Crystalline polymorphic forms of TRPV1 antagonists and formulations thereof

Abstract

The present disclosure provides polymorphs and formulations of 1-(6-fluoro-5-((((1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol (Compound I). [Formula 1] TIFF2025531367000028.tif4343

Description

[Technical Field]

[0001] The present invention relates to crystalline forms of 1-(6-fluoro-5-((((1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol (Formula I), processes and methods for its preparation. The present invention also relates to formulations of Compound I and methods for treating ocular surface disorders therewith. [Background technology]

[0002] Patients with ocular surface pain, especially chronic ocular surface pain, experience a significantly reduced quality of life, often developing depression, moderate to severe angina, dialysis, disabling hip fractures, and in some cases suicide. In many patients, ocular surface pain persists despite treatment of the underlying pathology (e.g., recent trauma or surgery, infection, inflammation), and no other known treatments are available for long-term treatment.

[0003] The transient receptor potential vanilloid 1 (TRPV1) receptor is involved in pain signaling, and antagonism of this receptor may be beneficial for pain symptoms. To reduce pain, especially chronic pain, it may be desirable to administer a formulation of a TRPV1 antagonist topically to the ocular surface. Summary of the Invention

[0004] Formulation of hydrophobic ophthalmic drugs can be particularly challenging due to their tendency to aggregate, especially in aqueous topical ophthalmic compositions. Aggregation can cause stability and other potential quality issues for the composition, and may arise from other interactions between the drug and excipients. Therefore, there is a need to identify various polymorphic forms that can be formulated into ophthalmic formulations for delivery to the ocular surface.

[0005] In one aspect, the present invention provides a compound having the structure:

[0006] [ka] The present invention relates to a crystalline form of 1-(6-fluoro-5-((((1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol (Compound I) having the formula:

[0007] In another aspect, the present invention provides (S)-1-(6-fluoro-5-((((1r,3S)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol (Compound I), described and identified herein as crystalline Form A of Compound I:

[0008] [ka] Crystalline Form A of Compound I may be characterized by an X-ray diffraction pattern having three or more peaks at 2θ values ​​selected from 14.3, 14.8, and 21.8±0.2 degrees 2θ. In some embodiments, crystalline Form A of Compound I is characterized by an X-ray diffraction pattern having three or more peaks at 2θ values ​​selected from 12.5, 14.3, 14.8, 21.8, and 22.6±0.2 degrees 2θ.

[0009] In some embodiments, crystalline Form A of Compound I is characterized by an X-ray diffraction pattern as shown in Figure 1. Crystalline Form A of Compound I is characterized by one or more of: 1) a DSC thermogram exhibiting an endotherm at about 131.5°C, 2) a water loss of about 0.13% by weight as determined by thermogravimetric analysis, and 3) a melting point of about 130.3°C.

[0010] In some embodiments, crystalline Form A of Compound I is characterized by an X-ray diffraction pattern having 3 or more, 4 or more, 5 or more, 6 or more, or 7 or more peaks at 2θ values ​​selected from 12.5, 14.3, 14.8, 20.1, 21.8, 22.6, and 23.2±0.2 degrees 2θ.

[0011] In some embodiments, crystalline Form A of Compound I is characterized by an X-ray diffraction pattern having 3 or more, 4 or more, 5 or more, 6 or more, or 7 or more peaks at 2θ values ​​selected from 7.1, 12.5, 14.3, 14.8, 18.7, 20.1, 21.8, 22.6, 23.2, 25.1, and 27.9±0.2 degrees 2θ.

[0012] In another aspect, the present invention provides a method for preparing crystalline Form A of Compound I, comprising cooling a hot saturated solution of the free base of Compound I in a solvent to crystallize Compound I as crystalline Form A.

[0013] In another aspect, the present invention provides a method for preparing crystalline Form A of Compound I, comprising crystallizing Form A from a solution of Compound I in a solvent, for example, at room temperature.

[0014] In another aspect, the present invention provides a method for preparing crystalline Form A of a compound, the method comprising adding an antisolvent to a solution of compound I in a solvent.

[0015] In certain embodiments of the process for preparing crystalline Form A of Compound I, Compound I is used as the free base.

[0016] In some embodiments, crystalline Form A of Compound I is characterized by a melting point of about 130.3° C. or a differential scanning calorimetry pattern as shown in FIG.

[0017] In another aspect, the present invention provides a pharmaceutical formulation comprising crystalline Form A in substantially pure form.

[0018] In a further aspect, a method of preparing a pharmaceutical formulation comprising crystalline Form A is provided, the method comprising dissolving crystalline Form A disclosed herein in an ophthalmologically acceptable carrier formulated for ophthalmic use (e.g., topical application to the ocular surface).

[0019] In yet another aspect, the present invention provides a method for treating a TRPV1-mediated disease or disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of a pharmaceutical formulation comprising Compound I, or a pharmaceutically acceptable salt, solvate, or co-crystal thereof prepared from crystalline Form A of Compound I, or crystalline Form A of Compound I, or a combination thereof.

[0020] In yet another aspect, the present invention provides a method for treating an ocular surface disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of a pharmaceutical formulation comprising Compound I, or a pharmaceutically acceptable salt, solvate, or co-crystal thereof prepared from crystalline Form A of Compound I, or crystalline Form A of Compound I, or a combination thereof.

[0021] In yet another aspect, the present invention provides a method for treating ocular surface pain in a subject in need thereof, the method comprising administering to the subject an effective amount of a pharmaceutical formulation comprising Compound I, or a pharmaceutically acceptable salt, solvate, or co-crystal thereof prepared from crystalline Form A of Compound I, or crystalline Form A of Compound I, or a combination thereof.

[0022] In some embodiments, the present disclosure provides a compound having the following structure:

[0023] [ka] or a pharmaceutically acceptable salt, solvate, polymorph, or co-crystal thereof, having the formula:

[0024] In some embodiments, the compound of Formula I has the following structure:

[0025] [ka] It has.

[0026] In some embodiments, the ocular surface pain is acute or sudden ocular surface pain. In some embodiments, the ocular surface pain is chronic ocular surface pain that lasts for at least 3 months. In some embodiments, the compound of Formula I is administered to the cornea of ​​the subject.

[0027] In some embodiments, the COSP is associated with dry eye disease. In some embodiments, administration reduces symptoms of dry eye disease. In some embodiments, administration reduces pain associated with dry eye disease. In some embodiments, administration reduces the incidence of one or more of ocular dryness, ocular discomfort, ocular redness, ocular burning or stinging, gritty or foreign body sensation, or photophobia by at least about 10%.

[0028] In some embodiments, the subject is diagnosed with dry eye disease, Sjogren's syndrome, conjunctivitis (including epidemic conjunctivitis, vernal keratoconjunctivitis, and allergic conjunctivitis), epithelial basement membrane corneal dystrophy, Acanthamoeba, fibromyalgia, meibomian gland dysfunction, thyroid eye disease, rosacea, ptosis, keratoconus, eye pain syndrome, Stevens-Johnson syndrome, corneal epitheliopathy, corneal neuropathy (including LASIK-induced corneal neuropathy), corneal dystrophy (including recurrent corneal dystrophy), epithelial basement membrane dystrophy, and the like. Patients have one or more of the following conditions: ocular surface disease, blepharitis, corneal erosion or abrasion (including recurrent corneal erosion or abrasion), ocular surface disease, blepharitis, graft-versus-host disease, meibomianitis, glaucoma, conjunctival laxity, keratopathy (including herpetic keratopathy, filamentous keratopathy, band or bullous keratopathy, and exposure keratopathy), keratitis (including herpes simplex virus keratitis), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, or xerophthalmia, or are recovering from neurotrophic keratitis.

[0029] In some embodiments, the method includes administering to the subject an additional therapeutic agent.

[0030] In some embodiments, administration results in a reduction in pain score on the Visual Acuity Scale (VAS) of at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, or at least about 10 compared to placebo. In some embodiments, the reduction in VAS score is due to the difference in VAS score before and after administration of Compound I to the subject. In the methods of the invention, the reduction in VAS score occurs within about 30 minutes, within about 1 hour, within about 2 hours, within about 4 hours, or within about 2-4 hours after administration of Compound I to the subject.

[0031] In some embodiments, administration of Compound I reduces hyperemia in a subject by at least about 1, at least about 2, at least about 3, at least about 4, or at least about 5 on the McMonnies scale.

[0032] In some embodiments, administration results in no change compared to placebo in one or more of best corrected visual acuity, intraocular pressure, slit lamp examination, mydriatic examination, blink rate, tear production, or corneal staining.

[0033] In some embodiments, the compound of Formula I is administered in the form of a formulation described herein. In some embodiments, the formulation is administered for at least about 1 month, about 2 months, or about 3 months. In some embodiments, the formulation is administered 1 to 4 times daily.

[0034] In some embodiments, the present disclosure provides a formulation described herein for use in treating ocular surface pain. In some embodiments of the described uses, the ocular surface pain is sudden (e.g., acute) ocular surface pain or chronic ocular surface pain lasting at least 3 months.

[0035] In some embodiments, the present disclosure provides a compound having the following structure:

[0036] [ka] and n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 12

[0037] In some embodiments, the compound of Formula I has the following structure:

[0038] [ka] It has.

[0039] In some embodiments, the ocular surface pain is episodic (e.g., acute) ocular surface pain and the ocular surface pain is chronic ocular surface pain (COSP). In some embodiments, the COSP is associated with dry eye disease.

[0040] In some embodiments, administration reduces the symptoms of dry eye disease. In some embodiments, administration reduces pain associated with dry eye disease. In some embodiments, administration reduces the incidence of one or more of ocular dryness, ocular discomfort, ocular redness, ocular burning or stinging, gritty or foreign body sensation, or photophobia by at least about 10%.

[0041] In some embodiments, the subject is diagnosed with dry eye disease, Sjogren's syndrome, conjunctivitis (including epidemic conjunctivitis, vernal keratoconjunctivitis, and allergic conjunctivitis), epithelial basement membrane corneal dystrophy, Acanthamoeba, fibromyalgia, meibomian gland dysfunction, thyroid eye disease, rosacea, ptosis, keratoconus, eye pain syndrome, Stevens-Johnson syndrome, corneal epitheliopathy, corneal neuropathy (including LASIK-induced corneal neuropathy), corneal dystrophy (including recurrent corneal dystrophy), epithelial basement membrane dystrophy, and the like. Patients have one or more of the following conditions, or are recovering from neurotrophic keratitis: ocular surface disease, blepharitis, corneal erosion or abrasion (including recurrent corneal erosion or abrasion), ocular surface disease, blepharitis, graft-versus-host disease, meibomianitis, glaucoma, conjunctival laxity, keratopathy (including herpetic keratopathy, filamentous keratopathy, band or bullous keratopathy, and exposure keratopathy), keratitis (including herpes simplex virus keratitis), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, or xerophthalmia.

[0042] In some embodiments, the method includes administering to the subject an additional therapeutic agent.

[0043] In some embodiments, administration results in a reduction in pain score on the Visual Analogue Scale (VAS) by at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, or at least about 10 compared to placebo. In some embodiments, administration results in a reduction in pain score on the VAS by at least about 6, at least about 7, at least about 8, at least about 9, or at least about 10 compared to placebo. In some embodiments, the reduction in pain score is due to the difference in pain score before and after administration of Compound I to the subject.

[0044] In some embodiments, administration reduces the subject's congestion by at least about 1, at least about 2, at least about 3, at least about 4, or at least about 5 on the McMonnie scale.

[0045] In some embodiments, administration results in a reduction in pain score on the visual acuity scale (VAS) of at least about 3 compared to the VAS score prior to administration of the compound.

[0046] In some embodiments of the methods described, the compound of formula I is administered in the form of a formulation described herein.

[0047] Certain preferred embodiments of the present invention will become apparent from the following more detailed description of certain preferred embodiments and the claims. [Brief explanation of the drawings]

[0048] [Figure 1] FIG. 1 shows the X-ray powder diffraction pattern of crystalline Form A of Compound I. [Figure 2] FIG. 1 shows a differential scanning calorimetry scan of crystalline Form A of Compound I. [Figure 3] FIG. 1 shows a thermogravimetric analysis of crystalline Form A of Compound I. [Figure 4] FIG. 1 shows XRPD patterns of Compound I Form A after milling and granulation: from bottom to top: starting material, after milling, after granulation with water, and after granulation with ethanol. DETAILED DESCRIPTION OF THE INVENTION

[0049] In the manufacture of pharmaceutical compounds and their formulations, it is important that the active compound is in a form that can be easily handled and processed in order to have a commercially viable, reliable and reproducible manufacturing process.

[0050] It has been surprisingly found that crystalline Form A of Compound I possesses favorable physicochemical properties that make it particularly useful as a drug substance for use in preparing ophthalmic dosage forms.

[0051] As used herein, the term "about" refers to a range of + / - 10% of the specified value.

[0052] "TRPV1 receptor" refers to transient receptor potential vanilloid 1, which has been characterized by molecular cloning and pharmacology. See, e.g., Caterina MJ, et al., Nature 1997;389:816-824. TRPV1 receptor activity is measured as described in WO 2005 / 120510, the entire contents of which are incorporated herein by reference.

[0053] The term "effective amount" of a compound described herein refers to the amount of a therapeutic compound necessary or sufficient to perform its intended function in a mammal. The effective amount of a therapeutic compound may vary depending on factors such as the amount of causative agent already present in the mammal, the age, sex, and weight of the mammal, and the ability of the therapeutic compound of the present disclosure to treat the ocular surface disorder and / or its symptoms in the mammal.

[0054] The phrase "ophthalmologically compatible" refers to formulations, polymers, and other materials and / or dosage forms that are suitable for use in contact with the ocular tissues of humans and animals without undue toxicity, irritation, allergic response, or other problems or complications, consistent with a reasonable benefit / risk ratio.

[0055] As used herein, the terms "treat," "treating," or "treatment" in connection with a disease or disorder refer, in some embodiments, to ameliorating the disease or disorder (i.e., delaying, preventing, or alleviating the onset of the disease or at least one of its clinical symptoms). In another embodiment, "treat," "treating," or "treatment" refers to alleviating or improving at least one physical parameter, including one that may not be discernible by the patient. In yet another embodiment, "treat," "treating," or "treatment" refers to modulating the disease or disorder, either physically (e.g., stabilizing a discernible symptom), physiologically (e.g., stabilizing a physical parameter), or both. In yet another embodiment, "treat," "treating," or "treatment" refers to preventing or delaying the onset or development or progression of the disease or disorder or its symptoms.

[0056] As used herein, the term "subject" or "patient" refers to humans and non-human mammals, including, but not limited to, primates, rabbits, pigs, horses, dogs, cats, sheep, and cattle. In certain embodiments, a subject or patient is a human. In some embodiments, the term "patient" or "subject" refers to a human suffering from a condition (i.e., a disease or disorder) described herein and who would benefit from treatment. As used herein, a subject is "in need" of treatment if such a subject (patient) would benefit biologically, medically, or in quality of life from such treatment. In certain embodiments, the subject is an adult at least about 18 years of age. In some embodiments, the subject is an adult between about 18 and about 75 years of age. In some embodiments, the subject is a child up to about 18 years of age.

[0057] As used herein, "ocular surface" refers to the outer surface of the eye, anatomically including the cornea (including the epithelium, Bowman's membrane, stroma, decidua, and endothelium), the conjunctiva, and the corneoscleral junction, i.e., the limbus.

[0058] As used herein, "pain" refers to a persistent or intermittent sensation of actual pain described, and may be described as, but not limited to, tingling, dull, sharp, or aching. Pain may also refer to similar related descriptors, such as, but not limited to, burning, stinging, rumbling, foreign body sensation, dryness, sandiness, fatigue, itching, irritation, sensitivity to light, etc.

[0059] As used herein, "ocular surface pain" refers to pain on the surface of the eye, for example, the cornea. Ocular pain can be nociceptive pain, which is usually caused by external physical or chemical injury stimuli, such as corneal surgery, inflammation, or other damage to the corneal surface. Ocular pain can also be caused by neuropathic pain, which can be caused by direct damage to the body's neurons, resulting in pain messages being sent to the central nervous system and the brain, regardless of the presence of noxious stimuli. As used herein, "ocular surface pain" includes both nociceptive pain and neuropathic pain.

[0060] As used herein, the term "visual analog scale" (VAS) refers to a measure of pain intensity, where subjects typically mark the spot on the scale that corresponds to their pain level. Pain is marked on a scale ranging from "no pain" (score 0) to "worst possible pain" or "worst imaginable pain" (score 100). See, e.g., Hawker, et al., Arthritis Care & Research 63(11), pp. S240-S252 (November 2011). There are several other well-designed pain scales available to help assess pain intensity. Numeric rating scales (NRS) are commonly used, where subjects rate pain using numbers. The numeric scale ranges from 1 to 10 or 1 to 100. The Wong-Baker FACES pain scale combines pictures and numbers to assess pain. It is available for use in children over 3 years of age and adults. Six faces represent a range of facial expressions, from happy to very upset. Each is assigned a numerical rating from 0 (smiling) to 10 (crying). The verbal pain intensity scale uses words on a scale to rate pain intensity: no pain / mild pain / moderate pain / severe pain / very severe pain / worst possible pain.

[0061] The Ocular Sensation Scale is a specific pain scale developed to measure the severity of ocular pain. See Caudle LE et al., Optom Vis Sci. 2007 Aug;84(8):752-62. This scale measures pain, discomfort, or light sensitivity, typically with five categorical labels: "extreme," "severe," "moderate," "mild," and "none."

[0062] The Ocular Pain Assessment Survey (OPAS) is a quantitative, multidimensional questionnaire specifically designed to assess changes in corneal and ocular surface pain and quality of life (QoL). The OPAS quantitatively assesses pain intensity, frequency of ocular and non-ocular pain, changes in QoL, exacerbating factors, associated factors, and symptom relief, allowing for monitoring of treatment response. See Qazi et al., Ophthalmology July 123(7):1458-1468(2016).

[0063] As used herein, ocular redness refers to the redness of the ocular surface.Ocular redness can be a clinical marker of inflammation and / or eye irritation.Ocular redness is typically measured using a McMonnie scale with a value of 0 to 5 based on standard photographs.

[0064] As used herein, "placebo" refers to an ophthalmic formulation that contains all of the components of the administered drug composition except the drug.

[0065] As used herein, "polymorph" refers to crystalline forms that have the same chemical composition but differ in the spatial arrangement of the molecules, atoms, and / or ions that form the crystals.

[0066] As used herein, "solvate" refers to a crystalline form of a molecule, atom, and / or ion that further contains solvent molecules or solvent incorporated into the crystal lattice structure. The solvent molecules in a solvate can be in an ordered and / or disordered arrangement. A solvate can contain either a stoichiometric or non-stoichiometric amount of solvent molecules. For example, a solvate with a non-stoichiometric amount of solvent molecules can be formed by partial loss of solvent from the solvate. A solvate can occur as a dimer or oligomer containing one or more molecules of Compound I within the crystal lattice structure.

[0067] As used herein, "amorphous" refers to a solid form of molecules, atoms, and / or ions that is not crystalline. Amorphous solids do not exhibit distinct X-ray diffraction patterns.

[0068] As used herein, "substantially pure," when used with respect to a form, refers to a compound having a purity of greater than 90% by weight of Compound I, based on the weight of the compound, including purities of greater than 90, 91, 92, 93, 94, 95, 96, 97, 98, and 99% by weight, and including purities equal to about 100% by weight. The remaining material includes other forms of the compound and / or reaction and / or processing impurities resulting from its preparation. For example, a crystalline form of Compound I can be considered substantially pure in that it has a purity of greater than 90% by weight as measured by means currently known and generally accepted in the art, with the remaining less than 10% by weight of the material including other forms of Compound I and / or reaction and / or processing impurities. In some embodiments, crystalline Form A of Compound I has a purity of greater than 92% by weight. In some embodiments, crystalline Form A of Compound I has a purity of greater than 95% by weight. In some embodiments, crystalline Form A of Compound I has a purity of greater than 97% by weight. In some embodiments, crystalline Form A of Compound I has a purity of greater than 99% by weight.

[0069] The term "substantially chloride-free" means that a compound, e.g., crystalline Form A of Compound I, does not contain significant amounts of extraneous chloride, e.g., undesirable chloride ions resulting from the formation of a hydrochloride salt during the manufacture of the drug substance. In some embodiments, crystalline Form A contains less than 0.5% by weight of chloride ions. In some embodiments, crystalline Form A contains less than 0.3% by weight of chloride ions. In some embodiments, crystalline Form A contains less than 0.1% by weight of chloride ions. In particular embodiments, crystalline Form A contains less than 0.05% by weight of chloride ions.

[0070] As used herein, "compound of Formula I," "compound I," "formula I," and "compound I" are used interchangeably and refer to the name 1-(6-fluoro-5-((((1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol, the structure shown below:

[0071] [ka] In certain embodiments, compound I refers to a compound having the name (S)-1-(6-fluoro-5-((((1r,3S)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol, the structure shown below:

[0072] [ka] It has.

[0073] As used herein, "crystal form," "crystalline form," "modification," or "polymorph," or "polymorphic form," used interchangeably in upper or lower case, refer to a crystalline or polymorphic form of Compound I.

[0074] As described herein, the compounds of Formula I are stereoisomers that can exist in either racemic or enantiomeric excess (ee) of said isomers, wherein said isomers are present in at least 90% ee, at least 95% ee, at least 96% ee, at least 97% ee, at least 98% ee, or at least 99% ee.

[0075] Any chemical formula described herein is also intended to represent the unlabeled form of the compound as well as the isotopically labeled form.Isotopically labeled compounds have the structure represented by the formula described herein, except that one or more atoms are replaced by atoms with selected atomic mass or mass number.The isotopes that can be incorporated into the compounds of the present disclosure include, for example: 2 H, 3 H, 11 C. 13 C. 14 C, and 15 Isotopes of hydrogen, carbon, nitrogen, and oxygen, such as N, are included. Thus, the methods of the present invention can be used, for example, 3 H and 14 radioactive isotopes such as C, or 2 H and 13 It is understood that compounds can or may contain compounds incorporating one or more of any of the aforementioned isotopes, including radioactive isotopes where non-radioactive isotopes exist, such as C. Such isotopically labeled compounds are useful for metabolic studies ( 14 C), reaction kinetic studies (e.g. 2 H or 3 H), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution assays, or for radiation treatment of patients. Isotopically labeled compounds can generally be prepared by conventional techniques known to those of skill in the art, e.g., by substituting an appropriate isotopically labeled reagent for a previously used non-labeled reagent.

[0076] The present invention encompasses embodiments that include all pharmaceutically acceptable salts of compounds useful according to the inventions provided herein. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds, where the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, and the like. Pharmaceutically acceptable salts include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts can be synthesized from parent compounds that contain a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of both (generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred). A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17 th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.For example, preferred pharmaceutically acceptable salts include, but are not limited to, mineral salts or organic acid salts of basic residues such as amines.For example, salts can be hydrochlorides.

[0077] As used herein, the phrase "pharmaceutically acceptable" refers to compounds, substances, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0078] Unless otherwise stated, all ingredient concentrations are expressed in % weight / volume (% w / v) units.

[0079] Unless otherwise specified, weights or dosages referred to herein for compounds of Formula I are weights or dosages of the compound itself, and not of its salts or prodrugs, which may vary to achieve the intended therapeutic effect. For example, the weight or dosage of a corresponding salt of a compound suitable for a method, composition, or combination disclosed herein can be calculated based on the ratio of the molecular weights of the salt and the compound itself.

[0080] Crystalline Form of Compound I Polymorphism is the ability of a solid substance to exist in two or more crystalline forms, with different arrangements or structures of components within the crystal lattice. Polymorphism and pseudomorphism are very common in drugs and account for many differences in their properties. Traditionally, the lowest-energy polymorph is selected for incorporation into a formulation for chemical stability; however, to achieve desirable chemical and physical stability, and therefore efficacy, excipients in the formulation must be considered. Crystalline Form A of Compound I can be advantageously used to prepare or be incorporated into ophthalmic formulations for the treatment of TRPV1-mediated disorders. Crystalline Form A of Compound I can be advantageously used to prepare or be incorporated into ophthalmic formulations for the treatment of ocular surface pain. In some embodiments, the subject is diagnosed with dry eye disease, Sjogren's syndrome, conjunctivitis (including epidemic conjunctivitis, vernal keratoconjunctivitis, and allergic conjunctivitis), epithelial basement membrane corneal dystrophy, Acanthamoeba, fibromyalgia, meibomian gland dysfunction, thyroid eye disease, rosacea, ptosis, keratoconus, eye pain syndrome, Stevens-Johnson syndrome, corneal epitheliopathy, corneal neuropathy (including LASIK-induced corneal neuropathy), corneal dystrophy (including recurrent corneal dystrophy), epithelial basement membrane dystrophy, and the like. Patients have one or more of the following conditions, or are recovering from neurotrophic keratitis: ocular surface disease, blepharitis, corneal erosion or abrasion (including recurrent corneal erosion or abrasion), ocular surface disease, blepharitis, graft-versus-host disease, meibomianitis, glaucoma, conjunctival laxity, keratopathy (including herpetic keratopathy, filamentous keratopathy, band or bullous keratopathy, and exposure keratopathy), keratitis (including herpes simplex virus keratitis), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, or xerophthalmia.

[0081] Thus, in one embodiment, the present invention provides crystalline Form A of Compound I. Crystalline Form A of Compound I is a non-solvated crystalline solid that crystallizes in two distinct forms. Non-aqueous solvent systems produce irregularly shaped tabular particles, while solvent mixtures containing water produce needle-like particles. Despite the change in form, both forms have the same physicochemical properties and the same XRD pattern and are therefore considered to be the same polymorph. Crystalline Form A can be characterized as such by powder X-ray diffraction (XRPD), and the pattern obtained from the analysis can include significant peaks at characteristic two-theta (2θ) angles. Form A can be characterized, for example, by an X-ray diffraction pattern with three or more peaks at 2θ values ​​selected from 14.3, 14.8, and 21.8±0.2°2θ. Form A can be characterized by an X-ray diffraction pattern having three or more peaks at 2θ values ​​selected from, for example, 12.5, 14.3, 14.8, 21.8, and 22.6±0.2°2θ. Further, Form A can be characterized by an X-ray diffraction pattern having three or more, four or more, five or more, six or more, or seven or more peaks at 2θ values ​​selected from, for example, 12.5, 14.3, 14.8, 20.1, 21.8, 22.6, and 23.2±0.2°2θ. As described herein, the X-ray diffraction peaks include CuK with a wavelength of 0.15418 nm. α The XRPD parameter can be used to analyze Compound I by XRPD, and can be described in the general test conditions of the Examples section disclosed herein. In some embodiments, crystalline Form A is characterized by a melting point of about 130.3°C.

[0082] Crystalline Form A can be prepared by cooling a hot saturated solution of Compound I in a solvent and crystallizing Compound I as Crystalline Form A. In another embodiment, crystalline Form A may be prepared by crystallizing crystalline Form A from a saturated solution of Compound I in a solvent.

[0083] In some embodiments, crystalline Form A of Compound I can be prepared by dissolving an appropriate amount of Compound I in a minimum amount of solvent to ensure saturation at a temperature of about 50°C to 70°C, e.g., 60°C. The solution can then be slowly cooled to a temperature of about 0°C to about 30°C, e.g., 20°C, while stirring. To facilitate the crystallization process, the solution can optionally be cooled to a temperature of about -20°C to about -5°C over a period of about 5 days. Crystalline Form A of Compound I can then be isolated, for example, by filtering the suspension and isolating the formed solid. The solid can then be dried, for example, under vacuum at about 45°C to about 55°C for about 4 hours to about 5 hours, to obtain Compound I in its polymorphic Form A crystalline form.

[0084] In some embodiments, the solvent is selected from the group consisting of acetone, acetonitrile, dichloromethane, ethanol, ethyl acetate, isopropyl acetate, 2-methyl-2-butanol, methyl tert-butyl ether, water, methanol / water, and acetone / heptane.

[0085] In some embodiments, crystalline Form A of Compound I can be prepared by suspending Compound I in a solvent and stirring the mixture. The mixture can be stirred for a period of time, for example, up to 28 days. The solid can optionally be filtered, and the mother liquor can be evaporated at about 25°C. In the case of a non-volatile solvent (e.g., benzyl alcohol), the solution can be stored at -20°C for at least 1 day (24 hours). Crystalline Form A of Compound I can then be isolated, for example, by filtering the reaction mixture and isolating the solid that forms. The solid can be dried, for example, under vacuum at about 45 to about 55°C for about 4 to about 5 hours to obtain polymorphic Form A crystalline form of Compound I.

[0086] In some embodiments, the solvent is selected from the group consisting of acetone, acetonitrile, benzyl alcohol, dichloromethane, dioxane, ethanol, ethyl acetate, isopropyl acetate, methanol, 2-methyl-2-butanol, methyl tert-butyl ether, tetrahydrofuran, 1-propanol / water, 2-propanol / water, and methanol / water.

[0087] Crystalline Form A of Compound I can also be prepared by suspending Compound I in a solvent and heating to a temperature not exceeding the boiling point of the solvent. The reaction mixture can be heated and stirred to obtain a solution. For example, if the solvent is acetonitrile, the suspension can be heated to a temperature of about 50-70°C, e.g., 55°C. Heating can be carried out until dissolution is achieved, e.g., for 2-3 hours. The starting material of Compound I can be in any form (e.g., crystalline, amorphous, solvated) to form the reaction mixture.

[0088] The solution can then be cooled to a lower temperature to obtain Compound I in polymorph Form A crystalline form.

[0089] Cooling can be carried out stepwise over a period of up to 1 day (24 hours). For example, a solution at 55°C can be slowly cooled to a temperature of about 50°C. Cooling can be carried out over a period of, for example, about 1 hour. The reaction mixture can be maintained at this temperature for an extended period of time, for example, about 3 to about 4 hours.

[0090] In a second step, the reaction mixture can be further cooled slowly to about 25° C. to about 30° C., for example, over a period of about 5 to about 6 hours. The reaction mixture can then be heated to 50° C. and maintained at this temperature for an extended period of time, for example, about 3 to about 4 hours, during which time the reaction mixture can be stirred.

[0091] In the third cooling step, the reaction mixture may then be further slowly cooled to about 10° C. to about 15° C. The reaction mixture may be maintained at this temperature for an extended period of time, for example, about 5 hours to about 6 hours, during which time the reaction mixture may be stirred.

[0092] Crystalline Form A of Compound I can then be isolated, for example, by filtering the reaction mixture and isolating the solid that forms. The solid can be dried, for example, under vacuum at about 45 to about 55°C for about 4 to about 5 hours to obtain Compound I in its polymorphic Form A crystalline form.

[0093] Optionally, a further purification step may be carried out to remove residual chloride ions. Crystalline Form A of Compound I is suspended in water and stirred at 25°C for an extended period of time, e.g., about 5 hours to about 7 hours, e.g., about 6 hours. In a next step, the reaction mixture is filtered and the solid is resuspended in water. This process can be repeated at least four times. The residual chloride content can be measured by ion chromatography. Crystalline Form A of Compound I can then be isolated, e.g., by filtering the reaction mixture and isolating the solid. The solid can be dried, e.g., under vacuum at about 45 to about 55°C, e.g., about 50°C, for about 5 hours to about 6 hours, to obtain substantially chloride-free Compound I in its Form A crystalline form.

[0094] Alternatively, crystalline Form A may be prepared by adding an anti-solvent to a solution of Compound I in a solvent. In certain embodiments, the anti-solvent is selected from heptane, toluene, and water. In further embodiments, the anti-solvent / solvent system is selected from the group consisting of heptane / acetone, water / acetone, heptane / dioxane, toluene / dioxane, water / dioxane, heptane / ethanol, water / ethanol, heptane / ethyl acetate, water / methanol, heptane / 2-methyl-2-butanol, heptane / tetrahydrofuran, and toluene / tetrahydrofuran.

[0095] Crystalline Form A of Compound I can be prepared by dissolving an appropriate amount of Compound I in a minimum amount of solvent to ensure saturation and then adding the solution to an excess of anti-solvent. The reaction mixture may be stirred during this time. If precipitation does not occur immediately, the reaction mixture may be stirred at room temperature for 1 day (24 hours). Crystalline Form A of Compound I can then be isolated, for example, by filtering the reaction mixture and isolating the solid that forms. The solid can be dried, for example, under vacuum at about 45 to about 55°C for about 4 to about 5 hours, to obtain polymorphic Form A of Compound I.

[0096] In some embodiments, compound I useful for preparing crystalline form A is substantially pure.

[0097] In one embodiment, crystalline Form A of Compound I is provided in a substantially pure form. This substantially pure form of crystalline Form A of Compound I may be used in pharmaceutical compositions, such as the ophthalmic formulations described herein. In some embodiments, the present disclosure provides pharmaceutical formulations comprising crystalline Form A of Compound I. In some embodiments, the present disclosure provides pharmaceutical formulations comprising Compound I or a pharmaceutically acceptable salt, solvate, or co-crystal thereof prepared from crystalline Form A of Compound I.

[0098] formulation Some embodiments herein relate to pharmaceutical formulations comprising crystalline Form A of Compound I or prepared from crystalline polymorphic Form A of Compound I.

[0099] In some embodiments, the formulation further comprises at least one ophthalmically acceptable excipient.

[0100] In some embodiments, the present invention provides the use of crystalline Form A of Compound I in the preparation of a pharmaceutical formulation.

[0101] In some embodiments, the present invention provides methods for preparing a pharmaceutical formulation comprising crystalline Form A of Compound I, the method comprising dissolving crystalline Form A of Compound I in an ophthalmologically acceptable carrier formulated for ophthalmic use, e.g., topical application to the ocular surface.

[0102] In some embodiments, the formulation includes a buffer. Examples of buffers include acetate, ascorbate, borate, bicarbonate, carbonate, citrate, edetate (EDTA), gluconate, lactate, phosphate, propionate, and TRIS (tromethamine) buffer. In certain embodiments, the buffer is a phosphate buffer system. In certain embodiments, the buffer is a tromethamine buffer. The amount of buffer substance added is typically an amount necessary to ensure and maintain a physiologically acceptable pH range. In some embodiments, the pH range is about 4 to about 9, about 4.5 to about 8.5, about 5.0 to about 8.0, about 5.5 to about 8.0, or about 6.4 to about 8.4. In some embodiments, the pH is about 6.0. In certain embodiments, the pH is about 7.4.

[0103] In some embodiments, the formulation may be self-preserving and does not contain a preservative. In other embodiments, the formulation contains a preservative. In some embodiments, the preservative includes, but is not limited to, polyhexylmethylene biguanidine (PHMB), polymeric quaternary ammonium compounds (e.g., polyquaternium-1), chlorine-containing preservatives such as benzalkonium chloride (BAK), chlorite preservatives, and the like.

[0104] In some embodiments, the preservative is an ophthalmically acceptable polymeric quaternary ammonium compound. Compounds of this type are described in U.S. Patent Nos. 3,931,319, 4,027,020, 4,407,791, 4,525,346, 4,836,986, 5,037,647, and 5,300,287, as well as PCT Publication WO 91 / 09523 (Dziabo et al.). In certain embodiments, the polymeric ammonium compound is polyquaternium 1 (also known as POLYQUAD® or ONAMERM®) having a number average molecular weight of 2,000 to 30,000. In even more specific embodiments, the number average molecular weight is 3,000 to 14,000.

[0105] When used, polymeric quaternary ammonium compounds are generally used in amounts greater than about 0.00001 w / v%, greater than about 0.0003 w / v%, or greater than about 0.0007 w / v% of the formulation. Furthermore, when used in a formulation, polymeric quaternary ammonium compounds are generally used at concentrations less than about 0.03 w / v%, less than about 0.003 w / v%, or less than about 0.0015 w / v% of the formulation. In some embodiments, the concentration of the polymeric quaternary ammonium compound in the formulation is as follows: greater than about 0.0003 w / v% but less than about 0.003 w / v%, greater than about 0.0003 w / v% but less than about 0.0015 w / v%, greater than about 0.0007 w / v% but less than about 0.003 w / v%, and greater than about 0.0007 w / v% but less than about 0.0015 w / v%. In certain embodiments, the formulation includes polyquaternium 1 at a concentration of about 0.001% w / v.

[0106] In some embodiments, the formulation comprises BAK at a concentration of at least about 0.0005%, about 0.001%, or about 0.007% w / v of the formulation, and at a concentration of less than about 0.1%, about 0.02%, or about 0.0035% w / v of the ophthalmic composition. It is specifically contemplated that any of the lower BAK concentration limits may be used in combination with any of the upper BAK concentration limits. In certain embodiments, the concentration of BAK in the composition is as follows: greater than about 0.001% w / v but less than about 0.02% w / v, greater than about 0.001% w / v but less than about 0.0035% w / v, greater than about 0.007% w / v but less than about 0.02% w / v, and greater than about 0.007% w / v but less than about 0.0035% w / v.

[0107] In some embodiments, the formulation of the present invention may contain additional therapeutic agents in addition to Compound I. Additional therapeutic agents may include, for example, other compounds and antibodies useful for treating ocular surface disorders. A non-limiting list of such agents includes nonsteroidal anti-inflammatory drugs such as ketorolac, nepafenac, bromfenac, corticosteroids, cyclosporine, lifitegrast, and other drugs for dry eye disease, or other TRPV1 inhibitors.

[0108] In some embodiments, the formulation is stored at refrigerated temperatures (e.g., 4° C.). In some embodiments, the formulation is allowed to warm to room temperature before administration.

[0109] In some embodiments, the suspension is packaged in a single-dose container. In some embodiments, the formulation is packaged in a multi-dose container.

[0110] The formulations described herein are delivered to the ocular surface from 1 to 6 times per day, depending on the routine judgment of a skilled clinician. In some embodiments, the formulations are administered 1, 2, 3, or 4 times per day.

[0111] In some embodiments, the pharmaceutical preparation of the present invention may contain additional therapeutic agents in addition to Compound (I).The additional therapeutic agents may include, for example, other compounds and antibodies that are useful for treating ocular surface disorders.The non-limiting list of such agents includes nonsteroidal anti-inflammatory drugs such as ketorolac, nepafenac, bromfenac, corticosteroids, cyclosporine, lifitegrast, and other drugs for dry eye disease, or other TRPV1 inhibitors.

[0112] How to use Without wishing to be bound by theory, it is hypothesized that blockers of the transient receptor potential vanilloid 1 (TRPV1) receptor may be useful in the treatment of pain, for example, chronic pain.

[0113] Thus, in some embodiments, the present invention provides a method of treating ocular surface pain in a subject, the method comprising administering to the subject an effective amount of Compound (I), or a pharmaceutically acceptable salt, solvate, or cocrystal thereof. In some embodiments, the method comprises administering Compound (I), or a pharmaceutically acceptable salt, solvate, or cocrystal thereof, as a pharmaceutical formulation, e.g., as disclosed herein. In some embodiments, the pharmaceutical formulation is prepared from crystalline polymorph Form A of Compound I, as disclosed herein. In some embodiments, the present invention provides a method of alleviating ocular surface pain in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound (I), or a pharmaceutically acceptable salt, solvate, or cocrystal thereof. In some embodiments, the method comprises administering Compound (I), or a pharmaceutically acceptable salt, solvate, or cocrystal thereof, as a pharmaceutical formulation, e.g., as disclosed herein. In some embodiments, the pharmaceutical formulation is prepared from crystalline polymorph Form A of Compound I, as disclosed herein. In some embodiments, the present invention provides a method for treating an ocular surface disorder in a subject, the method comprising administering to the subject an effective amount of Compound (I), or a pharmaceutically acceptable salt, solvate, or cocrystal thereof. In some embodiments, the method comprises administering Compound (I), or a pharmaceutically acceptable salt, solvate, or cocrystal thereof, as a pharmaceutical formulation, e.g., as disclosed herein. In some embodiments, the pharmaceutical formulation is prepared from crystalline polymorph Form A of Compound I, as disclosed herein.

[0114] In some embodiments, the present invention provides the use of a compound of formula I, or a pharmaceutically acceptable salt, solvate, or co-crystal thereof, in treating or alleviating ocular surface pain.

[0115] In some embodiments, the compound of formula I is polymorphic Form A.

[0116] In certain embodiments, the methods described herein are carried out by administering a formulation of Compound I described above. Accordingly, the present invention provides methods for treating ocular surface pain by administering a formulation of Compound I described herein. In some embodiments, the methods reduce ocular surface pain.

[0117] In some embodiments, the present invention provides a pharmaceutical formulation comprising Compound (I) or a pharmaceutically acceptable salt, solvate, or cocrystal thereof for use in treating ocular surface pain. In one embodiment, the formulation is prepared from crystalline polymorph Form A of Compound I disclosed herein.

[0118] In some embodiments, the present invention provides a pharmaceutical formulation comprising crystalline Form A of Compound I for use in treating ocular surface pain.

[0119] In some embodiments, the present invention provides a pharmaceutical formulation comprising Compound (I) or a pharmaceutically acceptable salt, solvate, or cocrystal thereof for use in treating ocular surface disorders. In one embodiment, the formulation is prepared from crystalline polymorph Form A of Compound I disclosed herein.

[0120] In some embodiments, the present invention provides a pharmaceutical formulation comprising crystalline Form A of Compound I for use in treating an ocular surface disorder.

[0121] In some embodiments, the present invention provides a pharmaceutical formulation comprising Compound (I) or a pharmaceutically acceptable salt, solvate, or cocrystal thereof for use in reducing ocular surface pain. In one embodiment, the formulation is prepared from crystalline polymorph Form A of Compound I disclosed herein.

[0122] In some embodiments, the present invention provides a pharmaceutical formulation comprising crystalline Form A of Compound I for use in the relief of ocular surface pain.

[0123] In some embodiments, the present invention provides the use of crystalline Form A of Compound I disclosed herein in the manufacture of a medicament for the treatment of ocular surface pain.

[0124] In some embodiments, the subject is suffering from sudden or acute ocular pain. In some embodiments, the subject is suffering from chronic ocular surface pain that has lasted for at least three months. In some embodiments, the subject is suffering from chronic ocular surface pain that has lasted for at least two months. In some embodiments, the subject is suffering from chronic ocular surface pain that has lasted for at least one month. In some embodiments, the subject is suffering from chronic ocular surface pain that has lasted for at least four months. In some embodiments, the subject is suffering from chronic ocular surface pain that has lasted for at least five months. Thus, in some embodiments, the present invention provides a method of treating chronic ocular surface pain in a subject by administering to the subject an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, polymorph, or cocrystal thereof. In some embodiments, the present invention provides a method of alleviating chronic ocular surface pain in a subject by administering to the subject an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, polymorph, or cocrystal thereof. The present invention provides the use of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, polymorph, or cocrystal thereof, in treating chronic ocular surface pain. In some embodiments, the compound of formula I is present in a formulation described herein.

[0125] In some embodiments, the formulation is administered to the ocular surface of a subject, for example, the cornea, any part of the conjunctiva, or the conjunctival sac of the eye.

[0126] In some embodiments, the present invention provides for administering a compound of Formula I to a subject in need of treatment in an ophthalmologically compatible formulation. In some embodiments, the compound of Formula I is administered to the subject from 1 to 6 times daily, for example, 1, 2, 3, or 4 times daily. In some embodiments, the compound of Formula I is administered to the subject for a period of at least about 1 month, at least about 2 months, or at least about 3 months. In some embodiments, the compound of Formula I is administered to the subject for a period of at least about 12 weeks.

[0127] In some embodiments, the ocular surface pain or chronic ocular surface pain is associated with dry eye disease, Sjogren's syndrome, conjunctivitis (including epidemic conjunctivitis, vernal keratoconjunctivitis, and allergic conjunctivitis), corneal epithelial basement membrane dystrophy, acanthamoeba, fibromyalgia, meibomian gland dysfunction, thyroid eye disease, rosacea, ptosis, keratoconus, eye pain syndrome, Stevens-Johnson syndrome, corneal epitheliopathy, corneal neuropathy (including LASIK-induced corneal neuropathy), corneal dystrophies (including recurrent corneal dystrophy), Associated with one or more of the following: epithelial basement membrane dystrophy, corneal erosion or abrasion (including recurrent corneal erosion or abrasion), ocular surface disease, blepharitis, graft-versus-host disease, meibomianitis, glaucoma, conjunctival laxity, keratopathy (including herpes keratopathy, filamentous keratopathy, band or bullous keratopathy, and exposure keratopathy), keratitis (including herpes simplex virus keratitis), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, or in patients recovering from neurotrophic keratitis.

[0128] In certain embodiments, the ocular surface pain or chronic ocular surface pain is associated with dry eye disease or Sjogren's syndrome. In some embodiments, the subject suffers from conjunctivitis, subconjunctival hemorrhage, subconjunctival scarring, membranous conjunctivitis, conjunctival ulcer, superficial punctate epithelial erosion, epithelial defect, eyelid margin ulcer, eyelid margin keratinization, synechiae, synechiae, trichiasis, anterior blepharitis, lacrimal punctum occlusion, meibomian gland disease, corneal opacity, dry eye, hypertrichosis, limbal stem cell failure, or corneal neovascularization.

[0129] In some embodiments, administration of a compound of Formula I reduces the subject's ocular pain compared to a placebo. In some embodiments, the reduction in the subject's ocular pain is at least about 3 compared to a placebo, as measured by a VAS score. In some embodiments, administration reduces the subject's ocular pain by at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, or at least about 10 compared to a placebo, as measured by a VAS score. In some embodiments, administration reduces the subject's pain by at least about 10%, at least about 15%, at least about 20%, or at least about 25% compared to a placebo.

[0130] In some embodiments, administration of a compound of Formula I reduces the subject's pain by at least about 2 compared to placebo, as measured by a VAS score, at about 30 minutes after administration, at about 1 hour after administration, at about 2 hours after administration, or at about 2-4 hours after administration.

[0131] In some embodiments, the reduction in pain score is due to the difference in pain score before and after administering Compound I to the subject. In some embodiments, the reduction in pain score as measured by VAS is due to the difference in pain score before and after administering Compound I to the subject. In some embodiments, the reduction in pain score occurs within about 30 minutes after administering Compound I to the subject. In some embodiments, the reduction in pain score occurs within about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, or about 6 hours after administering Compound I to the subject.

[0132] In some embodiments, administration of a compound of Formula I reduces ocular hyperemia (eye redness) compared to placebo, hi certain embodiments, administration of a compound of Formula I reduces Grade 1, Grade 2, Grade 3, or Grade 4 hyperemia compared to placebo.

[0133] In some embodiments, administration reduces the redness score in at least about 1, at least about 2, at least about 3, at least about 4, or at least about 5 eyes on the McMonnie scale.

[0134] Thus, in some embodiments, the present invention provides a method for treating or alleviating ocular redness in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, polymorph, or cocrystal thereof. In some embodiments, the present invention provides for the use of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, or cocrystal thereof, in treating ocular redness. In some embodiments, administration results in a reduction in ocular redness score of at least about 1, at least about 2, at least about 3, at least about 4, or at least about 5 on the McMonney scale. In some embodiments, the present invention provides for administering to a subject in need thereof a compound of Formula I at a concentration of about 0.5% w / v to about 3.5% w / v in an ophthalmologically compatible formulation. In some embodiments, the administration concentration ranges from about 0.5% to about 3.5% w / v, about 0.5% to about 2.5% w / v, about 0.5% to about 1.5% w / v, about 0.5% to about 3.0% w / v, about 1.0% to about 2.5% w / v, about 1.5% to about 3.0% w / v, or about 0.5% to about 2.5% w / v. In certain embodiments, the concentration of the compound of Formula I in a formulation for topical use is about 0.5% w / v, about 1.0% w / v, about 1.5% w / v, about 2.0% w / v, about 2.5% w / v, about 3.0% w / v, or about 3.5% w / v. In some embodiments, the dose per eye per administration is about 0.15 to about 1.15 mg, or about 0.15 mg, 0.2 mg, about 0.25 mg, 0.3 mg, about 0.35 mg, about 0.4 mg, about 0.45 mg, about 0.5 mg, about 0.55 mg, about 0.6 mg, about 0.65 mg, about 0.7 mg, about 0.75 mg, about 0.8 mg, about 0.85 mg, about 0.9 mg, about 0.95 mg, about 1.0 mg, about 1.05 mg, about 1.1 mg, or about 1.15 mg. In some embodiments, the dose per eye per administration is about 0.18 mg, about 0.37 mg, about 0.55 mg, about 0.74 mg, or about 0.92 mg. In some embodiments, the total daily dose per eye is about 0.5 to about 3.5 mg, or about 0.5 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 2.5 mg, about 3.0 mg, or about 3.5 mg.In some embodiments, the compound of Formula I is administered to the subject 1 to 6 times daily, for example, 1, 2, 3, or 4 times daily. In some embodiments, the compound of Formula I is administered to the subject for a period of at least about 1 month, at least about 2 months, or at least about 3 months. In certain embodiments, the compound of Formula I is administered in a formulation described herein.

[0135] In some embodiments, the ocular redness is caused by dry eye disease, Sjogren's syndrome, conjunctivitis (including epidemic conjunctivitis, vernal keratoconjunctivitis, and allergic conjunctivitis), corneal epithelial basement membrane dystrophy, acanthamoeba, fibromyalgia, meibomian gland dysfunction, thyroid eye disease, rosacea, ptosis, keratoconus, eye pain syndrome, Stevens-Johnson syndrome, corneal epitheliopathy, corneal neuropathy (including LASIK-induced corneal neuropathy), corneal dystrophies (including recurrent corneal dystrophy), epithelial basement membrane dystrophy, corneal keratoconjunctivitis, ... Associated with one or more of the following conditions: membrane dystrophy, corneal erosion or abrasion (including recurrent corneal erosion or abrasion), ocular surface disease, blepharitis, graft-versus-host disease, meibomianitis, glaucoma, conjunctival chalazion, keratopathy (including herpetic keratopathy, filamentous keratopathy, band or bullous keratopathy, and exposure keratopathy), keratitis (including herpes simplex virus keratitis), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, or patients recovering from neurotrophic keratitis.

[0136] In some embodiments, ocular surface pain or chronic ocular surface pain is associated with dry eye disease. In some embodiments, administration of a compound of Formula I alleviates the symptoms of dry eye disease. Dry eye disease is generally understood to be a complex, multifactorial condition characterized by inflammation of the ocular surface and lacrimal glands, and a decrease in tear quality and / or quantity. It is believed that up to 30% of patients with dry eye disease suffer from ocular surface pain, which may be chronic. Thus, in some embodiments, the present invention alleviates at least about 10%, at least about 15%, at least about 20%, or at least about 30% of the symptoms of dry eye disease, including one or more of dry eye, eye discomfort, eye redness, burning or stinging in the eye, gritty or foreign body sensation, or photophobia.

[0137] In some embodiments, the present invention relates to a method of treating dry eye disease in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, polymorph, or cocrystal thereof. In some embodiments, the present invention relates to a method of treating dry eye disease in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, polymorph, or cocrystal thereof, wherein the compound of Formula I is safe for administration over a period of at least 2 months, at least 3 months, at least 4 months, or at least 5 months. In certain embodiments, the present invention provides the use of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, or cocrystal thereof, in the treatment of dry eye disease. In some embodiments, the present invention provides at least about 10% relief of symptoms of dry eye disease, including one or more of dry eye, eye discomfort, eye redness, burning or stinging in the eye, a gritty or foreign body sensation, or photophobia. In some embodiments, the present invention provides for administering to a subject in need of treatment a compound of Formula I at a concentration of about 0.5% w / v to about 3.5% w / v in an ophthalmologically compatible formulation. In some embodiments, the administration concentration ranges from about 0.5% to about 3.5% w / v, about 0.5% to about 2.5% w / v, about 0.5% to about 1.5% w / v, about 0.5% to about 3.0% w / v, about 1.0% to about 2.5% w / v, about 1.5% to about 3.0% w / v, or about 0.5% to about 2.5% w / v. In certain embodiments, the concentration of the compound of Formula I in a formulation for topical use is about 0.5% w / v, about 1.0% w / v, about 1.5% w / v, about 2.0% w / v, about 2.5% w / v, about 3.0% w / v, or about 3.5% w / v. In some embodiments, the dose per eye per administration is about 0.15 to about 1.15 mg, or about 0.15 mg, 0.2 mg, about 0.25 mg, 0.3 mg, about 0.35 mg, about 0.4 mg, about 0.45 mg, about 0.5 mg, about 0.55 mg, about 0.6 mg, about 0.65 mg, about 0.7 mg, about 0.75 mg, about 0.8 mg, about 0.85 mg, about 0.9 mg, about 0.95 mg, about 1.0 mg, about 1.05 mg, about 1.1 mg, or about 1.15 mg.In some embodiments, the dose per eye per administration is about 0.18 mg, about 0.37 mg, about 0.55 mg, about 0.74 mg, or about 0.92 mg. In some embodiments, the total daily dose per eye is about 0.5 to about 3.5 mg, or about 0.5 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 2.5 mg, about 3.0 mg, or about 3.5 mg. In some embodiments, the compound of Formula I is administered to a subject from 1 to 6 times daily, e.g., once, twice, three times, or four times daily. In some embodiments, the compound of Formula I is administered to a subject for a period of at least about 1 month, at least about 2 months, or at least about 3 months. In some embodiments, the compound of Formula I is administered in a formulation described herein.

[0138] In some embodiments of the methods described herein, administration of a compound of Formula I does not result in a change (e.g., less than a 5% difference, less than a 4% difference, or less than a 3% difference) in one or more of best corrected visual acuity, slit lamp examination, mydriatic examination, blink rate, tear production, intraocular pressure, or corneal staining compared to a placebo. In some embodiments of the methods described herein, administration of a compound of Formula I does not result in a delay in wound healing in a patient in need of treatment compared to a placebo.

[0139] Patient population In certain embodiments, the subject treated by the methods provided herein suffers from an ocular surface disorder. Non-limiting examples of ocular surface disorders include chronic ocular surface pain (COSP), dry eye disease, Sjogren's syndrome, conjunctivitis (including epidemic conjunctivitis, vernal keratoconjunctivitis, and allergic conjunctivitis), corneal epithelial basement membrane dystrophy, acanthamoeba, fibromyalgia, meibomian gland dysfunction, thyroid eye disease, rosacea, ptosis, keratoconus, eye pain syndrome, Stevens-Johnson syndrome, corneal epitheliopathy, corneal neuropathy (including LASIK-induced corneal neuropathy), and corneal dystrophies (including recurrent corneal dystrophy). This includes patients recovering from epithelial basement membrane dystrophy, corneal erosion or abrasion (including recurrent corneal erosion or abrasion), ocular surface disease, blepharitis, graft-versus-host disease, meibomianitis, glaucoma, conjunctival chalazion, keratopathy (including herpes keratopathy, filamentous keratopathy, band or bullous keratopathy, and exposure keratopathy), keratitis (including herpes simplex virus keratitis), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, or neurotrophic keratitis.

[0140] In certain embodiments, the methods provided herein are for treating or alleviating ocular surface pain, such as acute ocular surface pain.

[0141] In certain embodiments, the methods provided herein are for treating or alleviating ocular surface pain, such as chronic ocular surface pain (COSP). In certain aspects, COSP is characterized as persistent ocular surface pain (e.g., persistent severe ocular surface pain) that can or may interfere with normal daily activities. In certain aspects, COSP results in a decrease in quality of life and may persist for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, or at least 6 months. In some aspects, COSP may persist for at least about 2 months or at least about 3 months. In other aspects, COSP may persist for at least 3 months or at least 4 months. In certain aspects, subjects with COSP remain symptomatic despite adherence to other treatments required for the underlying condition (e.g., ocular surface disorder such as dry eye disease or Sjögren's syndrome).

[0142] In some embodiments, the subject to be treated suffers from ocular neuropathic pain (ONP). ONP is a group of eye pain disorders that can be caused by damage or disease affecting nerves, such as corneal nerves. Symptoms of ONP can include one or more of the following: eye pain, light sensitivity, dryness, tingling, foreign body sensation, and other hyperalgesia or abnormal sensations (dysesthesia), and pain from stimuli that are not normally painful (allodynia). Gabapentin and other neuropathic pain medications can be used to reduce the perception of sensory nerve stimuli or nerve stimuli.

[0143] In some embodiments, the subject being treated suffers from exposure keratopathy. EK is corneal damage primarily caused by prolonged exposure of the ocular surface to the external environment. EK can lead to permanent vision loss due to ulcers, microbial keratitis, and scarring. Patients at risk for EK include those with conditions that interfere with the cornea's ability to protect itself, incomplete eyelid closure (e.g., lagophthalmos, exophthalmos, eyelid malposition), a poor blink reflex, a poor blink rate (e.g., caused by neurological disorders such as Parkinson's disease or neuromuscular disorders), and / or reduced protective corneal lubrication. Symptoms of EK include a foreign body sensation, burning sensation, increased tear production, and intermittent blurred vision (due to an unstable tear film), pain, and photophobia. Standard treatments include frequent use of artificial tears with lubricating ointments at night and punctal plugs.

[0144] In some embodiments, the subject to be treated suffers from keratoconjunctivitis.Keratoconjunctivitis is an inflammatory process that affects both the conjunctiva and the cornea.Superficial inflammation of the cornea (keratitis) generally occurs in association with viral and bacterial conjunctivitis, for example, in adults.The following types of keratoconjunctivitis are distinguished based on the underlying cause of inflammation: Keratoconjunctivitis sicca is caused by inflammation due to dryness; · Vernal keratoconjunctivitis (VKC) occurs seasonally and is thought to be allergen-induced; Atopic keratoconjunctivitis is a symptom of atopy; Epidemic keratoconjunctivitis or adenoviral keratoconjunctivitis is caused by adenovirus infection; Infectious bovine keratoconjunctivitis (IBK) is a disease of cattle caused by the bacterium Moraxella bovis; Conjunctivitis in sheep and goats is mainly caused by Chlamydophila pecorum; Superior limbal keratoconjunctivitis is thought to be caused by mechanical trauma; Snow blindness (arc eye) refers to inflammation caused by photosensitive ultraviolet rays.

[0145] In some embodiments, the subject being treated suffers from dry eye. As used herein, the term "dry eye" refers to insufficient tear production and / or abnormal tear composition. Dry eye syndrome (DEDS), also known as dry eye syndrome, keratoconjunctivitis sicca, keratitis sicca, lacrimal dysfunction syndrome, or burning eye syndrome, is caused by a deficiency in the tear film. Dry eye is a multifactorial disease of the tears and ocular surface that causes symptoms of discomfort, visual disturbance, and tear film instability with potential damage to the ocular surface. It is characterized by a loss of tear film homeostasis, with ocular symptoms in which tear film instability and hyperosmolality, ocular surface inflammation and damage, and neurosensory abnormalities play a pathogenetic role (Craig JP, et al., The Ocular Surface 2017;15:276-83). Dry eye may be accompanied by increased tear film osmolality and ocular surface inflammation. Dry eye disorders can range from mild to moderate to severe. Symptoms of dry eye syndrome include a gritty feeling, foreign body sensation, burning sensation, photophobia, decreased vision, tearing, stinging, itching, gritty or gritty sensation, discharge, frequent blinking, clumped or clotted eyelashes due to discharge (usually worse upon waking), redness, blurred or fluctuating vision (worsened while reading, using a computer, watching television, driving, or playing video games), light sensitivity, eye pain and / or headache, eyelid heaviness, and eye strain. Causes of dry eye disease include, but are not limited to, idiopathic, congenital alacrimal vein thrombosis, xerophthalmia, lacrimal gland resection, and desensoritis; collagen diseases including rheumatoid arthritis, Wegener's granulomatosis, and systemic lupus erythematosus; Sjogren's syndrome and autoimmune diseases associated with Sjogren's syndrome; lipid tear film abnormalities due to blepharitis or rosacea; mucin tear film abnormalities due to vitamin A deficiency; trachoma, diphtheria keratoconjunctivitis; mucocutaneous diseases; aging; menopause; and diabetes.The signs and / or symptoms of dry eye, as defined herein, may also be caused by other conditions, including, but not limited to, prolonged visual work; working at a computer; being in a dry environment; warm or cold wind or air currents; seasonal changes; eye irritation, contact lenses, LASIK and other refractive surgery; fatigue; and medications such as isotretinoin, sedatives, diuretics, tricyclic antidepressants, antihypertensives, oral contraceptives, antihistamines, antinasal congestion medications, beta-blockers, phenothiazines, atropine, analgesic opiates such as morphine, etc.

[0146] Diagnostic tests for dry eye include assessment of corneal sensation, for example, using a cotton-tipped applicator, or more precisely, a Cochet-Bonnet aesthesiometer (corneal hypersensitivity and / or hyposensitivity may be present in severe and chronic dry eye disease); measurement of tear breakup time, for example, using fluorescein-impregnated strips moistened with preservative-free saline or more objective computerized methods that do not require fluorescein eye drops; performing ocular surface staining, for example, with sodium fluorescein, rose bengal, or lissamine green; and performing a Schirmer test (for mild dry eye). These include: delayed tear clearance tests; tear meniscus height; measurement of MMP-9 levels (MMP-9 has been shown to be increased in tears of patients with dry eye disease, and levels correlate with test findings in patients with moderate to severe dry eye), measurement of tear osmolality and tear film interferometry; and performance of the Sjo test (detection of SS-A (anti-Ro) and SS-B (anti-La) autoantibodies, salivary gland protein 1 (SP-1), carbonic anhydrase 6 (CA6), and parotid secretory protein (PSP), SP-1, CA, and PSP in serum).

[0147] Initial treatment includes artificial tears, lubricating ointments, and corticosteroids (e.g., loteprednol 0.5% eye drops four times a day). Prescription medications include cyclosporine, lifitegrast, diquafosol, rebamipide, and corticosteroids (e.g., loteprednol 0.5% eye drops four times a day).

[0148] The term "tear film dysfunction" refers to a condition in which the tear film breaks down at various locations on the cornea and conjunctiva, leading to symptoms of irritation as well as unstable and intermittently changing vision. For example, dry eye syndrome is characterized by tear film dysfunction. Symptoms of tear film dysfunction include tearing, burning, stinging, itching, gritty or gritty sensation, abrasion or foreign body sensation, discharge, frequent blinking, clumping or clotted eyelashes due to discharge (usually worse upon waking), redness, blurred or fluctuating vision (worsened while reading, using a computer, watching television, driving, or playing video games), light sensitivity, eye pain and / or headache, eyelid heaviness, and eye strain.

[0149] Adenoviral keratoconjunctivitis, also known as epidemic keratoconjunctivitis, is a common and highly contagious viral infection of the eye. The clinical course of adenoviral keratoconjunctivitis is divided into an acute phase with conjunctivitis of varying intensity, with or without corneal involvement, and a chronic phase with corneal opacification.

[0150] Vernal keratoconjunctivitis (VKC) is an atopic condition of the external ocular surface characterized by symptoms consisting of intense itching, photophobia, foreign body sensation, mucous discharge (often described as "ropy"), blepharospasm, and blurred vision (Buckley, RJ, Int Ophthalmol Clin, 1988 28(4):p. 303-8; Kumar, S., Acta Ophthalmologica, 2009.87(2):p. 133-147). It is generally symmetrical, but may also be asymmetric. It typically occurs in young men during seasonally hot, dry weather. In 23% of cases, the condition may be perennial (Kumar, S., Acta Ophthalmologica, 2009.87(2):p. 133-147; Bonini, S., et al., Ophthalmology, 2000.107(6):p. 1157-63).

[0151] Signs of VKC can be classified into conjunctival signs, limbal signs, and corneal signs. Conjunctival signs include diffuse conjunctival hyperemia and isolated giant upper eyelid papillae measuring more than 1 mm in diameter. Limbal signs include thickening and opacification of the limbal conjunctiva and limbal papillae that have a gelatinous appearance and sometimes confluent. Perilibal Horner-Trantas spots include focal white limbal patches composed of degenerated epithelial cells and eosinophils (Buckley, RJ, Int Ophthalmol Clin, 1988.28(4):p. 303-8). Corneal signs vary with the severity of the disease progression and include macroerosions, corneal ulcers, and scarring (Buckley, RJ, Int OphthalmolClin, 1988.28(4):p. 303-8).

[0152] Patients with active VKC (defined as moderate to severe ocular discomfort on examination, including photophobia, papillae on the upper tarsal conjunctiva, or clearly visible limbal Horner-Trantas spots) showed a significant increase in symptoms and signs of ocular surface disease. Patients with inactive VKC (defined as asymptomatic or mild discomfort on examination and no corneal abnormalities) showed increased photophobia, elevated conjunctival Lissamine Green staining and Schirmer test scores, and decreased fluorescein break-up time (BUT) and corneal sensitivity. This syndrome affects the ocular surface at all stages (active and quiescent), and abnormalities in tear film stability, epithelial cell integrity, and corneal nerve function may be detected (Villani E. et al., Medicine (Baltimore). 2015 Oct;94(42):e1648).

[0153] The following factors are thought to play a role in VKC: IgE-mediated responses via the release of mast cells, activated eosinophils, monocytes, and neutrophils, as well as CD4+ T helper 2-induced type IV hypersensitivity by immune regulators such as IL-4, IL-5, and bFGF (Buckley, RJ, Int OphthalmolClin, 1988.28(4):pp.303-8; Kumar, S., Acta Ophthalmologica, 2009.87(2):pp.133-147; LaRosa, M., et al., Ital J Pediatr, 2013.39:pp.18).

[0154] Treatment consists of cold compresses and eyelid scrubs, saline eye drops, along with topical antihistamines, nonsteroidal anti-inflammatory drugs, or corticosteroids, such as poorly absorbed corticosteroids (e.g., fluoromethorone, loteprednol, remexolone), ophthalmic mast cell stabilizers (sodium cromoglycate, nedocromil sodium, and lodoxamide), topical cyclosporine A, or tacrolimus, which may help relieve symptoms. For example, Oray, M. and E. Toker, Cornea, 2013.32(8):p. 1149-54; Vichyanond, P. and P. Kosrirukvongs, Curr Allergy Asthma Rep, 2013.13(3):p.308-14; Barot, RK et al., J Clin Diagn Res. 2016 June; 10(6):NC05-9; see Wan Q et al., Ophthalmic Res. 2018;59(3):126-134.

[0155] Atopic keratoconjunctivitis (AKC) usually begins at an older age, between the ages of 20 and 50, whereas VKC typically begins before age 10. Conjunctival lesions classically appear on the upper tarsal plate in VKC and the lower tarsal plate in AKC. AKC is typically more chronic in nature and more commonly causes corneal and conjunctival scarring.

[0156] Sjögren's syndrome (Sjögren's syndrome with dry eye) is a chronic inflammatory disorder characterized by dysfunction of exocrine glands, including the salivary and lacrimal glands, often resulting in severe dry eye. Primary symptoms include dry eyes (keratitis sicca or keratoconjunctivitis sicca) and dry mouth (xerostomia). Severe dry eye can lead to corneal pain, corneal scarring, ulcers, infection, and even perforation. Differential diagnoses include conditions such as adult blepharitis, dry eye disease, and juvenile idiopathic arthritis, uveitis, and keratopathy (e.g., superficial punctate keratopathy, filamentous keratopathy, neurotrophic keratopathy, and exposure keratopathy). Treatment of Sjögren's syndrome aims to maintain tear film integrity through preservation, enhancement, and / or replacement of deficient tear secretion. Thus, treatments for Sjögren's syndrome include artificial tears and lubricating ointments; autologous serum eye drops; oral omega-6 essential fatty acids; liquid-ventilated, gas-permeable scleral lenses; topical corticosteroids; punctal closure to reduce tear drainage; small lateral tarsorrhaphy; environmental humidification; hydrophilic barrier lenses; bromhexine and 3-isobutyl-1-methylxanthine (IBMX) (to enhance tear production / secretion); drugs that stimulate muscarinic receptors (pilocarpine and cevimeline); immunosuppressants such as methotrexate; antimalarials, cyclophosphamide, leflunomide, or infliximab, a monoclonal antibody against tumor necrosis factor (TNF), such as TNF-α; cyclosporine A; and barrier contact lenses.

[0157] Stevens-Johnson syndrome (SJS) is a dermatological emergency or severe skin reaction characterized by the presence of vesicular lesions of the epidermis and mucous membranes covering less than 10% of the body surface area. Early symptoms of SJS include fever and flu-like symptoms, which may precede or occur with a macular rash on the trunk and face. As the disease progresses, the macular rash coalesces, blisters form in the affected areas, and eventually the epidermal layer peels off. During the acute phase of SJS-TEN, ocular involvement occurs in 80% of patients.

[0158] High fever (>102.2), fatigue, joint pain, a macular rash on the trunk, neck, and face, and a recent history of new medications or a recent increase in the dose of an existing medication are indicators used to diagnose SJS. A skin biopsy of the affected area can be performed to confirm the diagnosis. Granulolysin can be used as a diagnostic marker for SJS. The concentration of granulolysin in blister fluid correlates with the severity of the acute phase of SJS (Chung WH, et al. Nat Med. 2008;14(12):1343-50).

[0159] Ocular manifestations of SJS include conjunctivitis, subconjunctival hemorrhage, subconjunctival scarring, membranous conjunctivitis, conjunctival ulcers, superficial punctate epithelial erosions, epithelial defects, lid margin ulcers, lid margin keratinization, lid synechiae, ptosis, trichiasis, anterior blepharitis, lacrimal punctal obstruction, meibomian gland disease, corneal opacity, dry eye, hypertrichosis, limbal stem cell failure, and corneal neovascularization. Ocular treatment for SJS consists of saline eye drops, preservative-free artificial tears, and ointments to provide adequate lubrication and reduce epithelial damage. Patients with corneal or conjunctival epithelial defects are treated with prophylactic topical antibiotics, such as fourth-generation fluoroquinolones. Patients with mild or moderate ocular involvement (involvement of less than one-third of the lid margin, a conjunctival defect less than 1 cm in greatest diameter, and no corneal epithelial defect) are typically treated with topical moxifloxacin 0.5% four times daily, cyclosporine 0.05% twice daily, and topical steroids (prednisolone acetate 1% four to eight times daily or dexamethasone 0.1% twice daily). Patients with severe or very severe ocular involvement (involvement of more than one-third of the lid margin, a conjunctival defect greater than 1 cm, and a corneal epithelial defect) undergo amniotic membrane (AM) transplantation in addition to the above treatments.

[0160] In some embodiments, the subject being treated has corneal epitheliopathy, which is a disease associated with the corneal epithelium, for example, a disease that manifests as an alteration in the corneal epithelial barrier function.

[0161] In some embodiments, the subject being treated suffers from corneal neuropathy or corneal neuralgia. Corneal neuropathy or corneal neuralgia is a disorder associated with corneal pain caused by damage to the nerve and sensory fibers of the cornea. One example of corneal neuropathy is LASIK-induced corneal neuropathy. Corneal neuropathy can generally be identified and diagnosed by dry eye testing. Although the causes and risk factors are still unclear, patients who have dry eye-like symptoms, increased corneal sensitivity, and changes in corneal nerve morphology but no signs of dryness may suffer from corneal neuropathy.

[0162] In some embodiments, the subject being treated suffers from an ocular surface disease or disorder. The term "ocular surface disease" or "ocular surface disorder" encompasses disease entities and associated symptoms resulting from various abnormalities, including abnormal eyelid structure or function, abnormal or altered tear production or composition, and related subclinical manifestations. Many diseases can cause ocular surface disorders. Patients with ocular surface disorders may exhibit clinical signs common to several diseases, including chronic punctate keratopathy, filamentous keratopathy, recurrent corneal erosion, bacterial conjunctivitis, culture-negative conjunctivitis, cicatricial (scar-forming) conjunctivitis, persistent epithelial defects, infectious keratitis, corneal melt, and ocular surface failure. The most common ocular surface disorders are due to tear film abnormalities and / or palpebral gland dysfunction ("blepharitis").

[0163] In some embodiments, the subject being treated suffers from neurotrophic keratitis or neurotrophic keratopathy. Neurotrophic keratitis or neurotrophic keratopathy (NK) is a corneal degenerative disease characterized by reduced or absent corneal sensitivity. In NK, corneal innervation by the trigeminal nerve is impaired. Because corneal sensory innervation is impaired in NK, patients usually do not complain of ocular surface symptoms. However, blurred vision may be reported due to irregular epithelium or epithelial defects (PEDs), scarring, or edema. NK is usually classified into three different stages according to the "Mackie classification." Stage II NK is defined by recurrent or persistent epithelial defects, most commonly occurring in the upper half of the cornea. One treatment available for stage II NK includes topical nerve growth factor (NGF). Patients typically experience pain during treatment with NGF due to nerve remodeling.

[0164] In some embodiments, the subject being treated suffers from blepharitis. Blepharitis is an inflammatory condition of the eyelid margin that can lead to permanent changes in the eyelid margin or vision loss due to superficial keratopathy, corneal neovascularization, and ulcers. Depending on the anatomical location, blepharitis can be divided into anterior and posterior blepharitis. Anterior blepharitis affects the eyelid skin, the base of the eyelashes, and the eyelash follicles, and includes the traditional classifications of staphylococcal blepharitis and seborrheic blepharitis. Posterior blepharitis affects the meibomian glands and glandular orifices, and is primarily caused by meibomian gland dysfunction. Symptoms of chronic blepharitis may include redness, burning, irritation, tearing, crusting and sticking of the eyelids, and visual problems such as photophobia and blurred vision. Long-term management of symptoms may include a daily eyelid cleansing routine and the use of medications to reduce infection and inflammation. Treatments include topical or systemic antibiotics such as bacitracin or erythromycin; oral antibiotics, such as tetracyclines (tetracycline, doxycycline, minocycline) or macrolides (erythromycin, azithromycin); topical steroids, such as corticosteroids, e.g., loteprednol etabonate, fluorometholone; topical combinations of antibiotics and corticosteroids, such as tobramycin / dexamethasone or tobramycin / loteprednol; and topical cyclosporine 0.05%.

[0165] In some embodiments, the subject being treated suffers from meibomian gland dysfunction. Meibomian glands are holocrine exocrine glands located within the tarsal plate at the eyelid margin and responsible for the production of meibum, an oily substance that prevents the evaporation of the eye's tear film. Meibomian gland dysfunction (MGD), also known as meibomitis, posterior blepharitis, or meibomian gland inflammation, is a chronic, diffuse abnormality of the meibomian gland, generally characterized by end-duct obstruction and / or qualitative or quantitative changes in glandular secretion (Nelson JD, et al., Invest Ophthalmol Vis Sci 2011;52:1930-7). This can result in tear film changes, ocular irritation, clinically evident inflammation, and ocular surface disease. MGD often causes dry eye and may contribute to blepharitis. Topical steroids and topical / oral antibiotics may also be prescribed to reduce inflammation. Intense pulsed light (IPL) treatments, or other mechanical treatments that apply heat and pressure to express the glands (e.g., LipiFlow), have also been shown to reduce inflammation and improve glandular function in patients.

[0166] In some embodiments, the subject being treated suffers from graft-versus-host disease. Graft-versus-host disease (GVHD) is an inflammatory disease specific to allogeneic transplants. It occurs when transplanted white blood cells attack the recipient's tissues and can occur even when the donor and recipient are HLA-matched. Acute GVHD typically occurs within three months of transplantation and can affect the skin, intestines, or liver. Corticosteroids such as prednisone are the standard treatment. Chronic GVHD can also develop after allogeneic transplantation and is a major cause of late complications. In addition to inflammation, chronic GVHD can lead to the development of fibrosis or scar tissue similar to scleroderma or other autoimmune diseases, potentially causing functional impairment and requiring long-term immunosuppressive therapy.

[0167] In some embodiments, the subject being treated suffers from ocular graft-versus-host disease. GVHD occurs in patients who have undergone allogeneic hematopoietic stem cell transplantation. It can occur in patients with acute or chronic GVHD, but it is more common in patients with the chronic form. Approximately 40-90% of patients with chronic GVHD experience ocular symptoms. Ocular manifestations may include moderate to severe keratoconjunctivitis sicca, bilateral marginal keratitis, anterior uveitis, corneal ulcers, or neovascularization. Treatments include topical lubricants, including preservative-free artificial tears, autologous serum tears, and other topical and systemic immunosuppressive treatments; systemic steroids; and topical cyclosporine 0.5%. [Example]

[0168] The following examples are included to demonstrate non-limiting embodiments of the invention.

[0169] General Test Conditions The following procedures were used for each test condition.

[0170] [Table 1]

[0171] [Table 2]

[0172] Those skilled in the art will understand that X-ray diffraction patterns may be obtained with measurement errors that depend on the measurement conditions used. In particular, it is generally known that the intensities of X-ray diffraction patterns may vary depending on the measurement conditions used. It should also be understood that relative intensities may vary depending on the experimental conditions and the wavelength of the X-ray radiation used. The agreement of the 2-theta diffraction angles between the sample and the reference is within 0.2° for the same crystalline form, and the degree of such measurement error should be considered relative to the aforementioned diffraction angle. Therefore, it should be understood that the crystalline forms of the present invention are not limited to those that provide X-ray diffraction patterns that are completely identical to those shown in the accompanying figures disclosed herein. Any crystalline form that provides an X-ray diffraction pattern substantially identical to that disclosed in the accompanying figures is within the scope of the present invention. The ability to confirm the substantial identity of X-ray diffraction patterns is within the knowledge of those skilled in the art.

[0173] [Table 3]

[0174] [Table 4]

[0175] [Example 1] Preparation of Compounds of Formula I (S)-1-(6-fluoro-5-((((1r,3S)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol (or trans-(S)-1-(6-fluoro-5-(((3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol) and Synthesis of (R)-1-(6-fluoro-5-((((1r,3R)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol (or trans-(R)-1-(6-fluoro-5-(((3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol)

[0176] Step 1.1: Synthesis of 6-fluoro-8-(oxiran-2-yl)isoquinoline

[0177] [ka] To a solution of NaH (1.0 g, 41.45 mmol) and anhydrous DMSO (40 mL) at room temperature, trimethylsulfoxonium iodide (8.3 g, 37.68 mmol) was added and stirred for 30 min. Then, 6-fluoroisoquinoline-8-carbaldehyde (Step 6.5, 3.3 g, 18.84 mmol) dissolved in DMSO (20 mL) was added dropwise at room temperature. After 5 min, the reaction was quenched with ice-water and extracted three times with EtOAc. The combined organic portions were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (12 g SiliCycle column, eluted with 0–20% EtOAc in hexane) to give 6-fluoro-8-(oxiran-2-yl)isoquinoline (2.3 g, 64%). MS (ESI+) [Method 6A]: m / z 190.1 (M+H); Rt 0.79 min. 1H NMR (600 MHz, CDCl3) δ 9.55 (s, 1H), 8.58 (d, J = 5.4 Hz, 1H), 7.65 (d, J = 5.4 Hz, 1H), 7.37 (d, J = 9.0 Hz, 2H), 4.60 - 4.59 (m, 1H), 3.37 - 3.35 (m, 1H), 2.82 - 2.80 (m, 1H).

[0178] Step 1.2: Synthesis of 1-(6-fluoroisoquinolin-8-yl)ethane-1,2-diol

[0179] [ka] To a solution of 6-fluoro-8-(oxiran-2-yl)isoquinoline (2.1 g, 11.11 mmol) in THF-HO (12 mL, 2:1 v / v) was added HSO (5 mL) dropwise at room temperature and stirred at 60 °C for 16 h. The reaction mixture was made basic with saturated NaHCO solution and extracted twice with EtOAc. The combined organic portions were washed with brine, dried over anhydrous NaSO, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (12 g SiliCycle column, eluted with 0–5% MeOH in CHCl) to give 1-(6-fluoroisoquinolin-8-yl)ethane-1,2-diol (1.6 g, 69%). MS (ESI+) [Method 4A]: m / z 208.3 (M+H); Rt 0.40 min. 1 H NMR (600 MHz, CDCl3) δ 9.49 (s, 1H), 8.49 (d, J = 6.0 Hz, 1H), 7.67 - 7.63 (m, 2H), 7.35 (dd, J = 8.4, 1.8 Hz, 1H), 4.13 - 4.10 (m, 1H), 4.06 (dd, J = 12.6, 3.6 Hz, 1H), 3.76 (dd, J = 11.4, 3.6 Hz, 1H).

[0180] Step 1.3: Synthesis of 6-fluoro-8-(2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disiladecan-5-yl)isoquinoline

[0181] [ka] To a solution of 1-(6-fluoroisoquinolin-8-yl)ethane-1,2-diol (1.5 g, 7.24 mmol) and imidazole (3.4 g, 50.68 mmol) in DMF (15 mL), TBDMS-Cl (5.4 g, 36.17 mmol) was added portionwise at 0 °C and stirred at room temperature for 16 h. The reaction mixture was then diluted with water and extracted three times with EtOAc. The combined organic portions were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (12 g SiliCycle column, eluted with 0–10% EtOAc in hexanes) to give 6-fluoro-8-(2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disiladecan-5-yl)isoquinoline (2.7 g, 85%). MS (ESI+) [Method 6A]: m / z 436.3 (M+H); Rt 2.15 min. 1 H NMR (600 MHz, CDCl3) δ 9.59 (s, 1H), 8.52 (d, J = 4.8 Hz, 1H), 7.61 (d, J = 5.4 Hz, 1H), 7.55 (dd, J = 10.2, 1.8 Hz, 1H), 7.33 (dd, J = 9.0, 2.4 Hz, 1H), 5.54 (d, J = 6.0 Hz, 1H), 3.87 - 3.85 (m, 1H), 3.77 - 3.74 (m, 1H), 0.92 (s, 9H), 0.90 (s, 9H), 0.13 (s, 6H), 0.09 (s, 6H).

[0182] Step 1.4: Synthesis of 6-fluoro-8-(2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disiladecan-5-yl)isoquinoline-5-carbaldehyde

[0183] [ka] The title compound was prepared according to the procedure in Step 6.8. The residue was purified by flash chromatography (40 g SiliCycle column, eluting with 0–15% EtOAc in hexanes) to give 6-fluoro-8-(2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disiladecan-5-yl)isoquinoline-5-carbaldehyde (2.0 g, 62%). MS (ESI+) [Method 4A]: m / z 464.4 (M+H); Rt 1.77 min. 1 H NMR (600 MHz, CDCl3) δ 9.57 (s, 1H), 8.50 (d, J = 5.4 Hz, 1H), 7.60 (d, J = 5.4 Hz, 1H), 7.54 (dd, J = 7.8, 2.4 Hz, 1H), 7.32 (dd, J = 9.0, 2.4 Hz, 1H), 5.53 (d, J = 6.0 Hz, 1H), 3.87 - 3.84 (m, 1H), 3.76 - 3.73 (m, 1H), 0.89 (s, 9H), 0.75 (s, 9H), 0.12 (s, 6H), -0.05 (s, 6H).

[0184] Step 1.5: Synthesis of (1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)-N-((6-fluoro-8-(2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disiladecan-5-yl)isoquinolin-5-yl)methyl)cyclobutan-1-amine

[0185] [ka] The title compound was synthesized according to the procedure described in step 1.4 using (1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutan-1-amine, HCl (step 1.3, 1.0 g, 3.50 mmol) and 6-fluoro-8-(2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disiladecan-5-yl)isoquinoline-5-carbaldehyde (1.46 g, 3.15 mmol). The crude product was purified by flash chromatography (24 g SiliCycle column, eluting with 0–5% MeOH in CHCl) to give (1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)-N-((6-fluoro-8-(2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disiladecan-5-yl)isoquinolin-5-yl)methyl)cyclobutan-1-amine (1.5 g, 62%). MS (ESI+) [Method 6A]: m / z 697.3 (M+H); RT 1.63 min.

[0186] Step 1.6: Synthesis of (S)-1-(6-fluoro-5-((((1r,3S)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol and (R)-1-(6-fluoro-5-((((1r,3R)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol

[0187] [ka] To a solution of (1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)-N-((6-fluoro-8-(2,2,3,3,8,8,9,9-octamethyl-4,7-dioxa-3,8-disiladecan-5-yl)isoquinolin-5-yl)methyl)cyclobutan-1-amine (1.5 g, 2.15 mmol) in THF (25 mL) was added dropwise TBAF solution (1 M in THF) (5.4 mL, 5.38 mmol) at 0 °C and stirred for 2 h. The reaction mixture was diluted with water and extracted twice with EtOAc. The combined organic portions were washed with brine solution, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (24 g SiliCycle column, eluting with 0-10% MeOH in CHCl) to give 1-(6-fluoro-5-((((1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol (1.0 g, 98%). MS (ESI+) [Method 6A]: m / z 469.2 (M+H); Rt 1.29 min. 1 H NMR (400 MHz, CD3OD) δ 9.57 (s, 1H), 8.52 (d, J = 6.0 Hz, 1H), 8.12 (d, J = 5.6 Hz, 1H), 7.67 (d, J = 10.6 Hz, 1H), 7.22 (t, J = 9.6 Hz, 1H), 7.05 - 7.01 (m, 2H), 5.58 - 5.56 (m, 1H), 4.85 - 4.82 (m, 1H), 4.17 (d, J = 1.6 Hz, 2H), 3.86 - 3.82 (m, 1H), 3.78 - 3.74 (m, 1H), 3.60 - 3.57 (m, 1H), 2.36 - 2.33 (m, 4H).

[0188] Chiral preparative HPLC of the racemic compound (column: CHIRALPAK-IG (250 mm x 20 mm); mobile phase: hexane and IPA:MeOH (1:1); isocratic: 60 / 40; flow rate: 15 mL / min) yielded (S)-1-(6-fluoro-5-((((1r,3S)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol as a white solid, peak 1 (395 mg, 40%): Chiral HPLC: 99% (Rf 7.840 min, column: CHIRALPAK-IG (150 mm x 4.6 mm), 5.0 μL, mobile phase: n-hexane and EtOH, isocratic: 80 / 20; flow rate: 1 mL / min). MS (ESI+) [Method 1A]: m / z 469.2 (M+H); Rt 1.29 min. 1 H NMR (400 MHz, CD3OD) δ 9.57 (s, 1H), 8.53 (d, J = 6.0 Hz, 1H), 8.13 (d, J = 5.6 Hz, 1H), 7.68 (d, J = 10.6 Hz, 1H), 7.23 (t, J = 9.6 Hz, 1H), 7.06 - 7.01 (m, 2H), 5.59 - 5.56 (m, 1H), 4.85 - 4.82 (m, 1H), 4.19 (s, 2H), 3.87 - 3.83 (m, 1H), 3.78 - 3.74 (m, 1H), 3.60 - 3.57 (m, 1H), 2.37 - 2.34 (m, 4H); and (R)-1-(6-fluoro-5-((((1r,3R)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol white solid peak 2 (345 mg, 35%). Chiral HPLC: 97% (Rf 17.481 min; Column: CHIRALPAK-IG (150 mm × 4.6 mm), 5.0 μL; Mobile phase: n-hexane and EtOH; Isocratic: 80 / 20; Flow rate: 1 mL / min). MS (ESI+) [Method 3A]: m / z 469.0 (M+H); Rt 1.25 min. 1H NMR (400 MHz, CD3OD) δ 9.57 (s, 1H), 8.53 (d, J = 6.0 Hz, 1H), 8.13 (d, J = 5.6 Hz, 1H), 7.68 (d, J = 10.6 Hz, 1H), 7.23 (t, J = 9.6 Hz, 1H), 7.06 - 7.01 (m, 2H), 5.59 - 5.56 (m, 1H), 4.85 - 4.82 (m, 1H), 4.18 (d, J = 1.2 Hz, 2H), 3.87 - 3.83 (m, 1H), 3.78 - 3.74 (m, 1H), 3.60 - 3.57 (m, 1H), 2.37 - 2.34 (m, 4H). Example 1 was isolated in amorphous form.

[0189] [Example 2] Crystal form A Compound I was prepared as described in Example 1. Crystalline Form A is as follows:

[0190] [ka] It was obtained from the isomer shown in

[0191] The starting material for polymorphic studies was recrystallized from acetonitrile and washed with water. Approximately 28 g of this material was suspended in 500 ml of acetonitrile and heated to 55°C to obtain a clear solution. The solution was gradually cooled to 50°C within 1 hour and held isothermal for an additional 3 hours. A small amount of solid precipitate formed and adhered to the stirrer paddle. The mixture was cooled to 25°C within an additional 5 hours, held at 50°C for an additional 3 hours, then cooled to 10°C and held for 5 hours. The resulting solid was isolated by suction filtration and dried under vacuum at 50°C for 4 hours. A pale yellow solid was obtained in approximately 70% yield.

[0192] Approximately 13.8 g of Form A was suspended in 700 mL of water and stirred at 25° C. for 6 hours. The mixture was filtered and the solid was resuspended in 750 mL of water. This process was repeated four times. Residual chloride was confirmed by ion chromatography. The solid was collected and dried under vacuum at 50° C. for 5 hours. 12.8 g of an off-white solid was obtained in 92.7% yield (purity: 99.6%).

[0193] Preparation of Form A on a kg scale: A reactor was charged with 2.8 kg of Compound I, 10.8 kg of ethanol, and 8.54 kg of water and heated to 53°C until a clear solution was obtained. The solution was filtered through a 0.45 μm filter cloth and transferred to a crystallizer, where the solution was cooled to 40°C. 17.1 g of Form A seeds were introduced into the system, and the suspension was held at 40°C for 4 hours. 10.7 kg of water was introduced over 4 hours, and the suspension was held for 3 hours. The suspension was cooled to 20°C in 3 hours and then held at 20°C for 4 hours. The suspension was then filtered and washed with a mixture of 3.8 kg of ethanol and 1.5 kg of water. The wet cake was then dried under vacuum at 60°C for 12 hours to obtain 2.64 kg of crystalline Form A of Compound I.

[0194] Preparation of Form A from solution at 25°C: Approximately 100 mg of Compound I was equilibrated with 0.5 ml of solvent at 25°C for 28 days under stirring. The solution was filtered and dried in air for 10 minutes. Compound I precipitated in various solvents under these conditions to obtain Form A.

[0195] [Table 5]

[0196] Preparation of Form A from a hot saturated solution at 60° C.: Approximately 100-300 mg of Compound I (or an amount appropriate to ensure saturation) was dissolved in a minimum amount of solvent at approximately 60° C. The solution was allowed to cool slowly to ambient temperature under stirring.

[0197] If cooling to room temperature did not result in a suspension, or if the suspension was too light to collect sufficient material for analysis, the samples were stored at 5°C for at least 5 days or at −20°C for at least 72 h.

[0198] The solution was filtered and dried in air for 10 minutes. Compound I precipitated in various solvents under these conditions to give Form A.

[0199] [Table 6]

[0200] Preparation of Form A by reverse antisolvent addition: A nearly saturated solution of Compound I was added to excess antisolvent with vigorous stirring. If no immediate precipitation occurred, the mixture was allowed to stir at room temperature for up to 24 hours. The precipitate was used to characterize the polymorphic form.

[0201] [Table 7]

[0202] The X-ray powder diffraction pattern of crystalline form A is shown in FIG. 1 and the peak list is given in Table 4.

[0203] [Table 8]

[0204] Solvent Solubility: Compound I is soluble in many solvents, including acetone, 1,4-dioxane, ethanol, ethyl acetate, isopropyl acetate, methanol, 2-methyl-2-butanol, tetrahydrofuran, and solvent mixtures of 1-propanol / water (98.5:1.5), 2-propanol / water (96:4), and methanol / water (78:22), at a solubility of greater than 25 mg / mL. Compound I is sparingly soluble (1-25 mg / mL) in acetonitrile, dichloromethane, methyl tert-butyl ether, and nitromethane. Compound I is slightly soluble (<1 mg / mL) in toluene and heptane.

[0205] Compound I was not observed to form hydrates after 2 and 4 weeks when equilibrated in aqueous fluids such as water, methanol / water (33:67, 42:58) at 4°C.

[0206] Thermal studies: Compound I Form A exhibits a melting point of 130.3 °C and a melting enthalpy of 90 J / g, indicating a highly crystalline material (Figure 2). Varying the heating rate only changed the melting point by 0.3% and the enthalpy by approximately 3%. Cooling and reheating the melt resulted in a glass transition of the amorphous material at 40 °C, with a change in isobaric heat capacity of 0.54 J / g / K.

[0207] Using TGA, the loss on drying of Form A was determined to be 0.13% at 130°C (Figure 3). No change was observed when Form A was exposed to 92% RH for 24 hours.

[0208] Compound I Form A exhibited slight hygroscopicity with DVS, absorbing approximately 0.4% water vapor at 95% RH, but there was no change in its solid state. Furthermore, no change in the shape of Form A was observed after compression, although only slight peak broadening was observed. Furthermore, Form A did not exhibit any shape change under dry milling or wet granulation with water and ethanol (Figure 4).

[0209] [Example 3] Biological activity of Compound 1 (Example 1) Measurement of TRPV1 inhibition Chinese hamster ovary (CHO) cells transfected to express the human TrpV1 receptor (herein referred to as CHO-huTrpV1 cells) were cultured in F-12 Ham's Nutrient Mixture Medium (HyClone SH30026.01) supplemented with 10% fetal bovine serum (Invitrogen #26140-079), 1% antibiotic / antimycotic (Invitrogen #15240-062), and 500 μg / mL Geneticin (ThermoFisher Scientific #1031035). Cells were cultured in T-75 flasks in a 37°C incubator with 5% CO2. Cells were subcultured twice weekly at a ratio of 1:10 to 1:20 to maintain stable growth. For experiments, cells were harvested at approximately 80% confluence and seeded at 15,000 cells per well in 20 μl of medium into a 384-well black cell culture plate (cat#781091, Greinier Bio-One Inc.) and cultured overnight.

[0210] FLIPR calcium assay for detecting calcium influx in CHO-huTrpV1 cells Loading dye was prepared according to the instructions in the Calcium 6 Assay Kit (Molecular Probes, #R8190): 10 ml of buffer from bottle B was added to one vial of bottle A (adapted from -20°C to room temperature) and mixed well. 2.5 mM freshly prepared probenecid was then added and mixed well. 20 μl / well of loading dye was added to the top of the cells and incubated at 37°C for 1 hour and 30 minutes.

[0211] Assay buffer was prepared: 1x HBSS, 2mM HEPES, 0.1% BSA (Invitrogen, #P36400) supplemented with 2.5mM freshly prepared probenecid. 25µl of assay buffer per well in a 384-well clear plate (cat#782281, GreinerBio-one) was dispensed using a buffer distributor (Thermo Scientific Multidrop ComBi). Compounds were placed in a 384-Echo plate (cat#LPL0200, Labcyte) at a starting compound concentration of 10mM, followed by serial dilutions (8µl / well) of 100% DMSO. 125nl of compound was transferred to the 384-well plate containing 25µl of buffer per well using an Echo® 555 Liquid Handler (Labcyte) to ensure compound concentrations were 5x the final concentration. The plate was gently shaken at 40 rpm / min for 10 minutes to mix. Five-fold compound concentrations in 10 μl of buffer were transferred to the cell plate (containing 20 μl of cells and 20 μl of dye) using a Vertical Pipetting Station 384ST (Agilent Technologies). NADA (N-arachidonyl dopamine, cat#A8848, Sigma) in assay buffer was prepared at 6x the final concentration and dispensed at 30 μL / well into a 384-well clear plate.

[0212] Within 10–15 minutes of adding the compound to the cell plate along with the loading dye, the cell plate and the plate containing NADA were placed in a FLIPR (Fluorescence Imaging Plate Reader) instrument (Tetra System, Molecular Devices). TRPV1 receptors were stimulated by the application of 10 μl of NADA per well. To test the effect of compounds on antagonistic potential, 2.5 μM NADA was used at the EC concentration.

[0213] Antagonist IC 50To determine the IC value (the concentration of antagonist that inhibits the response to NADA by 50%), at least 10 antagonist concentrations were measured in triplicate. Responses in the presence of antagonist were calculated as a percentage of the control response to NADA and plotted against the antagonist concentration. 50 was estimated by nonlinear regression analysis of sigmoid logistic curves using the HELIOS (PROD2) system. These values ​​were averaged (mean and standard error of the mean) for at least three independent experiments.

[0214] [Table 9]

[0215] All publications and patent documents cited in this specification are herein incorporated by reference as if each such publication or document was specifically and individually indicated to be incorporated by reference herein. The present invention and its embodiments have been described in detail. However, the scope of the present invention is not intended to be limited to the particular embodiments of any process, manufacture, composition of matter, compound, means, method, and / or step described herein. Various modifications, substitutions, and changes can be made to the disclosed materials without departing from the spirit and / or essential characteristics of the invention. Accordingly, those skilled in the art will readily appreciate that subsequent modifications, substitutions, and / or variations can be utilized in accordance with related embodiments of the present invention that perform substantially the same function or achieve substantially the same result as the embodiments described herein. Therefore, the following claims are intended to include within their scope such modifications, substitutions, and variations of the processes, manufacture, compositions of matter, compounds, means, methods, and / or steps disclosed herein. The claims should not be construed as being limited to the described order or elements unless expressly stated to that effect. It should be understood that various changes in form and detail may be made therein without departing from the scope of the appended claims.

Claims

1. The following structure 【Chemical 1】 A crystalline form of 1-(6-fluoro-5-((((1r,3r)-3-(4-fluoro-3-(trifluoromethyl)phenoxy)cyclobutyl)amino)methyl)isoquinolin-8-yl)ethane-1,2-diol (Compound I) having the formula:

2. 2. The crystalline form of Compound I of claim 1, having the following structure: 【Chemistry 2】

3. 3. The crystalline form of compound I of claim 2 having crystalline form A.

4. 4. A crystalline form of the compound of formula I according to any one of claims 1 to 3, characterized by an X-ray diffraction pattern having three or more peaks at values ​​of 2θ selected from 14.3, 14.8, and 21.8±0.2 degrees 2θ.

5. 5. The crystalline form of Compound I of any one of claims 1 to 4, characterized by an X-ray diffraction pattern having three or more peaks at values ​​of 2θ selected from 12.5, 14.3, 14.8, 21.8, and 22.6±0.2 degrees 2θ.

6. 6. The crystalline form of Compound I of any one of claims 1 to 5, characterized by an X-ray diffraction pattern having 3 or more, 4 or more, 5 or more, 6 or more, or 7 or more peaks at 2θ values ​​selected from 12.5, 14.3, 14.8, 20.1, 21.8, 22.6, and 23.2±0.2 degrees 2θ.

7. 7. The crystalline form of Compound I according to any one of claims 1 to 6, characterized by the X-ray diffraction pattern shown in Figure 1.

8. 8. The crystalline form of Compound I according to any one of claims 1 to 7, characterized by the differential scanning calorimetry pattern shown in Figure 2.

9. 9. The crystalline form of Compound I of any one of claims 1 to 8, characterized by a DSC thermogram exhibiting an endotherm at about 131.5°C.

10. 10. The crystalline form of Compound I according to any one of claims 1 to 9, characterized by a water loss of about 0.13% by weight as measured by thermogravimetric analysis.

11. 11. The crystalline form of Compound I of claim 10, characterized by a water loss of about 0.13% by weight at 130°C as measured by thermogravimetric analysis.

12. 12. A method for preparing crystalline Form A of Compound I according to any one of claims 1 to 11, comprising cooling a hot saturated solution of Compound I in a solvent to crystallize Compound I as crystalline Form A.

13. 13. The method of claim 12, wherein the solution is cooled to a temperature of about 0°C to about 25°C.

14. 14. The method of claim 12 or 13, wherein the solvent is selected from the group consisting of acetone, acetonitrile, dichloromethane, ethanol, ethyl acetate, isopropyl acetate, methyl tert-butyl ether, acetone / heptane, and methanol / water.

15. A method for preparing crystalline Form A of Compound I according to any one of claims 1 to 11, comprising crystallizing Form A from a solution of Compound I in a solvent.

16. 16. The method of claim 15, wherein the solution is saturated.

17. 17. The method of any one of claims 15-16, wherein said Form A is crystallized at a temperature of about 25°C.

18. 18. The method of any one of claims 15 to 17, wherein the solvent is selected from the group consisting of acetone, acetonitrile, dichloromethane, ethyl acetate, isopropyl acetate, 2-methyl-2-butanol, and methyl tert-butyl ether.

19. 18. The method of any one of claims 15 to 17, wherein the solvent is selected from the group consisting of mixtures of water / methanol, water / 1-propanol, and water / 2-propanol.

20. 12. A method for preparing crystalline Form A of Compound I according to any one of claims 1 to 11, comprising adding an anti-solvent to a solution of Compound I in a solvent.

21. 21. The method of claim 20, wherein the solvent is selected from the group consisting of acetone, dioxane, ethanol, ethyl acetate, methanol, 2-methyl-2-butanol, and tetrahydrofuran.

22. 22. The method of claim 20 or claim 21, wherein the anti-solvent is selected from the group consisting of water, heptane, and toluene.

23. A composition comprising a crystalline form of Compound I according to any one of claims 1 to 11.

24. A composition comprising crystalline Form A of Compound I in substantially pure form.

25. Use of a crystalline form of Compound I according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment of ocular surface pain.

26. Use of a crystalline form of Compound I according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment of an ocular surface disorder.

27. Use of a crystalline form of compound I according to any one of claims 1 to 11 in the preparation of a pharmaceutical formulation.

28. 12. A method for preparing a pharmaceutical formulation comprising the crystalline form of Compound I of any one of claims 1 to 11, comprising dissolving the crystalline form of Compound I in an ophthalmologically acceptable carrier formulated for ophthalmic use, e.g., topical application to the ocular surface.

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