ophthalmic pharmaceutical formulation for humans is stable for at least 6 months, 7 months, 8 months, or 9 months under ambient conditions. The term "significantly" as used in this context means that the cromakalim, or levcromakalim, or pharmaceutically acceptable salt thereof, does not crystallize or separate to such an extent that the concentration drops significantly below ±5% from the initial concentration.

[0037] In certain embodiments, the present invention provides a topical ophthalmic formulation comprising cromakalim, such as levcromakalim (e.g., (lev)cromakalim) (which is taken to mean cromakalim which may be a mixture of enantiomers or which may be primarily or solely in the levo configuration), or a pharmaceutically acceptable salt thereof, and a mixture of selected pharmaceutically acceptable ingredients, which (i) provides at least about 0.05 mM, 0.5 mM, 1.0 mM, 1.5 mM, 2.0 mM, or 2.5 mM, and typically about 5 mM or less, of (lev)cromakalim, or a pharmaceutically acceptable salt thereof, and (ii) provides stability of cromakalim, such as levcromakalim, in the formulation for at least 4 months, typically 5 months, 6 months, or 7 months or more under ambient conditions (i.e., without significant crystallization or separation from the formulation in which the concentration of cromakalim, such as levcromakalim, falls below ±5% or below ±10% of the original amount). The components of the formulation are required to be stable with respect to each other over the same period of time under ambient conditions. In certain advantageous embodiments, topical formulations of the present invention using an effective amount of cromakalim, e.g., levcromakalim, do not cause significant or prolonged hyperemia (which may result in moderate to severe "red eye," vascular congestion, minor bleeding, petechiae, or microhemorrhage) when administered to a host in need thereof. When mM or mg / mL concentrations are used herein, the active compound is measured without regard to any salt form.

[0038] Additionally, the (rev)cromakalim formulations of the present invention optionally exhibit an osmolality of between about 200 Osm / L and 400 Osm / L, more typically between about 250 Osm / L and 350 Osm / L, which is a measure of the total solute concentration within a specific volume of solvent, expressed in osmoles per liter (Osm / L). Hyperosmotic conditions can damage ocular tissues and stimulate epithelial cell death, which can subsequently trigger an inflammatory cascade leading to cell death via apoptosis.

[0039] Thus, in one aspect, the present invention provides a pharmaceutical composition comprising cromakalim (e.g., levcromakalim), which may be a pharmaceutically acceptable salt thereof; Kolliphor™ (which may be, for example, but is not limited to, EL, HS, RH, ELP, etc.); polysorbate (which may be, for example, but is not limited to, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, etc.); PVP (polyvinylpyrrolidone, which may be, for example, but is not limited to, PVP-K30, PVP-K90, etc.); and poloxamer (which may be, for example, but is not limited to, poloxamer). and optionally benzalkonium chloride (BAK) and / or a pH adjusting agent, wherein the aqueous formulation satisfies (i) and (ii) above, i.e., has a concentration of cromakalim of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or at least 2.5 mM, and typically not more than about 5 mM; is stable at ambient conditions for at least 5 months; and has a pH of about 6-8, or more particularly 6.5-7.5.

[0040] In primary embodiments, the aqueous formulation is not an emulsion, and certainly not an emulsion with reduced transparency or even milky whiteness that may interfere with vision. For example, in certain embodiments, the aqueous formulation is clear (e.g., a solution with a transmittance percentage greater than 85%, 90%, or 95%). In additional aspects, the cromakalim formulation is a clear aqueous solution, including but not limited to, a micellar or nanomicellar aqueous solution. In certain embodiments, the topical formulation does not include an oil, as specifically defined below.

[0041] In another aspect, the present invention provides a pharmaceutical composition comprising cromakalim (e.g., levcromakalim) or a pharmaceutically acceptable salt thereof, glycerin, Kolliphor™ (which may be, for example, but is not limited to, EL, HS, RH, or ELP), a polysorbate (which may be, for example, but is not limited to, polysorbate 20, polysorbate 40, polysorbate 60, or polysorbate 80), a poloxamer (which may be, for example, but is not limited to, poloxamer 407), hypromellose, and mannitol. The present invention provides an aqueous formulation comprising, comprising, consisting essentially of, or consisting of ethanol, water, phosphate buffer, and optionally benzalkonium chloride and / or a pH adjuster, which satisfies (i) and (ii) above, i.e., has a concentration of cromakalim of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically not more than about 5 mM, is stable at ambient conditions for at least 5 months, and has a pH of about 6 to 8, or more particularly 6.5 to 7.5.

[0042] In yet another aspect, the present invention provides a pharmaceutical composition comprising (lev)cromakalim (e.g., levcromakalim) or a pharmaceutically acceptable salt thereof, glycerin, Kolliphor™ (which may be, for example, EL, HS, RH, or ELP), a polysorbate (which may be, for example, but is not limited to, polysorbate 20, polysorbate 40, polysorbate 60, or polysorbate 80), a poloxamer (which may be, but is not limited to, poloxamer 407), PVP, and mannitol. and water, a phosphate buffer, and optionally benzalkonium chloride and / or a pH adjuster, wherein the aqueous formulation satisfies (i) and (ii) above, i.e., has a cromakalim concentration of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically not more than about 5 mM; is stable at ambient conditions for at least 5 months; and has a pH of about 6 to 8, or 6.5 to 7.5.

[0043] In some embodiments, the ophthalmic formulations contain glycerin, polysorbate 80, poloxamer 407, Pluronic™ F127, tyloxapol, Kolliphor™ ELP, Kolliphor™ RH 40, Kolliphor™ HS. 15, polyoxyl-25 castor oil, carboxymethylcellulose (CMC), hypromellose, CMC sodium, PVP, Premulen™ (which may be, for example, TR-1 or TR-2), and octoxynol-40, which satisfies (i) and (ii) above, has a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically not more than about 5 mM, is stable at ambient conditions for at least 5 months, and has a pH of about 6 to 8, or 6.5 to 7.5.

[0044] In another aspect, the present invention provides an aqueous formulation of cromakalim or a pharmaceutically acceptable salt thereof, such as levcromakalim, at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, which formulation is stable under ambient conditions for at least 5 months, has a pH of about 6-8, or 6.5-7.5, and comprises, comprises, consists essentially of, or consists of cromakalim (e.g., levcromakalim) and a polyethoxylated polyhydroxy (e.g., glycerol) ester. Non-limiting examples of polyethoxylated polyhydroxy (e.g., glycerol) esters include Kolliphor™ EL, Kolliphor™ ELP, Kolliphor™ HS 15, Kolliphor™ RH 40, Kolliphor™ RH 60, Eumulgin™ B25, Polyoxyl 100 stearate, Polyoxyl 40 stearate, Polyoxyl 75 stearate, Polyoxyl 6 stearate, and Polyoxyl 32 stearate.

[0045] In yet another aspect, the present invention provides an aqueous formulation of cromakalim or a pharmaceutically acceptable salt thereof, such as levcromakalim, at a concentration of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically no more than about 5 mM, having a stability of at least 5 months under ambient conditions, having a pH of about 6-8, or 6.5-7.5, and comprising, comprising, consisting essentially of, or consisting of cromakalim (e.g., levcromakalim) and a polyethoxylated furanose fatty acid ester. Non-limiting examples of polyethoxylated furanose fatty acid esters include polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, and polysorbate 80.

[0046] In yet another aspect, the present invention provides an aqueous formulation of cromakalim or a pharmaceutically acceptable salt thereof, such as levcromakalim, having a concentration of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, which has a stability of at least 5 months under ambient conditions, has a pH of about 6-8, or 6.5-7.5, and comprises, comprises, or consists essentially of cromakalim (e.g., levcromakalim) and a polymeric lactam. Non-limiting examples of polymeric lactams include polyvinylpyrrolidone (PVP), such as Kollidon™ 12 PF, Kollidon™ 17 PF, Kollidon™ 25, and Kollidon™ 30, and polyvinylpyrrolidone (PVPP), such as Kollidon™ VA 64, Kollidon™ VA-fine, Kollidon™ CL, Kollidon™ CL-F, Kollidon™ CL-SF, and Kollidon™ CL-M.

[0047] In yet another aspect, the present invention provides an aqueous formulation of cromakalim, such as levcromakalim, or a pharmaceutically acceptable salt thereof, having a cromakalim, such as levcromakalim, concentration of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically no more than about 5 mM, and having a stability of cromakalim, such as levcromakalim, in the formulation under ambient conditions for at least 5 months, having a pH of about 6-8, or 6.5-7.5, and comprising, including, consisting essentially of, or consisting of cromakalim (e.g., levcromakalim) and a triblock polyalkylene glycol or poloxamer. Non-limiting examples of triblock polyalkylene glycols include poloxamer 407, poloxamer 188, poloxamer 237, poloxamer 338, Pluronic™ 127, and Synperonic 108.

[0048] In yet another aspect, the present invention provides an aqueous formulation of cromakalim or a pharmaceutically acceptable salt thereof, such as levcromakalim, having a concentration of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, which has a stability of at least 5 months under ambient conditions, has a pH of about 6-8, or 6.5-7.5, and comprises, encompasses, consists essentially of, or consists of cromakalim (e.g., levcromakalim) and a hydroxyalkyl cellulose. Non-limiting examples of hydroxyalkyl cellulose include hypromellose or hydroxypropyl methylcellulose, hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, (2-hydroxypropyl)-γ-cyclodextrin, hydroxypropyl guar gum, dextran, xanthan gum, and guar gum.

[0049] In yet another aspect, the present invention provides an aqueous formulation of cromakalim, such as levcromakalim, or a pharmaceutically acceptable salt thereof, having a cromakalim, such as levcromakalim, concentration of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically no more than about 5 mM, and having a stability of (lev)cromakalim in the formulation under ambient conditions for at least 5 months, having a pH of about 6-8, or 6.5-7.5, and comprising, encompassing, or consisting essentially of cromakalim (e.g., levcromakalim) and a polyol, such as a triol. Non-limiting examples of triols include glycerin, polyglycerol, and polyglycerol polyricinoleate.

[0050] In yet another aspect, the present invention provides an aqueous formulation of cromakalim or a pharmaceutically acceptable salt thereof, such as levcromakalim, at a concentration of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically not more than about 5 mM, wherein the stability of (lev)cromakalim in the formulation is at least 5 months under ambient conditions, the formulation has a pH of about 6 to 8, or 6.5 to 7.5, and the formulation comprises, comprises, or consists essentially of cromakalim (e.g., levcromakalim) and a polyol of a sugar alcohol subtype. Non-limiting examples of sugar alcohols include mannitol, ethylene glycol, glycerol, mannitol, sorbitol, erythritol, threitol, arabitol, xylitol, ribitol, galactitol, fucitol, iditol, inositol, volemitol, isomalt, maltitol, lactitol, maltotriitol, maltotetriitol, and polyglycitol.

[0051] In yet another aspect, the present invention provides an aqueous formulation of cromakalim, such as levcromakalim, or a pharmaceutically acceptable salt thereof, having a cromakalim, such as levcromakalim, concentration of at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, wherein the formulation has a stability under ambient conditions of at least 5 months, has a pH of about 6-8, or 6.5-7.5, and comprises, comprises, or consists essentially of cromakalim (e.g., levcromakalim) and a polyol of the subtype of polymeric alkyl or aryl polyol. Non-limiting examples of polymeric polyols include polyethylene glycol, polypropylene glycol, poly(tetramethylene ether) glycol, sorbitol polyether polyol, sucrose polyether polyol, tyloxapol, and polyvinyl alcohol.

[0052] In some aspects, the present invention provides an aqueous formulation of cromakalim, such as levcromakalim, having at least 0.01 mM, 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically no more than about 5 mM, of cromakalim, such as levcromakalim, or a pharmaceutically acceptable salt thereof, wherein the stability of (lev)cromakalim in the formulation is at least 5 months under ambient conditions, and the formulation has a pH of about 6-8, or 6.5-7.5, and optionally includes or comprises BAK (benzalkonium chloride).

[0053] An effective amount of a cromakalim formulation of the present invention can be delivered locally to treat glaucoma associated with elevated intraocular pressure, including, for example, but not limited to, primary open-angle glaucoma (POAG) (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), primary angle-closure glaucoma, pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, steroid-induced glaucoma, progressing cataract, and / or acute glaucoma resulting from intravitreal injection.

[0054] In certain embodiments, the levcromakalim formulations of the present invention can be used in effective amounts to treat elevated intraocular pressure associated with diabetic retinopathy or diabetic retinopathy-induced glaucoma.

[0055] In another embodiment, the formulations of the present invention are used to treat glaucoma without elevated intraocular pressure, including but not limited to normal tension glaucoma (NTG) (also known as low tension or normal tension glaucoma).

[0056] An effective amount of a topical formulation of cromakalim, e.g., levcromakalim, or a pharmaceutically acceptable salt thereof, as described herein, can also be used to treat a host in need thereof, either as a primary, secondary, or adjunctive treatment as part of a protocol for MIGS (Minimally Invasive Glaucoma Surgery), including, but not limited to, miniaturized forms of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, total intraocular or suprachoroidal shunts, gentler, more gentle forms of laser cyclophotocoagulation, and in alternative embodiments, Schlemm's canal stents to dilate Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty.

[0057] In another embodiment, the ophthalmic formulations of the present invention may be used in an effective amount to treat Sturge-Weber syndrome, including, but not limited to, glaucoma associated with elevated episcleral venous pressure (EVP). Sturge-Weber syndrome is a congenital disorder that affects the skin, nervous system, and sometimes the eyes. Sturge-Weber syndrome is also sometimes referred to as a neurocutaneous disorder. Sturge-Weber syndrome can lead to Sturge-Weber syndrome-induced glaucoma, which affects 30% to 70% of patients with this disorder.

[0058] In certain embodiments, the ophthalmic formulations of the present invention contain (rev)cromakalim at a concentration of at least 0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.15 mg / mL, 0.5 mg / mL, 1 mg / mL, or 1.4 mg / mL, and typically no more than 1.5 mg / mL.

[0059] Specific, non-limiting examples of the present invention are described herein. In certain embodiments, an ophthalmic formulation comprises at least (lev)cromakalim or a pharmaceutically acceptable salt thereof, polysorbate, Kolliphor, and PVP, and has a concentration of (lev)cromakalim of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and is stable at ambient conditions for at least 5 months.

[0060] In other aspects, the ophthalmic formulation comprises at least (lev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor, polysorbate, and poloxamer, and has a concentration of (lev)cromakalim of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically not more than about 5 mM, and is stable at ambient conditions for at least 5 months.

[0061] In additional embodiments, the ophthalmic formulation comprises at least Kolliphor, polysorbate, PVP, and poloxamer and has a concentration of cromakalim or a pharmaceutically acceptable salt thereof of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less (rev) and is stable at ambient conditions for at least 5 months.

[0062] In other embodiments, the ophthalmic formulation comprises at least Kolliphor™ EL or Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol, and has a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and is stable at ambient conditions for at least 5 months.

[0063] In a further embodiment, the ophthalmic formulation comprises at least glycerin, Kolliphor™ EL or Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, and PBS, and has a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and is stable at ambient conditions for at least 5 months.

[0064] In certain other embodiments, the ophthalmic formulation comprises at least Kolliphor™ EL or Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, mannitol, and benzalkonium chloride, and has a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and is stable at ambient conditions for at least 5 months.

[0065] In other embodiments, the ophthalmic formulation comprises at least Kolliphor™ EL or Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, mannitol, and benzalkonium chloride, and has a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and is stable at ambient conditions for at least 5 months.

[0066] Another embodiment is an ophthalmic formulation comprising at least glycerin, Kolliphor™ EL or Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, and mannitol, and having a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically no more than about 5 mM, and which is stable at ambient conditions for at least 5 months.

[0067] In certain non-limiting embodiments, the topical dosage can be administered once daily in the morning (QAM), once daily in the evening (QPM), every morning and at bedtime (QAM HS), every morning and evening (QAM PM), or once daily at bedtime (QHS).

[0068] In certain embodiments, the ophthalmic formulations may be prepared as aqueous solutions or clear emulsions through heating, cooling, incubation, centrifugation, vortexing, standing, stirring, shaking, sonication, or any combination thereof, including those described herein.

[0069] For example, in certain embodiments, the ophthalmic formulation is prepared using a method in which (lev)cromakalim or a salt thereof and other ingredients are first dissolved, individually or collectively, in ethanol or other low-volatility organic solvent(s), mixed, and then evaporated under vacuum to produce a film which is then resuspended in a buffer, typically a phosphate buffer and deionized water, and optionally autoclaved and filtered to obtain the formulation.

[0070] In certain embodiments, the ophthalmic formulation of (lev)cromakalim is delivered for retention and subsequent release within the ocular tissues, and release from the ocular tissues following administration of the immediate release formulation results in efficient absorption and pharmacological effects of levcromakalim.

[0071] Thus, in certain aspects, the present invention provides for the treatment of any of the disorders described herein, including, but not limited to, the reduction of intraocular pressure, or the treatment of glaucoma (such as normal-tension glaucoma), whether or not the condition involves elevated intraocular pressure, by effectively delivering (rev)cromakalim or a pharmaceutically acceptable salt thereof to a host, including a human, via topical administration of an effective amount of an ophthalmic formulation. In certain embodiments, efficient delivery of (rev)cromakalim to the eye is achieved by topical administration of one or more drops or one or more applications of the ophthalmic formulations described herein once daily, twice daily, or multiple times daily (or alternatively, every other day). As non-limiting exemplary embodiments, the ophthalmic formulations in Examples 5 and 6 can significantly reduce intraocular pressure during in vivo efficacy studies conducted in mammals (mice), and the ophthalmic formulations in Examples 8 and 9 are safely administered and well tolerated by human subjects, with reductions in intraocular pressure observed.

[0072] In certain embodiments, topical drops are administered via disposable (unit-dose) pharmaceutical packages. In certain embodiments, the unit-dose packages contain at least about 100 μL to about 500 μL, more typically about 300 μL, of the ophthalmic formulation. In certain embodiments, the delivery volume per drop of formulation delivered via the unit-dose packages is at least about 10 μL to about 50 μL, 20 μL to about 40 μL, more typically about 30 μL per eye. In certain specific embodiments, the concentration of (rev)cromakalim in the unit-dose packages is at least about 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, or 2.5 mM, and typically 5 mM or less. In certain specific embodiments, five unit-dose dropper bottles are dispensed within a single foil package. In certain embodiments, the concentration of (lev)cromakalim is between 0.5 mM and 2.5 mM, hi certain embodiments, the concentration of (lev)cromakalim is between 0.05 mM and 0.5 mM, between 0.5 mM and 1.5 mM, or between 1 mM and 2.5 mM.

[0073] In certain embodiments, the ophthalmic formulations of the present invention reduce intraocular pressure by reducing episcleral venous pressure (EVP). For example, as described in Example 7, the reduction in IOP in C57BL / 6J mice is the result of a reduction in EVP, with significant changes observed in IOP but no significant changes in outflow capacity, uveoscleral outflow, or aqueous humor flow.

[0074] In certain embodiments, the ophthalmic formulation of (lev)cromakalim or a pharmaceutically acceptable salt thereof is administered to the eye, e.g., as topical drops, to deliver (lev)cromakalim into the eye, e.g., to the sclera, conjunctiva, optic nerve, cornea, iris, ciliary body, trabecular meshwork, and / or retina.

[0075] In certain embodiments, the pH of the (rev)cromakalim ophthalmic formulation is adjusted using a pharmaceutically acceptable base to a pH level desirable for pharmaceutical administration, often between about 6 and 8, more typically between 6.5 and 7.5.

[0076] In certain embodiments, the ophthalmic formulation of (rev)cromakalim is not an emulsion, suspension, or gel. In certain embodiments, the ophthalmic formulation of (rev)cromakalim is a clear aqueous solution that is not an emulsion. For example, in certain embodiments, the ophthalmic formulation of (rev)cromakalim is a clear solution, a clear micellar solution, or a clear nanomicellar solution. In certain embodiments, the formulation does not include oil, as defined below.

[0077] Thus, the present invention encompasses at least the following exemplary embodiments: (i) have a concentration of (lev)cromakalim or a pharmaceutically acceptable salt of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and are stable at ambient conditions for at least 5 months, and are not particularly limited to triols or polyols (typically aliphatic, more typically alkyl) (e.g., glycerin), polyethoxylated furanose fatty acid esters (e.g., polysorbates), polyoxyethylene terephthalates (polyoxyethylene terephthalates), polyisobutylene terephthalates (polyisobutylene ... Nonionic triblock copolymers with a central hydrophobic chain of dipropylene (poly(propylene oxide)) flanked by two hydrophilic chains of polyoxyethylene (poly(ethylene oxide)) (e.g., poloxamer or Pluronic), alkylaryl polyols (e.g., tyloxapol), ethoxylated glycerol esters (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor™, Kolliphor™ RH 40, or Kolliphor™ HS 15), polyoxyalkylene castor oil, carboxymethylcellulose / CMC, hypromellose, polymeric lactams (e.g., PVP), polyacrylic acid and hydrophobic C 10 ~C 30A topical ophthalmic formulation comprising three or more ingredients selected from the group consisting of an oil-in-water polymeric emulsifier which is a block copolymer with an alkyl acrylate (e.g., Premulen® TR-1 or Premulen® TR-2), and an ethoxylated alkylphenol (e.g., Octoxynol-40) in an aqueous formulation having a pH of about 6 to 8, or 6.5 to 7.5; (ii) A topical ophthalmic formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof having a concentration of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, which is stable at ambient conditions for at least 5 months and which does not contain any of glycerin, polysorbate 80, poloxamer 407, Pluronic™ F127, tyloxapol, Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor™, Kolliphor™ RH 40, or Kolliphor™ HS). 15), a topical ophthalmic formulation comprising three or more ingredients selected from the group consisting of polyoxyl-25 castor oil, carboxymethylcellulose / CMC, hypromellose, PVP, Premulen™ (e.g., TR-1 or TR-2), and octoxynol-40 in an aqueous solution having a pH of about 6 to 8, or 6.5 to 7.5, (iii) an aqueous ophthalmic formulation comprising, consisting essentially of, or consisting of (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor™, Kolliphor™ RH 40, or Kolliphor™ HS 15), polysorbate (which may be, for example, 20, 40, 60, or 80), polyvinylpyrrolidone (PVP), poloxamer (which may be, for example, 407), mannitol, water or phosphate buffer, optionally benzalkonium chloride, and optionally a pH adjuster, wherein the aqueous ophthalmic formulation has a concentration of (rev)cromakalim of at least 3.5 mM, and typically not more than about 5 mM, and is stable for at least 5 months at ambient conditions, and has a pH of 6 to 8 or 6.5 to 7.5; (iv) (rev) Cromakalim or a pharmaceutically acceptable salt thereof, glycerin, and Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor™, Kolliphor™ RH 40, or Kolliphor™ HS). 15), a polysorbate (which may be, for example, 20, 40, 60, or 80), a poloxamer (which may be, for example, 407), hypromellose, mannitol, and optionally benzalkonium chloride and optionally a pH adjusting agent, wherein the aqueous ophthalmic formulation has a concentration of (lev)cromakalim of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and wherein the aqueous ophthalmic formulation is stable for at least 5 months at ambient conditions and has a pH of 6 to 8 or 6.5 to 7.5. (v) (rev) Cromakalim or a pharmaceutically acceptable salt thereof, glycerin, and Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor™, Kolliphor™ RH 40, or Kolliphor™ HS). 15), a polysorbate (which may be, for example, 20, 40, 60, or 80), a poloxamer (which may be, for example, 407), PVP, mannitol, and optionally benzalkonium chloride and optionally a pH adjuster, i.e., an aqueous ophthalmic formulation comprising, consisting essentially of, or consisting of (rev)cromakalim at a concentration of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically not more than about 5 mM, which aqueous ophthalmic formulation is stable for at least 5 months at ambient conditions and has a pH of 6 to 8 or 6.5 to 7.5; (vi) Treatment of a host in need of treatment for any of the disorders described herein, including but not limited to the general treatment of glaucoma and / or the reduction of intraocular pressure, comprising administering an effective amount of an aqueous topical formulation of (i)-(v) to treat an ocular disorder or any other disorder described herein in a host in need of treatment; (vii) an aqueous ophthalmic formulation of (i) to (v) for use in treating a host in need of treatment for any of the disorders described herein, including, but not limited to, the general treatment of glaucoma and / or the reduction of intraocular pressure; (viii) Use of the aqueous ophthalmic formulations of (i) to (v) to treat a host in need of treatment for any of the disorders described herein, including, but not limited to, the general treatment of glaucoma and / or the reduction of intraocular pressure; (ix) a method for producing a medicament comprising the aqueous formulation of (i) to (v) for treating a host for any of the disorders described herein; (x) a formulation as described herein for use in the manufacture of a medicament in the treatment of a disorder as described herein, including but not limited to the general treatment of glaucoma and / or the reduction of intraocular pressure; (xi) embodiments of (vi) to (x) for treating glaucoma with elevated intraocular pressure, including but not limited to primary open-angle glaucoma (POAG) (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), primary angle-closure glaucoma, pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, steroid-induced glaucoma, glaucoma associated with diabetic retinopathy, advanced cataract and / or acute glaucoma due to intravitreal injection, and glaucoma without elevated intraocular pressure, including but not limited to normal tension glaucoma (NTG) (also known as low tension glaucoma or normal tension glaucoma); (xii) treating Sturge-Weber syndrome, including but not limited to Sturge-Weber syndrome-induced glaucoma, in a host in need thereof; (xiii) embodiments (vi) to (x) that do not cause significant hyperemia (which may result in "red eye," vascular congestion, hemorrhage, petechiae, or microhemorrhage) in a host in need of treatment; (xiv) aspects (vi)-(xiii) as primary or secondary or adjunctive treatment as part of a protocol for MIGS (Minimally Invasive Glaucoma Surgery), including but not limited to use in connection with miniaturized forms of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, total intraocular or suprachoroidal shunts, gentler, more gentle forms of laser cyclophotocoagulation, and in alternative embodiments, Schlemm's canal stents to dilate Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty; (xv) embodiments (vi) to (xiii) for treating ocular disorders in a host, such as Graves' ophthalmopathy, thyroid-associated orbitopathy (TAO), Graves' orbitopathy (GO), retrobulbar tumors, cavernous sinus thrombosis, orbital vein thrombosis, episcleral / orbital vein vasculitis, superior vena cava occlusion, superior vena cava thrombosis, carotid-cavernous sinus fistula, dural-cavernous sinus shunt, orbital varicose veins, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), arterial occlusive / embolic disease and / or hypoperfusion disease, optic nerve damage due to ischemia (posterior and anterior ischemic optic neuropathy (NAION)), and (xvi) Embodiments (vi) to (xiii) that provide cytoprotection and / or neuroprotection to a host in need thereof. [Brief explanation of the drawings]

[0078] [Figure 1]

[0023] Figure 1 shows the reduction in intraocular pressure (IOP) in normotensive mice by Formulation 3f and Formulation 3d, using vehicle as a negative control and levcromakalim in DMSO / Cremophor EL as a positive control, as detailed in Example 5. Baseline indicates pre-dose IOP levels. The Y-axis indicates intraocular pressure in mmHg. IOP measurements were performed in triplicate at two daily time points (1 hour and 23 hours) and averaged to calculate daily IOP. The graph shows the mean and standard deviation of daily IOP on days 3 and 4. [Figure 2]

[0023] Figure 1 shows the reduction in intraocular pressure (IOP) in normotensive mice by specific formulations 2j and 3g, using vehicle as a negative control and levcromakalim in DMSO / Cremophor EL as a positive control, as detailed in Example 6. Baseline indicates pre-dose IOP levels. The Y-axis indicates intraocular pressure in mmHg. IOP measurements were performed in triplicate at two daily time points (1 hour and 23 hours) and averaged to calculate daily IOP. The graph shows the mean and standard deviation of daily IOP on days 3 and 4. [Figure 3]

[0023] Figure 1 shows the reduction in intraocular pressure (IOP) in normotensive mice with various concentrations of levcromakalim formulated in specific formulations 3f and 3d, and DMSO-Cremophor EL, as detailed in Example 5. Baseline indicates pre-dose IOP levels. The X-axis indicates levcromakalim concentration, and the Y-axis indicates intraocular pressure in mmHg. IOP measurements were performed in triplicate at two time points (1 hour and 23 hours) each day, and averaged to calculate daily IOP. The graph shows the mean and standard deviation of daily IOP on days 3 and 4. [Figure 4]

[0023] Figure 1 shows a high-performance liquid chromatography (HPLC) calibration curve for the levcromakalim standard listed in Table 1 in Section VIII. The graph shows the mean and standard deviation of triplicate measurements. The X-axis shows the concentration of the levcromakalim standard, and the Y-axis shows the area of ​​the peak showing the elution of levcromakalim. This calibration curve is used in Examples 1 to 4. [Figure 5]

[0023] Figure 1 shows the mean diurnal IOP (GAT) in 21 patients treated with Formulation 3d (2.5 mM and 0.5 mM) and vehicle control at baseline, day 7, and day 14. Baseline indicates the IOP level before treatment. The Y-axis indicates intraocular pressure in mmHg. Details are provided in Example 9. [Figure 6] (a) Baseline IOP (GAT) in OU before dosing with Formulation 3d or vehicle (n=7 [14 eyes] per treatment group), (b) IOP measured on day 7 after 7 days of QAM dosing with Formulation 3d or vehicle (n=7 [14 eyes] per treatment group), (c) IOP measurements on day 14 after 7 days of BID dosing with Formulation 3d or vehicle (n=7 [14 eyes] per treatment group). Details are provided in Example 9. [Figure 7] Figure 1 shows the reduction in mean diurnal IOP (GAT) compared to baseline in 21 patients treated with Formulation 3d (2.5 mM and 0.5 mM) and vehicle control on days 7 and 14. The Y-axis shows the reduction in intraocular pressure in mmHg. Details are provided in Example 9. [Figure 8] (a) Reduction in IOP measured on day 7 compared to baseline after 7 days of QAM dosing with Formulation 3d or vehicle (n=7 [14 eyes] per treatment group), (b) Reduction in IOP measurements on day 14 compared to baseline after 7 days of BID dosing with Formulation 3d or vehicle (n=7 [14 eyes] per treatment group). Details are provided in Example 9. DETAILED DESCRIPTION OF THE INVENTION

[0079] Surprisingly, it has been found that a parent cromakalim drug, e.g., levcromakalim (a compound without regard to stereochemistry or of either configuration (e.g., (-) or without regard to stereochemistry) is referred to as (rev)cromakalim) or a pharmaceutically acceptable salt thereof, can be efficiently delivered locally in the form of drops in an effective therapeutic amount to treat any of the anterior and posterior segment indications described herein that respond to (rev)cromakalim, including, but not limited to, ocular disorders associated with elevated intraocular pressure, ocular disorders not associated with elevated intraocular pressure such as normal-tension glaucoma, or any form of glaucoma in general. The (rev)cromakalim formulations of the present invention, using the combinations of excipients described herein, can be used to treat the anterior segment of the human eye without the use of covalent prodrug approaches, polymeric delivery systems, or high levels of toxic components. Nearly 40 years after the discovery of (rev)cromakalim, it is a major advancement that we finally have a stable topical pharmaceutical formulation that provides humans with an effective amount of this drug.

[0080] The ocular structure can be divided into two parts: the anterior and posterior. The anterior segment comprises the front one-third of the eye and includes structures such as the cornea, iris, ciliary body, and lens, which are anterior to the vitreous. The posterior segment comprises the back two-thirds of the eye and includes the sclera, choroid, retinal pigment epithelium, neural retina, optic nerve, and vitreous. The formulations of the present invention can be used to treat anterior or posterior ocular disorders that respond to (lev)cromakalim.

[0081] The topical formulations provided herein may mark a paradigm shift in the use of (rev)cromakalim for intraocular delivery to treat eye disorders that respond to (rev)cromakalim, including, but not limited to, lowering intraocular pressure that can damage the optic nerve, and may therefore be used to treat glaucoma and other diseases caused by or exacerbated by elevated intraocular pressure, as well as normal-tension glaucoma. The present invention does not require the production of microparticle or nanoparticle formulations or covalently linked prodrugs. Cromakalim formulations may advantageously be used as topical drops for ocular mucosal delivery.

[0082] The (rev)cromakalim formulations of the present invention are not subject to endogenous conversion by the phosphate ester (compared to the phosphate ester prodrug QLS-101).The (rev)cromakalim formulations of the present invention are well suited for topical ophthalmic administration due to their water-soluble nature.

[0083] The aqueous ophthalmic formulations of the present invention can deliver (lev)cromakalim at a concentration of at least about 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, to the intraocular chamber, where it can cross the mucosal barrier and reach the anterior segment. The aqueous formulations provided herein have a stability of (lev)cromakalim for at least 5 months, for example, 5, 6, or 7 months or more, under ambient conditions (i.e., without significant crystallization or separation from the formulation, where the concentration of (lev)cromakalim decreases by less than about ±5% or less than about ±10% from the initial concentration). The components of the formulation are required to be stable with each other under ambient conditions for the same period. Aqueous ophthalmic formulations have a pH in the range of about 6-8, more typically 6.5-7.5, and an osmolality of between about 200-400 or between 250-350.

[0084] In certain embodiments, the ophthalmic formulation of (rev)cromakalim is not an emulsion, suspension, or gel. In certain embodiments, the ophthalmic formulation of (rev)cromakalim is a clear aqueous solution that is not an emulsion. For example, in certain embodiments, the ophthalmic formulation of (rev)cromakalim is a clear solution, a clear micellar solution, or a clear nanomicelle solution. In certain embodiments, the formulation does not contain oil as defined below.

[0085] When reference is made herein to the following concentrations, such as 0.05 mM, 0.1 mM, 0.3 mM, 0.5 mM, 1 mM, 2 mM, 2.5 mM, 3 mM, or 3.5 mM (e.g., about 1 mg / mL), or even 4 mM (e.g., about 1.1 mg / mL), 4.5 mM, or 5 mM (e.g., about 1.4 mg / mL), it should be understood that each concentration is specifically and uniquely referred to as disclosing a species, and that the list of concentrations is not intended by the inventors and should not be considered as disclosing a genus that may or may not hold together. This list of concentrations includes, but is not limited to:

[0086] In certain embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 0.05 mM, or at least 0.05 mM and no more than 5 mM.

[0087] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 0.1 mM, or at least 0.1 mM and no more than 5 mM.

[0088] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 0.5 mM, or at least 0.5 mM and no more than 5 mM.

[0089] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 1 mM, or at least 1 mM and no more than 5 mM.

[0090] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 1.5 mM, or at least 1.5 mM and no more than 5 mM.

[0091] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 2 mM, or at least 2 mM and no more than 5 mM.

[0092] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 2.5 mM, or at least 2.5 mM and no more than 5 mM.

[0093] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 3 mM, or at least 3 mM and no more than 5 mM.

[0094] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 3.5 mM, or at least 3.5 mM and no more than 5 mM.

[0095] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 4 mM, or at least 4 mM and no more than 5 mM.

[0096] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 4.5 mM, or at least 4.5 mM and no more than 5 mM.

[0097] In certain other embodiments, the ophthalmic formulations described herein have a concentration of (rev)cromakalim or a pharmaceutically acceptable salt of 5 mM, or at least 5 mM and no more than 5 mM.

[0098] In one aspect, the present invention provides a pharmaceutical composition comprising a (lev)cromakalim or pharmaceutically acceptable salt thereof having a concentration of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, which is stable at ambient conditions for at least 5 months, and which is in the presence of a triol or polyol (typically aliphatic, more typically alkyl) (e.g., glycerin), a polyethoxylated furanose fatty acid ester (e.g., polysorbate 100), a hydroxybenzoate (hydroxybenzoate ... esters), nonionic triblock copolymers with a central hydrophobic chain of polyoxypropylene (poly(propylene oxide)) flanked by two hydrophilic chains of polyoxyethylene (poly(ethylene oxide)) (e.g., poloxamer or Pluronic), alkylaryl polyols (e.g., tyloxapol), ethoxylated glycerol esters (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15), polyoxylalkylene castor oil, carboxymethylcellulose / CMC, hypromellose, polymeric lactams (e.g., PVP), polyacrylic acid and hydrophobic C 10 ~C 30 Provided is a topical ophthalmic formulation of (lev)cromakalim, comprising three or more components selected from the group consisting of an oil-in-water polymeric emulsifier that is a block copolymer with an alkyl acrylate (e.g., Premulen™ (which may be, for example, TR-1 or TR-2)), and an ethoxylated alkylphenol (e.g., Octoxynol-40), in an aqueous formulation having a pH of about 6 to 8, or 6.5 to 7.5.

[0099] In another aspect, the present invention provides a pharmaceutical composition having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, which is stable at ambient conditions for at least 5 months, and which does not contain any of glycerin, polysorbate, poloxamer 407, Pluronic™ F127, tyloxapol, Kol A topical ophthalmic formulation of (rev)cromakalim comprising three or more ingredients selected from the group consisting of liphor™ (which may be, for example, EL, ELP, RH, or HS), polyoxyl-25 castor oil, carboxymethylcellulose / CMC, hypromellose, PVP, Premulen™ (e.g., TR-1 or TR-2), and octoxynol-40 in an aqueous solution having a pH of about 6 to 8, or 6.5 to 7.5.

[0100] In yet another embodiment, the present invention provides a pharmaceutical composition comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ (which may be, for example, EL, ELP, RH, or HS), polysorbate (which may be, for example, 20, 40, 60, or 80), PVP (polyvinylpyrrolidone), poloxamer (which may be, for example, 407 or another variant), mannitol, water or phosphate buffer, and optionally benzalkonium chloride. and optionally a pH adjusting agent, the aqueous ophthalmic formulation having a concentration of (rev)cromakalim of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically not more than about 5 mM, and which is stable at ambient conditions for at least 5 months, and which has a pH of 6-8 or 6.5-7.5.

[0101] Another aspect of the present invention comprises or consists essentially of (rev)cromakalim or a pharmaceutically acceptable salt thereof, glycerin, Kolliphor™ (which may be, for example, EL, ELP, RH, or HS), polysorbate (which may be, for example, 20, 40, 60, or 80), poloxamer (which may be, for example, 407 or another variation), hypromellose, mannitol, and optionally benzalkonium chloride and optionally a pH adjuster. or an aqueous ophthalmic formulation comprising the same, which satisfies exemplary embodiment (i) and embodiment (ii) described herein, i.e., has a (rev)cromakalim concentration of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically not more than about 5 mM, is stable at ambient conditions for at least 5 months, and has a pH of 6 to 8 or 6.5 to 7.5.

[0102] Further, the present invention provides a pharmaceutical composition comprising, consisting essentially of, or consisting of (rev)cromakalim or a pharmaceutically acceptable salt thereof, glycerin, Kolliphor™ (which may be, for example, EL, ELP, RH, or HS), polysorbate (which may be, for example, 20, 40, 60, or 80), poloxamer (which may be, for example, 407), PVP, mannitol, and optionally benzalkonium chloride and optionally a pH adjuster. The present invention provides an aqueous ophthalmic formulation comprising: a (rev)cromakalim solution containing at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and the aqueous ophthalmic formulation is stable for at least 5 months at ambient conditions; and the aqueous ophthalmic formulation has a pH of 6 to 8 or 6.5 to 7.5, and satisfies exemplary embodiment (i) and embodiment (ii) described herein.

[0103] In certain embodiments, the (rev)cromakalim formulation comprises a mixture of polysorbate 80, Kolliphor™ (which may be, for example, EL, ELP, RH, or HS), and PVP, has a (rev)cromakalim concentration of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, is stable at ambient conditions for at least 5 months, and has a pH of 6-8 or 6.5-7.5.

[0104] In certain embodiments, the (rev)cromakalim formulation comprises a mixture of Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15), polysorbate 80, phosphate buffer, and poloxamer 407, has a (rev)cromakalim concentration of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, is stable at ambient conditions for at least 5 months, and has a pH of 6-8 or 6.5-7.5.

[0105] In certain embodiments, the (rev)cromakalim formulation is Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15) (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15), polysorbate 80, PVP, and phosphate buffer, having a concentration of (lev)cromakalim of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and stable for at least 5 months at ambient conditions, and having a pH of 6-8 or 6.5-7.5.

[0106] In certain embodiments, the (rev)cromakalim formulation comprises a component mixture of Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and PBS, has a (rev)cromakalim concentration of at least 3.5 mM, and typically no more than 5 mM, is stable at ambient conditions for at least 5 months, and has a pH of 6-8 or 6.5-7.5.

[0107] In certain embodiments, the (rev)cromakalim formulation comprises a mixture of ingredients including Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15), polysorbate (e.g., 20, 40, 60, or 80), PVP, poloxamer 407, and mannitol, has a concentration of (rev)cromakalim of at least 3.5 mM, and typically no more than 5 mM, is stable at ambient conditions for at least 5 months, and has a pH of 6-8 or 6.5-7.5.

[0108] In certain embodiments, the (rev)cromakalim formulation comprises a component mixture of glycerin, Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15), polysorbate (e.g., 20, 40, 60, or 80), poloxamer (e.g., 407), hypromellose, and PBS, and has a concentration of (rev)cromakalim of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and is stable at ambient conditions for at least 5 months, and has a pH of 6-8 or 6.5-7.5.

[0109] In certain embodiments, the (rev)cromakalim formulation comprises a component mixture of Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15), polysorbate (e.g., 20, 40, 60, or 80), PVP, poloxamer 407, phosphate buffer, mannitol, and BAK, has a (rev)cromakalim concentration of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, is stable at ambient conditions for at least 5 months, and has a pH of 6-8 or 6.5-7.5.

[0110] In certain embodiments, the (rev)cromakalim formulation comprises a mixture of ingredients including Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15), polysorbate (e.g., 20, 40, 60, or 80), PVP, poloxamer (e.g., 407), phosphate buffer, and mannitol, has a concentration of (rev)cromakalim of at least 3.5 mM, and typically no more than 5 mM, is stable at ambient conditions for at least 5 months, and has a pH of 6-8 or 6.5-7.5.

[0111] In certain embodiments, the (rev)cromakalim formulation comprises a mixture of glycerin, Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15), polysorbate (e.g., 20, 40, 60, and 80), poloxamer (e.g., 407), mannitol, hypromellose, and phosphate buffer, has a (rev)cromakalim concentration of at least 3.5 mM, and typically no more than about 5 mM, is stable at ambient conditions for at least 5 months, and has a pH of 6-8 or 6.5-7.5.

[0112] In certain embodiments, the (rev)cromakalim formulation comprises a component mixture of glycerin, Kolliphor™ (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor, Kolliphor™ RH 40, or Kolliphor™ HS 15), polysorbate (which may be 20, 40, 60, or 80), poloxamer 407, mannitol, hypromellose, BAK, and phosphate buffer, and has a concentration of (rev)cromakalim of at least 0.01 mM, 0.05 mM, 0.4 mM, 0.5 mM, 0.8 mM, 0.9 mM, 1 mM, 1.5 mM, 2 mM, or 2.5 mM, and typically about 5 mM or less, and is stable at ambient conditions for at least 5 months, and has a pH of 6-8 or 6.5-7.5.

[0113] In certain embodiments, topical drops are administered via a disposable (unit-dose) pharmaceutical package. In certain embodiments, the unit-dose package contains at least about 100 μL to about 500 μL, more typically about 300 μL, of the ophthalmic formulation. In certain embodiments, the delivery volume per drop of the formulation delivered via the unit-dose package is at least about 10 μL to about 50 μL, more typically about 30 μL, per eye. In certain specific embodiments, the concentration of (lev)cromakalim in the unit-dose package is at least about 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, or 2.5 mM, and typically 5 mM or less. In certain specific embodiments, five unit-dose dropper bottles are dispensed within a single foil package. In certain embodiments, the concentration of (lev)cromakalim is between 0.5 mM and 2.5 mM. In certain embodiments, the concentration of (lev)cromakalim is between 0.05 mM and 0.5 mM, between 0.5 mM and 1.5 mM, or between 1 mM and 2.5 mM.

[0114] In certain non-limiting exemplary embodiments, (rev)cromakalim formulations for topical administration are provided in potencies of 0.015%, 0.030%, and 0.075%, containing 0.15 mg / mL, 0.30 mg / mL, and 0.75 mg / mL (rev)cromakalim, respectively. As described in Example 13, levcromakalim ophthalmic solution is provided in a form-fill-fill (BFS) disposable unit-dose container made of low-density polyethylene. Examples include, but are not limited to: 0.015% (0.15 mg / mL) BFS Unit Dose: A clear, colorless 300 μL unit dose BFS of (rev)cromakalim formulation ophthalmic solution at a concentration of 0.015% (0.15 mg / mL) in isotonic phosphate buffer solution at pH 6.5; 0.030% (0.30 mg / mL) BFS Unit Dose: A clear, colorless 300 μL unit dose BFS of (rev)cromakalim formulation ophthalmic solution at a concentration of 0.030% (0.30 mg / mL) in isotonic phosphate buffer solution at pH 6.5; 0.075% (0.75 mg / mL) BFS Unit Dose: A clear, colorless 300 μL unit dose BFS of (rev)cromakalim formulation ophthalmic solution at a concentration of 0.075% (0.75 mg / mL) in isotonic phosphate buffer solution at pH 6.5.

[0115] In certain embodiments, the (rev)cromakalim formulation is a clear micellar aqueous solution.

[0116] In certain embodiments, the (rev)cromakalim formulation is a clear aqueous solution.

[0117] In certain embodiments, the (rev)cromakalim formulation does not include oil as defined below.

[0118] In certain embodiments, the (rev)cromakalim formulation is not an emulsion.

[0119] In certain embodiments, the (rev)cromakalim formulation is not a gel.

[0120] In certain embodiments, the (rev)cromakalim formulation is not a topical gel.

[0121] In certain embodiments, the (rev)cromakalim formulation is a clear liquid.

[0122] In certain embodiments, the (rev)cromakalim formulation is a micellar solution.

[0123] In certain embodiments, the (rev)cromakalim formulation is a nanomicelle liquid formulation.

[0124] In certain embodiments, the (rev)cromakalim formulation has a percent transmittance of greater than 85% when tested with a UV-Visible spectrophotometer.

[0125] In certain embodiments, the (rev)cromakalim formulation has a percent transmittance of greater than 90% when tested with a UV-Visible spectrophotometer.

[0126] In certain embodiments, the (rev)cromakalim formulation has a percent transmittance of greater than 95% when tested with a UV-Visible spectrophotometer.

[0127] In certain embodiments, the (rev)cromakalim formulation has a percent transmittance of greater than 98% when tested with a UV-Visible spectrophotometer.

[0128] In certain embodiments, the (rev)cromakalim formulation does not contain long-chain triglycerides.

[0129] In certain embodiments, the (rev)cromakalim formulation does not contain medium chain triglycerides.

[0130] In certain embodiments, the (rev)cromakalim formulation does not contain short chain triglycerides.

[0131] In certain embodiments, the (rev)cromakalim formulation does not include oil as specifically defined below.

[0132] I. Definition As used herein, a "patient" or "host" or "subject" is typically a human, and the method is for human therapy. In appropriate circumstances, the scope may alternatively include non-human animals, such as mammals, primates (non-human), cattle, sheep, goats, horses, dogs, cats, etc., in need of treatment or prevention of any of the disorders specifically described herein.

[0133] Oil as an optional excluded ingredient In certain embodiments, the (rev)cromakalim formulation does not contain oil. The term "oil", when used as an excluded ingredient for the topical formulations described herein, refers to liquid organic compounds that are primarily hydrocarbons and can be used to prepare water-in-oil emulsions, such as long-chain triglycerides (including but not limited to castor oil, soybean oil, corn oil, linseed oil, cottonseed oil, coconut oil, canola oil, argan oil, palm oil, peanut oil, and other vegetable oils), and / or medium-chain triglycerides (including but not limited to caprylic / capric triglycerides (e.g., miglyol 812)), and / or mono- and diglycerides (including but not limited to caprylic / capric mono- and diglycerides (e.g., imwitor 742)). The term "oil," when used as an optionally excluded ingredient, does not include any of the optional ingredients listed below (in other words, the formulation ingredients listed below are excluded from the definition of oil herein).

[0134] Polysorbate Polysorbate as used herein is a kind of emulsifier derived from the esterification of ethoxylated sorbitan and fatty acid.Polysorbate as used herein is an oily liquid that is used to solubilize oil in water-based products.Examples of polysorbate include polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80.Common trade names of polysorbate are Tween, Scattics, Alkest and Canarcel.The number after polysorbate refers to the main fatty acid of the molecule, for example, monolaurate is represented by 20, monopalmitate is represented by 40, monostearate is represented by 60, and monooleate is represented by 80. Polysorbate 80 (polyoxyethylene (20) sorbitan monooleate), an example of a polysorbate component described herein, is a viscous, non-ionic hydrophilic surfactant derived from polyethoxylated sorbitan and oleic acid, with an average molecular weight of 1310 Da and an HLB value of 15.

[0135] [ka]

[0136] Polysorbate 80 is an emulsifying agent because it contains a polyoxyethylene hydrophilic component and a polysorbate lipophilic component. Polysorbate 80 is not carcinogenic or genotoxic, as it has not shown adverse effects at doses of 2500 mg / kg per day. In certain non-limiting embodiments, Polysorbate 80 is used in ophthalmic solutions at concentrations up to about 1% (weight / volume).

[0137] Kolliphor(TM) or Cremophor(TM) As used herein, Kolliphor™ or Cremophor™ (BASF) is a polyoxyethylated castor oil (ethoxylated glycerol ester) made from castor oil and ethylene oxide, where the fatty acid ester of glycerol represents the hydrophobic portion and polyethylene glycol represents the hydrophilic portion. Kolliphor™ or Cremophor™ is used as a surfactant, emulsifier, and solubilizer (HLB value = 12-14) in drug formulations because it is completely soluble in aqueous formulations and can emulsify and solubilize oils and water-insoluble active ingredients. One example of an ethoxylated glycerol ester, Kolliphor™ ELP (cremophor™ ELP or polyoxyl-35-castor oil), is a nonionic polyethoxylated detergent made by reacting castor oil with ethylene oxide in a 1:35 molar ratio.

[0138] [ka]

[0139] Kolliphor™ ELP is more viscous than Kolliphor™ EL (Cremophor™ EL). Kolliphor™ ELP forms a clear aqueous solution in water and is soluble in common organic solvents because it contains free polyethylene glycol and ethoxylated glycol as hydrophilic components, and glycerol polyethylene glycol ricinoleate and fatty acid esters of polyethylene glycol as lipophilic components. Kolliphor™ ELP is used as a purified solubilizer for paclitaxel formulations. In certain non-limiting embodiments, Kolliphor™ ELP is used at a concentration of up to about 5% (weight / volume).

[0140] polymeric lactams As used herein, lactams are cyclic amides derived from aminoalkanoic acids. Five common types of lactams are named based on ring size: α-lactam (3-atom ring), β-lactam (4-atom ring), γ-lactam (5-atom ring), δ-lactam (6-atom ring), and ε-lactam (7-atom ring). Polymeric lactams can be synthesized from lactam vinyl monomers with various molecular weights and viscosities. For example, PVP (polyvinylpyrrolidone, povidone, or polyvidone), derived from N-vinylpyrrolidone, is a biodegradable, water-soluble polymer. PVP is one of the most widely used ingredients in pharmaceutical compositions because it exhibits excellent solubility in solvents of various polarities and stabilizes suspensions and emulsions.

[0141] [ka]

[0142] TIFF2025538061000008.tif31170

[0143] PVP contributes to solubility due to its hydrophilic components, which bind to polar molecules such as phenolic compounds through hydrogen bonds. PVP is used as a thickener because it is available in a variety of molecular weights (2.5 x 10), as further explained in the table above. 3 Da ~ 2.5 × 10 6 PVP is used in ophthalmic solutions (eye drops or lens packaging solutions) because its availability in viscosities (Da) allows its viscosity to be adjusted. PVP functions as a lubricant or wetting agent. In certain non-limiting exemplary embodiments, PVP K-30 is used at up to about 2% (wt / vol).

[0144] Poloxamer As used herein, poloxamers are nonionic triblock copolymers composed of a central hydrophobic chain of polyoxypropylene flanked by two hydrophilic chains of polyoxyethylene. Pluronics, as described herein, are poloxamers composed of hydrophilic poly(ethylene oxide) (PEO) and hydrophobic poly(propylene oxide) (PPO) arranged in an ABA triblock structure, resulting in PEO-PPO-PEO. An important property of poloxamer solutions is their temperature-dependent self-assembly and thermogelation behavior. Concentrated aqueous solutions of poloxamers are liquid at low temperatures and form gels at higher temperatures in a reversible process. The transitions that occur in these systems depend on the molecular weight and hydrophilic / hydrophobic molar ratio. They can be used to increase the water solubility of hydrophobic oily substances or to increase the miscibility of two substances with otherwise different hydrophobic properties.

[0145] An example of a poloxamer is Poloxamer 407, a hydrophilic non-ionic surfactant that is a polyoxyethylene polymer. Poloxamer 407 can be used in ophthalmic solutions (drops) at concentrations up to about 0.1% (weight / volume).

[0146] [ka]

[0147] Poloxamer 407 is a triblock copolymer consisting of a central lipophilic block of polypropylene glycol (PPG) with an average repeating unit length of 56, flanked on both sides by hydrophilic polyethylene glycol (PEG) blocks with an average repeating unit length of 101. Poloxamer 407 is listed as an ingredient in the Inactive Ingredients Database (IID) and is approved by the FDA. Poloxamer 407 is used as a hydrophilic nonionic surfactant to solubilize oily substances in aqueous media and has an HLB value greater than 18. Poloxamer 407 is used as an emulsifier and solubilizer in ophthalmic irrigation solutions at concentrations up to 0.2%. Ophthalmic gel-forming poloxamer 407 and hydroxypropyl methylcellulose are used for the intraocular delivery of chloramphenicol.

[0148] Polyol As used herein, a polyol is an aliphatic, typically alkyl, organic compound containing multiple hydroxyl groups (-OH). Polyols containing two, three, and four hydroxyl groups, as well as diols, triols, and tetrols, can be used in the formulations of the present invention.

[0149] The triols described herein are alkyl polyols containing three hydroxyl groups. Glycerol (e.g., glycerin) is a viscous, non-toxic triol solvent containing hydrophilic components. Glycerol is derived from triglycerides (esters of glycerol and long-chain carboxylic acids) extracted from plants and animals by hydrolysis, saponification with sodium hydroxide, or transesterification.

[0150] [ka]

[0151] Glycerol is miscible in water or alcohol-based solvents and can solubilize emulsions and surfactants through polar interactions. Because glycerol has alcohol-like solvent properties but is highly viscous, it is used as a bulking agent and osmotic diuretic ophthalmic agent.

[0152] Sugar alcohols, as described herein, are polyols derived from the hydrogenation of sugars and contain one hydroxyl group (-OH) attached to each carbon atom. Sugar alcohols have the general formula HOCH(CHOH) n Sugar alcohols have a CHOH group and occur in various chain lengths, most commonly five- or six-carbon chains derived from pentoses (five-carbon sugars) and hexoses (six-carbon sugars), respectively. Sugar alcohols can typically be distinguished by the relative orientation (stereochemistry) of their -OH groups. For example, mannitol and sorbitol differ only in the orientation of the hydroxyl group on carbon 2. Mannitol, as described herein, is a type of sugar alcohol derived from the reduction of mannose, which produces sorbitol, the 2'-OH isomer of mannitol, as another product. In certain non-limiting exemplary embodiments, mannitol may be used in ophthalmic solutions at concentrations up to about 4.6% (weight / volume).

[0153] [ka]

[0154] Sugar alcohols are inert and non-hygroscopic. They are used as additives and bulking agents (vehicles) for lyophilized formulations. Other sugar alcohols include, but are not limited to, erythritol, ethylene glycol, glycerol, threitol, arabitol, xylitol, ribitol, sorbitol, galactitol, fucitol, iditol, inositol, volemitol, maltitol, and lactitol.

[0155] High molecular weight alkyl or aryl polyols As used herein, polymeric alkyl or aryl polyols are liquid polymers of alkyl or aryl alcohols prepared by the reaction of alkyl or aryl alcohols with an aldehyde, such as formaldehyde.

[0156] Tyloxapol is a non-ionic liquid polymer of alkylaryl polyether alcohols. It is formed by the reaction of 4-(1,1,3,3-tetramethylbutyl)phenol with formaldehyde, followed by reaction to form an oxirane.

[0157] [ka]

[0158] The hydrophilic component of polyether alcohols consists of hydrogen bond donors and acceptors, and the lipophilic alkyl component can emulsify the hydrophobic component in aqueous formulations. Tyloxapol has an HLB of 13 and is therefore used as a surfactant in ophthalmic formulations to balance the hydrophilic-lipophilic mixture. WO 1998 / 025620 describes the use of tyloxapol in the preparation of ophthalmic suspensions.

[0159] Hydroxyalkyl cellulose The hydroxyalkyl cellulose described herein is a hydroxyalkyl ether of cellulose produced by treating cellulose with sodium hydroxide and reacting with alkylene oxide.Hydroxyalkyl cellulose is used as a water-binding agent and thickener in pharmaceutical compositions to promote hydrophilization.Non-limiting examples of hydroxyalkyl cellulose include hydroxymethyl cellulose (HMC), hydroxyethyl cellulose (HEC), and hydroxypropyl methylcellulose (HPMC or hypromellose).

[0160] Hypromellose (hydroxypropyl methylcellulose) is a semi-synthetic polymer derived from non-ionic, partially O-methylated and O-(2-hydroxypropylated) cellulose ethers containing β-linked D-glucose.

[0161] [ka]

[0162] Hypromellose is produced by reacting alkali cellulose with methyl chloride and propylene oxide. Hypromellose is available in various substitution ratios and molecular weight grades, with HLB values ​​ranging from 10 to 11. Due to its film-forming ability, biocompatibility, and biodegradability, it is used as an emulsifier, hydrophilic thickener, and stabilizer. In ophthalmic solutions, hypromellose also functions as a lubricant.

[0163] Benzalkonium chloride (BAK) As used herein, benzalkonium chloride is a cationic surfactant containing a quaternary ammonium as the cationic head group. Benzalkonium chloride is composed of alkyldimethylbenzylammonium chloride with an 8-18 carbon paraffinic chain as the lipophilic alkyl residue.

[0164] [ka]

[0165] Benzalkonium chloride can be used to dissolve lipophilic ingredients in aqueous formulations at neutral to slightly alkaline pH, as aqueous solutions containing BAK have emulsifying properties due to their low surface tension.

[0166] Carbomer Copolymer A and Carbomer Copolymer B [ka]

[0167] Carbomer copolymer type A, such as Premulen™ TR-2, used herein, is a copolymer of hydrophilic acrylic acid and a hydrophobic alkyl acrylate comonomer. Specifically, Carbomer Copolymer A is a copolymer of acrylic acid crosslinked with pentaerythritol and a hydrophobic alkyl acrylate comonomer. 10 ~C 30 It is a high molecular weight copolymer with alkyl acrylate. It is a non-ethoxylated polymer for mild oil-in-water emulsions. Carbomer Copolymer A (Premulen™ TR-2) can produce low viscosity emulsions and, due to its higher level of hydrophobic groups, can stabilize 60% oil by weight. Carbomer Copolymer A forms polyethylene glycol-free formulations, and the resulting formulations do not require HLB calculations. Carbomer Copolymer A is used in Restasis (see below for information on Restasis).

[0168] Carbomer copolymer B used herein, e.g., Premulen™ TR-1, has a similar structure to carbomer copolymer A, but contains a lower level of hydrophobic groups compared to carbomer copolymer A. Carbomer copolymer B produces high viscosity emulsions and can incorporate up to 20% oil by weight in the pH range of 3-11, and up to 30% oil by weight in the pH range of 4-5.5.

[0169] Solubilizers or solubilizing agents are surfactants that increase the solubility of one agent in another.

[0170] Emulsifiers / emulsifying agents are agents that help prevent other agents from mixing and separating. Water-in-oil (w / o) emulsions keep water droplets enclosed in oil, while oil-in-water (o / w) emulsions keep oil droplets enclosed in water.

[0171] Wetting agents are surface-active molecules that can reduce the surface tension of water.

[0172] Antifoaming agents are molecules that reduce or prevent the formation of foam (formed by trapped gas pockets within a liquid).

[0173] When trade names of formulation ingredients are used, it should be understood that they are for illustrative purposes only and are not intended to limit the scope of the invention. Non-limiting examples of chemical names or chemical classes that can be directly substituted for the commercial product names used are set forth above and include, but are not limited to, triols or polyols (typically aliphatic, more typically alkyl) (e.g., glycerin), polyethoxylated furanose fatty acid esters (e.g., polysorbates), nonionic triblock copolymers of a central hydrophobic chain of polyoxypropylene (poly(propylene oxide)) flanked by two hydrophilic chains of polyoxyethylene (poly(ethylene oxide)) (e.g., poloxamers or Pluronics), alkylaryl polyols (e.g., tyloxapol), ethoxylated glycerol esters (e.g., Kolliphor™ EL or Kolliphor™ ELP, Cremophor™, Kolliphor™ RH 40, or Kolliphor™ HS), and the like. 15), polyoxyalkylene castor oil, carboxymethylcellulose / CMC, hypromellose, polymeric lactams (e.g., PVP), polyacrylic acid and hydrophobic C 10 ~C 30 These include polymeric oil-in-water emulsifiers that are block copolymers with alkyl acrylates (eg, Premulen™ such as TR-1 and TR-2), and ethoxylated alkylphenols (eg, Octoxynol-40).

[0174] In certain embodiments, a "micellar solution" is a solution containing a dispersion of micelles.

[0175] In certain embodiments, a "micelle" is an aggregate of amphiphilic lipid molecules dispersed in a liquid.

[0176] In certain embodiments, a "nanomicellar solution" is a micellar solution comprising micelles less than 100 nm in diameter.

[0177] II. Pharmaceutical Compositions and Dosage Forms for Ocular Delivery An aqueous pharmaceutically acceptable topical ophthalmic formulation comprising an effective amount of (rev)cromakalim or a pharmaceutically acceptable salt thereof can be administered to treat any ocular disorder that can be treated with (rev)cromakalim, including, but not limited to, any disorder described herein, including, but not limited to, reducing intraocular pressure in a host's eye in need of reduction, normal tension glaucoma, or any other indication that can be treated with (rev)cromakalim as described herein.

[0178] The ophthalmic formulations of (rev)cromakalim described herein are stable at ambient conditions for at least 5, 6, or 7 months or longer and can achieve effective concentrations without the use of pharmaceutically unacceptable "common organic solvents" such as DMSO or other excipients to the extent required for solubilization. Indeed, the formulations of (rev)cromakalim in the following examples demonstrate improved solubility over formulations of (rev)cromakalim in a mixture of DMSO and Cremophor™ (i.e., Kolliphor™). The improved solubility of the ophthalmic formulations described herein translates to improved bioavailability and efficacy, as shown in the study shown in Figure 3. Figure 3 shows intraocular pressure (IOP) at various concentrations of formulated (rev)cromakalim compared to formulations of the same concentration in DMSO and Cremophor™. This data demonstrates the improved performance of the (rev)cromakalim formulation described herein (approximately 33% IOP reduction) compared to DMSO and Cremophor™ formulations (approximately 23% IOP reduction).

[0179] Non-limiting examples of aqueous formulations of (lev)cromakalim according to the present invention that exhibit a concentration of (lev)cromakalim greater than 1 mg / mL include, but are not limited to:

[0180] Example 2a, an ophthalmic formulation comprising levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, mannitol, and BAK. For example, formulations of Formulation 2a containing this component mixture exhibited a levcromakalim concentration of 1.4 mg / mL, or 4.7 mM.

[0181] Example 2b, an ophthalmic formulation comprising levcromakalim, Kolliphor™ RH 40, Octoxynol-40, and Premulen™ TR-2. For example, formulation 2b, comprising this component mixture, exhibited a levcromakalim concentration of 1.2 mg / mL, or 4.3 mM.

[0182] Example 2c, an ophthalmic formulation comprising levcromakalim, Kolliphor® RH 40, Kolliphor® HS 15, and PVP. For example, formulation 2c containing this component mixture exhibited a levcromakalim concentration of 1.2 mg / mL, or 4.1 mM.

[0183] Example 2d, an ophthalmic formulation comprising levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, and hypromellose. For example, formulation 2d, comprising this component mixture, exhibited a levcromakalim concentration of 1.3 mg / mL, or 4.6 mM.

[0184] Example 2e, where the ophthalmic formulation comprises levcromakalim, Kolliphor™ ELP, polysorbate 80, and PVP. For example, formulation 2e, which comprises this mixture of ingredients, exhibited a levcromakalim concentration of 1.1 mg / mL, or 4 mM.

[0185] Example 2f, an ophthalmic formulation comprising levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose. For example, formulation 2f containing this component mixture exhibited a levcromakalim concentration of 1.3 mg / mL, or 4.4 mM.

[0186] Example 2g, an ophthalmic formulation comprising levcromakalim, glycerin, Kolliphor® RH 40, Premulen® TR-2, and PVP. For example, a formulation of Formulation 2g containing this component mixture exhibited a levcromakalim concentration of 1 mg / mL, or 3.7 mM.

[0187] Example 2h, an ophthalmic formulation comprising levcromakalim, Kolliphor™ HS 15, Premulen™ TR-2, polysorbate 80, octoxynol-40, and poloxamer 407. For example, formulation 2h containing this component mixture exhibited a levcromakalim concentration of 1.3 mg / mL, or 4.4 mM.

[0188] Example 2i, an ophthalmic formulation comprising levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, PVP, mannitol, octoxynol-40, and poloxamer 407. For example, formulations of Formulation 2i comprising this component mixture exhibited a levcromakalim concentration of 1.2 mg / mL, or 4.3 mM.

[0189] Example 2j, an ophthalmic formulation comprising levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, mannitol, and BAK. For example, formulation 2j, comprising this component mixture, exhibited a levcromakalim concentration of 1.3 mg / mL, or 4.6 mM.

[0190] Example 3a, where the ophthalmic formulation comprises levcromakalim, polysorbate 80, Kolliphor™ ELP, and PVP. For example, formulation 3a, which comprises this mixture of ingredients, exhibited a levcromakalim concentration of 1.7 mg / mL, or 5.8 mM.

[0191] Example 3b, where the ophthalmic formulation comprises levcromakalim, Kolliphor™ ELP, polysorbate 80, and poloxamer 407. For example, formulation 3b containing this component mixture exhibited a levcromakalim concentration of 2.1 mg / mL, or 7.3 mM.

[0192] Example 3c, an ophthalmic formulation comprising levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, and poloxamer 407. For example, formulation 3c containing this component mixture exhibited a levcromakalim concentration of 1.8 mg / mL, or 6.4 mM.

[0193] Example 3d, an ophthalmic formulation comprising levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol. For example, formulation 3d, comprising this component mixture, exhibited a levcromakalim concentration of 1.6 mg / mL, or 5.5 mM.

[0194] Example 3e, an ophthalmic formulation comprising levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose. For example, formulation 3e containing this component mixture exhibited a levcromakalim concentration of 1.9 mg / mL, or 6.5 mM.

[0195] Example 3f, an ophthalmic formulation comprising levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, mannitol, and BAK. For example, formulation 3f containing this component mixture exhibited a levcromakalim concentration of 1.5 mg / mL, or 5.4 mM.

[0196] Example 3g, an ophthalmic formulation comprising levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol. For example, a formulation of Formulation 3g containing this component mixture exhibited a levcromakalim concentration of 1.6 mg / mL, or 5.7 mM.

[0197] Example 3h, an ophthalmic formulation comprising levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, mannitol, and hypromellose. For example, formulation 3h containing this component mixture exhibited a levcromakalim concentration of 1.7 mg / mL, or 5.9 mM.

[0198] In certain embodiments, the ophthalmic formulation of (rev)cromakalim or a pharmaceutically acceptable salt is formed as a solution or emulsion, e.g., at a concentration of at least about 0.01 mg / mL, 0.05 mg / mL, 1 mg / mL, 1.1 mg / mL, 1.2 mg / mL, 1.3 mg / mL, or 1.4 mg / mL, and typically at or below about 1.5 mg / mL.

[0199] In certain embodiments, the ophthalmic formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof is formed as a solution or emulsion, e.g., at a concentration of at least about 0.05 mM, 0.5 mM, 1 mM, 1.5 mM, 2 mM, 2.5 mM, 3 mM, 3.5 mM, 4 mM, or 4.5 mM, and typically at a concentration of about 5 mM or less.

[0200] The average topical drop tends to be in the range of 20 to 50 microliters, but can be larger or smaller depending on viscosity and dropper size. In certain embodiments, the average drop may be approximately 30 milliliters ±25%. Typical dosing calls for patients to administer one drop per eye once daily, twice daily, three times daily, or four times daily.

[0201] The (rev)cromakalim ophthalmic formulations described herein can be provided in any dosage strength that achieves the desired results. In certain exemplary, non-limiting embodiments, the pharmaceutical composition is provided as a single drop per eye to administer a range of about 0.0005 mg to about 0.001 mg, about 0.001 mg to about 0.005 mg, about 0.005 mg to about 0.01 mg, about 0.01 mg to about 0.04 mg, or about 0.04 mg to about 0.09 mg of the (rev)cromakalim ophthalmic formulation compound. In certain embodiments, the dosage form contains at least about 0.001 mg, 0.005 mg, 0.01 mg, 0.02 mg, 0.025 mg, or 0.05 mg of the active compound or salt thereof.

[0202] Suitable pH active ingredients such as buffers or pH adjusters for use in pharmaceutical compositions according to the present invention include, but are not limited to, disodium phosphate (dibasic sodium phosphate), monobasic sodium phosphate (monosodium phosphate), boric acid, sodium borate, sodium citrate, hydrochloric acid, acetate buffers containing sodium hydroxide, borate buffers, carbonate buffers, citrate buffers, and phosphate buffers.

[0203] In certain embodiments, ophthalmic formulations of (rev)cromakalim have a pH between approximately 6 and 8 or between 6.5 and 7.5. In certain embodiments, the formulation comprises a citrate buffer with a pH of around 6.5 to 7. In alternative embodiments, the formulation comprises a phosphate buffer with a pH of around 6.5 to 7. Suitable osmotically active ingredients for use in pharmaceutical compositions according to the present invention include, but are not limited to, sodium chloride, mannitol, and glycerol.

[0204] Non-limiting examples of buffers, with or without additional components or other additives, that can be used as pharmaceutically acceptable formulations for the appropriate indications described herein include, for example (with illustrative, but not limiting, concentrations and pHs): acetate buffer (0.1 M, pH 5.0), acetate buffer (pH 3.6-5.6), BES buffered saline (2x) (0.05 M, pH 6.95), bicine (1 M, pH 8.26), CA 91100, ... PS (1M, pH 10.4), CHES (1M, pH 9.5), citrate buffer (0.1M, pH 6.0), citrate-phosphate buffer (0.15M, pH 5.0), citrate buffer (pH 3.0-6.2), carbonate-bicarbonate buffer (pH 9.2-10.6), diethanolamine (1M, pH 9.8), EBSS (magnesium, calcium, phenol red) (pH 7.0), glycine-HCl buffer (0.1M, pH 3.0), Lysine-sodium hydroxide buffer (0.08 M, pH 10), HBSS (Hank's Balanced Salt Solution), HEPPSO (1 M, pH 7.85), HHBS (Hank's buffer solution containing Hepes), hydrochloric acid-potassium chloride buffer (0.1 M, pH 2.0), imidazole-HCl buffer (0.05 M, pH 7.0), MES (0.5 M, pH 6), MOPS buffer (10x) (0.2 M, pH 7), PBS (phosphate buffered saline) (1x, pH Examples of suitable buffers include H7.4, sodium borate buffer (1 M, pH 8.5), TAE (1 M, pH 8.6), TAE buffer (50x) (0.04 M, pH 8.5), TBS (1 M, pH 7.4), TE buffer (10x), Tricine (1 M, pH 8.05), Tris buffer (1 M, pH 7.2), phosphate buffer (pH 5.8-8.0), potassium phosphate (pH 5.8-8.0), and Trizma™ buffer (pH 7.0-9.2).

[0205] In certain embodiments, levcromakalim may be administered as a salt in the ophthalmic formulation. Pharmaceutically acceptable salts of levcromakalim include, but are not limited to: [ka] (In the formula, X + and M 2+Z can be any pharmaceutically acceptable cation that achieves the desired result. + is X + (representing a mixed salt of

[0206] In certain embodiments, the cation is selected from sodium, potassium, aluminum, calcium, magnesium, lithium, iron, zinc, arginine, chloroprocaine, choline, diethanolamine, ethanolamine, lysine, histidine, meglumine, procaine, hydroxyethylpyrrolidine, ammonium, tetrapropylammonium, tetrabutylphosphonium, methyldiethanolamine, and triethylamine.

[0207] In certain embodiments, X + Na + or K + In certain embodiments, X + Li + In certain embodiments, X + is Cs + In certain embodiments, X + is an ammonium ion with a net positive charge of 1. Non-limiting examples of ammonium ions with a net positive charge of 1 include: [ka]

[0208] In an alternative embodiment, the ammonium ion having a net positive charge of 1 has the formula: [ka] (In the formula, R 1is C1-C6 alkyl, such as, but not limited to, methyl, ethyl, propyl, isopropyl, butyl, t-butyl, sec-butyl, isobutyl, —CH2C(CH3)3, —CH(CH2CH3)2, and —CH2CH(CH2CH3)2, cyclopropyl, CH2-cyclopropyl, cyclobutyl, and CH2-cyclobutyl, or aryl, such as phenyl or naphthyl, where the C1-C6 alkyl or aryl may be optionally substituted, for example, with a hydroxyl group. In certain embodiments, the ammonium ion is [ka] is.

[0209] M 2+ M can be, for example, but not limited to, an alkaline earth metal cation (magnesium, calcium, or strontium), a metal cation having an oxidation state of +2 (e.g., zinc or iron), or an ammonium ion having a net positive charge of 2 (e.g., benzathine, hexamethyldiammonium, and ethylenediamine). 2+ is Mg 2+ In certain embodiments, M 2+ Ca 2+ In certain embodiments, M 2+ is Sr 2+ In certain embodiments, M 2+ Zn 2+ In certain embodiments, M 2+ is Fe 2+ In certain embodiments, M 2+ is an ammonium ion with a net positive charge of 2. Non-limiting examples of ammonium ions with a net positive charge of 2 include: [ka]

[0210] In an alternative embodiment, the ammonium ion having a net positive charge of two has the formula: [ka] (In the formula, R 1 is C1-C6 alkyl, such as, but not limited to, methyl, ethyl, propyl, isopropyl, butyl, t-butyl, sec-butyl, isobutyl, —CH2C(CH3)3, —CH(CH2CH3)2, and —CH2CH(CH2CH3)2, cyclopropyl, CH2-cyclopropyl, cyclobutyl, and CH2-cyclobutyl, or aryl, such as phenyl or naphthyl, wherein the C1-C6 alkyl or aryl may be optionally substituted, for example, with a hydroxyl group; and and y is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8.

[0211] III. Method of Administration The ophthalmic formulations of (rev)cromakalim or its pharmaceutically acceptable salts of the present invention described herein can be administered in an effective amount to a host, typically a human, in need of administration for any of the indications described herein. These immediate-release formulations are administered as pharmaceutical compositions containing an effective amount for a host, typically a human, in need of such treatment. Thus, the present disclosure provides pharmaceutical compositions comprising an effective amount of an ophthalmic formulation of (rev)cromakalim. The pharmaceutical composition may contain the compound or a salt thereof as the only active agent in a formulation, or may contain the compound or a salt thereof and at least one additional active agent, as described in detail herein.

[0212] The exact amounts and dosages of the active compounds in the ophthalmic formulations described herein to be delivered to a host in need thereof, typically a human, will be determined by a healthcare provider to achieve the desired clinical benefit.

[0213] In certain embodiments, the levcromakalim ophthalmic formulation containing levcromakalim is administered for at least about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 2 weeks, 3 weeks, 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months or more, including indefinitely during therapy. In certain embodiments, the levcromakalim ophthalmic formulation containing levcromakalim is administered once daily, twice daily, three times daily, or more times.

[0214] The ophthalmic formulation of (rev)cromakalim described herein is administered locally.For local delivery, the topical dosage form can be administered as needed, for example, once a day (QD), twice a day (BID), three times a day (TID), four times a day (QID), once every two days (Q2D), once every three days (Q3D), or any other dosing schedule that results in the treatment of the disorders described herein.Alternatively, the immediate release formulation can be prepared for long-term delivery, such as every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks or more, or every 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months or more or less, including indefinitely indefinitely.

[0215] Ophthalmic formulations of (rev)cromakalim may also be used for ocular therapy using alternative routes, i.e., intravitreal, intrastromal, intracameral, subtenon, subretinal, retrobulbar, periocular, suprachoroidal, subchoroidal, choroidal, conjunctival, subconjunctival, episcleral, periocular, transscleral, posterior juxtascleral, pericorneal, or lacrimal injection, or through a mucus, mucin, or mucosal barrier, in the immediate release manner described, or via an ophthalmic device or injection. In certain embodiments, the ophthalmic device is a contact lens that releases an immediate release formulation of (rev)cromakalim.

[0216] Patient compliance and adherence are important issues, and the fewer daily doses required, the more likely compliance is achieved. Dosing an ophthalmic formulation once a day for glaucoma is advantageous for maintaining intraocular pressure within a desirable range to minimize optic nerve damage, while also optimizing compliance and adherence. In certain embodiments, a selected effective dosage of the ophthalmic formulation of (rev)cromakalim can be administered once a day (QD), twice a day (BID), or three times a day (TID) as topical drops or other convenient methods.

[0217] In certain embodiments, the formulations described herein comprising a mixture of cromakalim, such as levcromakalim, and selected pharmaceutically acceptable ingredients described herein are administered by daily dosing. Non-limiting examples of (lev)cromakalim formulations used for daily dosing include the following:

[0218] Example 2j, wherein the ophthalmic formulation comprises levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, mannitol, and BAK.

[0219] Example 3a, wherein the ophthalmic formulation comprises levcromakalim, polysorbate 80, Kolliphor™ ELP, and PVP.

[0220] Example 3b, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™ ELP, polysorbate 80, and poloxamer 407.

[0221] Example 3c, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, and poloxamer 407.

[0222] Example 3d, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol.

[0223] Example 3d, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol.

[0224] Example 3e, wherein the ophthalmic formulation comprises levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose.

[0225] Example 3f, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, mannitol, and BAK.

[0226] Example 3g, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol.

[0227] Example 3h, wherein the ophthalmic formulation comprises levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, mannitol, and hypromellose.

[0228] IV. Use of Cromakalim Ophthalmic Preparations The present invention provides a method of using the (rev)cromakalim formulation of the present invention to deliver an effective amount of an ophthalmic formulation of (rev)cromakalim or a salt thereof to treat any disorder that can be treated with (rev)cromakalim or a pharmaceutically acceptable salt thereof.

[0229] In certain embodiments, ophthalmic formulations suitable for treating high IOP glaucoma, including but not limited to primary open-angle glaucoma (POAG), primary angle-closure glaucoma, pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, and iridocorneal endothelial glaucoma, are provided (primary open-angle glaucoma is also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma). In alternative embodiments, the formulations are used to treat acute high-pressure glaucoma resulting from a developing cataract. In another embodiment, the formulations are used to treat steroid-induced glaucoma, uveitic glaucoma, or acute high-pressure glaucoma resulting from intravitreal injection, or glaucoma without elevated intraocular pressure, including but not limited to normal-tension glaucoma (NTG) (also known as low-tension or normal-tension glaucoma). In non-limiting exemplary embodiments, the formulation is administered with an ophthalmic formulation of a compound described herein, including, for example, a formulation including Kolliphor™, polysorbate, polyvinylpyrrolidone, i.e., PVP, poloxamer, mannitol, and benzalkonium chloride (BAK).

[0230] Intraocular pressure (IOP) IOP is determined by the rates of aqueous humor (AH) production and outflow. One way to specifically quantify these factors is via the modified Goldman equation (Lee SS, Robinson MR, Weinreb RN. Episcleral venous pressure and the ocular hypotensive effects of topical and intracameral prostaglandin analogs. J Glaucoma.2019;28(9):846-57). In this equation, IOP is equal to the episcleral venous pressure (EVP) plus the AH inflow fraction (Q) minus the uveoscleral outflow fraction (U), divided by the conventional trabecular outflow fraction (C) and EVP: IOP=EVP+[(QU) / C]

[0231] Currently approved pharmacological therapies for managing IOP primarily affect "Q," "C," and "U" by acting on the ciliary body, trabecular meshwork (TM), and uveoscleral tissues of the eye, respectively. For example, beta-blockers such as timolol, betaxolol, levobunolol, and metipranolol, alpha-agonists such as brimonidine and apraclonidine, and carbonic anhydrase inhibitors such as brinzolamide, dorzolamide, acetazolamide, and methazolamide affect the AH inflow rate (Q). Therapeutics targeting the uveoscleral outflow rate (U) include prostaglandin analogs such as latanoprost, bimatoprost, travoprost, tafluprost, and latanoprost bunod, and alpha-agonists such as brimonidine and apraclonidine. Traditional trabecular outflow capacity (C) is affected by cholinergic or mitotic drugs such as pilocarpine and carbachol, and Rho kinase inhibitors such as netarsudil. In contrast, none of the available pharmacological therapies primarily reduce EVP or specifically target tissues and vessels distal to the TM, such as the collecting duct, deep scleral and intrascleral venous plexus, or episcleral veins. In some embodiments, the present invention is administered with one or a combination of these standard or (all) drugs.

[0232] Of the four components of IOP, EVP is typically the largest component, equaling approximately 50%–60% of total IOP (Lee SS, Robinson MR, Weinreb RN. Episcleral venous pressure and the ocular hypotensive effects of topical and intracameral prostaglandin analogs. J Glaucoma. 2019;28(9):846-57). Importantly, EVP defines the lower IOP limit (typically 8–12 mmHg), which sets a “floor” for maximal therapy. For example, a higher EVP limits the therapeutic potential of pharmacological therapy, i.e., for NTG, and certain surgical therapies, such as minimally invasive glaucoma surgery (MIGS) devices, for the treatment of POAG. In particular, EVP has been found to be higher in untreated NTG and POAG compared with age-matched controls (Selbach JM, Posielek K, Steuhl KP, Kremmer S. Episcleral venous pressure in untreated primary open-angle and normal-tension glaucoma. Ophthalmologica. 2005;219(6):357-61). Therefore, both NTG and POAG patients may benefit from EVP-lowering agents.

[0233] In certain embodiments, the ophthalmic formulations of the present invention reduce IOP by reducing EVP at certain specific concentrations. For example, as shown in Example 7, formulation 3d reduces IOP by reducing episcleral venous pressure (EVP) in normotensive C57 / BL6J mice.

[0234] glaucoma Glaucoma is a complex, multifactorial optic neuropathy and a leading cause of irreversible blindness, affecting more than 80 million people worldwide (Quigley HA, Broman AT. The number of people with glaucoma worldwide in 2010 and 2020. Br J Ophthalmol. 2006;90(3):262-7; Weinreb RN, Aung T, Medeiros FA. The pathophysiology and treatment of glaucoma: a review. JAMA. 2014;311(18):1901-11; and Tham YC, Li X, Wong TY, et al. Global Prevalence of Glaucoma and Projections of Glaucoma Burden through 2040: A Systematic Review and Meta-Analysis. Ophthalmology. 2014;121(11):2081-90). Historically, elevated intraocular pressure (IOP) was the primary and only modifiable risk factor for glaucoma. Lowering IOP is essential to slow disease progression and vision loss (Weinreb RN, Aung T, Medeiros FA. The pathophysiology and treatment of glaucoma: a review. JAMA. 2014;311(18):1901-11).

[0235] Conventionally, an IOP below 21 mmHg is considered "normal," while a pressure above 21 mmHg is considered a clinical warning sign (Dielemans I, Vingerling JR, Algra D, et al. Primary open-angle glaucoma, intraocular pressure, and systemic blood pressure in the general elderly population. The Rotterdam Study. Ophthalmology. 1995;102(1):54-60). Ocular hypertension (OHT) is a condition in which IOP is elevated without any of the findings of glaucoma (e.g., structural changes in the cup-to-disc ratio or visual field defects). Economic analyses have revealed that subjects with OHT have an overall 10% risk of developing glaucoma over a 5-year period (Mansberger SL, Medeiros FA, Gordon M. Diagnostic tools for calculation of glaucoma risk. Surv Ophthalmol. 2008;53(SUPPL1):S11-6), so prophylactic treatment is often undertaken as a preventative measure. Results from central ocular hypertension treatment studies have shown that medical treatments that lower IOP can reduce the risk of developing POAG. Patients are considered to have POAG only after clinical signs of glaucoma, such as a change in cup-to-disc ratio or observation of visual field changes (Kass MA, Heuer DK, Higginbotham EJ, et al. The Ocular Hypertension Treatment Study: a randomized trial determines that topical ocular hypotensive medication delays or prevents the onset of primary open-angle glaucoma. Arch Ophthalmol. 2002;120(6):701-13).

[0236] IOP is maintained by balancing the production and outflow of aqueous humor (AH), a clear fluid that serves as a nutrient source for anterior avascular tissues such as the lens and cornea (Goel M, Picciani RG, Lee RK, Bhattacharya SK. Aqueous humor dynamics: a review. Open Ophthalmol J. 2010;4:52-9). AH is produced at a rate of 2.4 μL / min in humans and must be drained at approximately the same rate to maintain IOP within a "normal" and stable homeostatic range. AH can exit the eye through one of two drainage pathways: the conventional outflow pathway, which involves the trabecular meshwork (TM) and Schlemm's canal, or the uveoscleral pathway, in which AH permeates through spaces within the extracellular matrix and ciliary muscle. In middle-aged and older adults, up to 80% of AH exits the eye through the conventional pathway, while the remaining 20% ​​or less exits via the uveoscleral pathway. Disruption of this balance between AH production and conventional outflow can result in elevated IOP.

[0237] Open-angle glaucoma is the most common type in the United States, accounting for 9 out of 10 glaucoma cases in the United States. Over time, pressure damages the optic nerve, affecting vision. This can ultimately lead to blindness. In primary open-angle glaucoma (POAG), the drainage angle formed by the cornea and iris remains open, but there is increased resistance to outflow through the traditional outflow pathway, causing elevated IOP (Kwon YH, Fingert JH, Kuehn MH, Alward WL. Primary open-angle glaucoma. N Engl J Med. 2009;360(11):1113-24 and Weinreb RN, Khaw, PT. Primary open-angle glaucoma. Lancet. 2004;363(9422):1711-20). In most patients, increases in IOP over time correlate closely with damage to the retinal nerve fiber layer and optic nerve head, and subsequently with gradual but persistent vision loss due to loss of retinal ganglion nerve fibers that converge at the optic nerve head to form the optic nerve (Quigley HA, Broman AT. The number of people with glaucoma worldwide in 2010 and 2020. Br J Ophthalmol. 2006;90(3):262-7 and Weinreb RN, Aung T, Medeiros FA. The pathophysiology and treatment of glaucoma: a review. JAMA. 2014;311(18):1901-11). Therefore, lowering IOP remains the primary treatment goal for POAG and may even be “neuroprotective” given the ultimate preservation of optic nerve fibers (Weinreb RN, Leung CK, Crowston JG, et al. Primary open-angle glaucoma. Nat Rev Dis Primers. 2016;2:16067).

[0238] Normal-tension glaucoma (NTG) is a type of open-angle glaucoma that occurs in individuals with normal intraocular pressure. In NTG, IOP is within the normal range, but the optic nerve remains damaged, and subjects develop progressive glaucomatous visual field loss despite "normal" IOP. Although IOP-independent risk factors are thought to be involved in the pathogenesis of NTG, IOP reduction remains the core of treatment, as demonstrated in the Collaborative Normal Tension Glaucoma Study (Anderson DR, Normal Tension Glaucoma Study. Collaborative normal tension glaucoma study. Curr Opin Ophthalmol. 2003; 14(2):86-90). NTG is more common in Asians.

[0239] Angle-closure glaucoma, also known as narrow-angle glaucoma or acute glaucoma, is a severe form of glaucoma that constitutes a medical emergency. In this type of glaucoma, the outer edge of the iris blocks drainage from the front of the eye. Fluid builds up quickly, causing a sudden increase in intraocular pressure. If untreated, angle-closure glaucoma can cause blindness in just a few days. Another type of angle-closure glaucoma is sometimes called slow-angle-closure or chronic angle-closure glaucoma. In another type of glaucoma, congenital glaucoma, babies are born with eye problems that prevent fluid from draining properly.

[0240] Glaucoma caused by another medical condition is called secondary glaucoma. Neovascular glaucoma is often caused by diabetes or high blood pressure, which causes the eye to develop extra blood vessels that cover areas of the eye that normally drain fluid. Pigmentary glaucoma is a secondary glaucoma in which pigment (color) from the iris (the colored part of the eye) peels off, blocking drainage from the eye. Exfoliation glaucoma (sometimes called pseudoexfoliation) is a type of open-angle glaucoma that occurs in some patients with exfoliation syndrome, a condition in which extra material deposits in parts of the eye, blocking drainage. Uveitic glaucoma can occur in patients with uveitis, a condition that causes swelling and inflammation of the eye. Uveitis can cause inflammation and scar tissue in the center of the eye. This can damage or block drainage areas of the eye, causing high intraocular pressure and potentially leading to uveitic glaucoma and vision loss.

[0241] High intraocular pressure is a common symptom of the most common form of glaucoma, and treatments that lower intraocular pressure slow the onset and progression of glaucoma.

[0242] Glaucoma can be treated using the formulations described herein, which include cromakalim, such as levcromakalim, and a mixture of selected pharmaceutically acceptable ingredients as described herein. In certain embodiments, the formulations of the present invention can be used to treat glaucoma in which the reduction in IOP is due to a reduction in EVP. Non-limiting examples of (lev)cromakalim formulations used to treat glaucoma (elevated or normal intraocular pressure) include:

[0243] Example 2j, wherein the ophthalmic formulation comprises levcromakalim, glycerin, Kolliphor™, polysorbate, poloxamer, hypromellose, mannitol, and BAK.

[0244] Example 3a, wherein the ophthalmic formulation comprises levcromakalim, polysorbate, Kolliphor™, and PVP.

[0245] Example 3b, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™, polysorbate, and poloxamer 407.

[0246] Example 3c, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™, polysorbate, PVP, and poloxamer.

[0247] Example 3d, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™, polysorbate, PVP, poloxamer, and mannitol.

[0248] Example 3e, wherein the ophthalmic formulation comprises levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose.

[0249] Example 3f, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, mannitol, and BAK.

[0250] Example 3g, wherein the ophthalmic formulation comprises levcromakalim, Kolliphor™, polysorbate, PVP, poloxamer, hypromellose, and mannitol.

[0251] Example 3h, wherein the ophthalmic formulation comprises levcromakalim, glycerin, Kolliphor™, polysorbate, poloxamer, mannitol, and hypromellose.

[0252] Hyperemia In certain embodiments, an ocular therapy using an effective amount of an ophthalmic formulation of (rev)cromakalim is provided that does not result in significant redness. Redness is an excessive and / or noticeable increase in blood supply to an organ in the blood vessels. Ocular redness, also known as "red eye," can include or cause vascular congestion, excessive vasodilation, small hemorrhages, small petechiae, and / or microhemorrhages. Ocular redness can have a variety of causes, including, but not limited to, exogenous irritants, contact lenses, inflammation, vascular disruption, conjunctivitis (including infectious or allergic), trauma, endogenous ocular injury, subconjunctival hemorrhage, conjunctival hemorrhage, blepharitis, anterior uveitis, glaucoma, or irritating medications and environmental stimuli (i.e., sunlight and wind).

[0253] Certain ophthalmic medications either do not reduce or cause redness. According to the present invention, the use of an ophthalmic formulation of (rev)cromakalim does not cause significant redness in patients when used during therapy. In certain embodiments, significant redness causes sufficient discoloration or discomfort in the patient that the patient perceives it as a side effect of treatment, which, if sufficiently severe and / or prolonged, may lead to poor compliance or even discontinuation of therapy. The present invention provides an advancement in the art by supporting patient compliance and comfort. In certain embodiments, administration of an ophthalmic formulation of (rev)cromakalim does not significantly induce expression of at least one protein independently selected from CD31 and VE-cadherin.

[0254] In certain embodiments, the administered ophthalmic formulation that does not produce significant hyperemia comprises a mixture of cromakalim, such as levcromakalim, and selected pharmaceutically acceptable ingredients described herein.

[0255] Sturge-Weber syndrome Sturge-Weber syndrome is a congenital disorder that affects the skin, nervous system, and sometimes the eyes. It is sometimes referred to as a neurocutaneous disorder. Sturge-Weber syndrome is a condition that affects the development of certain blood vessels, causing abnormalities in the brain, skin, and eyes from birth. Sturge-Weber syndrome is characterized by three major features: a red or pink birthmark called a port-wine stain, a brain abnormality called a leptomeningeal angioma, and elevated intraocular pressure (IOP) in the eye (glaucoma). Patients with Sturge-Weber syndrome typically develop glaucoma during infancy or early adulthood, which can cause vision problems. In some affected infants, intraocular pressure can become so high that the eyeball appears enlarged and bulging (buphthalmos). Patients with Sturge-Weber syndrome may have abnormal vascular plexuses (angiomas) in various parts of the eye. When these abnormal blood vessels develop into the vascular network at the back of the eye (choroid), this condition is called diffuse choroidal hemangioma, which occurs in approximately one-third of patients with Sturge-Weber syndrome. Diffuse choroidal hemangioma can cause vision loss. When present, the eye abnormality typically occurs in the same temporal region as the port-wine stain.

[0256] Another aspect of the present invention is the use of an ophthalmic formulation for the treatment of glaucoma associated with Sturge-Weber syndrome. Sturge-Weber syndrome-induced glaucoma affects 30% to 70% of patients. Management of Sturge-Weber syndrome-induced glaucoma can be complex, with many patients requiring surgery or drainage devices. According to the present invention, Sturge-Weber syndrome-induced glaucoma can be treated by administering an effective amount of an ophthalmic formulation of (lev)cromakalim.

[0257] In certain embodiments, glaucoma can be treated with the formulations described herein that include a mixture of cromakalim, such as levcromakalim, and selected pharmaceutically acceptable ingredients described herein.

[0258] diabetic retinopathy Diabetic retinopathy (DR) is a complication of diabetes caused by damage to the blood vessels in the light-sensitive tissue at the back of the eye (retina). DR has two stages: nonproliferative and proliferative. In the nonproliferative stage, blood vessels swell and leak, causing macular edema and potentially affecting vision due to loss of blood supply. In proliferative / advanced DR, abnormal new blood vessels grow on the retinal surface, which can rupture and bleed into the vitreous, resulting in vision loss. DR can be caused by hyperglycemia in type 1 or type 2 diabetes.

[0259] Another aspect of the present invention is the use of an ophthalmic preparation for the treatment of elevated intraocular pressure and glaucoma associated with diabetic retinopathy. According to the present invention, elevated intraocular pressure and glaucoma associated with DR can be treated by administering an effective amount of an ophthalmic preparation of (lev)cromakalim.

[0260] In certain embodiments, DR can be treated with the formulations described herein that include a mixture of cromakalim, such as levcromakalim, and selected pharmaceutically acceptable ingredients described herein.

[0261] Mitochondrial disorders associated with optic neuropathy Mitochondrial diseases are often medically induced, genetic, and inherited. They are a clinically heterogeneous group of disorders resulting from dysfunction in the mitochondrial respiratory chain. The mitochondrial respiratory chain is the final common pathway essential for aerobic metabolism, and tissues and organs highly dependent on aerobic metabolism are preferentially affected by mitochondrial disorders. While some mitochondrial disorders affect only a single organ, many affect multiple organ systems and often present with prominent neurological and myopathic features. Mitochondria contain ATP-sensitive potassium-specific channels (mitoKATP channels). Mitochondrial KATP channels play important roles in mitochondrial volume control and regulating components of the proton motive force.

[0262] Inside the mitochondria are a group of proteins that transport electrons along a four-step chain (Complex I-Complex IV), resulting in the generation of energy. This chain is known as the electron transport chain. A fifth group (Complex V) produces large amounts of ATP. Together, the electron transport chain and ATP synthase form the respiratory chain, a process known as oxidative phosphorylation, or OXPHOS. Complex I, the first step in this chain, is the most common site of mitochondrial abnormalities, accounting for as many as one-third of respiratory chain deficiencies.

[0263] Several specific mitochondrial disorders have been associated with complex I deficiency, including Leber's hereditary optic neuropathy, mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS), myoclonic epilepsy with ragged-red fibers (MERRF), and Leigh syndrome.

[0264] There are many other types of mitochondrial diseases. For example, dominant optic atrophy (DOA) is a hereditary optic neuropathy characterized by degeneration of the optic nerve, which typically begins within the first decade of life. Affected individuals usually develop moderate vision loss and color vision deficiency. Severity varies, and vision can range from normal to legally blind. Autosomal dominant optic atrophy plus syndrome (ADOA plus) is a rare syndrome that causes vision loss, hearing loss, and muscle-affecting symptoms. This syndrome is associated with optic atrophy. Other symptoms of ADOA plus include sensorineural hearing loss and muscle-affecting symptoms such as muscle pain and weakness. ADOA plus is caused by mutations in the OPA1 gene. Both DOA and ADOA are inherited in an autosomal dominant manner. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered to treat dominant optic atrophy (DOA) or autosomal dominant optic atrophy plus syndrome (ADOA plus).

[0265] Chronic progressive external ophthalmoplegia (CPEO) is a condition characterized by the loss of muscle function, primarily involved in eye and eyelid movement. Signs and symptoms tend to begin in early adulthood and most commonly include weakness or paralysis of the muscles that move the eye (ophthalmoplegia) and drooping of the eyelids (ptosis). Some affected individuals also have a myopathy, which may be particularly noticeable during movement. CPEO can be caused by mutations in any of several genes that can be located in mitochondrial DNA or nuclear DNA. CPEO can occur as part of other underlying conditions, such as the ataxia neuropathy spectrum and Kearns-Sayre syndrome (KSS). KSS is a slowly progressive, multisystem mitochondrial disorder that often begins with ptosis. Eventually, other eye muscles are involved, causing paralysis of eye movements. Retinal degeneration causes visual difficulty, usually in dimly lit environments. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered to treat chronic progressive external ophthalmoplegia or Kearns-Sayre syndrome.

[0266] Leber's hereditary optic neuropathy (LHON) is a condition characterized by vision loss. Some affected individuals may exhibit features similar to multiple sclerosis. LHON is caused by mutations in the MT-ND1, MT-ND4, MT-ND4L, and MT-ND6 genes. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered to treat Leber's hereditary optic neuropathy.

[0267] Mitochondrial enoyl-CoA reductase protein-associated neurodegeneration (MEPAN) is caused by two mutations in the MECR gene (encoding the mitochondrial trans-2-enoyl-coenzyme A reductase protein). Characteristics of MEPAN include optic atrophy and childhood-onset dystonia. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered to treat mitochondrial enoyl-CoA reductase protein-associated neurodegeneration (MEPAN).

[0268] POLG-related disorders comprise a range of overlapping phenotypes with onset from infancy to late adulthood. Mutations in POLG can cause mitochondrial DNA (mtDNA) depletion syndrome in early childhood or late-onset syndromes resulting from mtDNA deletions. POLG mutations are the most common cause of inherited mitochondrial disorders, with as many as 2% of the population carrying these mutations. The six major disorders caused by POLG mutations are Alpers-Hutten-Locker syndrome, one of the most severe phenotypes; childhood myoencephalohepatopathy spectrum, which manifests within the first three years of life; myoclonic epilepsy-myopathy-sensory ataxia; ataxic neuropathy spectrum (including phenotypes previously termed mitochondrial recessive ataxia syndrome (MIRAS) and sensory ataxic neuropathy-dysarthria-ophthalmoplegia (SANDO)); autosomal recessive progressive external ophthalmoplegia; and autosomal dominant progressive external ophthalmoplegia. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered to treat a POLG-related disorder.

[0269] In certain embodiments, formulations described herein comprising a mixture of cromakalim, such as levcromakalim, and selected pharmaceutically acceptable ingredients described herein can be used to treat optic neuropathy.

[0270] Ophthalmic Neuroprotection Neuroprotection is a therapeutic strategy that aims to maximize neuronal recovery and minimize neuronal death due to injury. Injury can be mechanical, ischemic, degenerative, or caused by radiation. Many neurodegenerative disorders are associated with aging and can be detrimental to the elderly. For example, glaucoma, often characterized by loss of retinal ganglion cells, is a leading cause of vision loss and blindness in the elderly.

[0271] In certain embodiments, an ophthalmic formulation of (lev)cromakalim is administered to a host in need of treatment to treat an ocular-related neurodegenerative disorder, which is any disorder associated with the dysfunction or degeneration of neurons or cells, including nerve cells such as retinal ganglion cells.

[0272] In certain embodiments of the present invention, an ophthalmic formulation of (rev)cromakalim is administered as a method of reducing neuronal or cellular damage in the eye of a host in need thereof. In certain embodiments, an ophthalmic formulation of (rev)cromakalim is administered as a method of reducing neuronal or cellular damage in the eye of a host in need thereof, the eye suffering from glaucoma.

[0273] In certain embodiments, ophthalmic formulations of (rev)cromakalim promote the survival, growth, regeneration, and / or neurite outgrowth of retinal ganglion cells, hi certain embodiments, ophthalmic formulations of (rev)cromakalim prevent the death of damaged nerve cells.

[0274] Because neuronal cell death can be a result of retinal ischemia, in certain embodiments, an ophthalmic formulation of (lev)cromakalim is administered as a method of reducing neuronal or cellular damage in the eye following retinal ischemia in a host in need thereof.

[0275] Optic neuropathy, damage to the optic nerve that is often characterized by vision loss, results in the loss of retinal ganglion cells. There are many types of optic neuropathy, including ischemic optic neuropathy, optic neuritis, compressive optic neuropathy, infiltrative optic neuropathy, and traumatic optic neuropathy. Nutritional deficiencies and / or vitamin B 12 Deficiency may also result in nutritional optic neuropathy. Exposure to ethylene glycol, methanol, ethambutol, amiodarone, tobacco, or certain drugs, such as chloramphenicol or digitalis, may result in toxic optic neuropathy. Certain forms of optic neuropathy, including Leber's hereditary optic neuropathy (LHON), dominant optic atrophy, Beer's syndrome, and Berk-Tabatznik syndrome, may be inherited. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered as a method of reducing neuronal or cellular damage in the eye of a host in need thereof associated with optic neuropathy.

[0276] Additional non-limiting examples of neurodegenerative diseases associated with the eye include lattice dystrophy, retinitis pigmentosa, age-related macular degeneration (wet or dry), photoreceptor degeneration associated with wet or dry age-related macular degeneration, and optic nerve drusen.

[0277] Integrated or adjunctive therapy with minimally invasive glaucoma surgery (MIGS) Minimally invasive glaucoma surgery (MIGS) has become a revolutionary technique in the evolution of glaucoma surgery. Because glaucoma is a disease in which the optic nerve is damaged primarily due to elevated IOP, the goal of glaucoma surgery is to lower IOP and prevent or reduce damage to the optic nerve.

[0278] Standard glaucoma surgery is still considered major surgery and requires trabeculectomy, an Express shunt, or external tube shunts such as Ahmed-style, Molteno-style, and Baerveldt-style valve implants. While these procedures are often effective in lowering intraocular pressure and halting the progression of glaucoma, they are associated with many potential complications, including double vision, severe eye infections, exposure of the drainage implant, corneal swelling, and excessively low IOP.

[0279] According to Saheb and Ahmed, minimally (or minimally) invasive glaucoma surgery has five favorable qualities: 1. ab interno and / or ab externo approach via a clear corneal incision, which may eliminate the need for a conjunctival incision; 2. Minimally traumatic procedure to target tissue, 3. IOP-lowering effect that justifies the approach; 4. A high safety profile avoiding serious complications compared to other glaucoma surgeries and given lower potential for ocular hypotension, and 5. Effective recovery with minimal impact on the patient's quality of life, Refers to a group of surgical procedures that share a common theme.

[0280] MIGS procedures have been developed in recent years to reduce some of the complications of most standard glaucoma surgeries. Thus, in certain embodiments, ophthalmic formulations of (rev)cromakalim are used as an adjunct in combination with minimally invasive glaucoma surgery (MIGS).

[0281] MIGS aims to achieve lower IOP in patients with glaucoma through a less invasive surgical procedure, ideally with medication-sparing benefits. MIGS procedures work by using microscopic instruments and small incisions, allowing for outflow control and are often performed at the time of cataract surgery. They reduce complication rates, but some effectiveness comes at the expense of improved safety (Pillunat, LE, et al., Clin Ophthalmol. 2017; 11: 1583-1600).

[0282] MIGS surgery falls into several categories: 1. Trabecular bypass surgery (i.e., angle-based devices and / or subconjunctival shunt devices), 2. Microtrabeculectomy (a miniaturized form of trabeculectomy), 3. Complete internal or suprachoroidal shunt, and 4. A kinder, gentler form of laser photocoagulation, It is classified as follows.

[0283] Trabecular meshwork surgery (trabeculectomy) involves dissecting the trabecular meshwork with a small device under high-powered microscopic control using a special contact lens on the eye. This is done without damaging other tissues in the intraocular drainage pathway. The trabecular meshwork may be disrupted (Trabectome, Trab360, or OMNI Surgical System) or bypassed using a small snorkel-like device (iStent) or a plug-shaped stent device (iStent Inject). Both procedures are FDA-approved but generally do not lower intraocular pressure as much, making them useful in early to moderate stages of glaucoma. With these devices, the resistance of the trabecular meshwork is obviated, leaving distal outflow capacity and episcleral venous pressure as the primary limitations to further aqueous humor drainage. In certain embodiments, ophthalmic formulations of (rev)cromakalim are used as additives in combination with Trabectome or Trab360 and / or iStent / iStent Inject to treat glaucoma by additively lowering IOP via increasing distal outflow or reducing episcleral venous pressure before or after treatment in acute or chronic use settings.

[0284] Microtrabeculectomy works by inserting a small, microscopic tube into the eye and draining fluid from inside the eye under the outer membrane (conjunctiva) of the eye. Xen Gel Stent and Preserflo are two new devices that may make trabeculectomy procedures safer. Results have shown superior pressure reduction with improved safety compared to trabeculectomy in studies conducted outside the United States. In certain embodiments, the compounds of the present invention are used as part of a protocol using Xen Gel Stent and / or Preserflo to treat glaucoma by additively lowering IOP via increasing distal outflow or reducing episcleral venous pressure before or after the procedure in acute or chronic use settings.

[0285] Suprachoroidal shunts, including the Gold Micro shunt, iStent Supra, Aquashunt, and STARflo, work by using a small tube with a very small internal opening to connect the anterior segment of the eye to the suprachoroidal space between the retina and the wall of the eye, increasing drainage from the eye. This procedure has relatively few serious complications and reduces pressure enough to be useful in even moderate glaucoma. In certain embodiments, ophthalmic formulations of (rev)cromakalim are used in combination with a suprachoroidal shunt procedure to treat glaucoma by additively lowering IOP via increasing distal outflow or reducing episcleral venous pressure before or after the procedure in acute or chronic use settings.

[0286] Trabecular meshwork bypass stents and shunts are investigational devices that function to dilate Schlemm's canal. These procedures promote aqueous humor inflow into Schlemm's canal by inserting a shunt (Eyepass Glaucoma Implant, GMP Companies, Inc., Fort Lauderdale, Florida), inserting a stent into Schlemm's canal itself (iStent, Glaukos Corp., Laguna Hills, California), or dilating Schlemm's canal using a viscoelastic material (OMNI Surgical System, Sight Sciences, Menlo Park, California). Other devices, such as the Solx Gold Micro-Shunt (OccuLogix, Inc., Mississauga, Ontario, Canada), divert aqueous humor into the suprachoroidal space. In certain embodiments, ophthalmic formulations of (rev)cromakalim are used in combination with trabecular meshwork bypass stent or shunt procedures to treat glaucoma by additively lowering IOP via increased distal outflow or reduced episcleral venous pressure before or after the procedure in acute or chronic use settings.

[0287] Selective laser trabeculoplasty (SLT) is used during the management of glaucoma to help lower IOP. Since the implementation of the LiGHT study, SLT is now more frequently used as a first-line treatment to help lower IOP, effectively acting at the level of the trabecular meshwork to improve outflow. In certain embodiments, ophthalmic formulations of (rev)cromakalim are used together with or in addition to SLT to treat glaucoma by additively lowering IOP via increasing distal outflow and / or reducing episcleral venous pressure before or after treatment in acute or chronic use settings.

[0288] Laser photocoagulation was previously reserved for advanced glaucoma that had not progressed despite trabeculectomy or tube shunting. Endoscopic cyclophotocoagulation and micropulse diode cyclophotocoagulation are two recent advancements in the use of laser photocoagulation and have proven useful in cases where glaucoma has not yet progressed. In certain embodiments, ophthalmic formulations of (rev)cromakalim are used in endoscopic cyclophotocoagulation and micropulse cyclophotocoagulation protocols to treat glaucoma by additively lowering IOP via increasing distal outflow and / or reducing episcleral venous pressure before or after the procedure in acute or chronic use settings.

[0289] In recent years, endoscopic cyclophotocoagulation (ECPC) has become a widely accepted and popular treatment for refractory and pediatric glaucoma, both as an adjunct to cataract surgery in both medically controlled and uncontrolled glaucoma, and in combination with phacoemulsification with intraocular lens placement. Following lens removal and intraocular lens implantation, endoscopic cyclophotocoagulation is performed by inserting an endolaser unit through the cataract incision, across the anterior segment, and into the posterior chamber on the nasal side of the eye. Laser energy is applied to the ciliary processes to destroy the ciliary epithelial cells that produce aqueous humor. In certain embodiments, ophthalmic formulations of (rev)cromakalim are used in endoscopic cyclophotocoagulation protocols to treat glaucoma by additively lowering IOP via increasing distal outflow and / or reducing episcleral venous pressure before or after the procedure in acute or chronic use settings.

[0290] Micropulse cyclophotocoagulation delivers the laser in short bursts, allowing the surgeon to target specific areas of the ciliary body while allowing the tissue time to cool between bursts, minimizing damage. Both the MicroPulse P3 probe and the new Cyclo G6 Glaucoma Laser System (Iridex) have been successfully used in retinal diseases, demonstrating excellent safety and efficacy rates. In certain embodiments, ophthalmic formulations of (rev)cromakalim are used in micropulse cyclophotocoagulation surgical protocols to treat glaucoma by additively lowering IOP via increasing distal outflow and / or reducing episcleral venous pressure before or after the procedure in acute or chronic use settings.

[0291] Other devices include gonioscopy-assisted transluminal trabeculotomy (GATT), Kafuk Dual Blade, Ab interno trabeculoplasty, and Hydrus Microstent, iStent Supra, Xen Glaucoma Treatment System, and InnFocus MicroShunt. In certain embodiments, ophthalmic formulations of (rev)cromakalim are used in the surgical protocols of these devices to treat the above-mentioned glaucoma.

[0292] Laser trabeculoplasty, including selective laser trabeculoplasty (SLT), argon laser trabeculoplasty (ALT), excimer laser trabeculoplasty, and micropulse laser trabeculoplasty (MLT), is a surgical laser procedure that serves to reduce resistance in the trabecular meshwork by ablating cells of the trabecular meshwork and improving outflow in a manner similar to other forms of trabeculoplasty and certain MIGS devices. In certain embodiments, excimer laser trabeculoplasty is used as an additive in combination with laser trabeculoplasty to treat glaucoma by additively lowering IOP via increased distal outflow or reduced episcleral venous pressure before or after the procedure in an acute or chronic use setting.

[0293] In certain embodiments, ophthalmic formulations of (rev)cromakalim are used as second-line therapy to prostaglandin analogs such as latanoprost (Xalatan), bimatoprost (Lumigan), travoprost (Travatan or Travatan Z), latanoprost bunod (Vyzulta), or tafluprost (Zioptan), and as an adjunct to minimally invasive glaucoma surgery (MIGS) described herein. In further embodiments, the MIGS is trabeculectomy. In further embodiments, the MIGS is microtrabeculectomy. In further embodiments, the MIGS is a suprachoroidal shunt. In further embodiments, the MIGS is a trabecular bypass stent or shunt. In further embodiments, the MIGS is selective laser trabeculoplasty (SLT). In further embodiments, the MIGS is laser photocoagulation. In further embodiments, the MIGS is endoscopic cyclophotocoagulation. In a further embodiment, the MIGS is laser trabeculoplasty.

[0294] In certain embodiments, ophthalmic formulations of (rev)cromakalim are used as a second-line therapy to latanoprost (Xalatan) and as an adjunct to the minimally (or minimally) invasive glaucoma surgeries described herein. In further embodiments, the MIGS is trabeculectomy. In further embodiments, the MIGS is microtrabeculectomy. In further embodiments, the MIGS is a suprachoroidal shunt. In further embodiments, the MIGS is a trabecular bypass stent or shunt. In further embodiments, the MIGS is selective laser trabeculoplasty (SLT). In further embodiments, the MIGS is laser photocoagulation. In further embodiments, the MIGS is endoscopic cyclophotocoagulation. In further embodiments, the MIGS is laser trabeculoplasty.

[0295] In certain embodiments, ophthalmic formulations of (rev)cromakalim are used as second-line therapy to alpha adrenergic agonists such as brimonidine (Alphagan™), epinephrine, dipivefrin (Propine™), or apraclonidine (Lopidine™), and as an adjunct to minimally invasive glaucoma surgery (MIGS) as described herein. In further embodiments, the MIGS is trabeculectomy. In further embodiments, the MIGS is microtrabeculectomy. In further embodiments, the MIGS is a suprachoroidal shunt. In further embodiments, the MIGS is a trabecular bypass stent or shunt. In further embodiments, the MIGS is selective laser trabeculoplasty (SLT). In further embodiments, the MIGS is laser photocoagulation. In further embodiments, the MIGS is endoscopic cyclophotocoagulation. In a further embodiment, the MIGS is laser trabeculoplasty.

[0296] In certain embodiments, ophthalmic formulations of (lev)cromakalim are used as second-line therapy to beta-blockers such as timolol, betaxolol, levobunolol, metipranolol, or carteolol, and as an adjunct to minimally (or minimally) invasive glaucoma surgery (MIGS) as described herein.

[0297] In a further embodiment, the MIGS is a trabeculectomy. In a further embodiment, the MIGS is a microtrabeculectomy. In a further embodiment, the MIGS is a suprachoroidal shunt. In a further embodiment, the MIGS is a trabecular bypass stent or shunt. In a further embodiment, the MIGS is selective laser trabeculoplasty (SLT). In a further embodiment, the MIGS is laser photocoagulation. In a further embodiment, the MIGS is endoscopic cyclophotocoagulation. In a further embodiment, the MIGS is laser trabeculoplasty. In a further embodiment, the MIGS is a trabeculectomy. In a further embodiment, the MIGS is a microtrabeculectomy. In a further embodiment, the MIGS is a suprachoroidal shunt. In a further embodiment, the MIGS is a trabecular bypass stent or shunt. In a further embodiment, the MIGS is selective laser trabeculoplasty (SLT). In a further embodiment, the MIGS is laser photocoagulation. In a further embodiment, the MIGS is endoscopic cyclophotocoagulation.In a further embodiment, the MIGS is laser trabeculoplasty.

[0298] In certain embodiments, ophthalmic formulations of (rev)cromakalim are used as second-line therapy to ROCK inhibitors such as ripasudil, netarsudil (Rhopressa), fasudil, RKI-1447, GSK429286A, or Y-30141, and as an adjunct to minimally invasive glaucoma surgery (MIGS) described herein. In further embodiments, the MIGS is trabeculectomy. In further embodiments, the MIGS is microtrabeculectomy. In further embodiments, the MIGS is a suprachoroidal shunt. In further embodiments, the MIGS is a trabecular bypass stent or shunt. In further embodiments, the MIGS is selective laser trabeculoplasty (SLT). In further embodiments, the MIGS is laser photocoagulation. In further embodiments, the MIGS is endoscopic cyclophotocoagulation. In further embodiments, the MIGS is laser trabeculoplasty.

[0299] In certain embodiments, ophthalmic formulations of (rev)cromakalim are used as a second-line therapy to a second ATP-sensitive potassium channel opener, such as minoxidil, diazoxide, nicorandil, or pinacidil, and as an adjunct to minimally (or minimally) invasive glaucoma surgery (MIGS) described herein. In further embodiments, the MIGS is trabeculectomy. In further embodiments, the MIGS is microtrabeculectomy. In further embodiments, the MIGS is a suprachoroidal shunt. In further embodiments, the MIGS is a trabecular bypass stent or shunt. In further embodiments, the MIGS is selective laser trabeculoplasty (SLT). In further embodiments, the MIGS is laser photocoagulation. In further embodiments, the MIGS is endoscopic cyclophotocoagulation. In further embodiments, the MIGS is laser trabeculoplasty.

[0300] In certain embodiments, ophthalmic formulations of (rev)cromakalim are used as second-line therapy to carbonic anhydrase inhibitors such as dorzolamide (Trusopt™), brinzolamide (Azopt™), acetazolamide (Diamox™), or methazolamide (Neptazane™), and as an adjunct to minimally invasive glaucoma surgery (MIGS) as described herein. In further embodiments, the MIGS is trabeculectomy. In further embodiments, the MIGS is microtrabeculectomy. In further embodiments, the MIGS is a suprachoroidal shunt. In further embodiments, the MIGS is a trabecular bypass stent or shunt. In further embodiments, the MIGS is selective laser trabeculoplasty (SLT). In further embodiments, the MIGS is laser photocoagulation. In further embodiments, the MIGS is endoscopic cyclophotocoagulation. In a further embodiment, the MIGS is laser trabeculoplasty.

[0301] Other exemplary eye disorders Orbital tumors are benign or malignant space-occupying lesions of the orbit that often cause ocular heterotopia, movement disorders, diplopia, visual field defects, and sometimes complete vision loss. Orbital tumors are often removed by surgery, making drug therapy an advantageous treatment option. In certain embodiments, an effective amount of an ophthalmic formulation of (rev)cromakalim is administered to treat or alleviate orbital tumors. In certain embodiments, (rev)cromakalim is administered topically once daily, twice daily, three times daily, or more. In certain embodiments, (rev)cromakalim is administered before or after surgery to remove or alleviate orbital tumors.

[0302] Episcleral / orbital venous vasculitis is inflammation of the blood vessel wall. Clinical features of ocular vasculitis can vary depending on the location and distribution of affected vessels, from conjunctivitis, episcleritis, scleritis, marginal ulcerative keratitis, exophthalmos, retinal vasculitis, orbititis, to uveitis. In certain embodiments, an effective amount of an ophthalmic formulation of (rev)cromakalim is administered to treat episcleral / orbital venous vasculitis. In certain embodiments, the (rev)cromakalim formulation is administered as a topical drop.

[0303] A carotid-cavernous fistula is an abnormal connection between the arteries in the neck and the venous network at the back of the eye. The fistula can increase pressure within the cavernous sinus, compressing the cranial nerves surrounding the sinus. This compression can impair nerve function controlling eye movement. A carotid-cavernous fistula can be direct or indirect. Direct carotid-cavernous fistulas are often caused by an accident or injury that tears the carotid artery wall, while indirect carotid-cavernous fistulas often occur without warning and are associated with hypertension, arteriosclerosis, pregnancy, and connective tissue disorders. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered to treat a carotid-cavernous fistula.

[0304] A dural cavernous shunt is a vascular connection in which blood flows through a small meningeal branch of the carotid artery and enters the venous circulation near the cavernous sinus. In many cases, the disorder is congenital, and the onset of clinical abnormalities may be associated with the occurrence of intracranial venous thrombosis. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered to treat a dural cavernous shunt.

[0305] Orbital varicose veins are vascular hamartomas typified by a thin-walled, distensible vascular plexus with low pressure and low flow that intermingles with normal orbital vessels. Most patients experience postural proptosis with head-down positioning and intermittent proptosis exacerbated by coughing, straining, the Valsalva maneuver, or jugular vein compression. In certain embodiments, an ophthalmic formulation of (lev)cromakalim is administered to treat orbital varicose veins.

[0306] Branch retinal vein occlusion (BRVO) occurs when a branch of the retinal vein becomes blocked, causing blood and fluid to spill over into the retina. Risk factors for BRVO include diabetes, elevated IOP, and high blood pressure. This fluid can cause the macula to swell and affect central vision. Ultimately, the lack of blood circulation can cause nerve cells in the eye to die, potentially resulting in vision loss. In certain embodiments, an effective amount of an ophthalmic formulation of (rev)cromakalim is administered to treat branch retinal vein occlusion (BRVO). In certain embodiments, the (rev)cromakalim formulation is administered as topical drops, given once daily, twice daily, three times daily, or more frequently.

[0307] Non-arteritic anterior ischemic optic neuropathy (NAION) refers to the loss of blood flow to the optic nerve due to impaired blood circulation at the optic nerve head. Non-arteritic anterior ischemic optic neuropathy is associated with diabetes, hypertension, atherosclerosis, small optic nerve, elevated IOP, and sleep apnea. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered to treat non-arteritic anterior ischemic optic neuropathy. In certain embodiments, the (lev)cromakalim formulation is administered as topical drops, given once daily, twice daily, three times daily, or more times.

[0308] In an additional aspect of the invention, ophthalmic formulations of (lev)cromakalim are used to treat selected ocular disorders, as described below.

[0309] Graves' ophthalmopathy or Graves' orbitopathy (or thyroid eye disease or thyroid-associated orbitopathy) is an autoimmune inflammatory disorder of the orbit and periorbital tissues, and typical symptoms of this disease include upper eyelid retraction, eyelid lag, swelling, and bulging eyes. These disorders are orbital autoimmune disorders caused by hyperthyroidism. An effective amount of an ophthalmic formulation of (rev)cromakalim can be administered to treat Graves' ophthalmopathy, Graves' orbitopathy, or thyroid-associated orbitopathy. The compound can be administered in any manner that achieves the desired effect, including as topical drops used as needed to reduce swelling and redness. In certain embodiments, an ophthalmic formulation of (rev)cromakalim is given in combination with a corticosteroid or an immunosuppressant drug (rituximab or mycophenolate).

[0310] Cavernous sinus thrombosis is the formation of a blood clot in the cavernous sinus, a cavity at the base of the brain that drains deoxygenated blood from the brain and returns it to the heart. It is a rare disorder and can be classified into two types: septic cavernous thrombosis and aseptic cavernous thrombosis. Its cause is often secondary to infection in the nose, sinuses, ears, or teeth. A common disorder secondary to cavernous sinus lesions is superior ophthalmic vein thrombosis, a rare orbital pathology that can present with sudden-onset exophthalmos, conjunctival hyperemia, and visual impairment. In certain embodiments, an effective amount of an ophthalmic formulation of (lev)cromakalim is administered to treat cavernous sinus thrombosis or superior ophthalmic vein thrombosis. In certain embodiments, the effective amount is administered in combination with or alternating with an antibiotic, heparin, or steroid. In one embodiment, the compound is administered orally and is given at least once a day, twice a day, three times a day, or more times as needed.

[0311] Central retinal vein occlusion, also known as CRVO, is a condition in which the main vein that drains blood from the retina is partially or completely blocked. This can lead to blurred vision and other eye-related problems. Risk factors for CRVO include diabetes, elevated IOP, and high blood pressure. This fluid can cause the macula to swell and affect central vision. Ultimately, the lack of blood circulation can cause nerve cells in the eye to die, potentially resulting in vision loss. In certain embodiments, an effective amount of an ophthalmic formulation of (rev)cromakalim is administered to treat central retinal vein occlusion. In certain embodiments, the compound is administered as topical drops, given once daily, twice daily, or three times daily.

[0312] V. Combination Therapy In certain embodiments, an effective amount of the (rev)cromakalim ophthalmic formulation described herein is administered to a host in need of treatment in combination with an effective amount of a second active agent to treat the patient's ocular disorder.

[0313] Non-limiting examples of exemplary additional active agents that may be used in accordance with the present invention include, but are not limited to, effective amounts of the following: 1) Latanoprost or latanoprostene bunod, 2) cyclosporine, 3) nitric oxide donors, including, but not limited to, NO-donating prostaglandin analog eye drops (NCX-470), NO-donating PDE5 inhibitors (NCX-1728), fluticanose propionate nanocrystal suspension (NCX-4251), or sodium nitroprusside (SNP); 4) Prostaglandin analogues such as latanoprost (Xalatan), bimatoprost (Lumigan), travoprost (Travatan or Travatan Z), or tafluprost (Zioptan); 5) alpha-2 adrenergic agonists such as brimonidine (Alphagan™), epinephrine, dipivefrin (Propine™), or apraclonidine (Lopidine™); 6) β-blockers such as timolol, levobunolol, metipranolol, or carteolol; 7) ROCK inhibitors such as ripasudil, netarsudil (Rhopressa™), fasudil, RKI-1447, GSK429286A, or Y-30141; 8) Prostaglandin and ROCK inhibitor combination products such as Rocklatan™ (latanoprost and netarsudil); 9) a second ATP-sensitive potassium channel opener such as minoxidil, diazoxide, nicorandil, or pinacidil; 10) Carbonic anhydrase inhibitors such as dorzolamide (Trusopt™), brinzolamide (Azopt™), acetazolamide (Diamox™), or methazolamide (Neptazane™); 11) Combination products of carbonic anhydrase inhibitors and beta-blockers, such as Cosopt™ (dorzolamide and timolol), or 12) Anti-VEGF inhibitors such as bevacizumab (Avastin™), ranibizumab (Lucentis™), aflibercept (Eylea™), and brolucizumab (Beovu™).

[0314] Embodiments of the present invention 1. An ophthalmic topical formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof having a concentration of between about 0.01 mM and about 5 mM, wherein the ophthalmic topical formulation is stable at ambient conditions for at least 4 months; and a. polyol, b. polyethoxylated furanose fatty acid esters, c. Nonionic triblock copolymers (poloxamers) with a central hydrophobic chain of polyoxypropylene flanked by two hydrophilic chains of polyoxyethylene; d. polymeric alkyl or aryl polyols; e. ethoxylated glycerol esters, f. polymeric lactams, g. hydroxyalkyl cellulose, h. Polyacrylic acid and hydrophobic C 10 ~C 30 an oil-in-water polymeric emulsifier that is a block copolymer with an alkyl acrylate; and i. ethoxylated alkylphenols, A topical ophthalmic formulation comprising three or more ingredients selected from the group consisting of: in an aqueous formulation having a pH of about 6 to about 8.

[0315] 2. The topical ophthalmic formulation of embodiment 1, wherein the topical ophthalmic formulation comprises four or more ingredients.

[0316] 3. The topical ophthalmic formulation of embodiment 1, wherein the topical ophthalmic formulation comprises five or more ingredients.

[0317] 4. The topical ophthalmic formulation of embodiment 1, wherein the topical ophthalmic formulation comprises six or more ingredients.

[0318] 5. The topical ophthalmic formulation of any one of embodiments 1-4, wherein the composition comprises a polyol.

[0319] 6. The topical ophthalmic formulation of embodiment 5, wherein the polyol is an alkyl polyol.

[0320] 7. The topical ophthalmic formulation of embodiment 6, wherein the alkyl polyol is a triol.

[0321] 8. The topical ophthalmic formulation of embodiment 6, wherein the alkyl polyol is a sugar alcohol.

[0322] 9. The ophthalmic topical formulation of embodiment 7, wherein the triol is glycerin.

[0323] 10. The ophthalmic topical formulation of embodiment 6, wherein the alkyl polyol is mannitol.

[0324] 11. The topical ophthalmic formulation of any one of embodiments 1-10, wherein the composition comprises a polyethoxylated furanose fatty acid ester.

[0325] 12. The ophthalmic topical formulation of embodiment 11, wherein the polyethoxylated furanose fatty acid ester is polysorbate 80.

[0326] 13. The topical ophthalmic formulation of any of embodiments 1-12, wherein the composition comprises a poloxamer.

[0327] 14. The topical ophthalmic formulation of embodiment 13, wherein the poloxamer is poloxamer 407 or Pluronic 127.

[0328] 15. The topical ophthalmic formulation of any of embodiments 1-14, wherein the composition comprises a polymeric alkyl or aryl polyol.

[0329] 16. The ophthalmic topical formulation of embodiment 15, wherein the polymeric alkyl or aryl polyol is tyloxapol.

[0330] 17. The topical ophthalmic formulation of any of embodiments 1-16, wherein the composition comprises an ethoxylated glycerol ester.

[0331] 18. The topical ophthalmic formulation of embodiment 17, wherein the ethoxylated glycerol ester is Kolliphor™ or Cremophor™.

[0332] 19. The topical ophthalmic formulation of embodiment 18, wherein Kolliphor™ or Cremophor™ is selected from the group consisting of Kolliphor™ ELP, Kolliphor™ RH 40, Kolliphor™ HS 15.

[0333] 20. The topical ophthalmic formulation of embodiment 19, wherein Kolliphor™ or Cremophor™ is Kolliphor™ ELP.

[0334] 21. The topical ophthalmic formulation of embodiment 19, wherein Kolliphor™ or Cremophor™ is Kolliphor™ RH 40.

[0335] 22. The topical ophthalmic formulation of embodiment 19, wherein the Kolliphor™ or Cremophor™ is Kolliphor™ HS 15.

[0336] 23. The topical ophthalmic formulation of any of embodiments 1-22, wherein the composition comprises a polymeric lactam.

[0337] 24. The topical ophthalmic formulation of embodiment 23, wherein the polymeric lactam is PVP.

[0338] 25. The topical ophthalmic formulation of embodiment 24, wherein the PVP is selected from the group of PVP K-30 and PVP K-90.

[0339] 26. The topical ophthalmic formulation of embodiment 25, wherein the PVP is PVP K-30.

[0340] 27. The topical ophthalmic formulation of embodiment 25, wherein the PVP is PVP K-90.

[0341] 28. The topical ophthalmic formulation of any of embodiments 1-27, wherein the composition comprises a hydroxyalkyl cellulose.

[0342] 29. The ophthalmic topical formulation of embodiment 28, wherein the hydroxyalkyl cellulose is hypromellose.

[0343] 30. The composition is a mixture of polyacrylic acid and hydrophobic C 10 ~C 30 30. The topical ophthalmic formulation of any of embodiments 1-29, comprising an oil-in-water polymeric emulsifier which is a block copolymer with an alkyl acrylate.

[0344] 31. Polyacrylic acid and hydrophobic C 10 ~C 30The ophthalmic topical formulation of embodiment 30, wherein the oil-in-water polymeric emulsifier of a block copolymer with an alkyl acrylate is Premulen.

[0345] 32. The topical ophthalmic formulation of embodiment 31, wherein Premulen is Premulen™ TR-1 or Premulen™ TR-2.

[0346] 33. The topical ophthalmic formulation of any of embodiments 1-32, wherein the composition comprises an ethoxylated alkylphenol.

[0347] 34. The ophthalmic formulation of embodiment 33, wherein the ethoxylated alkylphenol is octoxynol.

[0348] 35. The topical ophthalmic formulation of embodiment 34, wherein the octoxynol is octoxynol-40.

[0349] 36. The topical ophthalmic formulation of any of embodiments 1-35, wherein the formulation is stable at ambient conditions for at least 5 months.

[0350] 37. The topical ophthalmic formulation of embodiment 36, wherein the formulation is stable at ambient conditions for at least 6 months.

[0351] 38. The topical ophthalmic formulation of embodiment 37, wherein the formulation is stable at ambient conditions for at least 7 months.

[0352] 39. The topical ophthalmic formulation of any of embodiments 1-38, wherein the formulation is an aqueous formulation with a pH of about 6.5 to about 7.5.

[0353] 40. The topical ophthalmic formulation of any of embodiments 1-39, wherein the concentration of (rev)cromakalim is between about 0.05 mM and about 5 mM.

[0354] 41. The topical ophthalmic formulation of any of embodiments 1-40, wherein the concentration of (rev)cromakalim is between about 0.5 mM and about 5 mM.

[0355] 42. The ophthalmic topical formulation of any of embodiments 1-41, wherein the concentration of (rev)cromakalim is between about 1 mM and about 5 mM.

[0356] 43. The topical ophthalmic formulation of any of embodiments 1-42, wherein the concentration of (rev)cromakalim is between about 1.5 mM and about 5 mM.

[0357] 44. The ophthalmic topical formulation of any of embodiments 1-43, wherein the concentration of (rev)cromakalim is between about 2 mM and about 5 mM.

[0358] 45. The ophthalmic topical formulation of any of embodiments 1-44, wherein the concentration of (rev)cromakalim is between about 2.5 mM and about 5 mM.

[0359] 46. ​​The ophthalmic topical formulation of any of embodiments 1-45, wherein the concentration of (rev)cromakalim is between about 3 mM and about 5 mM.

[0360] 47. The topical ophthalmic formulation of any of embodiments 1-46, wherein the concentration of (rev)cromakalim is between about 3.5 mM and about 5 mM.

[0361] 48. The topical ophthalmic formulation of any of embodiments 1-47, wherein the concentration of (rev)cromakalim is between about 4 mM and about 5 mM.

[0362] 49. A topical ophthalmic formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof having a concentration of between about 0.01 mM and about 5 mM, which is stable at ambient conditions for at least 5 months, and which comprises three or more ingredients selected from the group consisting of glycerin, polysorbate 80, poloxamer 407 or Pluronic™ F127, tyloxapol, Kolliphor™ ELP, Kolliphor™ RH 40, Kolliphor™ HS 15, hypromellose, PVP, Premulen™ TR-1 or Premulen™ TR-2, and octoxynol-40, in an aqueous formulation having a pH of about 6 to about 8.

[0363] 50. The topical ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of three or more ingredients selected from the group consisting of glycerin, polysorbate 80, poloxamer 407 or Pluronic™ F127, tyloxapol, Kolliphor™ ELP, Kolliphor™ RH 40, Kolliphor™ HS 15, hypromellose, PVP, Premulen™ TR-1 or Premulen™ TR-2, and octoxynol-40, in an aqueous formulation having a pH of about 6 to about 8.

[0364] 51. The topical ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ RH 40, Octoxynol-40, and Premulen™ TR-2 in an aqueous formulation having a pH of about 6 to about 8.

[0365] 52. The topical ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of Kolliphor® RH 40, Kolliphor® HS 15, and PVP in an aqueous formulation having a pH of about 6 to about 8.

[0366] 53. The ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor™ ELP, polysorbate 80, and hypromellose in an aqueous formulation having a pH of about 6 to about 8.

[0367] 54. The ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, Polysorbate 80, and PVP in an aqueous formulation having a pH of about 6 to about 8.

[0368] 55. The ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose in an aqueous formulation having a pH of about 6 to about 8.

[0369] 56. The ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor® RH 40, Premulen® TR-2, and PVP in an aqueous formulation having a pH of about 6 to about 8.

[0370] 57. The ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ HS 15, Premulen™ TR-2, polysorbate 80, octoxynol-40, and poloxamer 407 in an aqueous formulation having a pH of about 6 to about 8.

[0371] 58. The ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists of Polysorbate 80, Kolliphor™ ELP, and PVP in an aqueous formulation having a pH of about 6 to about 8.

[0372] 59. The ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, and poloxamer 407 in an aqueous formulation having a pH of about 6 to about 8.

[0373] 60. The ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, PVP, and poloxamer 407 in an aqueous formulation having a pH of about 6 to about 8.

[0374] 61. The ophthalmic formulation of embodiment 49, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose in an aqueous formulation having a pH of about 6 to about 8.

[0375] 62. A topical ophthalmic formulation comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP), poloxamer 407, mannitol, water, and phosphate buffer, and having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 0.01 mM and about 5 mM, and stable for at least 5 months at ambient conditions, and having a pH of about 6 to about 8.

[0376] 63. The topical ophthalmic formulation of embodiment 62, having a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof of between about 0.05 mM and about 5 mM.

[0377] 64. The ophthalmic topical formulation of embodiment 62 or 63, having a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof of between about 0.5 mM and about 5 mM.

[0378] 65. The topical ophthalmic formulation of any one of embodiments 62-64, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 1 mM and about 5 mM.

[0379] 66. The topical ophthalmic formulation of any one of embodiments 62-65, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 1.5 mM and about 5 mM.

[0380] 67. The ophthalmic formulation of any one of embodiments 62-66, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 2 mM and about 5 mM.

[0381] 68. The ophthalmic formulation of any one of embodiments 62-67, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 2.5 mM and about 5 mM.

[0382] 69. The ophthalmic formulation of any one of embodiments 62-68, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 3 mM and about 5 mM.

[0383] 70. The ophthalmic formulation of any one of embodiments 62-69, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 3.5 mM and about 5 mM.

[0384] 71. The ophthalmic formulation of any one of embodiments 62-70, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 4 mM and about 5 mM.

[0385] 72. The ophthalmic formulation of any one of embodiments 62-71, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol in an aqueous formulation having a pH of about 6 to about 8.

[0386] 73. The ophthalmic formulation of any one of embodiments 62-72, wherein the ophthalmic formulation consists of Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol in an aqueous formulation having a pH of about 6 to about 8.

[0387] 74. The ophthalmic formulation of any one of embodiments 1-73, wherein the ophthalmic formulation optionally comprises benzalkonium chloride (BAK) and optionally a pH adjuster.

[0388] 75. The ophthalmic formulation of any one of embodiments 62-71, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, mannitol, and BAK in an aqueous formulation having a pH of about 6 to about 8.

[0389] 76. The ophthalmic formulation of any one of embodiments 62-71, wherein the ophthalmic formulation consists of Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, mannitol, and BAK in an aqueous formulation having a pH of about 6 to about 8.

[0390] 77. A topical ophthalmic formulation comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, and having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof up to about 5 mM, and stable for at least 5 months at ambient conditions, and having a pH of about 6.5.

[0391] 78. A topical ophthalmic formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof having a concentration of between about 0.01 mM and about 5 mM, comprising Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, and stable for at least 5 months at ambient conditions, and having a pH of about 6.5.

[0392] 79. An aqueous ophthalmic topical formulation having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof between about 0.05 mM and about 5 mM, having a pH of about 6 to about 8, and stable at ambient conditions for at least 5 months, and comprising glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, and mannitol.

[0393] 80. The ophthalmic formulation of embodiment 78 or 79, wherein the ophthalmic formulation optionally comprises BAK and a pH adjuster.

[0394] 81. The ophthalmic formulation of embodiment 80, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, mannitol, optionally benzalkonium chloride, and optionally a pH adjuster, in an aqueous formulation having a pH of about 6 to about 8.

[0395] 82. The topical ophthalmic formulation of embodiment 80, wherein the ophthalmic formulation consists of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, mannitol, and BAK in an aqueous formulation having a pH of about 6 to about 8.

[0396] 83. An ophthalmic topical formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof having a concentration of between about 0.05 mM and about 5 mM, having a pH of about 6 to about 8 and stable at ambient conditions for at least 4 months, and comprising Kolliphor™ ELP, polysorbate 80, poloxamer 407, PVP K-30, mannitol, and optionally a pH adjuster.

[0397] 84. The topical ophthalmic formulation of embodiment 83, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, poloxamer 407, PVP K-30, and mannitol in an aqueous formulation having a pH of about 6 to about 8.

[0398] 85. The topical ophthalmic formulation of embodiment 83, wherein the ophthalmic formulation consists of Kolliphor™ ELP, polysorbate 80, poloxamer 407, PVP K-30, and mannitol in an aqueous formulation having a pH of about 6 to about 8.

[0399] 86. The topical ophthalmic formulation of any one of embodiments 1-85, wherein the phosphate buffer is selected from dibasic sodium phosphate and monobasic sodium phosphate.

[0400] 87. A pharmaceutical composition comprising (lev)cromakalim or a pharmaceutically acceptable salt thereof, having a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof between about 0.05 mM and about 5 mM, and stable at ambient conditions for at least 4 months; and a. polyol, b. polyethoxylated furanose fatty acid esters, c. A nonionic triblock copolymer with a central hydrophobic chain of polyoxypropylene flanked by two hydrophilic chains of polyoxyethylene; d. polymeric alkyl or aryl polyols; e. ethoxylated glycerol esters, f. polymeric lactams, g. hydroxyalkyl cellulose, h. Polyacrylic acid and hydrophobic C 10 ~C 30 an oil-in-water polymeric emulsifier that is a block copolymer with an alkyl acrylate; and i. ethoxylated alkylphenols, A pharmaceutical composition comprising three or more ingredients selected from the group consisting of the above in an aqueous formulation having a pH of about 6 to about 8.

[0401] 88. A topical formulation for eye drops comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, wherein the formulation has a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof between about 0.01 mM and about 5 mM, and is stable at ambient conditions for at least 5 months, and has a pH of about 6 to about 8.

[0402] 89. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 0.05 mM to 5 mM.

[0403] 90. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 0.1 mM to 5 mM.

[0404] 91. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 0.5 mM to 5 mM.

[0405] 92. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 1 mM to 5 mM.

[0406] 93. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 1.5 mM to 5 mM.

[0407] 94. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 2 mM to 5 mM.

[0408] 95. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 2.5 mM to 5 mM.

[0409] 96. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 3 mM to 5 mM.

[0410] 97. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 3.5 mM to 5 mM.

[0411] 98. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 4 mM to 5 mM.

[0412] 99. The topical formulation of embodiment 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 5 mM.

[0413] 100. A topical formulation for use as an eye drop comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, wherein the formulation has a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of about 0.4 mM, is stable at ambient conditions for at least 5 months, and has a pH of about 6.5.

[0414] 101. A topical formulation that can be used as an eye drop comprising (lev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, wherein the topical formulation has a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof of about 0.8 mM and is stable at ambient conditions for at least 5 months, and has a pH of about 6.5.

[0415] 102. A topical formulation that can be used as an eye drop comprising (lev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, wherein the topical formulation has a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof of about 2 mM and is stable at ambient conditions for at least 5 months, and has a pH of about 6.5.

[0416] 103. The topical ophthalmic formulation of any one of embodiments 87-102, comprising Kolliphor™ ELP in a concentration of about 5% (weight / volume).

[0417] 104. The topical ophthalmic formulation of embodiment 103, comprising Kolliphor™ ELP in a concentration of about 4% (weight / volume).

[0418] 105. The topical ophthalmic formulation of any one of embodiments 87-104, comprising polysorbate 80 in a concentration of about 1% (weight / volume).

[0419] 106. The topical ophthalmic formulation of any one of embodiments 87-105, comprising PVP K-30 in a concentration of about 2% (weight / volume).

[0420] 107. The topical ophthalmic formulation of any one of embodiments 87-106, comprising poloxamer 407 in a concentration of about 0.1% (weight / volume).

[0421] 108. The topical ophthalmic formulation of any one of embodiments 87-107, comprising mannitol in a concentration of about 4.6% (weight / volume).

[0422] 109. The topical ophthalmic formulation of embodiment 103, comprising mannitol at a concentration of about 3.3%.

[0423] 110. A topical ophthalmic formulation that can be used as an eye drop comprising between about 0.05 mM and about 5 mM (rev)cromakalim or a pharmaceutically acceptable salt thereof in an aqueous liquid, which is stable for at least 5 months under ambient conditions without significant crystallization or excessive separation of cromakalim from the liquid, and which does not contain DMSO, DMF, or NMP, or other topical carriers that are not acceptable for human use, and which contains an optional buffer in addition to at least two excipients.

[0424] 111. A topical ophthalmic formulation that can be used as an eye drop comprising between about 0.1 mM and about 5 mM (rev)cromakalim or a pharmaceutically acceptable salt thereof in an aqueous liquid, which is stable for at least 5 months under ambient conditions without significant crystallization or excessive separation of cromakalim from the liquid, and which does not contain DMSO, DMF, or NMP, or other topical carriers that are not acceptable for human use, and which contains an optional buffer in addition to at least two excipients.

[0425] 112. A topical ophthalmic formulation that can be used as an eye drop comprising between about 0.5 mM and about 5 mM (rev)cromakalim or a pharmaceutically acceptable salt thereof in an aqueous liquid, which is stable for at least 5 months under ambient conditions without significant crystallization or excessive separation of cromakalim from the liquid, and which does not contain DMSO, DMF, or NMP, or other topical carriers that are not acceptable for human use, and which contains an optional buffer in addition to at least two excipients.

[0426] 113. The topical ophthalmic formulation of any one of embodiments 1-112, wherein the formulation is not an emulsion.

[0427] 114. The topical ophthalmic formulation of any one of embodiments 1-112, wherein the formulation is not a gel.

[0428] 115. The topical ophthalmic formulation of any one of embodiments 1-112, wherein the formulation is not a topical gel.

[0429] 116. The topical ophthalmic formulation of any one of embodiments 1-112, wherein the formulation is a clear solution.

[0430] 117. The topical ophthalmic formulation of any one of embodiments 1-112, wherein the formulation is a micellar or nanomicellar solution.

[0431] 118. The topical ophthalmic formulation of any one of embodiments 1-112, wherein the formulation does not comprise an oil as specifically defined above.

[0432] 119. The topical ophthalmic formulation of any one of embodiments 1-118, wherein the formulation has a percent transmittance of greater than 85% when tested with a UV-Visible spectrophotometer.

[0433] 120. The topical ophthalmic formulation of any one of embodiments 1-118, wherein the formulation has a percent transmittance of greater than 90% when tested with a UV-Visible spectrophotometer.

[0434] 121. The topical ophthalmic formulation of any one of embodiments 1-118, wherein the formulation has a percent transmittance of greater than 95% when tested with a UV-Visible spectrophotometer.

[0435] 122. The topical ophthalmic formulation of any one of embodiments 1-121, wherein the formulation comprises levcromakalim.

[0436] 123. The topical ophthalmic formulation of any one of embodiments 1-121, wherein the formulation comprises cromakalim.

[0437] 124. A method for treating an ocular disorder affecting the anterior or posterior segment of the eye, comprising administering to a host in need of treatment an effective amount of (lev)cromakalim or a pharmaceutically acceptable salt thereof in a topical ocular formulation of any one of embodiments 1 to 123.

[0438] 125. The method of embodiment 124, wherein the host is a human.

[0439] 126. The method of embodiment 124, wherein the use of the topical preparation results in lower intraocular pressure.

[0440] 127. The method of embodiment 124, wherein the eye disorder is glaucoma.

[0441] 128. The method of embodiment 127, wherein the glaucoma is glaucoma with normal intraocular pressure or normal tension glaucoma (NTG).

[0442] 129. The method of embodiment 127, wherein the glaucoma is glaucoma associated with elevated intraocular pressure selected from the group consisting of primary open-angle glaucoma (POAG), primary angle-closure glaucoma (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, glaucoma associated with diabetic retinopathy, glaucoma associated with Sturge-Weber syndrome, steroid-induced glaucoma, and acute glaucoma due to progressive cataract and / or intravitreal injection.

[0443] 130. The method of embodiment 129, wherein the eye disorder is glaucoma induced by Sturge-Weber syndrome or Sturge-Weber syndrome-induced glaucoma.

[0444] 131. The method of embodiment 129, wherein the eye disorder is diabetic retinopathy.

[0445] 132. The method of embodiment 129, wherein the local treatment is a primary or secondary or adjunctive treatment as part of a protocol for MIGS (minimally invasive glaucoma surgery) selected from the group consisting of miniaturized forms of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, complete intraocular or suprachoroidal shunts, gentler, more gentle forms of laser cyclophotocoagulation, and in alternative embodiments, Schlemm's canal stents to dilate Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty.

[0446] 133. The method of embodiment 129, wherein the ocular disorder is Graves' ophthalmopathy, thyroid-associated orbitopathy (TAO), Graves' orbitopathy (GO), retrobulbar tumor, cavernous sinus thrombosis, orbital vein thrombosis, episcleral / orbital vein vasculitis, superior vena cava occlusion, superior vena cava thrombosis, carotid-cavernous sinus fistula, dural cavernous shunt, orbital varicose veins, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), arterial occlusive / embolic-hypoperfusion disease, or optic nerve damage due to ischemia (posterior and anterior ischemic optic neuropathy (NAION)).

[0447] 134. The method of embodiment 129, wherein treatment with the topical formulation provides cytoprotection and / or neuroprotection to a host in need of protection.

[0448] 135. Use of (lev)cromakalim or a pharmaceutically acceptable salt thereof, as a formulation of any of embodiments 1-123, in the manufacture of a medicament for topically treating an ocular disorder in a host in need thereof.

[0449] 136. The use of embodiment 135, wherein the eye disorder is glaucoma.

[0450] 137. The use of embodiment 136, wherein the glaucoma is glaucoma with normal intraocular pressure or normal tension glaucoma (NTG).

[0451] 138. The use of embodiment 136, wherein the glaucoma is glaucoma associated with elevated intraocular pressure selected from the group consisting of primary open-angle glaucoma (POAG), primary angle-closure glaucoma (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, glaucoma associated with diabetic retinopathy, glaucoma associated with Sturge-Weber syndrome, steroid-induced glaucoma, and acute glaucoma due to progressive cataract and / or intravitreal injection.

[0452] 139. The use of embodiment 138, wherein the eye disorder is glaucoma induced by Sturge-Weber syndrome or Sturge-Weber syndrome-induced glaucoma.

[0453] 140. The use of embodiment 138, wherein the eye disorder is diabetic retinopathy.

[0454] 141. The use of embodiment 138, wherein the local treatment is a primary or secondary or adjunctive treatment as part of a protocol for MIGS (Minimally Invasive Glaucoma Surgery) selected from the group consisting of miniaturized forms of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, complete intraocular or suprachoroidal shunts, gentler, more gentle forms of laser cyclophotocoagulation, and in alternative embodiments, Schlemm's canal stents to dilate Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty.

[0455] 142. The use of embodiment 138, wherein the ocular disorder is Graves' ophthalmopathy, thyroid-associated orbitopathy (TAO), Graves' orbitopathy (GO), retrobulbar tumor, cavernous sinus thrombosis, orbital vein thrombosis, episcleral / orbital vein vasculitis, superior vena cava occlusion, superior vena cava thrombosis, carotid-cavernous fistula, dural-cavernous shunt, orbital varicose veins, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), arterial occlusive / embolic-hypoperfusion disease, or optic nerve damage due to ischemia (posterior and anterior ischemic optic neuropathy (NAION)).

[0456] 143. The use of embodiment 138, wherein treatment with the local preparation provides cytoprotection and / or neuroprotection to a host in need of protection.

[0457] 144. (Rev) A topical ophthalmic formulation comprising cromakalim or a pharmaceutically acceptable salt thereof (0.015% wt / vol), Kolliphor™ ELP (4% wt / vol), polysorbate 80 (1% wt / vol), polyvinylpyrrolidone (PVP) K-30 (2% wt / vol), poloxamer 407 (0.1% wt / vol), mannitol (3.3% wt / vol), dibasic sodium phosphate buffer, monobasic sodium phosphate buffer, and water, and having a pH of about 6.5.

[0458] 145. (Rev) A topical ophthalmic formulation comprising cromakalim or a pharmaceutically acceptable salt thereof (0.03% wt / vol), Kolliphor™ ELP (4% wt / vol), polysorbate 80 (1% wt / vol), polyvinylpyrrolidone (PVP) K-30 (2% wt / vol), poloxamer 407 (0.1% wt / vol), mannitol (3.3% wt / vol), dibasic sodium phosphate buffer, monobasic sodium phosphate buffer, and water, and having a pH of about 6.5.

[0459] 146. (Rev) A topical ophthalmic formulation comprising cromakalim or a pharmaceutically acceptable salt thereof (0.075% wt / vol), Kolliphor™ ELP (4% wt / vol), polysorbate 80 (1% wt / vol), polyvinylpyrrolidone (PVP) K-30 (2% wt / vol), poloxamer 407 (0.1% wt / vol), mannitol (3.3% wt / vol), dibasic sodium phosphate buffer, monobasic sodium phosphate buffer, and water, and having a pH of about 6.5.

[0460] 147. Use of (lev)cromakalim or a pharmaceutically acceptable salt thereof, as a formulation of any of embodiments 1-123, for topically treating an ocular disorder in a host in need thereof.

[0461] 148. The use of embodiment 147, wherein the eye disorder is glaucoma.

[0462] 149. The use of embodiment 148, wherein the glaucoma is glaucoma with normal intraocular pressure or normal tension glaucoma (NTG).

[0463] 150. The use of embodiment 148, wherein the glaucoma is glaucoma associated with elevated intraocular pressure selected from the group consisting of primary open-angle glaucoma (POAG), primary angle-closure glaucoma (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, glaucoma associated with diabetic retinopathy, glaucoma associated with Sturge-Weber syndrome, steroid-induced glaucoma, and acute glaucoma due to progressive cataract and / or intravitreal injection.

[0464] 151. The use of embodiment 147, wherein the eye disorder is glaucoma induced by Sturge-Weber syndrome or Sturge-Weber syndrome-induced glaucoma.

[0465] 152. The use of embodiment 147, wherein the eye disorder is diabetic retinopathy.

[0466] 153. The use of embodiment 147, wherein the local treatment is a primary or secondary or adjunctive treatment as part of a protocol for MIGS (Minimally Invasive Glaucoma Surgery) selected from the group consisting of miniaturized forms of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, complete intraocular or suprachoroidal shunts, gentler, more gentle forms of laser cyclophotocoagulation, and in alternative embodiments, Schlemm's canal stents to dilate Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty.

[0467] 154. The use of embodiment 147, wherein the ocular disorder is Graves' ophthalmopathy, thyroid-associated orbitopathy (TAO), Graves' orbitopathy (GO), retrobulbar tumor, cavernous sinus thrombosis, orbital vein thrombosis, episcleral / orbital vein vasculitis, superior vena cava occlusion, superior vena cava thrombosis, carotid-cavernous fistula, dural-cavernous shunt, orbital varicose veins, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), arterial occlusive / embolic-hypoperfusion disease, or optic nerve damage due to ischemia (posterior and anterior ischemic optic neuropathy (NAION)).

[0468] 155. The use of embodiment 147, wherein treatment with the local preparation provides cytoprotection and / or neuroprotection to a host in need of protection.

[0469] 156. A topical formulation of (lev)cromakalim or a pharmaceutically acceptable salt thereof of any of embodiments 1-123, for use in treating an ocular disorder in a host in need thereof.

[0470] 157. The use of embodiment 156, wherein the eye disorder is glaucoma.

[0471] 158. The use of embodiment 156, wherein the glaucoma is glaucoma with normal intraocular pressure or normal tension glaucoma (NTG).

[0472] 159. The use of embodiment 156, wherein the glaucoma is glaucoma associated with elevated intraocular pressure selected from the group consisting of primary open-angle glaucoma (POAG), primary angle-closure glaucoma (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, glaucoma associated with diabetic retinopathy, glaucoma associated with Sturge-Weber syndrome, steroid-induced glaucoma, and acute glaucoma due to progressive cataract and / or intravitreal injection.

[0473] 160. The use of embodiment 156, wherein the eye disorder is glaucoma induced by Sturge-Weber syndrome or Sturge-Weber syndrome-induced glaucoma.

[0474] 161. The use of embodiment 156, wherein the eye disorder is diabetic retinopathy.

[0475] 162. The use of embodiment 156, wherein the local treatment is a primary or secondary or adjunctive treatment as part of a protocol for MIGS (Minimally Invasive Glaucoma Surgery) selected from the group consisting of miniaturized forms of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, complete intraocular or suprachoroidal shunts, gentler, more gentle forms of laser cyclophotocoagulation, and in alternative embodiments, Schlemm's canal stents to dilate Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty.

[0476] 163. The use of embodiment 156, wherein the ocular disorder is Graves' ophthalmopathy, thyroid-associated orbitopathy (TAO), Graves' orbitopathy (GO), retrobulbar tumor, cavernous sinus thrombosis, orbital vein thrombosis, episcleral / orbital vein vasculitis, superior vena cava occlusion, superior vena cava thrombosis, carotid-cavernous fistula, dural-cavernous shunt, orbital varicose veins, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), arterial occlusive / embolic-hypoperfusion disease, or optic nerve damage due to ischemia (posterior and anterior ischemic optic neuropathy (NAION)).

[0477] 164. The use of embodiment 156, wherein treatment with the local preparation provides cytoprotection and / or neuroprotection to a host in need of protection.

[0478] VI. General Synthesis of (Lev)cromakalim Below is a general synthesis of (lev)cromakalim.

[0479] [ka]

[0480] The reaction of 4-cyanophenol (4-hydroxybenzonitrile) with 2-hydroxy-2-methyl-3-butyne under phase transfer catalysis almost certainly proceeds with the initial formation of a propargyl carbocation. This then attacks the aromatic ring, and the resulting allylic cation can capture the adjacent phenolic oxygen, leading to the formation of the observed product (3). Treatment of this product with aqueous N-bromosuccinimide results in the addition of hypobromous acid to form the bromohydrin (4) as a mixture of trans enantiomers. This then cyclizes in the presence of sodium hydroxide to form the epoxide 5 (5). Ring-opening of the oxirane with ammonia gives a mixture of trans amino alcohols (6). These are almost certainly resolved at this stage, with the 3S,4R-enantiomer being used in the next step. This isomer is then acylated with 4-chlorobutyl chloride to give the chloroamide (7). The anion resulting from the reaction of this amide with sodium hydride then displaces the chlorine at the end of the chain to form a pyrrolidine ring, thus giving levcromakalim (8).

[0481] VII. Non-Limiting Exemplary Methods of Making (Lev)cromakalim Formulations of the Invention The topical ophthalmic formulation of (rev)cromakalim of the present invention can be prepared according to various methods to achieve the desired results. In certain embodiments, the topical ophthalmic formulation of (rev)cromakalim is prepared by first dissolving (rev)cromakalim and other ingredients, individually or collectively, in ethanol or other low-volatile organic solvent(s), mixing, and then evaporating under vacuum to produce a film, which is then resuspended in phosphate buffer and / or deionized water, homogenized, or mixed, and then incubated and filtered to obtain the formulation. Optionally, the resuspension can be autoclaved and optionally filtered.

[0482] In certain embodiments, the ophthalmic formulation is prepared by dissolving (rev)cromakalim in a volatile solvent, followed by evaporation and drying under high vacuum to obtain a dry residue, which is then resuspended in an aqueous solution combined with excipients, followed by incubation and filtration to obtain the formulation. Optionally, the resuspension can be autoclaved and filtered.

[0483] In certain aspects of certain embodiments, therapeutic levels of (rev)cromakalim in the topical ophthalmic formulations of the present invention can be achieved by encapsulating (rev)cromakalim in nanomicelles using various methods. In certain embodiments, a two-step encapsulation method can be used to prepare a (rev)cromakalim nanomicelle formulation. First, the (rev)cromakalim base formulation and non-aqueous excipients are separately dissolved in ethanol or other compatible, low-volatility organic solvent(s). These solutions are combined in a glass round-bottom flask and vortexed or stirred to form a homogenous solution, and the solvent is removed by rotary evaporation followed by high vacuum to produce a thin film. This film is then hydrated with double-distilled water, aqueous excipients, and phosphate buffer and / or deionized water, homogenized or mixed, incubated, and then filtered through a 0.22 μm filter to obtain the final formulation. Optionally, the fully hydrated formulation can be autoclaved before filtration.

[0484] In certain embodiments, the topical ophthalmic formulation of (rev)cromakalim is prepared by direct dissolution of a solution of (rev)cromakalim in ethanol into a vial or centrifuge, and the ethanol is removed under vacuum to leave a fine solid coating. An aqueous solution of excipients is then separately added to the dried (rev)cromakalim with rapid stirring, sonication, or homogenization. This solution is optionally autoclaved and then filtered through a 0.22 μm filter to obtain the final formulation.

[0485] In certain embodiments, nanomicelle formulations of (rev)cromakalim are prepared using a dialysis method that uses a semipermeable membrane to selectively allow certain molecules to pass through. A solution of (rev)cromakalim and excipients is prepared in a water-miscible organic solvent, placed in a dialysis bag, and then immersed in a larger volume of deionized water. During this process, the organic solvent is replaced by water, which causes the formation of micelles that entrap (rev)cromakalim. Unincorporated excipients and unloaded (rev)cromakalim then diffuse from the bag into the surrounding solution over time.

[0486] In certain embodiments, the volatile solvent used to dissolve (lev)cromakalim is selected from the group of acetonitrile, acetone, methylene chloride, chloroform, methanol, propanol, or other alcohol- or ether-based solvents. Other solvents can be used that do not impart excessive toxicity or leave unacceptable residues to the final product.

[0487] In certain embodiments, the ophthalmic formulation is homogenized or mixed through techniques including, but not limited to, heating, centrifugation, filtration, incubation, vortexing, settling, stirring, shaking, sonication, or any combination thereof.

[0488] In certain embodiments, the (rev)cromakalim formulations described herein are emulsions or solutions formed by stirring at a temperature of at least about 120°C, at least about 100°C, at least about 80°C, at least about 60°C, at least about 40°C, or at least about ambient temperature.

[0489] In certain embodiments, the ophthalmic formulation can be formed by mixing the components together (each or together may be pre-dissolved in a low volatility solvent, including but not limited to, ethanol), and then heating to remove the solvent, where heating can include autoclaving. In certain embodiments, the ophthalmic formulation can be formed by heating the solution or emulsion using a heating block.

[0490] In certain aspects of the embodiment, the present invention provides solutions or emulsions from which undissolved components have been removed by centrifugation at speeds of at least about 1000 rpm, at least about 1250 rpm, at least about 1500 rpm, at least about 1800 rpm, at least about 2000 rpm, at least about 2500 rpm, and at least about 3000 rpm.

[0491] In certain embodiments, the present invention provides solutions or emulsions filtered using filters with pore sizes of at least about 0.2 μm, at least about 0.45 μm, at least about 0.7 μm, and at least about 1.2 μm. In certain non-limiting embodiments, the filter membrane may be made from nylon, polytetrafluoroethylene (PTFE), polyvinylidene difluoride (PVDF), polyethersulfone (PES), cellulose acetate (CA), polypropylene (PP), and / or glass fiber.

[0492] The concentration of (lev)cromakalim in the formulations described herein can be extrapolated using the calibration curve method. Table 1 shows the concentrations of levcromakalim standards and the measured area of ​​the peak showing the elution of levcromakalim. Figure 4 shows the calibration curve for levcromakalim obtained using a plot of concentration versus area.

[0493] TIFF2025538061000023.tif56170

[0494] The following examples are provided to further illustrate aspects of the present invention, including non-limiting examples of (rev)cromakalim formulations having concentrations of less than 1 mg / mL, 1 mg / mL to 1.5 mg / mL, and greater than 1.5 mg / mL, and their manufacture. These illustrative examples are not limiting and should not be construed as limiting any aspect of the present invention.

[0495] Definition: PBS stock solution: Phosphate buffered saline, NaCl: 1.37M, KCl: 27 mM, Na2HPO4: 100mM, KH2PO4: 18mM, It was prepared at a concentration of

[0496] Non-saline phosphate buffer stock solution: Non-saline phosphate buffer Na2HPO4: 70mM, NaH2PO4: 25mM, It was prepared at a concentration of

[0497] Example 1 - Ophthalmic formulations of levcromakalim at concentrations less than 1 mg / mL, i.e., less than 3.5 mM Example 1a TIFF2025538061000024.tif25170

[0498] A 0.6% solution of levcromakalim in acetonitrile was added to a 5 mL vial. The solvent was removed via evaporation under a stream of nitrogen, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. The film was then hydrated with 2 mL of deionized H2O. 300 μL of PBS stock solution was then added, and the final volume was brought to 3 mL using deionized H2O. The mixture was stirred for 20 minutes while heated at 80°C. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.38 mg / mL, or 1.3 mM.

[0499] Example 1b TIFF2025538061000025.tif31170

[0500] A 0.6% solution of levcromakalim in acetonitrile was added to a 5 mL vial. After the solvent was removed via evaporation under a stream of nitrogen, the vial was placed under high vacuum for 24 hours to allow further evaporation and obtain a thin film. Next, 300 μL of PBS stock solution and polysorbate 80 were added, and the final volume was brought to 3 mL using deionized H2O, resulting in a polysorbate 80 concentration of approximately 1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and then filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.46 mg / mL, or 1.6 mM.

[0501] Example 1c TIFF2025538061000026.tif31170

[0502] A solution of levcromakalim in acetonitrile (1 mL of 6 mg / mL stock) was added to a 5 mL vial. After the solvent was removed via evaporation under a stream of nitrogen, the vial was placed under high vacuum for 24 hours to allow further evaporation and obtain a thin film. Next, 300 μL of PBS stock solution and glycerin were added, and the final volume was brought to 3 mL using deionized HO, resulting in a glycerin concentration of approximately 5%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature before being filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.34 mg / mL, or 1.2 mM.

[0503] Example 1d TIFF2025538061000027.tif31170

[0504] A solution of levcromakalim in acetonitrile (1 mL of 6 mg / mL stock) was added to a 5 mL vial. After the solvent was removed via evaporation under a stream of nitrogen, the vial was placed under high vacuum for 24 hours to allow further evaporation and obtain a thin film. 300 μL of PBS stock solution and poloxamer 407 were then added. The final volume was brought to 3 mL using deionized H2O, resulting in a poloxamer 407 concentration of approximately 0.1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.37 mg / mL, or 1.3 mM.

[0505] Example 1e TIFF2025538061000028.tif31170

[0506] A solution of levcromakalim in acetonitrile (1 mL of 6 mg / mL stock) was added to a 5 mL vial. After the solvent was removed via evaporation under a stream of nitrogen, the vial was placed under high vacuum for 24 hours to allow further evaporation and obtain a thin film. 300 μL of PBS stock solution and tyloxapol were then added. The final volume was brought to 3 mL using deionized H2O, resulting in a tyloxapol concentration of approximately 0.1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.38 mg / mL, or 1.3 mM.

[0507] Example 1f TIFF2025538061000029.tif31170

[0508] A solution of levcromakalim in acetonitrile (1 mL of 6 mg / mL stock) was added to a 5 mL vial. After the solvent was removed via evaporation under a stream of nitrogen, the vial was placed under high vacuum for 24 hours to allow further evaporation and obtain a thin film. 300 μL of PBS stock solution and Kolliphor™ ELP were then added. The final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ ELP concentration of approximately 5%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.39 mg / mL, or 1.4 mM.

[0509] Example 1g TIFF2025538061000030.tif30170

[0510] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of Kolliphor™ RH 40 and Premulen™ TR-1 in a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to allow further evaporation, resulting in a thin film. The final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ RH 40 concentration of approximately 2% and a Premulen™ TR-1 concentration of approximately 0.2%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.59 mg / mL, or 2.1 mM.

[0511] Example 1h TIFF2025538061000031.tif30170

[0512] A solution of levcromakalim in acetonitrile (1 mL of 6 mg / mL stock) was added to a 5 mL vial. The solvent was removed via evaporation under a stream of nitrogen, and the vial was placed under high vacuum for 24 hours to allow further evaporation and obtain a thin film. 300 μL of PBS stock solution and hypromellose were then added, and the final volume was brought to 3 mL using deionized HO, resulting in a hypromellose concentration of approximately 0.5%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.7 mg / mL, or 2.4 mM.

[0513] Example 1i TIFF2025538061000032.tif35170

[0514] A solution of levcromakalim in acetonitrile (1 mL of a 6 mg / mL stock) was dispensed into a 5 mL vial. The solvent was removed via evaporation under a stream of nitrogen, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. A water-based solution of polysorbate 80, glycerin, and EDTA was then added, and the final volume was brought to 3 mL using deionized H2O, resulting in a polysorbate 80 concentration of approximately 1%, a glycerin concentration of approximately 5%, and an EDTA concentration of approximately 0.1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.43 mg / mL, or 1.5 mM.

[0515] Example 1j TIFF2025538061000033.tif35170

[0516] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of tyloxapol, polysorbate 80, and hypromellose in a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. The final volume was brought to 3 mL using deionized H2O, resulting in a tyloxapol concentration of approximately 2%, a polysorbate 80 concentration of approximately 5%, and a hypromellose concentration of approximately 0.1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.89 mg / mL, or 3.1 mM.

[0517] Example 1k TIFF2025538061000034.tif35170

[0518] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of Kolliphor™ ELP, polysorbate 80, and Premulen™ TR-1 in a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. The final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ ELP concentration of approximately 2%, a polysorbate 80 concentration of approximately 1%, and a Premulen™ TR-1 concentration of approximately 0.2%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.79 mg / mL, or 2.8 mM.

[0519] Example 1l TIFF2025538061000035.tif36170

[0520] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of Kolliphor™ ELP, polysorbate 80, and Premulen™ TR-2 in a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. The final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ ELP concentration of approximately 2%, a polysorbate 80 concentration of approximately 1%, and a Premulen™ TR-2 concentration of approximately 0.2%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.68 mg / mL, or 2.4 mM.

[0521] Example 1m TIFF2025538061000036.tif45170

[0522] A 0.6% acetonitrile solution of levcromakalim was combined with an EtOH-based solution of glycerin, Kolliphor™ ELP, polysorbate 80, and hypromellose. The solvent was removed by evaporation under a stream of nitrogen and dried under high vacuum for 24 hours to yield a thin coating. 300 μL of PBS stock solution was added and the volume adjusted with deionized H2O, resulting in a glycerin concentration of approximately 5%, a Kolliphor™ ELP concentration of approximately 5%, a polysorbate 80 concentration of approximately 1%, and a hypromellose concentration of approximately 0.5%. The coating was dissolved or suspended by rapid stirring or sonication. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.5 mg / mL, or 1.7 mM.

[0523] Example 1n TIFF2025538061000037.tif45170

[0524] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of glycerin, castor oil, polysorbate 80, and Premulen™ TR-1 and dispensed into a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate and obtain a thin film. 300 μL of PBS stock solution was then added, and the final volume was brought to 3 mL using deionized H2O, resulting in a glycerin concentration of approximately 2.2%, a castor oil concentration of approximately 1.25%, a polysorbate 80 concentration of approximately 1%, and a Premulen™ TR-1 concentration of approximately 0.05%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim determined using the calibration curve method was approximately 0.56 mg / mL, or 2 mM.

[0525] Example 1o TIFF2025538061000038.tif31170

[0526] A solution of levcromakalim in acetonitrile (1 mL of 6 mg / mL stock) was added to a 5 mL vial. After the solvent was removed via evaporation under a stream of nitrogen, the vial was placed under high vacuum for 24 hours to allow further evaporation and obtain a thin film. 300 μL of PBS stock solution and sodium carboxymethylcellulose (NaCMC) were then added. The final volume was brought to 3 mL using deionized H2O, resulting in a NaCMC concentration of approximately 0.5%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 0.99 mg / mL, or 3.4 mM.

[0527] Example 2 - Ophthalmic formulations of levcromakalim at concentrations of 1 mg / mL to 1.5 mg / mL, i.e., 3.5 mM to 5.2 mM Example 2a TIFF2025538061000039.tif60170

[0528] A 0.6% acetonitrile solution of levcromakalim was added to a 3 mL vial. The solvent was removed by evaporation under a nitrogen stream and dried under high vacuum for 24 hours to yield a thin coating. 300 μL of a non-saline phosphate buffer stock solution and a water-based solution of the ingredients were then added, and the coating was dissolved or suspended by rapid stirring or sonication to yield a formulation with a glycerin concentration of approximately 4%, Kolliphor™ ELP concentration of approximately 2%, polysorbate 80 concentration of approximately 1%, poloxamer 407 concentration of approximately 0.1%, hypromellose concentration of approximately 0.1%, mannitol concentration of approximately 3.5%, and benzalkonium chloride concentration of approximately 0.02%. The concentration of levcromakalim determined using the calibration curve method was approximately 1.36 mg / mL, or 4.7 mM.

[0529] Example 2b TIFF2025538061000040.tif37170

[0530] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of Kolliphor™ RH 40, Octoxynol-40, and Premulen™ TR-2 in a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. The final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ RH 40 concentration of approximately 2%, Octoxynol-40 concentration of approximately 1%, and Premulen™ TR-2 concentration of approximately 0.2%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 1.23 mg / mL, or 4.3 mM.

[0531] Example 2c TIFF2025538061000041.tif35170

[0532] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of Kolliphor™ RH 40, Kolliphor™ HS 15, and PVP in a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. The final volume was adjusted to 3 mL using deionized H2O, resulting in a Kolliphor™ RH 40 concentration of approximately 5%, a Kolliphor™ HS 15 concentration of approximately 1%, and a PVP concentration of approximately 2%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 1.17 mg / mL, or 4.1 mM.

[0533] Example 2d TIFF2025538061000042.tif40170

[0534] A solution of levcromakalim in acetonitrile (1 mL of a 6 mg / mL stock) was added to a 5 mL vial. The solvent was removed via evaporation under a stream of nitrogen, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. A water-based solution of glycerin, Kolliphor™ ELP, polysorbate 80, and hypromellose was then added and brought to a final volume of 3 mL using deionized H2O, resulting in a glycerin concentration of approximately 5%, a Kolliphor™ ELP concentration of approximately 5%, a polysorbate 80 concentration of approximately 1%, and a hypromellose concentration of approximately 0.05%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 1.31 mg / mL, or 4.6 mM.

[0535] Example 2e TIFF2025538061000043.tif40170

[0536] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) and an EtOH-based solution of Kolliphor™ ELP, polysorbate 80, and PVP were added to a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. 300 μL of non-saline phosphate buffer stock solution was then added, and the final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ ELP concentration of approximately 5%, a polysorbate 80 concentration of approximately 1%, and a PVP concentration of approximately 2%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 1.1 mg / mL, or 4 mM.

[0537] Example 2f TIFF2025538061000044.tif45170

[0538] An ethanolic solution of levcromakalim (1 mL of a 6 mg / mL stock) was combined with an EtOH-based solution of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose and dispensed into 5 mL vials. The solvent was removed via evaporation under a nitrogen stream, and the vials were placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. The final volume was adjusted to 3 mL using deionized H2O, resulting in a glycerin concentration of approximately 5%, Kolliphor™ ELP concentration of approximately 2.5%, polysorbate 80 concentration of approximately 1%, poloxamer 407 concentration of approximately 1%, and hypromellose concentration of approximately 0.1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim determined using the calibration curve method was approximately 1.27 mg / mL, or 4.4 mM.

[0539] Example 2g TIFF2025538061000045.tif45170

[0540] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with EtOH-based Kolliphor™ RH 40, glycerin, PVP, and Premulen™ TR-2 and dispensed into a 5 mL vial. The solvent was removed via evaporation under a stream of nitrogen, and the vial was placed under high vacuum for 24 hours to further evaporate and obtain a thin film. 300 μL of PBS stock solution was then added, and the final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ RH 40 concentration of approximately 4%, a glycerin concentration of approximately 5%, a PVP concentration of approximately 1%, and a Premulen™ TR-2 concentration of approximately 0.1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim determined using the calibration curve method was approximately 1.05 mg / mL, or 3.7 mM.

[0541] Example 2h TIFF2025538061000046.tif50170

[0542] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of Kolliphor™ HS 15, octoxynol-40, polysorbate 80, Premulen™ TR-2, and poloxamer 407 and dispensed into 5 mL vials. The solvent was removed via evaporation under a nitrogen stream, and the vials were placed under high vacuum for 24 hours to allow further evaporation and yield a thin film. 300 μL of PBS stock solution was then added, and the final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ HS 15 concentration of approximately 4%, octoxynol-40 concentration of approximately 1%, polysorbate 80 concentration of approximately 1%, Premulen™ TR-2 concentration of approximately 0.1%, and poloxamer 407 concentration of approximately 1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim determined using the standard curve method was approximately 1.26 mg / mL, or 4.4 mM.

[0543] Example 2i TIFF2025538061000047.tif55170

[0544] An ethanol (EtOH)-based solution of levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and PVP was combined in a 50 mL round-bottom flask, stirred, and heated (if necessary) until homogeneous. The solvent was evaporated under a stream of nitrogen to yield a coating, which was further dried under high vacuum for 24 hours. The coating was reformulated by adding a water-based solution of mannitol. The mixture was brought to a volume of 10 mL with 1 mL of non-saline phosphate buffer stock solution and deionized H2O, resulting in a levcromakalim concentration of approximately 0.15%, a glycerin concentration of approximately 1%, a Kolliphor™ ELP concentration of approximately 2%, a polysorbate 80 concentration of approximately 1%, a poloxamer 407 concentration of approximately 0.1%, a PVP concentration of approximately 1%, and a mannitol concentration of approximately 2%. The solution was transferred to a septa vial and heated in an autoclave for 15 minutes at 120°C. The solution was incubated overnight at room temperature and filtered through a 0.22 μm sterile filter into a sterile vial. The concentration of levcromakalim determined using the calibration curve method was approximately 1.23 mg / mL, or 4.3 mM.

[0545] Example 2j TIFF2025538061000048.tif60170

[0546] An ethanol (EtOH)-based solution of levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose was combined in a 50 mL round-bottom flask, stirred, and heated (if necessary) until homogeneous. The solvent was evaporated under a stream of nitrogen to yield a coating, which was further dried under high vacuum for 24 hours. The coating was reformulated by adding the aqueous components. The mixture was brought to a volume of 10 mL with 1 mL of non-saline phosphate buffer stock solution and deionized H2O, resulting in a levcromakalim concentration of approximately 0.15%, a glycerin concentration of approximately 4%, a Kolliphor™ ELP concentration of approximately 2%, a polysorbate 80 concentration of approximately 1%, a poloxamer 407 concentration of approximately 0.1%, a hypromellose concentration of approximately 0.1%, a mannitol concentration of approximately 3.5%, and a benzalkonium chloride concentration of approximately 0.02%. The solution was transferred to a septa vial and heated in an autoclave for 15 minutes at 120°C. The solution was incubated overnight at room temperature and filtered through a 0.22 µm sterile filter into a sterile vial. The concentration of levcromakalim determined using the calibration curve method was approximately 1.31 mg / mL, or 4.6 mM.

[0547] Example 3 - Ophthalmic formulations of levcromakalim at concentrations greater than 1.5 mg / mL, i.e., greater than 5.2 mM Example 3a TIFF2025538061000049.tif35170

[0548] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) and an EtOH-based solution of Kolliphor™ ELP, polysorbate 80, and PVP were combined in a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin coating. The final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ ELP concentration of approximately 5%, a polysorbate 80 concentration of approximately 2%, and a PVP concentration of approximately 2%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 1.7 mg / mL, or 5.8 mM.

[0549] Example 3b TIFF2025538061000050.tif40170

[0550] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of Kolliphor™ ELP, polysorbate 80, and poloxamer 407 in a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin film. 300 μL of non-saline phosphate buffer stock solution was then added, and the final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ ELP concentration of approximately 5%, a polysorbate 80 concentration of approximately 1%, and a poloxamer 407 concentration of approximately 1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim, determined using the calibration curve method, was approximately 2.1 mg / mL, or 7.3 mM.

[0551] Example 3c TIFF2025538061000051.tif45170

[0552] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was combined with an EtOH-based solution of Kolliphor™ ELP, polysorbate 80, PVP, and poloxamer 407 and dispensed into a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate and obtain a thin film. 300 μL of PBS stock solution was then added, and the final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ ELP concentration of approximately 4%, polysorbate 80 concentration of approximately 1%, PVP concentration of approximately 2%, and poloxamer 407 concentration of approximately 0.1%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim determined using the calibration curve method was approximately 1.8 mg / mL, or 6.4 mM.

[0553] Example 3d TIFF2025538061000052.tif50170

[0554] An ethanol (EtOH)-based solution of levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, and poloxamer 407 was combined in a 50 mL round-bottom flask, stirred, and heated (if necessary) until homogeneous. The solvent was evaporated under a stream of nitrogen to yield a coating, which was further dried under high vacuum for 24 hours. The coating was reformulated by adding the aqueous components. The mixture was brought to a volume of 10 mL with 1 mL of non-saline phosphate buffer stock solution and deionized H2O, resulting in a levcromakalim concentration of approximately 0.15%, Kolliphor™ ELP concentration of approximately 4%, polysorbate 80 concentration of approximately 1%, PVP concentration of approximately 2%, poloxamer 407 concentration of approximately 0.1%, and mannitol concentration of approximately 3.5%. The solution was transferred to a septa vial and heated to 120°C in an autoclave for 15 minutes. The solution was incubated overnight at room temperature and filtered through a 0.22 μm sterile filter into a sterile vial. The concentration of levcromakalim determined using the standard curve method was approximately 1.6 mg / mL, or 5.5 mM.

[0555] Example 3e TIFF2025538061000053.tif50170

[0556] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) was added to a 3 mL vial. An EtOH-based solution of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose was dispensed into a 3 mL vial, vortexed, and heated (if necessary) until homogeneous. The solvent was removed by evaporation under a nitrogen stream and dried under high vacuum for 24 hours to yield a coating. 300 μL of PBS stock solution and water were then added, and the coating was dissolved or suspended by rapid vortexing or sonication to yield a formulation with a glycerin concentration of approximately 4%, a Kolliphor™ ELP concentration of approximately 2%, a polysorbate 80 concentration of approximately 1%, a poloxamer 407 concentration of approximately 0.1%, and a hypromellose concentration of approximately 0.1%. The concentration of levcromakalim, as determined using the calibration curve method, was approximately 1.9 mg / mL, or 6.5 mM.

[0557] TIFF2025538061000054.tif54170

[0558] An ethanol (EtOH)-based solution of levcromakalim, Kolliphor™ ELP, polysorbate 80, PVP, and poloxamer 407 was combined in a 50 mL round-bottom flask, stirred, and heated (if necessary) until homogeneous. The solvent was evaporated under a stream of nitrogen to yield a coating, which was further dried under high vacuum for 24 hours. The coating was reformulated by adding a water-based solution of mannitol and BAK. The mixture was brought to a volume of 10 mL with 1 mL of non-saline phosphate buffer stock solution and deionized H2O, resulting in a levcromakalim concentration of approximately 0.15%, Kolliphor™ ELP concentration of approximately 4%, polysorbate 80 concentration of approximately 1%, PVP concentration of approximately 2%, poloxamer 407 concentration of approximately 0.1%, mannitol concentration of approximately 3.5%, and benzalkonium chloride (BAK) concentration of approximately 0.02%. The solution was transferred to a septa vial and heated in an autoclave for 15 minutes at 120°C. The solution was incubated overnight at room temperature and filtered through a 0.22 μm sterile filter into a sterile vial. The concentration of levcromakalim determined using the calibration curve method was approximately 1.5 mg / mL, or 5.4 mM.

[0559] Example 3g TIFF2025538061000055.tif55170

[0560] An ethanol (EtOH)-based solution of levcromakalim, glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose was combined in a 50 mL round-bottom flask, stirred, and heated (if necessary) until homogeneous. The solvent was evaporated under a stream of nitrogen to yield a coating, which was further dried under high vacuum for 24 hours. The coating was reformulated by adding a water-based solution of mannitol. The mixture was brought to a volume of 10 mL with 1 mL of non-saline phosphate buffer stock solution and deionized H2O, resulting in a levcromakalim concentration of approximately 0.15%, a glycerin concentration of approximately 4%, a Kolliphor™ ELP concentration of approximately 2%, a polysorbate 80 concentration of approximately 1%, a poloxamer 407 concentration of approximately 0.1%, a hypromellose concentration of approximately 0.1%, and a mannitol concentration of approximately 3.5%. The solution was transferred to a septa vial and heated in an autoclave for 15 minutes at 120°C. The solution was incubated overnight at room temperature and filtered through a 0.22 µm sterile filter into a sterile vial. The concentration of levcromakalim determined using the calibration curve method was approximately 1.6 mg / mL, or 5.7 mM.

[0561] Example 3h TIFF2025538061000056.tif54170

[0562] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) and an EtOH-based solution of Kolliphor™ ELP, polysorbate 80, PVP, and poloxamer 407 were combined in a 5 mL vial. The solvent was removed via evaporation under a nitrogen stream, and the vial was placed under high vacuum for 24 hours to further evaporate, resulting in a thin coating. A water-based solution of mannitol, BAK, and 300 μL of non-saline phosphate buffer stock solution was then added, and the final volume was brought to 3 mL using deionized H2O, resulting in a Kolliphor™ ELP concentration of approximately 4%, polysorbate 80 concentration of approximately 1%, PVP concentration of approximately 2%, poloxamer 407 concentration of approximately 1%, mannitol concentration of approximately 3.5%, and benzalkonium chloride concentration of approximately 0.02%. The mixture was stirred in an 80°C heating block for 20 minutes. The mixture was then incubated overnight at room temperature and filtered through a 0.45 μm filter to obtain a saturated solution. The concentration of levcromakalim determined using the standard curve method was approximately 1.7 mg / mL, or 5.9 mM.

[0563] Example 4 Additional Formulations Example 4a TIFF2025538061000057.tif55170

[0564] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) and an EtOH-based solution of hydrogenated castor oil-40, octoxynol-40, PVP, and disodium EDTA were combined in a round-bottom flask and mixed to homogenize. The solvent was removed by evaporation and further dried under high vacuum to obtain a thin film. The film was rehydrated by adding a water-based solution of sodium chloride, BAK, PBS, and approximately 2.5 mL of deionized water and stirred overnight. The pH of the formulation was adjusted to neutral with additional phosphate buffer, and the final volume was brought to 3 mL using deionized H2O. The formulation was then filtered through a 0.2 μm nylon filter. The concentration of levcromakalim was measured using the calibration curve method.

[0565] Example 4b TIFF2025538061000058.tif59170

[0566] An ethanol-based solution of levcromakalim and an EtOH-based solution of hydrogenated castor oil-40, octoxynol-40, PVP, and disodium EDTA were combined in a round-bottom flask and mixed to homogenize. The solvent was removed by evaporation and further dried under high vacuum to yield a thin film. The film was rehydrated by adding an aqueous solution of sodium chloride, BAK, PBS, and approximately 2.5 mL of deionized water and stirred overnight. The pH of the formulation was adjusted to neutral with additional phosphate buffer, and the final volume was brought to 3 mL using deionized H2O. The formulation was then filtered through a 0.2 μm nylon filter. The concentration of levcromakalim was measured using a calibration curve method.

[0567] Example 4c TIFF2025538061000059.tif31170

[0568] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) and an EtOH-based solution of hydrogenated castor oil-40 and octoxynol-40 were combined in a round-bottom flask and mixed to homogenize. The solvent was removed by evaporation and further dried under high vacuum to obtain a thin film. The film was rehydrated with approximately 2.5 mL of deionized water and stirred overnight. The pH of the formulation was adjusted to neutral using phosphate buffer, and the final volume was brought to 3 mL using deionized H2O. The formulation was then filtered through a 0.2 μm nylon filter. The concentration of levcromakalim was measured using the calibration curve method.

[0569] Example 4d TIFF2025538061000060.tif55170

[0570] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) and an EtOH-based solution of hydrogenated castor oil-60, octoxynol-40, PVP, and disodium EDTA were combined in a round-bottom flask and mixed to homogenize. The solvent was removed by evaporation and further dried under high vacuum to obtain a thin film. The film was rehydrated by adding a water-based solution of sodium chloride, BAK, PBS, and approximately 2.5 mL of deionized water and stirred overnight. The pH of the formulation was adjusted to neutral with additional phosphate buffer, and the final volume was brought to 3 mL using deionized H2O. The formulation was then filtered through a 0.2 μm nylon filter. The concentration of levcromakalim was measured using the calibration curve method.

[0571] Example 4e TIFF2025538061000061.tif54170

[0572] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) and an EtOH-based solution of hydrogenated castor oil-80, octoxynol-40, PVP, and disodium EDTA were combined in a round-bottom flask and mixed to homogenize. The solvent was removed by evaporation and further dried under high vacuum to obtain a thin film. The film was rehydrated by adding a water-based solution of sodium chloride, BAK, PBS, and approximately 2.5 mL of deionized water and stirred overnight. The pH of the formulation was adjusted to neutral with additional phosphate buffer, and the final volume was brought to 3 mL using deionized H2O. The formulation was then filtered through a 0.2 μm nylon filter. The concentration of levcromakalim was measured using the calibration curve method.

[0573] Example 4f TIFF2025538061000062.tif54170

[0574] An ethanolic solution of levcromakalim (1 mL of 6 mg / mL stock) and an EtOH-based solution of hydrogenated castor oil-100, octoxynol-40, PVP, and disodium EDTA were combined in a round-bottom flask and mixed to homogenize. The solvent was removed by evaporation and further dried under high vacuum to obtain a thin film. The film was rehydrated by adding a water-based solution of sodium chloride, BAK, PBS, and approximately 2.5 mL of deionized water and stirred overnight. The pH of the formulation was adjusted to neutral with additional phosphate buffer, and the final volume was brought to 3 mL using deionized H2O. The formulation was then filtered through a 0.2 μm nylon filter. The concentration of levcromakalim was measured using the calibration curve method.

[0575] VIII. In vivo evaluation Example 5 In vivo evaluation of ophthalmic formulations 3f and 3d in mice TIFF2025538061000063.tif56170

[0576] TIFF2025538061000064.tif50170

[0577] Levcromakalim (DMSO + Cremophor) formulation Levcromakalim (>98%, molecular weight 286.33 g / mol, molecular weight of disodium salt -332.3 g / mol) was purchased from MilliporeSigma (catalog number C1055). A 100 mM stock solution was prepared by mixing 28.63 mg of levcromakalim into 1 mL of DMSO (MilliporeSigma, catalog number D2650) to a concentration of 28.63 mg / mL. The stock solution was diluted 20-fold with 10% Cremophor EL (MilliporeSigma, catalog number 238470) in sterile PBS (Corning, catalog number 21-040-CV) to make a 5 mM working solution (1.43 mg per mL in 5% DMSO / 10% Cremophor EL / 85% PBS).

[0578] Formulation 3f, an ophthalmic formulation of levcromakalim containing benzalkonium chloride (BAK) (osmolality = 245 Osm / L), and its analogous formulation 3d without BAK (osmolality = 280 Osm / L) were administered in efficacy experiments to determine their ability to reduce intraocular pressure (IOP) in groups of normotensive mice. A levcromakalim formulation (0.5 mM) made by serial dilution of the levcromakalim (DMSO + Cremophor) formulation was used as a positive control, and the vehicle for each formulation was used as a negative control.

[0579] Dosing: Mice (n=5 per group, 3 females and 2 males) received 5 μL drops of 0.5 mM (0.015% levcromakalim) formulated in one of two formulations (Formulation 3f and Formulation 3d) in one eye once daily in the morning. The contralateral eye received the formulation vehicle alone (no levcromakalim) as a control. Positive control mice (n=5, 2 females and 3 males) received a levcromakalim (0.5 mM) formulation made by serial dilution of the levcromakalim (DMSO + Cremophor) formulation in one eye, and the contralateral eye received only a similar serial dilution of DMSO + Cremophor in sterile PBS as a vehicle control.

[0580] Timing: Baseline IOP was measured for three days prior to dosing using an iCare TonoPen rebound tonometer. Mice were dosed with the above formulations for five days to determine efficacy. After a three-day washout period, mice (n=5 per group) received doses of 0.005 mM, 0.05 mM, and 5.0 mM for five additional days, with the contralateral eye receiving the corresponding vehicle control. These animals were compared with positive control mice that received 0.005 mM, 0.05 mM, and 5.0 mM levcromakalim formulated by serial dilution of the levcromakalim (DMSO+Cremophor) formulation in one eye, and the contralateral eye received only a similar serial dilution of DMSO+Cremophor in sterile PBS as the vehicle control. IOP was measured and analyzed daily at 1 and 23 hours after dosing. The comparison of IOP reduction is summarized in the tables provided herein.

[0581] Analysis: IOP was measured daily at 1 hour and 23 hours after dosing at approximately 10:30 AM. Data shown represent the mean IOP from days 3 and 4 of dosing (per dosing period). Figure 1 shows the IOP reduction data for Formulations 3f and 3d, using vehicle as the negative control and the DMSO + Cremophor formulation as the positive control. Figure 3 shows a comparison of IOP reduction with various concentrations of levcromakalim formulated in Formulation 3f, Formulation 3d, and levcromakalim formulated in DMSO and Cremophor EL (>98% levcromakalim).

[0582] Statistics: Daily IOP was calculated by averaging the 1-hour and 23-hour time points (i.e., calculating the mean ± SD). Baseline IOP was calculated for each eye by calculating the mean ± SD daily IOP of the three daily pre-treatment measurements. Intraocular pressure during the treatment period was determined for each eye by calculating the mean ± SD daily IOP on days 3 and 4 of treatment.

[0583] TIFF2025538061000065.tif35170

[0584] Example 6 In vivo evaluation of ophthalmic formulation 2j and ophthalmic formulation 3g in mice TIFF2025538061000066.tif60170

[0585] TIFF2025538061000067.tif55170

[0586] An ophthalmic formulation of levcromakalim containing benzalkonium chloride (BAK), Formulation 2j (osmolality = 723 Osm / L), and its analogous formulation without BAK, Formulation 3g (osmolality = 762 Osm / L), were administered in efficacy experiments to determine their ability to reduce intraocular pressure (IOP) in groups of normotensive mice. Levcromakalim formulations in DMSO and Cremophor EL were used as positive controls, and the vehicle for each formulation was used as a negative control.

[0587] Dosing: Mice (n=5 per group, 3 females and 2 males) received 5 μL drops of 0.5 mM (0.015% levcromakalim) formulated in one of two formulations (Formulation 2j and Formulation 3g) in one eye once daily in the morning. The contralateral eye received the formulation vehicle alone (no levcromakalim) as a control. Positive control mice (n=5, 2 females and 3 males) received a levcromakalim (0.5 mM) formulation made by serial dilution of the levcromakalim (DMSO + Cremophor) formulation in one eye, and the contralateral eye received only a similar serial dilution of DMSO + Cremophor in sterile PBS as a vehicle control.

[0588] Timing: Baseline IOP was measured for three days prior to dosing using an iCare TonoPen rebound tonometer. Mice were dosed with the above formulations for five days to determine efficacy. These animals were compared to positive control mice that received levcromakalim (0.5 mM) formulated by serial dilution of the levcromakalim (DMSO + Cremophor) formulation in one eye, and the contralateral eye received only a similar serial dilution of DMSO + Cremophor in sterile PBS as a vehicle control. IOP was measured and analyzed daily at 1 and 23 hours after dosing. The tables provided herein summarize the comparison of IOP reduction.

[0589] Analysis: IOP was measured daily at 1 hour and 23 hours after dosing at approximately 10:30 AM. Data shown represent the mean IOP from days 3 and 4 of dosing (per dosing period). Figure 2 shows the IOP reduction data for Formulation 2j and Formulation 3g, using vehicle as the negative control and the DMSO + Cremophor formulation as the positive control.

[0590] Statistics: Daily IOP was calculated by averaging the 1-hour and 23-hour time points (i.e., calculating the mean ± SD). Baseline IOP was calculated for each eye by calculating the mean ± SD daily IOP of the three daily pre-treatment measurements. Intraocular pressure during the treatment period was determined for each eye by calculating the mean ± SD daily IOP on days 3 and 4 of treatment.

[0591] TIFF2025538061000068.tif31170

[0592] Example 7 Aqueous Humor Dynamics (AHD) in C57BL / 6J Mice After Treatment with Formulation 3d in Mice TIFF2025538061000069.tif50170

[0593] Study Objective: The objective of this study was to determine the effect of topical ophthalmic administration of Formulation 3d on aqueous humor dynamics in normotensive mammals (mice).

[0594] Study design: C57BL / 6J mice were used in this study. The data can be compared with studies published in Non-Patent Document 2 (Roy Chowdhury et al., 2015, PLOS ONE) and Non-Patent Document 10 (Roy Chowdhury et al., 2017, IOVS).

[0595] Methods: Wild-type C57BL / 6J mice (n = 7) were obtained from Jackson Laboratories (Bar Harbor, ME, USA). One day after baseline IOP measurements, a 5 μL bolus of Formulation 3d (0.5 mM) was applied topically to both eyes of each animal (n = 4) once daily for three consecutive days. In a separate cohort (n = 3), a 5 μL bolus of vehicle was applied topically to both eyes once daily for three consecutive days. On the third day after treatment, IOP was measured and animals were anesthetized with an intraperitoneal injection of ketamine (100 mg / kg), xylazine (10 mg / kg), and acepromazine (1 mg / kg). AHD (aqueous humor dynamics) was assessed by constant-flow infusion as previously described (Millar et al., 2011, IOVS; Roy Chowdhury et al., 2017, IOVS). All data were recorded and analyzed using LabScribe4 software (World Precision Instruments). Outflow capacity and episcleral venous pressure were measured, and uveoscleral outflow and aqueous humor flow were calculated. Drug tolerability was assessed daily by visual observation.

[0596] Results: Mice treated with 0.5 mM Formulation 3d demonstrated a reduction in IOP from 16.46 ± 0.5 mmHg to 12.25 ± 0.89 mmHg. In contrast, mice treated with vehicle demonstrated no change from baseline IOP (16.33 ± 0.30 mmHg to 16.39 ± 0.49 mmHg). Mice treated with Formulation 3d demonstrated a reduction in episcleral venous pressure (4.05 ± 0.42 mmHg) compared to vehicle-treated controls (9.81 ± 1.12 mmHg). The reduction in episcleral venous pressure was statistically significant (p = 0.0002). No significant changes were observed in outflow capacity, uveoscleral outflow, or aqueous humor flow. Daily clinical assessments showed no differences between drug-treated and vehicle-treated control mice.

[0597] TIFF2025538061000070.tif69170

[0598] The data in Table 4 show that formulation 3d reduces IOP by reducing episcleral venous pressure (EVP) in normotensive C57 / BL6J mice. No significant changes were observed in outflow capacity, uveoscleral outflow, or aqueous humor flow.

[0599] Example 8 Exploratory Study to Assess the Safety and Tolerability of Formulation 3d in Humans An exploratory study of levcromakalim Formulation 3d was conducted in Mexico under the supervision of a board-certified ophthalmologist practicing in Mexico City, in accordance with GCP and ICH guidelines. A randomized, blinded study was initially conducted to examine the ocular and systemic safety and tolerability of two concentrations of Formulation 3d (0.5 mM and 5 mM) delivered topically as a topical intraocular pressure (IOP)-lowering agent. Of the 14 subjects receiving Formulation 3d, two patients experienced adverse events. None of the adverse events were considered serious, and no patients discontinued the study due to an adverse event. The onset of IOP reduction was rapid, occurring within 4 hours after the first topical administration of Formulation 3d. Both concentrations of Formulation 3d were well tolerated, and IOP-lowering effects were observed throughout the 14-day treatment period.

[0600] Study Objective: The primary objective of this study was to evaluate the ocular and systemic safety and tolerability of two blinded concentrations of Formulation 3d when administered once daily (QD) to both eyes (OU) for 14 days in 14 healthy humans. A secondary objective of this study was to evaluate the IOP-lowering ability of Formulation 3d in humans.

[0601] Study Design: This study screened subjects without significant comorbidities or significant ocular pathology. The first three enrolled study subjects had visual impairments and were administered Formulation 3d QAM OU (0.5 mM or 5 mM) for 7 days. After acceptable safety and tolerability were confirmed, 14 subjects were enrolled and administered Formulation 3d (0.5 mM or 5 mM) for 14 days.

[0602] Inclusion criteria for the first three enrollments: 1) 20 years of age or older, 2) Visual impairment of BCVA 20 / 200 or less, ocular and / or cortical (CNS) pathology, i.e., amblyopia, retinal or corneal pathology, and / or advanced cataract in at least one eye. Only the visually impaired eye will be treated. If both eyes are eligible, both may be treated. Visual impairment should not be due to severe corneal pathology that may affect accurate IOP measurement.

[0603] Inclusion criteria for volunteers: After screening, 14 volunteers were selected. 1) 20 years of age or older, 2) Having an IOP of 17 mmHg or higher at the morning screening visit.

[0604] TIFF2025538061000071.tif63170

[0605] As shown in Table 5, the first three study subjects enrolled had visual impairment (as defined in inclusion / exclusion) with a best-corrected visual acuity (BCVA) of 20 / 200 or less in at least one eye. After safety and tolerability were confirmed after study completion for the first three enrolled subjects, 14 volunteers were enrolled, randomized, and blinded to receive one of two concentrations of Formulation 3d (0.5 mM or 5 mM).

[0606] Methods: Formulation 3d was provided in low and high concentrations of 0.5 mM and 5 mM (all treatment groups received blinded bottles labeled A or B). Bottle A contained 0.5 mM Formulation 3d, and Bottle B contained 5 mM Formulation 3d. Formulation 3d was administered once daily in both eyes (OU) at 8:00 AM ± 90 minutes QAM for 14 days (+2 days). Safety and tolerability were monitored by collecting adverse events (AEs). IOP was determined by Goldmann applanation tonometry (GAT) and iCare rebound tonometry. Clinical assessments included vital signs (blood pressure / BP and heart rate / HR), IOP, BCVA (using the Early Treatment Diabetic Retinopathy Study / ETDRS chart), slit lamp biomicroscopy and fundus examination, review of concomitant medications, and review of AEs. Because the primary objective of this study was to evaluate the ocular and systemic safety and tolerability of the two strengths of Formulation 3d, exploratory endpoints included AEs, BCVA, slit lamp, and mydriatic fundus examination.

[0607] Safety: Two AEs were recorded during the study. One AE occurred in a visually impaired patient, and the other AE was reported in a generally healthy study participant. No subjects discontinued the study due to an AE. The AE of systemic hypotension in Subject 1, a participant in the 5 mM treatment group, is a finding that correlates with previously published findings when the active substance is given orally (Hamilton TC, Beerahee A, Moen JS, et al. Levcromakalim. Cardiovasc Drug Rev. 1993;11(2)199-222).

[0608] Other than the two reported AEs, no serious or adverse safety signals were noted on ophthalmologic assessment or vital signs. Several incidents of transient mild redness were noted on slit lamp assessment in both treatment groups; however, these generally resolved during the day and were deemed to be of no clinical significance. In most cases, the investigators attributed the redness to the use of the local anesthetic used during study assessment.

[0609] Formulation 3d was shown to have an acceptable safety and tolerability profile for daily ophthalmic administration. Only two AEs were observed, and the hypotensive event in subject 1 correlates with published findings for the active substance when given orally (Hamilton 1993). No other adverse safety findings were observed in this study.

[0610] Efficacy: Table 6 shows the IOP-lowering efficacy of Formulation 3d. Mean baseline IOP was 19.3 mmHg across all patients, 19.4 mmHg in the 5 mM treatment group, and 19.2 mmHg in the 0.5 mM treatment group.

[0611] In the 5 mM higher treatment group, the greatest IOP reduction was observed by Goldmann applanation tonometry (GAT) at the Day 7 visit, with a reduction of -5.7 mmHg (29% reduction from baseline) observed pre-dose (morning) and 8 hours post-dose (late afternoon). iCare rebound tonometry confirmed a maximum IOP reduction of -5.64 mmHg (29% reduction from baseline) at 4 hours post-dose (noon) on Day 7. In the 0.5 mM lower treatment group, the greatest IOP reduction was observed by Goldmann applanation tonometry (GAT) at the Day 1 visit, with a reduction of -5.67 mmHg (29% reduction from baseline) observed 8 hours post-dose (late afternoon). iCare rebound tonometry confirmed a maximum IOP reduction of -5.39 mmHg (27% reduction from baseline) at 4 hours post-dose (noon) on Day 14. The onset of IOP reduction was rapid, occurring within 4 hours of the first dose of study drug on Day 1. Both treatment groups (5 mM and 0.5 mM) demonstrated IOP-lowering efficacy.

[0612] TIFF2025538061000072.tif112170

[0613] Of the 14 people who received Formulation 3d, two patients experienced AEs. None of the AEs were considered serious, and no patients discontinued the study due to an AE. Several incidents of transient mild hyperemia were observed in both treatment groups during slit lamp evaluation; however, these generally resolved by the next time point and were deemed to be of no clinical significance. In most cases, the investigator attributed the hyperemia to the use of the local anesthetic used during the study evaluations. The onset of IOP reduction was rapid, occurring within 4 hours after the first topical administration of the study drug. Both concentrations (5 mM and 0.5 mM) had IOP-lowering effects throughout the 14-day treatment period.

[0614] Example 9 Blinded Clinical Study to Evaluate the Safety, Tolerability, and Intraocular Pressure-Lowering Efficacy of Formulation 3d in Humans A single-center, randomized, blinded, vehicle-controlled study was conducted in human subjects to investigate the ocular and systemic safety and tolerability of two concentrations of Formulation 3d (0.5 mM and 2.5 mM) administered QAM for 7 days followed by BID for 7 days in 21 subjects without any significant comorbidities or significant ocular pathology. No ocular or systemic AEs occurred in any of the 21 patients. Both the 0.5 mM and 2.5 mM Formulation 3d treatment groups demonstrated IOP-lowering efficacy in human subjects.

[0615] Study Objective: The primary objective of this study was to evaluate the ocular and systemic safety and tolerability of two blinded strengths of Formulation 3d when administered once daily (QD) in both eyes (OU) for 7 days followed by twice daily (BID) dosing in the OU for 7 days. A secondary objective of this study was to evaluate the IOP-lowering ability of Formulation 3d.

[0616] Study Design: Subjects without any significant comorbidities or significant ocular pathology were screened on Day 0 and subsequently visited on Days 1 (start of medication), 7, and 14 (end of study treatment).

[0617] Inclusion criteria: Subjects were screened and 21 subjects who met the following criteria were selected. 1) 20 years of age or older, 2) Having an IOP of 17 mmHg or higher at the morning screening visit.

[0618] TIFF2025538061000073.tif72170

[0619] Twenty-one enrolled subjects were randomized to either Formulation 3d or vehicle, as shown in Table 7. Formulation 3d (blinded concentrations of 0.5 mM and 2.5 mM) or vehicle was administered QAM OU for 7 days according to randomization, followed by dosing BID OU for an additional 7 days.

[0620] Methods: Formulation 3d was provided to human subjects at concentrations of 0.5 mM and 2.5 mM, i.e., low and high concentrations (all treatment groups received blinded bottles labeled A, B, or C), in preservative-free dropper bottles and administered once daily at 8:00 AM ± 90 minutes QAM OU for 7 days (+2 days), followed by BID for an additional 7 days (+2 days). Bottles of study drug were distributed at Visit 2 (Day 1) and Visit 3 (Day 7). Patients received the same assigned treatment (A, B, or C) throughout the study. Safety and tolerability were monitored through data collection, including adverse events (AEs), vital signs, and ophthalmologic examinations, including best-corrected visual acuity (BCVA), slit lamp examination, and fundus examination. At screening, and on days 1, 7, and 14, subjects' IOP was measured by Goldmann applanation tonometry (GAT) and iCare rebound tonometry at noon and 4:00 PM (pre-treatment on day 1).

[0621] Clinical assessments included vital signs (blood pressure / BP and heart rate / HR), IOP, BCVA (using the Early Treatment Diabetic Retinopathy Study / ETDRS chart), slit lamp biomicroscopy and fundus examination, review of concomitant medications, and review of AEs. Because the primary objective of this study was to evaluate the ocular and systemic safety and tolerability of two concentrations and two dosing regimens of Formulation 3d, exploratory endpoints included AEs, BCVA, slit lamp, and dilated fundus examination.

[0622] Safety: No serious ocular or systemic AEs, such as hyperemia (eye redness), pain, burning, or stinging upon instillation, superficial corneal events, or systemic adverse events, were observed in human subjects. Mild, transient hyperemia after the morning dose during the study visit was observed in 4 of 7 subjects in the 2.5 mM cohort only. This typically resolved within 20 minutes and always by the noon assessment. No changes were observed by slit lamp examination in the 0.5 mM formulation 3d cohort or vehicle cohort.

[0623] Efficacy: Table 8, Table 9, Figures 5, 6, 7, and 8 show the IOP-lowering efficacy of Formulation 3d. Mean baseline IOP (GAT) was 17.0 mmHg across all subjects, 17.0 mmHg in the 2.5 mM Formulation 3d treatment group, 16.7 mmHg in the lower 0.5 mM Formulation 3d treatment group, and 17.2 mmHg in the vehicle control treatment group.

[0624] On Day 7 (after the QD dosing period), the mean diurnal IOP reductions measured by GAT were -2.1 mmHg (-12%), -2.1 mmHg (-13%), and -1.6 mmHg (-9%) compared to baseline in the 2.5 mM Formulation 3d, 0.5 mM Formulation 3d, and vehicle treatment groups, respectively (Figure 5). On Day 14 (after the BID dosing period), the mean diurnal IOP reductions were -2.6 mmHg (-15%), -3.5 mmHg (-21%), and -1.5 mmHg (-9%) compared to baseline in the 2.5 mM Formulation 3d, 0.5 mM Formulation 3d, and vehicle treatment groups, respectively (Figures 5 and 7).

[0625] The mean diurnal IOP reduction (GAT) for the 0.5 mM Formulation 3d group was significantly lower than the vehicle control group (p=0.003). Both Formulation 3d treatment groups (2.5 mM and 0.5 mM) demonstrated IOP-lowering efficacy (GAT and iCare). IOP in the 2.5 mM treatment group was significantly lower than the vehicle control group at 4 PM on Day 7 (QAM dosing regimen) and at 4 PM on Day 14 (BID dosing regimen). IOP in the 0.5 mM treatment group was significantly lower than the vehicle control at noon and 4 PM on Day 14 (BID dosing regimen) (Table 8, Figures 6 and 8).

[0626] TIFF2025538061000074.tif75170

[0627] TIFF2025538061000075.tif243170TIFF2025538061000076.tif61170

[0628] No ocular or systemic AEs occurred. Mild, transient hyperemia was observed sporadically in the 2.5 mM group but was not perceived as an AE. This occurred in 4 of 7 subjects in the 2.5 mM cohort after the morning dose during the study visit. No changes were observed by slit lamp examination in the 0.5 mM formulation 3d cohort or vehicle cohort. Compared to vehicle control, the 2.5 mM treatment group demonstrated significantly lower IOP at 4 PM on Day 7 (QAM dosing regimen) and Day 14 (BID dosing regimen), whereas the 0.5 mM group demonstrated significantly lower IOP at noon and 4 PM on Day 14 (BID dosing regimen). Both concentrations (2.5 mM and 0.5 mM) had IOP-lowering effects throughout the 7- and 14-day treatment periods.

[0629] Example 10 Five-day tolerability study of formulation 3d via intraocular topical instillation in rabbits The toxicokinetic properties of Formulation 3d were evaluated in Dutch-Belted (DB) rabbits. The effects of Formulation 3d on ocular safety and systemic PK after bilateral dosing BID were evaluated in DB rabbits treated with two doses: 0.02 mg per eye per dose and 0.06 mg per eye per dose.

[0630] dosage Formulation 3d was administered to rabbits via topical ophthalmic (TO) instillation twice daily (BID) for 5 consecutive days in both eyes (OU, both eyes). Formulation 3d was administered via TO at two doses (0.05% [0.02 mg per eye per dose, 0.04 mg per eye per day] and 0.15% [0.06 mg per eye per dose, 0.12 mg per eye per day]) for 5 consecutive days.

[0631] observation Both doses were well tolerated in rabbits, with no drug-related adverse events. There were no deaths, no clinical signs or changes in body weight or food intake related to Formulation 3d, and no ocular changes considered test-related as assessed by gross eye examination, ophthalmology, or intraocular pressure measurements. There were no macroscopic findings in the tissues examined.

[0632] result Peak plasma exposure in this study was low, with measured concentrations below 3.29 ng / mL (lower limit of quantification = 0.5 ng / mL), and values ​​were not quantifiable in samples collected more than 4 hours after dosing. In male rabbits, the observed AUC tlast value and maximum plasma concentration (C max ) values ​​increased less than dose-proportionally with increasing dose administered, whereas in females, C max and AUC tlast were similar in both dose groups. Although AUC and AUC / dose values ​​were slightly higher on day 5 compared with day 1, dose accumulation over the 5 days was unlikely due to the high variability (Table).

[0633] TIFF2025538061000077.tif162170

[0634] Both doses of Formulation 3d (0.05% [1.7 mM], 0.02 mg per eye per dose [Group 1] and 0.15% [5 mM], 0.06 mg per eye per dose [Group 2]) were well tolerated in rabbits, and no significant changes were observed during ophthalmologic and macroscopic examinations over the 5-day dosing period. Levcromakalim was detectable in plasma for only up to 4 hours after dosing at both concentrations (1.7 mM and 5 mM). Area under the drug concentration-time curve (AUC) and AUC / dose values ​​were slightly higher on Day 5 compared with Day 1, but the wide variability made it difficult to draw any definitive conclusions regarding accumulation over the 5-day dosing period.

[0635] This study demonstrates the ability of a topical solution of Formulation 3d to measurably deliver the active pharmaceutical ingredient (API), levcromakalim, to rabbits. No serious ocular or systemic AEs, such as hyperemia (eye redness), were observed in this study.

[0636] Example 11 Evaluation of the Tolerability, Safety, and Pharmacokinetics of Formulation 3d in a 15-Day Non-GLP Study in Dutch-Belted Rabbits The effect of Formulation 3d on ocular safety, tolerability, and systemic PK after 14 days of QD dosing was evaluated in DB rabbits treated with levcromakalim Formulation 3d at doses of 0 mM (formulation buffer), 0.05 mM (0.0015%), 0.5 mM (0.015%), and 5 mM (0.15%).

[0637] method The left eyes of rabbits were topically dosed once daily with Formulation 3d at 0 mM (formulation buffer), [0.0015% (0.05 mM, 0.0006 mg per eye per dose), 0.015% (0.5 mM, 0.006 mg per eye per dose), 0.15% (5 mM, 0.06 mg per eye per dose)], whereas the right eyes received the formulation buffer.

[0638] Ocular-related safety endpoints included ocular examinations (baseline, days 3, 7, 10, and 14), intraocular pressure measurements (BID for 5 days prior to the start of dosing and 4 and 23 hours after dosing throughout the study, excluding weekend days and day 15), and histopathology. In high-dose animals, plasma was collected from three animals each at 30 minutes and 1, 2, 4, 6, 8, and 23 hours after dosing on days 1 and 15. Animals were euthanized on day 15, and left eyes (n=2 eyes per group) from all treatment groups were collected and processed for PK analysis of ocular tissues.

[0639] observation At all pre- and post-dose time points, IOP remained within the normal range for this strain and species. No other signs of histological abnormalities were observed. Ophthalmic examination scores were "0" in all animals at all time points, indicating the absence of inflammation. Ophthalmic examinations on various days during treatment did not reveal any signs of adverse events (AEs) (score of 0 in all animals and at all time points), except for two incident cases of conjunctivitis (one in control and one treated at the lowest dose).

[0640] result Bioanalysis results showed that low levels of levcromakalim were detected 30 minutes, 1 hour, 2 hours, and 4 hours after a single dose of Formulation 3d (high dose, 5 mM). After 4 hours, no levcromakalim was detected in plasma. Plasma levels of levcromakalim on Day 15, after 14 days of once-daily dosing, were similar to Day 1, where levcromakalim was detectable at low plasma levels up to 4 hours after dosing and did not accumulate. An exception was seen in one animal at the 2-hour time point, which showed a concentration of 991.60 ng / mL. This appears to be anomalous, as the levcromakalim concentrations from other time points for this and all other animals were within the normal range. The cause of this high reading is unknown.

[0641] Treatment with Formulation 3d resulted in low systemic levels of levcromakalim. Levcromakalim was detectable in ocular tissues up to 4 hours after dosing, with no evidence of accumulation. Low levels of levcromakalim were detected in the aqueous humor (AH) (0.88 ng / mL to 1.78 ng / mL), iris ciliary body (ICB) (0.75 ng / mL to 0.92 ng / mL), retina (0.69 ng / mL), and retinal pigment epithelium (RPE) / choroid (0.50 ng / mL to 6.63 ng / mL) up to 4 hours after dosing on Day 15. Overall, Formulation 3d was well tolerated without AEs when dosed QD at three different concentrations.

[0642] This study demonstrates that formulation 3d exhibits a surprisingly benign toxicity profile in rabbits up to its maximum formulateable concentration (5 mM) without significant adverse events. No serious ocular or systemic AEs, such as hyperemia (redness of the eye), were observed in this study.

[0643] Example 12: 28-day repeated dose toxicology study of formulation 3d via intraocular topical administration in Dutch-Belted rabbits Plasma concentrations of levcromakalim were evaluated in DB rabbits after topical ophthalmic (TO) administration of Formulation 3d to both eyes (OU, both eyes) twice daily (BID) for 28 days at dose levels of 0 mg, 0.012 mg, 0.03 mg, and 0.06 mg per eye per dose.

[0644] method Plasma concentrations of levcromakalim were evaluated in DB rabbits after 28 days of TO administration of Formulation 3d at dose levels of 0 mg, 0.012 mg, 0.03 mg, and 0.06 mg per eye per dose (0 mg, 0.024 mg, 0.06 mg, and 0.12 mg per eye per day, respectively, or 0.030% [1.0 mM], 0.075% [2.5 mM], or 0.15% [5 mM], respectively). Plasma was collected from three animals per dose group on days 1 and 28 at pre-dose and 0.5, 1, 2, 3, 4, 6, and 12 hours after the first dose.

[0645] observation Formulation 3d was well tolerated in rabbits at dose levels of 0.012 mg, 0.03 mg, and 0.06 mg per eye per dose (0.024 mg, 0.06 mg, and 0.12 mg per eye per day), respectively, and no adverse changes were observed during the 28-day dosing period or after a 2-week recovery period. No findings were observed at any endpoint, including clinical observations, body weight, food intake, ophthalmology, ERG, intraocular pressure measurements, clinical pathology, macroscopic and / or microscopic evaluations, and histopathology of a comprehensive list of systemic tissues.

[0646] result Peak plasma exposure was low, with measured concentrations below 4.942 ng / mL (lower limit of quantification = 0.500 ng / mL), and values ​​were not quantifiable in samples taken more than 6 hours after dosing (Table). tlast and C max increased at a rate that was less than dose-proportional with increasing dose levels. Due to the wide variability in the studies, AUC and C values ​​differed between females and males. max It was difficult to conclude whether there were any differences in the values. AUC tlast Value and C max Values ​​are for males in group 4 (R AUC =1.083, R Cmax = 0.986), was slightly lower on day 28 compared to day 1, indicating that no systemic accumulation of levcromakalim was observed over 28 days of TO dosing with formulation 3d BID.

[0647] TIFF2025538061000078.tif188170

[0648] The no-observed-adverse-effect level (NOAEL) was determined to be 0.06 mg per eye per dose (0.12 mg per eye per day). Note that this highest dose of 0.12 mg per eye per day was also the maximum administrable dose based on dose volume and solubility. At the NOAEL, mean C max The mean C value and AUC value were 2.58 ng / mL and 6.11 h ng / mL, respectively, and the mean C value in females was 2.58 ng / mL and 6.11 h ng / mL. max The RI and AUC values ​​were 2.06 ng / mL and 6.05 h ng / mL, respectively. Similar to previous studies, plasma exposure levels of levcromakalim were low and quantifiable only up to 6 hours after dosing, demonstrating no accumulation over 28 days of dosing with Formulation 3d.

[0649] This study demonstrates that this novel formulation, Formulation 3d, exhibits a surprisingly benign toxicity profile in rabbits up to its maximum formulateable concentration (5 mM) without significant adverse events. No serious ocular or systemic AEs, such as hyperemia (redness of the eye), were observed in this study.

[0650] Example 13 Exemplary Topical Ophthalmic Compositions of Levcromakalim in Humans Levcromakalim ophthalmic solution for topical administration was manufactured under cGMP conditions at potencies of 0.015%, 0.030%, and 0.075%, containing 0.0 mg / mL, 0.15 mg / mL, 0.30 mg / mL, and 0.75 mg / mL levcromakalim, respectively.

[0651] Levcromakalim ophthalmic solution was packaged in low-density polyethylene form-fill-seal (BFS) disposable unit-dose containers. A description of the formulation is provided in Table 12.

[0652] TIFF2025538061000079.tif75170

[0653] The compositions of Formulation 3d ophthalmic solutions at potencies of 0.0% (vehicle control), 0.015%, 0.030%, and 0.075% are shown in Table 13. A unit dose consists of 310 mg of Formulation 3d ophthalmic solution in a BFS container closure system. For human application, the dose consists of one drop of approximately 30 μL intended for topical administration to each eye.

[0654] TIFF2025538061000080.tif238170

[0655] The specific gravity of the formulation was measured during formulation development and converted to density. The density of the formulation was 1.02 g / mL.

[0656] The formulation components in levcromakalim ophthalmic solution (0.0% (vehicle control), 0.015%, 0.030%, and 0.075%) are shown in Table 14.

[0657] TIFF2025538061000081.tif114170

[0658] All excipients used in levcromakalim ophthalmic solution are present within the limits for the topical ophthalmic drug (drops) route of administration.

[0659] This specification has been described with reference to embodiments of the present invention. However, those skilled in the art will recognize that various modifications and changes can be made without departing from the scope of the invention as set forth herein. Accordingly, this specification is to be considered in an illustrative rather than a restrictive sense, and all such modifications are intended to be included within the scope of the invention.

Claims

1. 1. A topical ophthalmic formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof having a concentration of between about 0.01 mM and about 5 mM, wherein the topical ophthalmic formulation is stable at ambient conditions for at least 4 months; and a. a polyol, b. polyethoxylated furanose fatty acid esters; c. Nonionic triblock copolymers of a central hydrophobic chain of polyoxypropylene flanked by two hydrophilic chains of polyoxyethylene (poloxamers); d. polymeric alkyl or aryl polyols; e. ethoxylated glycerol esters, f. polymeric lactams, g. hydroxyalkyl cellulose, h. Polyacrylic acid and hydrophobic C １０ ~C ３０ an oil-in-water polymeric emulsifier that is a block copolymer with an alkyl acrylate; and i. ethoxylated alkylphenols, A topical ophthalmic formulation comprising three or more ingredients selected from the group consisting of:

2. 10. The topical ophthalmic formulation of claim 1, wherein the topical ophthalmic formulation comprises four or more ingredients.

3. 10. The topical ophthalmic formulation of claim 1, wherein the topical ophthalmic formulation comprises five or more ingredients.

4. 10. The topical ophthalmic formulation of claim 1, wherein the topical ophthalmic formulation comprises six or more ingredients.

5. The topical ophthalmic formulation of any one of claims 1 to 4, wherein the composition comprises a polyol.

6. 6. The topical ophthalmic formulation of claim 5, wherein the polyol is an alkyl polyol.

7. 7. The topical ophthalmic formulation of claim 6, wherein the alkyl polyol is a triol.

8. 7. The topical ophthalmic formulation of claim 6, wherein the alkyl polyol is a sugar alcohol.

9. 8. The topical ophthalmic formulation of claim 7, wherein the triol is glycerin.

10. 7. The topical ophthalmic formulation of claim 6, wherein the alkyl polyol is mannitol.

11. The topical ophthalmic formulation of any one of claims 1 to 10, wherein the composition comprises a polyethoxylated furanose fatty acid ester.

12. 12. The topical ophthalmic formulation of claim 11, wherein the polyethoxylated furanose fatty acid ester is polysorbate 80.

13. The topical ophthalmic formulation of any one of claims 1 to 12, wherein the composition comprises a poloxamer.

14. 14. The topical ophthalmic formulation of claim 13, wherein the poloxamer is poloxamer 407 or Pluronic 127.

15. The topical ophthalmic formulation of any one of claims 1 to 14, wherein the composition comprises a polymeric alkyl or aryl polyol.

16. 16. The topical ophthalmic formulation of claim 15, wherein the polymeric alkyl or aryl polyol is tyloxapol.

17. The topical ophthalmic formulation of any one of claims 1 to 16, wherein the composition comprises an ethoxylated glycerol ester.

18. 18. The topical ophthalmic formulation of claim 17, wherein the ethoxylated glycerol ester is Kolliphor™ or Cremophor™.

19. 20. The topical ophthalmic formulation of claim 18, wherein the Kolliphor™ or Cremophor™ is selected from the group consisting of Kolliphor™ ELP, Kolliphor™ RH 40, and Kolliphor™ HS 15.

20. 20. The topical ophthalmic formulation of claim 19, wherein the Kolliphor™ or Cremophor™ is Kolliphor™ ELP.

21. 20. The topical ophthalmic formulation of claim 19, wherein the Kolliphor™ or Cremophor™ is Kolliphor™ RH 40.

22. 20. The topical ophthalmic formulation of claim 19, wherein the Kolliphor™ or Cremophor™ is Kolliphor™ HS 15.

23. The topical ophthalmic formulation of any one of claims 1 to 22, wherein the composition comprises a polymeric lactam.

24. 24. The topical ophthalmic formulation of claim 23, wherein the polymeric lactam is PVP.

25. 25. The topical ophthalmic formulation of claim 24, wherein the PVP is selected from the group of PVP K-30 and PVP K-90.

26. 26. The topical ophthalmic formulation of claim 25, wherein the PVP is PVP K-30.

27. 26. The topical ophthalmic formulation of claim 25, wherein the PVP is PVP K-90.

28. The topical ophthalmic formulation of any one of claims 1 to 27, wherein the composition comprises a hydroxyalkyl cellulose.

29. 29. The topical ophthalmic formulation of claim 28, wherein the hydroxyalkyl cellulose is hypromellose.

30. The composition comprises polyacrylic acid and hydrophobic C １０ ~C ３０ 30. The topical ophthalmic formulation of any one of claims 1 to 29, comprising an oil-in-water polymeric emulsifier which is a block copolymer with an alkyl acrylate.

31. The polyacrylic acid and hydrophobic C １０ ~C ３０ 31. The topical ophthalmic formulation of claim 30, wherein the polymeric oil-in-water emulsifier of a block copolymer with an alkyl acrylate is Premulen.

32. 32. The topical ophthalmic formulation of claim 31, wherein the Premulen is Premulen™ TR-1 or Premulen™ TR-2.

33. The topical ophthalmic formulation of any one of claims 1 to 32, wherein the composition comprises an ethoxylated alkylphenol.

34. 34. The ophthalmic preparation of claim 33, wherein the ethoxylated alkylphenol is an octoxynol.

35. 35. The topical ophthalmic formulation of claim 34, wherein the octoxynol is octoxynol-40.

36. 36. The topical ophthalmic formulation of any one of claims 1 to 35, wherein the formulation is stable at ambient conditions for at least 5 months.

37. 37. The topical ophthalmic formulation of claim 36, wherein the formulation is stable at ambient conditions for at least six months.

38. 38. The topical ophthalmic formulation of claim 37, wherein the formulation is stable at ambient conditions for at least 7 months.

39. 39. The topical ophthalmic formulation of any one of claims 1 to 38, wherein the formulation is an aqueous formulation at a pH of about 6.5 to about 7.

5.

40. 40. The topical ophthalmic formulation of any one of claims 1 to 39, wherein the concentration of (rev)cromakalim is between about 0.05 mM and about 5 mM.

41. 41. The topical ophthalmic formulation of any one of claims 1 to 40, wherein the concentration of (rev)cromakalim is between about 0.5 mM and about 5 mM.

42. 42. The topical ophthalmic formulation of any one of claims 1 to 41, wherein the concentration of (rev)cromakalim is between about 1 mM and about 5 mM.

43. 43. The topical ophthalmic formulation of any one of claims 1 to 42, wherein the concentration of (rev)cromakalim is between about 1.5 mM and about 5 mM.

44. 44. The topical ophthalmic formulation of any one of claims 1 to 43, wherein the concentration of (rev)cromakalim is between about 2 mM and about 5 mM.

45. 45. The topical ophthalmic formulation of any one of claims 1 to 44, wherein the concentration of (rev)cromakalim is between about 2.5 mM and about 5 mM.

46. 46. ​​The topical ophthalmic formulation of any one of claims 1 to 45, wherein the concentration of (rev)cromakalim is between about 3 mM and about 5 mM.

47. 47. The topical ophthalmic formulation of any one of claims 1 to 46, wherein the concentration of (rev)cromakalim is between about 3.5 mM and about 5 mM.

48. 48. The topical ophthalmic formulation of any one of claims 1 to 47, wherein the concentration of (rev)cromakalim is between about 4 mM and about 5 mM.

49. 1. A topical ophthalmic formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof having a concentration of between about 0.01 mM and about 5 mM, which is stable at ambient conditions for at least 5 months, and which comprises three or more ingredients selected from the group consisting of glycerin, polysorbate 80, poloxamer 407 or Pluronic™ F127, tyloxapol, Kolliphor™ ELP, Kolliphor™ RH 40, Kolliphor™ HS 15, hypromellose, PVP, Premulen™ TR-1 or Premulen™ TR-2, and octoxynol-40, in an aqueous formulation having a pH of about 6 to about 8.

50. 50. The topical ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of three or more ingredients selected from the group consisting of glycerin, polysorbate 80, poloxamer 407 or Pluronic™ F127, tyloxapol, Kolliphor™ ELP, Kolliphor™ RH 40, Kolliphor™ HS 15, hypromellose, PVP, Premulen™ TR-1 or Premulen™ TR-2, and octoxynol-40 in an aqueous formulation having a pH of about 6 to about 8.

51. 50. The topical ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ RH 40, Octoxynol-40, and Premulen™ TR-2 in an aqueous formulation having a pH of about 6 to about 8.

52. 50. The topical ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ RH 40, Kolliphor™ HS 15, and PVP in an aqueous formulation having a pH of about 6 to about 8.

53. 50. The ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor™ ELP, polysorbate 80, and hypromellose in an aqueous formulation having a pH of about 6 to about 8.

54. 50. The ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, Polysorbate 80, and PVP in an aqueous formulation having a pH of about 6 to about 8.

55. 50. The ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose in an aqueous formulation having a pH of about 6 to about 8.

56. 50. The ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor™ RH 40, Premulen™ TR-2, and PVP in an aqueous formulation having a pH of about 6 to about 8.

57. 50. The ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ HS 15, Premulen™ TR-2, Polysorbate 80, Octoxynol-40, and Poloxamer 407 in an aqueous formulation having a pH of about 6 to about 8.

58. 50. The ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists of Polysorbate 80, Kolliphor™ ELP, and PVP in an aqueous formulation having a pH of about 6 to about 8.

59. 50. The ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, and poloxamer 407 in an aqueous formulation having a pH of about 6 to about 8.

60. 50. The ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, PVP, and poloxamer 407 in an aqueous formulation having a pH of about 6 to about 8.

61. 50. The ophthalmic formulation of claim 49, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, and hypromellose in an aqueous formulation having a pH of about 6 to about 8.

62. A topical ophthalmic formulation comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP), poloxamer 407, mannitol, water, and a phosphate buffer, and having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 0.01 mM and about 5 mM, and stable for at least 5 months at ambient conditions, and having a pH of about 6 to about 8.

63. 63. The topical ophthalmic formulation of claim 62, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 0.05 mM and about 5 mM.

64. 64. The topical ophthalmic formulation of claim 62 or 63, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 0.5 mM and about 5 mM.

65. 65. The topical ophthalmic formulation of any one of claims 62 to 64, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 1 mM and about 5 mM.

66. 66. The topical ophthalmic formulation of any one of claims 62 to 65, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 1.5 mM and about 5 mM.

67. 67. The ophthalmic formulation of any one of claims 62 to 66, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 2 mM and about 5 mM.

68. 68. The ophthalmic formulation of any one of claims 62 to 67, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 2.5 mM and about 5 mM.

69. 69. The ophthalmic formulation of any one of claims 62 to 68, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 3 mM and about 5 mM.

70. 70. The ophthalmic formulation of any one of claims 62 to 69, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 3.5 mM and about 5 mM.

71. 71. The ophthalmic formulation of any one of claims 62 to 70, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 4 mM and about 5 mM.

72. 72. The ophthalmic formulation of any one of claims 62-71, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol in an aqueous formulation having a pH of about 6 to about 8.

73. 73. The ophthalmic formulation of any one of claims 62-72, wherein the ophthalmic formulation consists of Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, and mannitol in an aqueous formulation having a pH of about 6 to about 8.

74. 74. The ophthalmic formulation of any one of claims 1 to 73, wherein the ophthalmic formulation optionally comprises benzalkonium chloride (BAK) and optionally a pH adjuster.

75. 72. The ophthalmic formulation of any one of claims 62-71, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, mannitol, and BAK in an aqueous formulation having a pH of about 6 to about 8.

76. 72. The ophthalmic formulation of any one of claims 62-71, wherein the ophthalmic formulation consists of Kolliphor™ ELP, polysorbate 80, PVP, poloxamer 407, mannitol, and BAK in an aqueous formulation having a pH of about 6 to about 8.

77. A topical ophthalmic formulation comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof up to about 5 mM, and stable for at least 5 months at ambient conditions, and having a pH of about 6.

5.

78. A topical ophthalmic formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof having a concentration of between about 0.01 mM and about 5 mM, comprising Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, and which is stable at ambient conditions for at least 5 months and has a pH of about 6.

5.

79. An aqueous ophthalmic topical formulation having a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 0.05 mM and about 5 mM, having a pH of about 6 to about 8, and being stable at ambient conditions for at least 5 months, and comprising glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, and mannitol.

80. 80. The ophthalmic formulation of claim 78 or 79, wherein the ophthalmic formulation optionally comprises BAK and a pH adjuster.

81. 81. The ophthalmic formulation of claim 80, wherein the ophthalmic formulation consists essentially of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, mannitol, optionally benzalkonium chloride, and optionally a pH adjuster in an aqueous formulation having a pH of about 6 to about 8.

82. 81. The topical ophthalmic formulation of claim 80, wherein the ophthalmic formulation consists of glycerin, Kolliphor™ ELP, polysorbate 80, poloxamer 407, hypromellose, mannitol, and BAK in an aqueous formulation having a pH of about 6 to about 8.

83. A topical ophthalmic formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof having a concentration of between about 0.05 mM and about 5 mM, the formulation having a pH of about 6 to about 8 and stable at ambient conditions for at least 4 months, and comprising Kolliphor™ ELP, polysorbate 80, poloxamer 407, PVP K-30, mannitol, and optionally a pH adjuster.

84. 84. The topical ophthalmic formulation of claim 83, wherein the ophthalmic formulation consists essentially of Kolliphor™ ELP, polysorbate 80, poloxamer 407, PVP K-30, and mannitol in an aqueous formulation having a pH of about 6 to about 8.

85. 84. The topical ophthalmic formulation of claim 83, wherein the ophthalmic formulation consists of Kolliphor™ ELP, polysorbate 80, poloxamer 407, PVP K-30, and mannitol in an aqueous formulation having a pH of about 6 to about 8.

86. 86. The topical ophthalmic formulation of any one of claims 1 to 85, wherein the phosphate buffer is selected from dibasic sodium phosphate and monobasic sodium phosphate.

87. A pharmaceutical composition comprising (lev)cromakalim or a pharmaceutically acceptable salt thereof, having a concentration of (lev)cromakalim or a pharmaceutically acceptable salt thereof between about 0.05 mM and about 5 mM, and stable at ambient conditions for at least 4 months; and a. a polyol, b. polyethoxylated furanose fatty acid esters; c. A non-ionic triblock copolymer of a central hydrophobic chain of polyoxypropylene flanked by two hydrophilic chains of polyoxyethylene; d. polymeric alkyl or aryl polyols; e. ethoxylated glycerol esters, f. polymeric lactams, g. hydroxyalkyl cellulose, h. Polyacrylic acid and hydrophobic C １０ ~C ３０ an oil-in-water polymeric emulsifier that is a block copolymer with an alkyl acrylate; and i. ethoxylated alkylphenols, A pharmaceutical composition comprising three or more ingredients selected from the group consisting of:

88. A topical formulation for eye drops comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, wherein the formulation has a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of between about 0.01 mM and about 5 mM, and is stable at ambient conditions for at least 5 months, and has a pH of about 6 to about 8.

89. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 0.05 mM to 5 mM.

90. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 0.1 mM to 5 mM.

91. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 0.5 mM to 5 mM.

92. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 1 mM to 5 mM.

93. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 1.5 mM to 5 mM.

94. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 2 mM to 5 mM.

95. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 2.5 mM to 5 mM.

96. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 3 mM to 5 mM.

97. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 3.5 mM to 5 mM.

98. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 4 mM to 5 mM.

99. 89. The topical formulation of claim 88, wherein the concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof is about 5 mM.

100. A topical formulation for use as an eye drop comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, wherein the topical formulation has a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of about 0.4 mM, is stable at ambient conditions for at least 5 months, and has a pH of about 6.

5.

101. A topical formulation that can be used as an eye drop, comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, wherein the topical formulation has a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of about 0.8 mM, is stable at ambient conditions for at least 5 months, and has a pH of about 6.

5.

102. A topical formulation that can be used as an eye drop, comprising (rev)cromakalim or a pharmaceutically acceptable salt thereof, Kolliphor™ ELP, polysorbate 80, polyvinylpyrrolidone (PVP) K-30, poloxamer 407, mannitol, water, and phosphate buffer, wherein the topical formulation has a concentration of (rev)cromakalim or a pharmaceutically acceptable salt thereof of about 2 mM, is stable at ambient conditions for at least 5 months, and has a pH of about 6.

5.

103. 103. The topical ophthalmic formulation of any one of claims 87-102, comprising Kolliphor™ ELP in a concentration of about 5% (weight / volume).

104. 104. The topical ophthalmic formulation of claim 103, comprising Kolliphor™ ELP at a concentration of about 4% (weight / volume).

105. 105. The topical ophthalmic formulation of any one of claims 87 to 104, comprising polysorbate 80 at a concentration of about 1% (weight / volume).

106. 106. The topical ophthalmic formulation of any one of claims 87 to 105, comprising PVP K-30 at a concentration of about 2% (weight / volume).

107. 107. The topical ophthalmic formulation of any one of claims 87 to 106, comprising poloxamer 407 in a concentration of about 0.1% (weight / volume).

108. 108. The topical ophthalmic formulation of any one of claims 87 to 107, comprising mannitol at a concentration of about 4.6% (weight / volume).

109. 104. The topical ophthalmic formulation of claim 103, comprising mannitol at a concentration of about 3.3%.

110. A topical ophthalmic formulation that can be used as an eye drop comprising between about 0.05 mM and about 5 mM (rev)cromakalim or a pharmaceutically acceptable salt thereof in an aqueous liquid, which is stable for at least 5 months under ambient conditions without significant crystallization or excessive separation of cromakalim from said liquid, and which does not contain DMSO, DMF, or NMP, or other topical carriers that are not acceptable for human use, and which contains an optional buffer in addition to at least two excipients.

111. A topical ophthalmic formulation that can be used as an eye drop comprising between about 0.1 mM and about 5 mM (rev)cromakalim or a pharmaceutically acceptable salt thereof in an aqueous liquid, which is stable for at least 5 months under ambient conditions without significant crystallization or excessive separation of cromakalim from said liquid, and which does not contain DMSO, DMF, or NMP, or other topical carriers that are not acceptable for human use, and which contains an optional buffer in addition to at least two excipients.

112. A topical ophthalmic formulation that can be used as an eye drop comprising between about 0.5 mM and about 5 mM (rev)cromakalim or a pharmaceutically acceptable salt thereof in an aqueous liquid, which is stable for at least 5 months under ambient conditions without significant crystallization or excessive separation of cromakalim from said liquid, and which does not contain DMSO, DMF, or NMP or other topical carriers that are not acceptable for human use, and which contains an optional buffer in addition to at least two excipients.

113. 113. The topical ophthalmic formulation of any one of claims 1 to 112, wherein the formulation is not an emulsion.

114. 113. The topical ophthalmic formulation of any one of claims 1 to 112, wherein the formulation is not a gel.

115. 113. The topical ophthalmic formulation of any one of claims 1 to 112, wherein the formulation is not a topical gel.

116. 113. The topical ophthalmic formulation of any one of claims 1 to 112, wherein the formulation is a clear liquid.

117. 113. The topical ophthalmic formulation of any one of claims 1 to 112, wherein the formulation is a micellar or nanomicelle solution.

118. 113. The topical ophthalmic formulation of any one of claims 1 to 112, wherein the formulation does not comprise an oil as specifically defined above.

119. 119. The topical ophthalmic formulation of any one of claims 1 to 118, wherein the formulation has a percent transmittance of greater than 85% when tested with a UV-Visible spectrophotometer.

120. 119. The topical ophthalmic formulation of any one of claims 1 to 118, wherein the formulation has a percent transmittance of greater than 90% when tested with a UV-Visible spectrophotometer.

121. 119. The topical ophthalmic formulation of any one of claims 1 to 118, wherein the formulation has a percent transmittance of greater than 95% when tested with a UV-Visible spectrophotometer.

122. 122. The topical ophthalmic formulation of any one of claims 1 to 121, wherein the formulation comprises levcromakalim.

123. 122. The topical ophthalmic formulation of any one of claims 1 to 121, wherein the formulation comprises cromakalim.

124. 124. A method of treating an ocular disorder affecting the anterior or posterior segment of the eye, comprising administering to a host in need thereof an effective amount of (lev)cromakalim or a pharmaceutically acceptable salt thereof in a topical ophthalmic formulation according to any one of claims 1 to 123.

125. 125. The method of claim 124, wherein the host is a human.

126. 125. The method of claim 124, wherein use of the topical formulation results in lower intraocular pressure.

127. 125. The method of claim 124, wherein the eye disorder is glaucoma.

128. 128. The method of claim 127, wherein the glaucoma is glaucoma with normal intraocular pressure or normal tension glaucoma (NTG).

129. The method of claim 127, wherein the glaucoma is a glaucoma associated with elevated intraocular pressure selected from the group consisting of primary open-angle glaucoma (POAG), primary angle-closure glaucoma (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, glaucoma associated with diabetic retinopathy, glaucoma associated with Sturge-Weber syndrome, steroid-induced glaucoma, and acute glaucoma due to progressive cataract and / or intravitreal injection.

130. 130. The method of claim 129, wherein the eye disorder is glaucoma induced by Sturge-Weber syndrome or Sturge-Weber syndrome induced glaucoma.

131. 130. The method of claim 129, wherein the eye disorder is diabetic retinopathy.

132. The method of claim 129, wherein the local treatment is a primary or secondary or adjunctive treatment as part of a protocol for MIGS (Minimally Invasive Glaucoma Surgery) selected from the group consisting of a miniaturized form of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, a complete intraocular or suprachoroidal shunt, a gentler, more gentle form of laser cyclophotocoagulation, and in alternative claims, a Schlemm's canal stent to dilate the Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty.

133. 130. The method of claim 129, wherein the ocular disorder is Graves' ophthalmopathy, thyroid-associated orbitopathy (TAO), Graves' orbitopathy (GO), retrobulbar tumor, cavernous sinus thrombosis, orbital vein thrombosis, episcleral / orbital vein vasculitis, superior vena cava occlusion, superior vena cava thrombosis, carotid-cavernous sinus fistula, dural cavernous shunt, orbital varicose veins, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), arterial occlusive / embolic-hypoperfusion disease, or optic nerve damage due to ischemia (posterior and anterior ischemic optic neuropathy (NAION)).

134. 130. The method of claim 129, wherein said treatment with the topical formulation provides cytoprotection and / or neuroprotection to a host in need thereof.

135. 124. Use of (lev)cromakalim or a pharmaceutically acceptable salt thereof, as a formulation according to any one of claims 1 to 123, in the manufacture of a medicament for the topical treatment of an ocular disorder in a host in need thereof.

136. 136. The use of claim 135, wherein the eye disorder is glaucoma.

137. 137. The use of claim 136, wherein the glaucoma is glaucoma with normal intraocular pressure or normal tension glaucoma (NTG).

138. The use of claim 136, wherein the glaucoma is glaucoma associated with elevated intraocular pressure selected from the group consisting of primary open-angle glaucoma (POAG), primary angle-closure glaucoma (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, glaucoma associated with diabetic retinopathy, glaucoma associated with Sturge-Weber syndrome, steroid-induced glaucoma, and acute glaucoma due to progressive cataract and / or intravitreal injection.

139. 139. The use of claim 138, wherein the eye disorder is glaucoma induced by Sturge-Weber syndrome or Sturge-Weber syndrome induced glaucoma.

140. 139. The use of claim 138, wherein the eye disorder is diabetic retinopathy.

141. The use described in claim 138, wherein the local treatment is a primary or secondary or adjunctive treatment as part of a protocol for MIGS (Minimally Invasive Glaucoma Surgery) selected from the group consisting of miniaturized forms of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, complete intraocular or suprachoroidal shunts, gentler and more gentle forms of laser cyclophotocoagulation, and in alternative claims, Schlemm's canal stents for dilating Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty.

142. 139. The use of claim 138, wherein the ocular disorder is Graves' ophthalmopathy, thyroid-associated orbitopathy (TAO), Graves' orbitopathy (GO), retrobulbar tumor, cavernous sinus thrombosis, orbital vein thrombosis, episcleral / orbital vein vasculitis, superior vena cava occlusion, superior vena cava thrombosis, carotid-cavernous sinus fistula, dural cavernous sinus shunt, orbital varicose veins, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), arterial occlusive / embolic-hypoperfusion disease, or optic nerve damage due to ischemia (posterior and anterior ischemic optic neuropathy (NAION)).

143. 139. The use of claim 138, wherein said treatment with a topical formulation provides cytoprotection and / or neuroprotection to a host in need thereof.

144. (Rev) A topical ophthalmic formulation comprising cromakalim or a pharmaceutically acceptable salt thereof (0.015% w / v), Kolliphor™ ELP (4% w / v), polysorbate 80 (1% w / v), polyvinylpyrrolidone (PVP) K-30 (2% w / v), poloxamer 407 (0.1% w / v), mannitol (3.3% w / v), dibasic sodium phosphate buffer, monobasic sodium phosphate buffer, and water, and having a pH of about 6.

5.

145. (Rev) A topical ophthalmic formulation comprising cromakalim or a pharmaceutically acceptable salt thereof (0.03% w / v), Kolliphor™ ELP (4% w / v), polysorbate 80 (1% w / v), polyvinylpyrrolidone (PVP) K-30 (2% w / v), poloxamer 407 (0.1% w / v), mannitol (3.3% w / v), dibasic sodium phosphate buffer, monobasic sodium phosphate buffer, and water, and having a pH of about 6.

5.

146. (Rev) A topical ophthalmic formulation comprising cromakalim or a pharmaceutically acceptable salt thereof (0.075% w / v), Kolliphor™ ELP (4% w / v), polysorbate 80 (1% w / v), polyvinylpyrrolidone (PVP) K-30 (2% w / v), poloxamer 407 (0.1% w / v), mannitol (3.3% w / v), dibasic sodium phosphate buffer, monobasic sodium phosphate buffer, and water, and having a pH of about 6.

5.

147. 124. Use of (rev)cromakalim or a pharmaceutically acceptable salt thereof, as a formulation according to any one of claims 1 to 123, for the topical treatment of an ocular disorder in a host in need thereof.

148. 148. The use of claim 147, wherein the eye disorder is glaucoma.

149. 149. The use of claim 148, wherein the glaucoma is glaucoma with normal intraocular pressure or normal tension glaucoma (NTG).

150. The use of claim 148, wherein the glaucoma is glaucoma associated with elevated intraocular pressure selected from the group consisting of primary open-angle glaucoma (POAG), primary angle-closure glaucoma (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, glaucoma associated with diabetic retinopathy, glaucoma associated with Sturge-Weber syndrome, steroid-induced glaucoma, and acute glaucoma due to progressive cataract and / or intravitreal injection.

151. 148. The use of claim 147, wherein the eye disorder is glaucoma induced by Sturge-Weber syndrome or Sturge-Weber syndrome induced glaucoma.

152. 148. The use of claim 147, wherein the eye disorder is diabetic retinopathy.

153. The use described in claim 147, wherein the local treatment is a primary or secondary or adjunctive treatment as part of a protocol for MIGS (minimally invasive glaucoma surgery) selected from the group consisting of miniaturized forms of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, complete intraocular or suprachoroidal shunts, gentler and more gentle forms of laser cyclophotocoagulation, and in alternative claims, Schlemm's canal stents for dilating Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty.

154. 148. The use of claim 147, wherein the ocular disorder is Graves' ophthalmopathy, thyroid-associated orbitopathy (TAO), Graves' orbitopathy (GO), retrobulbar tumor, cavernous sinus thrombosis, orbital vein thrombosis, episcleral / orbital vein vasculitis, superior vena cava occlusion, superior vena cava thrombosis, carotid-cavernous fistula, dural cavernous shunt, orbital varicose veins, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), arterial occlusive / embolic-hypoperfusion disease, or optic nerve damage due to ischemia (posterior and anterior ischemic optic neuropathy (NAION)).

155. 148. The use of claim 147, wherein said treatment with a topical formulation provides cytoprotection and / or neuroprotection to a host in need thereof.

156. 124. A topical formulation of (rev)cromakalim or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 123 for use in the treatment of an ocular disorder in a host in need thereof.

157. 157. The use of claim 156, wherein the eye disorder is glaucoma.

158. 157. The use of claim 156, wherein the glaucoma is glaucoma with normal intraocular pressure or normal tension glaucoma (NTG).

159. The use of claim 156, wherein the glaucoma is glaucoma associated with elevated intraocular pressure selected from the group consisting of primary open-angle glaucoma (POAG), primary angle-closure glaucoma (also known as chronic open-angle glaucoma, chronic simple glaucoma, and simple glaucoma), pediatric glaucoma, pseudoexfoliation glaucoma, pigmentary glaucoma, traumatic glaucoma, neovascular glaucoma, iridocorneal endothelial glaucoma (ICE), uveitic glaucoma, glaucoma associated with diabetic retinopathy, glaucoma associated with Sturge-Weber syndrome, steroid-induced glaucoma, and acute glaucoma due to progressive cataract and / or intravitreal injection.

160. 157. The use of claim 156, wherein the eye disorder is glaucoma induced by Sturge-Weber syndrome or Sturge-Weber syndrome induced glaucoma.

161. 157. The use of claim 156, wherein the eye disorder is diabetic retinopathy.

162. The use described in claim 156, wherein the local treatment is a primary or secondary or adjunctive treatment as part of a protocol for MIGS (minimally invasive glaucoma surgery) selected from the group consisting of miniaturized forms of trabeculectomy (microtrabeculectomy), trabecular bypass surgery, complete intraocular or suprachoroidal shunts, gentler and more gentle forms of laser cyclophotocoagulation, and in alternative claims, Schlemm's canal stents for dilating Schlemm's canal, goniotomy, canaloplasty, and laser trabeculoplasty.

163. 157. The use of claim 156, wherein the ocular disorder is Graves' ophthalmopathy, thyroid-associated orbitopathy (TAO), Graves' orbitopathy (GO), retrobulbar tumor, cavernous sinus thrombosis, orbital vein thrombosis, episcleral / orbital vein vasculitis, superior vena cava occlusion, superior vena cava thrombosis, carotid-cavernous sinus fistula, dural cavernous sinus shunt, orbital varicose veins, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), arterial occlusive / embolic-hypoperfusion disease, or optic nerve damage due to ischemia (posterior and anterior ischemic optic neuropathy (NAION)).

164. 157. The use of claim 156, wherein said treatment with a topical formulation provides cytoprotection and / or neuroprotection to a host in need thereof.

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