RU2850452C2 - Methods of treating pain on the surface of the eye - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: the group of inventions relates to a method for treating pain on the surface of the eye in a subject in need thereof, comprising administering to the subject an effective amount of 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)benzonitrile (Formula I) having the structure:

, or a pharmaceutically acceptable salt thereof, also relates to a method for reducing the intensity of pain on the surface of the eye in a subject in need thereof, comprising administering to the subject 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (formula I) having the structure: , or its pharmaceutically acceptable salt; wherein the compound of formula I is administered in a concentration of 0.5% w/v to 3.5% w/v, 0.5% w/v to 2.5% w/v, or 0.5% w/v to 1.5% w/v, from 0.5% to 3.0% w/v, from 1.0% to 2.5% w/v, from 1.5% to 3.0% w/v or from 0.5% to 2.5% w/v, also relates to a method for reducing the intensity of ocular hyperaemia in a subject in need thereof, comprising administering to the subject 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)benzonitrile (Formula I) having the structure: , or a pharmaceutically acceptable salt thereof; wherein the compound of formula I is administered at a concentration of 0.5% w/v to 3.5% w/v, 0.5% w/v to 2.5% w/v, or 0.5% w/v to 1.5% w/v, from 0.5% to 3.0% w/v, from 1.0% to 2.5% w/v, from 1.5% to 3.0% w/v, or from 0.5% to 2.5% w/v.

EFFECT: to provide treatment of pain on the surface of the eye in a subject, reduction of the intensity of pain on the surface of the eye in a subject, reduction of the intensity of eye hyperemia in a subject, ensures that compound I does not demonstrate delayed wound healing compared to the carrier medium and does not cause significant side effects.

15 cl, 14 dwg, 14 tbl, 2 ex

Description

Link to related applications

This application claims priority to U.S. Provisional Patent Application No. 62/806,682, which is incorporated herein in its entirety.

Field of technology to which the invention relates

The present invention relates to methods of treating diseases or disorders of the ocular surface using 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (compound I).

Background of the invention

The ocular surface, particularly the cornea, is richly innervated by sensory nerves. Corneal nerve activity can be modified by inflammation caused by a number of factors, including osmotic stress and tissue injury, as well as lesions of the ocular surface nerves. Ocular surface symptoms represent an "alarm system" indicating an imbalance in ocular surface homeostasis, leading to chronic ocular surface pain due to continuous exposure to stimuli that cause stress and hypersensitivity of the ocular surface.

Patients suffering from ocular surface pain, particularly chronic ocular surface pain, experience a significant reduction in quality of life. Utility studies to date have linked the burden of severe chronic ocular surface pain to moderate to severe angina, dialysis, and disabling hip fracture. Severe chronic ocular surface pain has also been associated with depression and suicidal ideation. In many patients, ocular surface pain remains unrelieved despite treatment of the underlying pathological condition (e.g., recent trauma or surgery, infection, or inflammation). Furthermore, treatments used for short-term ocular pain control (e.g., nonsteroidal anti-inflammatory drugs, steroids) are not suitable for long-term therapy. Thus, there is a pressing and unmet need for safe, effective treatments for ocular surface pain, particularly chronic ocular surface pain, when there are no other options to improve patients' quality of life or complement existing treatments.

Brief description of the invention

In one embodiment, the present invention provides a method of treating ocular surface pain in a subject in need thereof, comprising ocularly administering to the subject an effective amount of 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (formula I) having the structure:

Formula I,

or its salt, solvate, polymorph or cocrystal.

In some embodiments, the ocular surface pain is episodic or acute pain. In some embodiments, the ocular surface pain is chronic ocular surface pain (COSP), which lasts for at least about three months.

In some embodiments, the compound of Formula I is administered to the cornea of a subject. In some embodiments, the compound of Formula I is administered to the subject at a concentration of from about 0.5% w/v to about 3.5% w/v, from about 0.5% w/v to about 2.5% w/v, or from about 0.5% w/v to about 1.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, or from about 0.5% to about 2.5% w/v. In particular embodiments, the compound of Formula I is administered at a concentration of 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 compound of Formula I is administered to the subject at a concentration of about 5 mg/mL to about 35 mg/mL, about 5 mg/mL to about 25 mg/mL, or about 5 mg/mL to about 15 mg/mL, about 5 mg/mL to about 30 mg/mL, about 10 mg/mL to about 25 mg/mL, about 15 mg/mL to about 30 mg/mL, or about 5 mg/mL to about 25 mg/mL. In particular embodiments, the compound of Formula I is administered at a concentration of about 5 mg/mL, about 10 mg/mL, about 15 mg/mL, about 20 mg/mL, about 25 mg/mL, about 30 mg/mL, or about 35 mg/mL.

In some embodiments of the methods for treating ocular surface pain described herein, COSP is associated with dry eye disease. In some embodiments, administration results in a reduction in the severity of dry eye disease symptoms. In specific embodiments, administration results in a reduction in the severity of ocular pain associated with dry eye disease. In some embodiments, administration results in a reduction in the incidence of one or more of dry eye, ocular discomfort, ocular hyperemia, ocular burning or sharp pain, grittiness or foreign body sensation, or photophobia by at least about 10%.

In some embodiments of the methods for treating ocular surface pain described herein, the subject suffers from one or more of dry eye disease, Sjogren's syndrome, conjunctivitis (including keratoconjunctivitis, vernal keratoconjunctivitis, allergic conjunctivitis), punctate map-like "fingerprint" dystrophy, acanthamoebiasis, fibromyalgia, meibomian dysfunction, thyroid ophthalmopathy, acne rosacea, ptosis, keratoconus, ocular pain syndrome, Stevens-Johnson syndrome, forms of corneal epitheliopathy, forms of corneal neuropathy (including forms of LASIK-induced corneal neuropathy), forms of corneal dystrophy (including forms of recurrent corneal dystrophy), epithelial basement membrane dystrophy, forms of erosion corneal abrasions or abrasions (including recurrent forms of corneal abrasions or abrasions), ocular surface diseases, blepharitis, graft-versus-host disease, meibomitis, glaucoma, conjunctivochalasis, forms of keratopathy (including herpetic keratopathy, filiform keratopathy, band or bullous keratopathy, exposure keratopathy), keratitis (including keratitis due to herpes simplex virus), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, or refers to patients who are experiencing recovery from neurotrophic keratitis. In some embodiments of the methods described herein, the subject suffers from ocular pain that persists for at least three months following photorefractive keratectomy (PRK) surgery or laser in situ keratomileusis (LASIK) surgery.

In some embodiments, the methods for treating ocular surface pain described herein comprise administering to the subject an additional therapeutic agent.

In some embodiments of the methods for treating ocular surface pain described herein, administration of compound I to a subject results in a reduction in pain score 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 as measured by a visual analog scale (VAS). In further embodiments, administration of compound I to a subject results in a reduction in the subject's pain score 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 as measured by a VAS.

In some embodiments of the methods for treating ocular surface pain described herein, administration of compound I to a subject results in a reduction in pain intensity in the subject by at least about 10%, at least about 15%, at least about 20%, or at least about 25% compared to placebo. In some embodiments, the reduction in pain score results from the difference in pain scores before and after administration of compound I to the subject. In some embodiments of the methods described herein, upon administration of compound I to a subject, the reduction in pain score occurs within about half an hour after administration of compound I to the subject.

In some embodiments of the methods for treating ocular surface pain described herein, administration of compound I to a subject results in a decrease in the intensity of hyperemia in the 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 McMonney scale.

In some embodiments of the methods for treating ocular surface pain described herein, administration of compound I to a subject does not result in a change in one or more of best-corrected visual acuity, slit lamp biomicroscopy, dilated ophthalmoscopy, blink rate, tear production, or intraocular pressure compared to placebo.

In some embodiments of the methods for treating ocular surface pain described herein, the compound of Formula I is administered for at least about one, at least about two, or at least about three months. In some embodiments, the compound of Formula I is administered for more than 3 months. In some embodiments, the compound of Formula I is administered one to four times per day. In specific embodiments, the compound of Formula I is administered once or twice per day.

In some embodiments, the present invention relates to a compound of Formula I:

Formula I,

or to its salt, solvate, polymorph or cocrystal,

for use in the treatment or reduction of pain on the surface of the eye.

In some embodiments, the ocular surface pain is acute or episodic ocular surface pain. In some embodiments, the ocular surface pain is chronic ocular surface pain that has persisted for at least 1 month, at least 2 months, or at least 3 months.

In some embodiments of the routes of use described herein, the compound of Formula I is administered at a concentration of from about 0.5% w/v to about 3.5% w/v, from about 0.5% w/v to about 2.5% w/v, or from about 0.5% w/v to about 1.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, or from about 0.5% to about 2.5% w/v.

In some embodiments of the routes of use described herein, the compound of Formula I is administered at a concentration of 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 present invention relates to a medicament for treating or reducing the intensity of ocular surface pain, comprising an active ingredient that is a compound of formula I:

Formula I,

or a salt, solvate, polymorph, or cocrystal thereof.

In some embodiments, the ocular surface pain is acute or episodic ocular surface pain. In some embodiments, the ocular surface pain is chronic ocular surface pain that has lasted for at least 1 month, at least 2 months. In some embodiments, the ocular surface pain is chronic ocular surface pain that has lasted for at least 3 months. In some embodiments, the compound of Formula I is administered at a concentration of from about 0.5% w/v to about 3.5% w/v, from about 0.5% w/v to about 2.5% w/v, or from about 0.5% w/v to about 1.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, or from about 0.5% to about 2.5% w/v. In some embodiments, the compound of Formula I is administered at a concentration of 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 present invention relates to the use of a compound of formula I:

Formula I,

or a salt, solvate, polymorph or cocrystal thereof in the manufacture of a medicinal product for the treatment or reduction of pain on the surface of the eye.

In some embodiments, the use of Compound I is for acute or episodic ocular surface pain. In some embodiments, the use of Compound I is for chronic ocular surface pain that has lasted for at least 1 month, at least 2 months, or at least 3 months.

In some embodiments of the routes of use described herein, the compound of Formula I is administered at a concentration of from about 0.5% w/v to about 3.5% w/v, from about 0.5% w/v to about 2.5% w/v, or from about 0.5% w/v to about 1.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, or from about 0.5% to about 2.5% w/v.

In some embodiments of the routes of use described herein, the compound of Formula I is administered at a concentration of 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 present invention relates to a method for reducing the intensity of ocular surface pain in a subject in need thereof, comprising ocularly administering to the subject 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (formula I) having the structure:

Formula I,

or a salt, solvate, polymorph, or co-crystal thereof; wherein the compound of formula I is administered at a concentration of from about 0.5% w/v to about 3.5% w/v, from about 0.5% w/v to about 2.5% w/v, or from about 0.5% w/v to about 1.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, or from about 0.5% to about 2.5% w/v.

In some embodiments, the ocular surface pain is acute or episodic ocular surface pain. In some embodiments, the ocular surface pain is chronic ocular surface pain that has persisted for at least 1 month, at least 2 months, or at least 3 months.

In some embodiments of the methods for reducing ocular pain intensity described herein, COSP is associated with dry eye disease.

In some embodiments of the methods for reducing ocular surface pain intensity described herein, administration of compound I to a subject results in a reduction in the intensity of dry eye disease symptoms.

In some embodiments of the methods for reducing ocular surface pain described herein, administering Compound I to a subject results in a reduction in the intensity of ocular pain associated with dry eye disease. In some embodiments of the methods described herein, administering Compound I to a subject results in a reduction in the incidence of one or more of dry eye, ocular discomfort, ocular hyperemia, ocular burning or sharp pain, grittiness or foreign body sensation, or photophobia by at least about 10%.

In some embodiments, the subject in need of ocular surface pain relief suffers from one or more of dry eye disease, Sjogren's syndrome, conjunctivitis (including keratoconjunctivitis, vernal keratoconjunctivitis, allergic conjunctivitis), punctate map-like "fingerprint" dystrophy, acanthamoebiasis, fibromyalgia, meibomian dysfunction, thyroid ophthalmopathy, acne rosacea, ptosis, keratoconus, ocular pain syndrome, Stevens-Johnson syndrome, forms of corneal epitheliopathy, forms of corneal neuropathy (including forms of LASIK-induced corneal neuropathy), forms of corneal dystrophy (including forms of recurrent corneal dystrophy), epithelial basement membrane dystrophy, forms of erosion corneal abrasions or abrasions (including recurrent forms of corneal abrasions or abrasions), ocular surface diseases, blepharitis, graft-versus-host disease, meibomitis, glaucoma, conjunctivochalasis, forms of keratopathy (including herpetic keratopathy, filiform keratopathy, band or bullous keratopathy, exposure keratopathy), keratitis (including keratitis due to herpes simplex virus), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, or refers to patients who are experiencing recovery from neurotrophic keratitis.

In some embodiments, the methods for reducing ocular surface pain intensity described herein comprise administering to the subject an additional therapeutic agent.

In some embodiments of the methods for reducing ocular surface pain intensity described herein, administering compound I to a subject results in a decrease in the subject's pain score by at least about 3 compared to the pain score before administering the compound, as measured by a visual analog scale (VAS). In some embodiments of the methods described herein, administering compound I to a subject results in a decrease in the subject's pain score 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 a placebo, as measured by a visual analog scale (VAS). In some embodiments of the methods described herein, administering compound I to a subject results in a decrease in the subject's pain score by 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.

In some embodiments of the methods for reducing ocular surface pain intensity described herein, administration of compound I to a subject results in a reduction in pain intensity in the subject by at least about 10%, at least about 15%, at least about 20%, or at least about 25% compared to placebo. In some embodiments of the methods described herein, the reduction in pain score results from the difference in pain scores before and after administration of compound I to the subject.

In some embodiments of the methods for reducing ocular surface pain intensity described herein, administering compound I to a subject results in a decrease in hyperemia intensity in the 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 McMonney scale.

In some embodiments, the present invention relates to a method for reducing the intensity of ocular hyperemia in a subject in need thereof, comprising ocularly administering to the subject 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (formula I) having the structure:

Formula I,

or a salt, solvate, polymorph, or co-crystal thereof; wherein the compound of formula I is administered at a concentration of from about 0.5% w/v to about 3.5% w/v, from about 0.5% w/v to about 2.5% w/v, or from about 0.5% w/v to about 1.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, or from about 0.5% to about 2.5% w/v.

In some embodiments of the methods described herein, administration of compound I to a subject results in a reduction in the intensity of ocular hyperemia by 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 of the methods described herein, administration of compound I to a subject results in a reduction in the incidence of one or more of dry eye, eye discomfort, burning or sharp pain in the eye, grittiness or foreign body sensation, or photophobia by at least about 10%.

In some embodiments of the methods for reducing the intensity of ocular hyperemia, the subject suffers from one or more of dry eye disease, Sjogren's syndrome, conjunctivitis (including keratoconjunctivitis, vernal keratoconjunctivitis, allergic conjunctivitis), punctate map-like dystrophy of the "fingerprint" type, acanthamoebiasis, fibromyalgia, meibomian gland dysfunction, thyroid ophthalmopathy, acne rosacea, ptosis, keratoconus, ocular pain syndrome, Stevens-Johnson syndrome, forms of corneal epitheliopathy, forms of corneal neuropathy (including forms of LASIK-induced corneal neuropathy), forms of corneal dystrophy (including forms of recurrent corneal dystrophy), epithelial basement membrane dystrophy, forms of corneal erosions or scratches corneal lesions (including recurrent corneal erosions or corneal abrasions), ocular surface diseases, blepharitis, graft-versus-host disease, meibomitis, glaucoma, conjunctivochalasis, keratopathy forms (including herpetic keratopathy, filiform keratopathy, band or bullous keratopathy, exposure keratopathy), keratitis (including herpes simplex keratitis), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, or refers to patients who are recovering from neurotrophic keratitis.

Some embodiments of the methods for reducing the intensity of ocular hyperemia described herein further comprise administering to the subject an additional therapeutic agent.

Specific preferred embodiments of the present invention will become apparent from the following more detailed description of certain preferred embodiments and the claims.

Brief description of graphic materials

Figure 1 shows the model-estimated mean (+/− SE) for visual analogue scale (VAS) pain scores over time (primary PD analysis set).

Figure 2 shows a histogram of the incidence of oral rescue medication use (the number of patients who did not use the oral rescue medication) (the secondary PD analysis sample). For each time period, the right column represents subjects who received Compound I, and the left column represents subjects who received vehicle. The percentage of patients who did not use the oral rescue medication (ORM) is presented on the y-axis. n/N represents the number/total number of patients for each treatment.

Figure 3 presents the mean (+/− SD) for questions 4, 5, and 6 of the Ocular Pain Assessment Questionnaire (OPAS) over time (secondary PD analysis sample). Figure 3A presents the results for question 4: worst eye pain at 24 h (pain level), Figure 3B presents the results for question 5: least severe eye pain at 24 h (pain level), and Figure 3C presents the results for question 6: average eye pain score at 24 h (pain level). In each of Figures 3A, 3B, and 3C, the dashed line with "×" represents the vehicle, and the solid line with circles (o) represents compound I.

Figure 4 presents the mean (+/− SD) for questions 22, 23, 24, and 25 of the Ocular Pain Assessment Survey (OPAS) over time (secondary PD analysis sample). Figure 4A presents the results for question 22: incidence of eye pain with redness (%). Figure 4B presents the results for question 23: incidence of eye pain with burning (%). Figure 4C presents the results for question 24: incidence of eye pain with sensitivity (%). Figure 4D presents the results for question 25: incidence of eye pain with tearing (%). In each of Figures 4A, 4B, 4C, and 4D, the dotted line with "×" represents the vehicle, and the solid line with circles (o) represents compound I.

Figure 5A shows the mean (SD) arithmetic mean (SD) for Compound I plasma concentrations following topical ocular administration of 2.5% Compound I (PK assay sample) on day 1. Figure 5B shows the mean (SD) for Compound I plasma concentrations following topical ocular administration of 2.5% Compound I (PK assay sample) on day 4.

Figure 6A shows a bar graph of the intensity of ocular hyperemia over time for compound I, and Figure 6B shows a bar graph of the intensity of ocular hyperemia over time for the vehicle. The quadrants are as follows: S: superior, N: nasal, I: inferior, and T: temporal.

Figure 7 shows the mean (+/− SD) for epithelial defect size over time (safety analysis set). Parameter (units): epithelial wound size (mm), subcategory: area (mm ² ). The dotted line with "×" represents the vehicle, and the solid line with circles (o) represents compound I.

Detailed description

"TRPV1 receptor" refers to the vanilloid receptor with transient receptor potential type 1, the characteristics of which have been determined by molecular cloning and pharmacological analysis. See, e.g., Caterina MJ, et al., Nature 1997; 389:816-824. TRPV1 receptor activity was measured as described in WO 2005/120510, which is incorporated herein by reference in its entirety.

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

The phrase "ophthalmologically compatible" refers to formulations, polymers and other materials and/or dosage forms that are suitable for use in contact with ocular tissues in humans and animals without undue toxicity, irritation, allergic reaction or other problems or complications consistent with an acceptable benefit-risk ratio.

As used herein, the terms "treat," "treat," or "treatment" with respect to a disease or disorder refer in some embodiments to reducing the intensity of the manifestations of the disease or disorder (i.e., slowing, or stopping, or attenuating the progression of a disease or at least one of its clinical symptoms). In another embodiment, "treat," "treat," or "treating" refers to alleviating or reducing the intensity of at least one physical parameter, including those that may not be perceived by the patient. In yet another embodiment, "treat," "treating," or "treating" refers to modulating a disease or disorder by physical means (e.g., by stabilizing the perceptible symptom), physiologically (for example, by stabilizing a physical parameter), or by both. In another embodiment, "treating," "treating," or "treating" refers to preventing or delaying the onset, development, or progression of a disease or disorder or a symptom thereof.

As used herein, the terms "subject" or "patient" refer to humans and non-human mammals, including, but not limited to, primates, rabbits, pigs, horses, dogs, cats, sheep, and cows. In certain embodiments, the subject or patient is a human. In some embodiments, the term "patient" or "subject" refers to a human being who is afflicted with a condition (i.e., A subject who has a disease or disorder described herein and who would benefit from treatment. As used herein, a subject "in need of" treatment if the subject (patient) would benefit from such treatment from a biological, medical, or quality-of-life perspective. In specific embodiments, the subject is an adult human being at least about 18 years of age. In specific embodiments, the subject is an adult human being between about 18 and about 75 years of age. In specific embodiments, the subject is a human child under about 18 years of age.

As used in this document, "ocular surface" refers to the outer surface of the eye, which anatomically includes the cornea (with epithelium, Bowman's layer, stroma, Descemet's membrane, endothelium), conjunctiva, conjunctival sac, and corneoscleral junction, i.e., the corneal limbus.

As used herein, ocular administration includes administration to all parts of the eye, including all parts of the ocular surface such as the cornea, conjunctiva, conjunctival sac, and corneoscleral junction, i.e., the corneal limbus.

As used in this document, "pain" refers to a constant or intermittent sensation of actual pain, described, but not limited to, stabbing, aching, sharp, or dull. Pain may also refer to similar related descriptors, such as, but not limited to, burning, stabbing, gritty, foreign body sensation, dryness, grainy sensation, fatigue, itching, irritation, and sensitivity to light.

As used in this document, "ocular surface pain" refers to pain localized to the surface of the eye, such as the cornea. Ocular pain can be nociceptive pain, which is typically caused by external physical or chemical injurious stimuli, such as corneal surgery, inflammation, or other damage to the corneal surface. Eye pain can also be caused by neuropathic pain, which can result from direct damage to neurons in the body, resulting in pain messages being transmitted to the central nervous system and brain regardless of the presence of painful stimuli. As used in this document, "ocular surface pain" includes both nociceptive pain and neuropathic pain.

As used herein, the term "visual analogue scale" (VAS) is a measure of pain intensity in which the subject typically marks a spot on the scale that corresponds to their level of pain. Pain is rated on a range from "no pain" (score of 0) to "the worst possible pain" or "the worst pain imaginable" (score of 100). See, for example, Hawker et al., Arthritis Care & Research 63(11), pp. S240-S252 (November 2011). Several other well-established pain rating scales exist that can be used to assist in assessing pain severity. A commonly used numeric rating scale (NRS), in which subjects use numbers to rate pain, is often used. The numeric scale can range from 1-10 or 1-100. The Wong-Baker Face Pain Scale combines pictures and numbers to rate pain. It can be used with children aged 3 years and older and with adults. Six faces depict different facial expressions, ranging from happy to extremely upset. Each is assigned a numerical rating from 0 (smiling) to 10 (crying). The Verbal Pain Intensity Scale uses verbal statements on a scale to rate the intensity of pain: no pain/mild pain/moderate pain/severe pain/very severe pain/worst possible pain.

The Ocular Sensation Scale is a specialized pain rating scale designed to measure the severity of ophthalmic pain. See Caudle L.E.et al.,Optom Vis Sci. 2007 Aug; 84(8):752-62. This scale typically measures pain, discomfort, or sensitivity to light using 5 categories of "extreme," "severe," "moderate," "mild," or "none."

The Ocular Pain Assessment Questionnaire (OPAS) is a quantitative, multidimensional questionnaire specifically designed to assess corneal and ocular surface pain and changes in quality of life (QoL). The OPAS assesses pain intensity, frequency of ocular and non-ocular pain, changes in QoL, aggravating factors, associated factors, and quantitative symptom relief, enabling the monitoring of treatment responses. See also Qazi et al.,Ophthalmology July 123(7):1458-1468 (2016).

The term "visual task performance questionnaire" used in this document refers to a questionnaire in which subjects are asked to subjectively rate the amount of difficulty they experience performing certain activities that require staring or looking at something for long periods of time, which may exacerbate eye pain. This questionnaire also asks about coping mechanisms associated with the difficulties they experience during visual task performance.

As used in this document, ocular hyperemia refers to redness of the ocular surface. Ocular hyperemia can be a clinical marker of ocular inflammation and/or irritation. Ocular hyperemia can be measured using the McMonney scale, which ranges from 0 to 5 and is based on standardized photographs.

As used in this document, "placebo" refers to an ophthalmic formulation that contains all the components of the administered drug-based composition, without the drug itself.

The term "approximately" as used in this document refers to a range of values +10% of the stated value.

As used herein, a pharmaceutical composition is a composition suitable for pharmaceutical use. A composition suitable for pharmaceutical use may be sterile, homogeneous, and/or isotonic. Pharmaceutical compositions may be prepared in certain embodiments in aqueous form, for example, in a pre-filled syringe or other single- or multi-dose container. In certain embodiments of the present invention, the pharmaceutical composition is ophthalmologically compatible and suitable for ophthalmic administration to a human subject, for example, via topical or other known delivery methods.

As used herein, "compound of formula I", "compound I", "formula I" and "compound I" are used interchangeably and mean a compound that has the name 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile with the structure shown below and can be synthesized using procedures known in the art and described in WO 2005/120510 and in U.S. Patent No. 8,349,852 ("Quinazolinone derivatives useful as vanilloid antagonists") to Chen et al. , both of which are incorporated herein by reference in their entireties.

Various crystalline and polymorphic forms of compound I can be used as a supplement or alternative.

In some embodiments, compound I is in polymorph form B as described in U.S. Patent No. 8,349,852, incorporated herein by reference. In some embodiments, polymorph B is characterized by an X-ray diffraction pattern that has three or more peaks at 2θ values selected from 9.3, 10.6, and 14.4 ± 0.2°2θ. In some embodiments, polymorph B is characterized by an X-ray diffraction pattern that has three or more peaks at 2θ values selected from 9.3, 10.6, 14.4, 15.5, 17.9, 19.9, 23.4 ± 0.2°2θ.

Any chemical formula given in this document is also intended to represent unlabeled forms, as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have structures depicted by the formulas given in this document, except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Isotopes that may be included in the compounds of the present invention include, for example, isotopes of hydrogen, carbon, nitrogen, and oxygen, such as ³ H, ¹¹ C, ¹³ C, ¹⁴ C, and ¹⁵ N. Therefore, it should be understood that the methods of the present invention may encompass compounds that include one or more of any of the aforementioned isotopes, including, for example, radioactive isotopes such as ³ H and ¹⁴ C, or compounds in which non-radioactive isotopes such as ² H and ¹³ C are present. Such isotopically labeled compounds are useful in metabolism studies (using ¹⁴ C), reaction kinetics studies (using, for example, ² H or ³ H), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including in drug or substrate tissue distribution assays, or in radioisotope treatment of patients. Isotopically labeled compounds can generally be prepared using conventional techniques known to those skilled in the art, for example, by using suitable isotopically labeled reagents in place of the unlabeled reagent previously used.

The present invention encompasses embodiments that include all pharmaceutically acceptable salts of the compounds useful in the present invention as provided herein. As used herein, "pharmaceutically acceptable salt" refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acidic or basic moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, salts of mineral or organic acids and 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 a parent compound that contains a basic or acidic moiety using conventional chemical methods. Generally, such salts can be prepared by reacting the compounds in their free acid or base forms with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of both; In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences , ^17th 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, salts of mineral or organic acids and basic residues such as amines. For example, the salt can be a hydrochloride salt. Other examples of suitable salts can be found in U.S. Patent No. 8,349,852, the contents of which are incorporated herein by reference in their entirety.

As used herein, the phrase "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic reaction or other problems or complications, consistent with an acceptable balance of benefit and risk.

Compound (I) or its pharmaceutically acceptable salt can be incorporated into various delivery formulations. For example, topical formulations can be used, and they can contain ophthalmologically acceptable preservatives, surfactants, viscosity enhancers, buffers, sodium chloride, and water to form aqueous ophthalmically compatible solutions and suspensions.

The compound of Formula I will typically be present in these formulations in an amount of from about 0.1% to about 5.0% w/v. In some embodiments, the concentrations of Compound I for topical administration range from about 0.5% to about 1.5% w/v, from about 0.5% to about 2.5% w/v, from about 0.5% to about 3.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, from about 0.5% to about 2.5% w/v. In some embodiments, the concentration of the compound of Formula I in the topical formulation is at least about 0.5% w/v, at least about 1.0% w/v, at least about 1.5% w/v, at least about 2.0% w/v, or at least about 2.5% w/v. In some embodiments, the concentration of the compound of Formula I in the topical formulation is no more than about 5.0% w/v, no more than about 4.5% w/v, no more than about 4.0% w/v, no more than about 3.5% w/v, or no more than about 3.0% w/v. In particular embodiments, the concentration of the compound of Formula I in the topical formulation 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. It is generally understood that equivalent concentrations can be expressed in different units. For example, a concentration of 0.1% w/v can also be expressed as a 1 mg/mL solution. When expressed in mg/mL units, in some embodiments, the compound of Formula I is administered to a subject at a concentration of from about 5 mg/mL to about 35 mg/mL, from about 5 mg/mL to about 25 mg/mL, or from about 5 mg/mL to about 15 mg/mL, from about 5 mg/mL to about 30 mg/mL, from about 10 mg/mL to about 25 mg/mL, from about 15 mg/mL to about 30 mg/mL, or from about 5 mg/mL to about 25 mg/mL. In some embodiments, the concentration of the compound of Formula I in the topical formulation is at least about 5 mg/mL, at least about 10 mg/mL, at least about 15 mg/mL, at least about 20 mg/mL, or at least about 25 mg/mL. In some embodiments, the concentration of the compound of Formula I in the topical formulation is no more than about 50 mg/mL, no more than about 45 mg/mL, no more than about 40 mg/mL, no more than about 35 mg/mL, or no more than about 30 mg/mL. In particular embodiments, the compound of Formula I is administered at a concentration of about 5 mg/mL, about 10 mg/mL, about 15 mg/mL, about 20 mg/mL, about 25 mg/mL, about 30 mg/mL, or about 35 mg/mL.

In some embodiments, the formulations are delivered to the ocular surface one to six times per day, depending on the usual discretion of the qualified treating physician. In some embodiments, the formulations are administered once, twice, three, or four times per day.

Unless otherwise stated, the mass or dose referred to herein for a compound of Formula I represents the mass or dose of the compound itself, not its salt or prodrug, which may vary to achieve the intended therapeutic effect. For example, the mass or dose of a corresponding salt of the compound suitable for the methods, compositions, or combinations disclosed herein can be calculated based on the ratio of the molecular masses of the salt to the compound itself.

Compound (I) and/or its pharmaceutically acceptable salts can be incorporated into ophthalmologically compatible formulations for delivery to the eye. The compounds can be combined with ophthalmologically acceptable preservatives, surfactants, viscosity enhancers, permeation enhancers, buffers, sodium chloride, and water to form aqueous sterile ophthalmic suspensions or solutions.

The pharmaceutical compositions of the present invention may comprise an additional therapeutic agent in addition to compound (I). Additional therapeutic agents may include, for example, other compounds and antibodies useful for the treatment of ocular surface disorders. A non-limiting list of such agents includes non-steroidal anti-inflammatory drugs such as ketorolac, nepafenac, bromfenac, corticosteroids; drugs for the treatment of dry eye disease such as cyclosporine, lifitegrast, autologous serum, or other TRPV1 inhibitors. In specific embodiments, the additional therapeutic agent is an ophthalmic steroid such as dexamethasone, fluocinolone, loteprednol, difluprednate, fluorometholone, prednisolone, prednisone, medrysone, triamcinolone, betamethasone, rimexolone, or pharmaceutically acceptable salts thereof. Additional non-limiting examples of such additional therapeutic agents that may be included in the pharmaceutical composition include Xiidra® (lifitegrast), Restasis® (cyclosporine), minocycline, doxycycline, or other tetracycline antibiotics. Other examples include keratolytic agents such as selenium disulfide, salicylic acid, glycolic acid, and the like, or pharmaceutically acceptable salts thereof.

Without being limited by theory, it is hypothesized that transient receptor potential vanilloid receptor type 1 (TRPV1) blockers may be useful in the treatment of pain, such as chronic pain.

Accordingly, in some embodiments, the present invention provides a method of treating ocular surface pain in a subject in need thereof, said method comprising administering to the subject an effective amount of Compound (I) or a pharmaceutically acceptable salt, solvate, or co-crystal thereof. In some embodiments, the present invention provides a method of reducing ocular surface pain in a subject in need thereof, said method comprising administering to the subject an effective amount of Compound (I) or a pharmaceutically acceptable salt, solvate, or co-crystal thereof. 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 the treatment or reduction of ocular surface pain. In some embodiments, the compound of Formula I is in polymorphic form B.

In some embodiments, the ocular surface pain is episodic, i.e., acute.

In some embodiments, the subject suffers from chronic ocular surface pain that has lasted for at least three months. In some embodiments, the subject suffers from chronic ocular surface pain that has lasted for at least two months. In some embodiments, the subject suffers from chronic ocular surface pain that has lasted for at least one month. In some embodiments, the subject suffers from chronic ocular surface pain that has lasted for at least four months. In some embodiments, the subject suffers 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 salt, solvate, polymorph, or co-crystal thereof. In some embodiments, the present invention provides a method of reducing the intensity of chronic ocular surface pain in a subject by administering to the subject an effective amount of a compound of Formula I or a salt, solvate, polymorph, or co-crystal thereof. The present invention provides the use of a compound of formula I or a pharmaceutically acceptable salt, solvate, polymorph or co-crystal thereof in the treatment of chronic ocular surface pain.

In some embodiments, administration is made to the surface of the eye of the subject, such as any part of the cornea, conjunctiva, or conjunctival sac of the eye.

In some embodiments, the present invention provides for administering a compound of Formula I to a subject in need thereof in an ophthalmologically compatible formulation at a concentration of from about 0.5% w/v to about 3.5% w/v. In some embodiments, the concentrations for administration range from about 0.5% to about 3.5% w/v, from about 0.5% to about 2.5% w/v, from about 0.5% to about 1.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, from about 0.5% to about 2.5% w/v. In particular embodiments, the concentration of the compound of Formula I in the topical formulation 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 compound of Formula I is administered to a subject at a concentration of from about 5 mg/mL to about 35 mg/mL, from about 5 mg/mL to about 25 mg/mL, or from about 5 mg/mL to about 15 mg/mL, from about 5 mg/mL to about 30 mg/mL, from about 10 mg/mL to about 25 mg/mL, from about 15 mg/mL to about 30 mg/mL, or from about 5 mg/mL to about 25 mg/mL. In particular embodiments, the compound of Formula I is administered at a concentration of about 5 mg/mL, about 10 mg/mL, about 15 mg/mL, about 20 mg/mL, about 25 mg/mL, about 30 mg/mL, or about 35 mg/mL. In some embodiments, the dose per administration to each eye is from about 0.15 to about 1.15 mg, or about 0.15 mg, 0.2 mg, about 0.25 mg, about 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 administration to each eye 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 for each eye is from 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 one to six times per day, such as one, two, three, or four times per day. In some embodiments, the compound of Formula I is administered to the subject for a period of at least about one month, at least about two months, or at least about three months. In some embodiments, the compound of Formula I is administered to the subject for a period of at least about 12 weeks.

In some embodiments, the ocular surface pain or chronic ocular surface pain is associated with one or more of dry eye disease, Sjogren's syndrome, conjunctivitis (including keratoconjunctivitis, vernal keratoconjunctivitis, allergic conjunctivitis), punctate map-like "fingerprint" dystrophy, acanthamoebiasis, fibromyalgia, meibomian dysfunction, thyroid ophthalmopathy, acne rosacea, ptosis, keratoconus, ocular pain syndrome, Stevens-Johnson syndrome, forms of corneal epitheliopathy, forms of corneal neuropathy (including forms of LASIK-induced corneal neuropathy), forms of corneal dystrophy (including forms of recurrent corneal dystrophy), epithelial basement membrane dystrophy, forms of erosion corneal abrasions or abrasions (including recurrent forms of corneal abrasions or abrasions), ocular surface diseases, blepharitis, graft-versus-host disease, meibomitis, glaucoma, conjunctivochalasis, forms of keratopathy (including herpetic keratopathy, filiform keratopathy, band or bullous keratopathy, exposure keratopathy), keratitis (including herpes simplex keratitis), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, or in patients recovering from neurotrophic keratitis.

In specific embodiments, ocular surface pain or chronic ocular surface pain is associated with dry eye disease or Sjogren's syndrome. In some embodiments of the methods described herein, the subject suffers from ocular pain that persists for at least three months following photorefractive keratectomy (PRK) surgery or laser in situ keratomileusis (LASIK) surgery.

In some embodiments, the subject suffers from conjunctivitis, subconjunctival hemorrhage, subconjunctival scarring, conjunctival membranes, conjunctival ulceration, superficial punctate epithelial erosions, epithelial defects, eyelid margin ulcerations, eyelid margin keratinization, symblepharon, ankyloblepharon, trichiasis, anterior blepharitis, punctal autoocclusion, meibomian gland disease, corneal opacity, dry eye, districhiasis, limbal stem cell deficiency, or corneal vascularization.

In some embodiments, administration of a compound of Formula I results in a reduction in the intensity of ocular pain in the subject, compared to placebo. In some embodiments, the reduction in the intensity of ocular pain in the subject is at least about 3, as measured by a VAS, compared to placebo. In some embodiments, administration results in a reduction in the intensity of ocular pain in the subject 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, as measured by a VAS, compared to placebo. In some embodiments, administration results in a reduction in the intensity of pain in the subject by at least about 10%, at least about 15%, at least about 20%, or at least about 25%, compared to placebo. In some embodiments, the administration results in a reduction in pain intensity in the subject as measured 7 days after administration of the compound of Formula I. In some embodiments, the administration results in a reduction in pain intensity in the subject as measured 14 days after administration of the compound of Formula I.

In some embodiments, administration of a compound of Formula I results in a reduction in pain intensity in a subject by at least about 2% compared to placebo as measured by a VAS score at about half an hour after administration, about one hour, about 2 hours, or about 2-4 hours after administration.

In some embodiments, the reduction in pain score results from the difference in pain scores before and after administration of compound I to the subject. In some embodiments, the reduction in pain score, as measured by a VAS, results from the difference in pain scores before and after administration of compound I to the subject. In some embodiments, the reduction in pain score occurs within about half an hour after administration of 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 administration of compound I to the subject. In other embodiments, the reduction in pain score occurs about 7 days after administration of compound I to the subject. In some embodiments, the reduction in pain score occurs about 14 days after administration of compound I to the subject.

In some embodiments, administration of a compound of Formula I results in a reduction in the intensity of ocular hyperemia (eye redness) compared to placebo. In specific embodiments, administration of a compound of Formula I results in a reduction in the intensity of grade 1, grade 2, grade 3, or grade 4 hyperemia compared to placebo. In some embodiments, administration results in a reduction in the ocular hyperemia score by 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, administration of a compound of Formula I results in an improvement in the score on at least one question of the OPAS by at least about 10%, at least about 20%, or at least about 30%.

In some embodiments, administration of a compound of Formula I results in an improvement in the score on at least one question of a visual task performance questionnaire by at least about 10%, at least about 20%, or at least about 30%.

Thus, in some embodiments, the present invention provides a method of treating ocular hyperemia in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula I or a salt, solvate, polymorph, or co-crystal thereof. In some embodiments, the present invention provides a method of reducing the intensity of ocular hyperemia in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula I or a salt, solvate, polymorph, or co-crystal thereof. In some embodiments, administration of Compound I results in a reduction in the intensity of ocular hyperemia by at least 1, at least 2, at least 3, at least 4, or at least 5 on the McMonney scale. 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 for the treatment of ocular hyperemia. In some embodiments, the present invention provides the administration of a compound of Formula I to a subject in need thereof in an ophthalmologically compatible formulation at a concentration of from about 0.5% w/v. to about 3.5% w/v. In some embodiments, the concentrations for administration range from about 0.5% to about 3.5% w/v, from about 0.5% to about 2.5% w/v, from about 0.5% to about 1.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, from about 0.5% to about 2.5% w/v. In particular embodiments, the concentration of the compound of Formula I in the topical formulation 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. It should be understood that the percentages may be expressed in units of mg/mL. In some embodiments, the dose per administration to each eye is from 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 administration to each eye 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 for each eye is from 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 one to six times per day, such as one, two, three, or four times per day. In some embodiments, the compound of Formula I is administered to the subject for a period of at least about one month, at least about two months, or at least about three months.

In some embodiments, the ocular hyperemia is associated with one or more of dry eye disease, Sjogren's syndrome, conjunctivitis (including keratoconjunctivitis, vernal keratoconjunctivitis, allergic conjunctivitis), punctate map-like dystrophy of the "fingerprint" type, acanthamoebiasis, fibromyalgia, meibomian gland dysfunction, thyroid ophthalmopathy, acne rosacea, ptosis, keratoconus, eye pain syndrome, Stevens-Johnson syndrome, forms of corneal epitheliopathy, forms of corneal neuropathy (including forms of LASIK-induced corneal neuropathy), forms of corneal dystrophy (including forms of recurrent corneal dystrophy), epithelial basement membrane dystrophy, forms of corneal erosions or scratches cornea (including recurrent forms of corneal erosions or corneal abrasions), ocular surface diseases, blepharitis, graft-versus-host disease, meibomitis, glaucoma, conjunctivochalasis, forms of keratopathy (including herpetic keratopathy, filiform keratopathy, band or bullous keratopathy, exposure keratopathy), keratitis (including keratitis caused by the herpes simplex virus), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, or is present in patients who are recovering from neurotrophic keratitis. In specific embodiments, the ocular hyperemia is associated with dry eye disease. In some embodiments of the methods described herein, ocular hyperemia persists for at least three months following photorefractive keratectomy (PRK) surgery or laser in situ keratomileusis (LASIK) surgery.

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 results in a reduction in the severity of dry eye disease symptoms. Dry eye disease is generally understood as a complex, multifactorial condition characterized by inflammation of the ocular surface and lacrimal glands, as well as a decrease in the quality and/or quantity of tears. It is believed that up to 30% of patients with dry eye disease suffer from ocular surface pain, which can be chronic, i.e., lasting at least 12 weeks or three months. Thus, in some embodiments, the present invention results in a reduction in the intensity of dry eye disease symptoms, including one or more of dry eyes, eye discomfort, eye hyperemia, burning or sharp pain in the eyes, a gritty or foreign body sensation, or photophobia, by at least about 10%, at least about 15%, at least about 20%, or at least about 30%.

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 salt, solvate, polymorph, or co-crystal 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 salt, solvate, polymorph, or co-crystal thereof, wherein the compound of Formula I is safe for administration for a period of at least 2 months, at least 3 months, at least 4 months, or at least 5 months. In particular embodiments, the present invention provides the use of a compound of Formula I or a pharmaceutically acceptable salt, solvate, or co-crystal thereof in the treatment of dry eye disease. In some embodiments, the present invention results in a reduction in the intensity of dry eye disease symptoms, including one or more of dry eyes, eye discomfort, eye hyperemia, eye burning or stinging, grittiness or foreign body sensation, or photophobia, by at least about 10%. In some embodiments, the present invention provides administration of a compound of Formula I to a subject in need thereof in an ophthalmologically compatible formulation at a concentration of from about 0.5% w/v to about 3.5% w/v. In some embodiments, the concentrations for administration range from about 0.5% to about 3.5% w/v, from about 0.5% to about 2.5% w/v, from about 0.5% to about 1.5% w/v, from about 0.5% to about 3.0% w/v, from about 1.0% to about 2.5% w/v, from about 1.5% to about 3.0% w/v, from about 0.5% to about 2.5% w/v. In particular embodiments, the concentration of the compound of Formula I in the topical formulation 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 administration to each eye is from 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 administration to each eye 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 for each eye is from 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 one to six times per day, such as one, two, three, or four times per day. In some embodiments, the compound of Formula I is administered to the subject for a period of at least about one month, at least about two months, or at least about three months.

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 biomicroscopy, dilated ophthalmoscopy, blink rate, tear production, or intraocular pressure compared to 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 compared to placebo in a patient in need thereof.

Patient population

In specific embodiments, the subject to be treated using 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 keratoconjunctivitis, vernal keratoconjunctivitis, allergic conjunctivitis), punctate map-fingerprint dystrophy, acanthamoebiasis, fibromyalgia, meibomian gland dysfunction, thyroid ophthalmopathy, acne rosacea, ptosis, keratoconus, eye pain syndrome, Stevens-Johnson syndrome, forms of corneal epitheliopathy, forms of corneal neuropathy (including forms of LASIK-induced corneal neuropathy), forms of corneal dystrophy (including forms of recurrent corneal dystrophy), epithelial basement membrane dystrophy, forms of corneal abrasions or scratches corneal abrasions (including recurrent corneal erosions or corneal abrasions), ocular surface diseases, blepharitis, graft-versus-host disease, meibomitis, glaucoma, conjunctivochalasis, keratopathy forms (including herpetic keratopathy, filiform keratopathy, band or bullous keratopathy, exposure keratopathy), keratitis (including herpes simplex keratitis), iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, or in patients recovering from neurotrophic keratitis. In some embodiments, the subject suffers from ocular pain that persists for at least three months following photorefractive keratectomy (PRK) surgery or laser in situ keratomileusis (LASIK) surgery.

In certain embodiments, provided herein are methods for treating or reducing the intensity of ocular surface pain, such as acute ocular surface pain.

In certain embodiments, provided herein are methods for treating or reducing the intensity of ocular surface pain, such as chronic ocular surface pain (COSP). In particular aspects, COSP is characterized as persistent ocular surface pain (e.g., persistent severe ocular surface pain) that can distract from or interfere with normal daily activities. In particular aspects, COSP can lead to poor quality of life and can 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 can persist for at least about 2 months or at least about 3 months. In other aspects, COSP can persist for at least 3 months or at least 4 months. In specific aspects, a subject with COSP continues to have symptoms despite adherence to other therapies indicated for their underlying condition (e.g., an ocular surface disorder such as dry eye disease or Sjogren's syndrome).

In some embodiments, the subject to be treated suffers from ocular neuropathic pain (ONP). ONP is a spectrum of disorders characterized by eye pain that can be caused by injury or disease affecting nerves, such as the corneal nerves. Symptoms of ONP may include one or more of ocular pain, sensitivity to light, hyperalgesia, or dysesthesia (abnormal sensations), such as a sensation of dryness, sharp pain, or a foreign body sensation, and pain due to exposure to normally non-painful stimuli (allodynia). Gabapentin and other medications for neuropathic pain can be used to blunt stimulation of sensory nerves or the perception of nerve stimulation.

In some embodiments, the subject to be treated suffers from exposure keratopathy. EK is a corneal injury that occurs primarily as a result of prolonged exposure of the ocular surface to the external environment. EK can lead to ulceration, microbial keratitis, and permanent vision loss due to scarring. Patients at risk for developing EK include those with conditions that interfere with the ability to protect the cornea; due to incomplete eyelid closure (e.g., lagophthalmos, proptosis, ectopic eyelid), impaired blink reflex, inappropriate blink rate (e.g., caused by a neurological disease, such as Parkinson's disease, neuromuscular disease), and/or decreased amount of protective corneal lubrication. Symptoms of EK include foreign body sensation, burning, increased lacrimation, intermittent blurred vision (due to tear film instability), pain, and photophobia. Standard treatments include frequent use of artificial tears, nightly application of lubricating ointment, and punctal plugs.

In some embodiments, the subject to be treated suffers from keratoconjunctivitis. Keratoconjunctivitis is an inflammatory process involving both the conjunctiva and cornea. Superficial corneal inflammation (keratitis) typically occurs in combination with viral and bacterial conjunctivitis, for example, in adults. The following types of keratoconjunctivitis are differentiated based on the potential cause of the inflammation:

Keratoconjunctivitis sicca is caused by inflammation due to dryness;

Vernal keratoconjunctivitis (VKC) occurs seasonally and is thought to be caused by allergens;

Atopic keratoconjunctivitis is one of the manifestations of atopy;

Epidemic keratoconjunctivitis or adenoviral keratoconjunctivitis is caused by adenovirus infection;

Infectious bovine keratoconjunctivitis (IBK) is a disease affecting cattle caused by the bacterium Moraxella bovis;

Acute epidemic conjunctivitis in sheep and goats is mainly caused by Chlamydophila pecorum;

Superior limbal keratoconjunctivitis is thought to be caused by mechanical trauma;

Photoelectric keratoconjunctivitis (electrophthalmia) refers to inflammation caused by photoelectric UV radiation.

In some embodiments, the subject being treated suffers from dry eye. The term "dry eye," as used herein, refers to insufficient tear production and/or abnormal tear composition. Destructive dry eye syndrome (DEDS), also known as dry eye syndrome, keratoconjunctivitis sicca, keratitis sicca, tear film dysfunction syndrome, or burning eye syndrome, is caused by a deficiency in one of the layers of the tear film. Dry eye is a multifactorial disorder affecting the lacrimal fluid and ocular surface, resulting in symptoms of discomfort, visual impairment, and tear film instability with potential ocular surface damage. Dry eye is characterized by a loss of tear film homeostasis and is accompanied by ocular symptoms in which tear film instability and hyperosmolarity, ocular surface inflammation and damage, and neurosensory abnormalities play an etiologic role (Craig JP, et al., The Ocular Surface 2017;15:276-83). It may be accompanied by increased tear film osmolarity and ocular surface inflammation. Dry eye disorders can range from mild to moderate to severe. Symptoms of painful dry eye include a grittiness, foreign body sensation, burning, photophobia and decreased visual acuity, tearing, sharp pain, itching, a grainy or gritty feeling, discharge, frequent blinking, pus or clumping of eyelashes (usually worse on awakening), redness, blurred or fluctuating vision (worse when reading, using a computer, watching TV, driving, or playing video games), sensitivity to light, eye pain and/or headache, heaviness of the eyelids, and eye fatigue. Causes of dry eye disease include, but are not limited to, idiopathic congenital alacrima, xerophthalmia, lacrimal gland ablation, and sensory denervation; collagen vascular diseases, including rheumatoid arthritis, Wegener's granulomatosis, and systemic lupus erythematosus; Sjogren's syndrome and autoimmune diseases associated with Sjogren's syndrome; abnormalities of the lipid layer of the tear film caused by blepharitis or rosacea; abnormalities of the mucin layer of the tear film caused by vitamin A deficiency; trachoma, diphtheritic keratoconjunctivitis; skin and mucous membrane disorders; aging; menopause and diabetes mellitus. The signs and/or symptoms of dry eye defined in this document may also be caused by other circumstances, including, but not limited to, the following: prolonged visual tasks; computer work; dry environments; exposure to warm or cold winds or air currents; changes in seasons; eye irritation; use of contact lenses, LASIK and other refractive surgeries; fatigue and medications such as isotretinoin, sedatives, diuretics, tricyclic antidepressants, antihypertensives, oral contraceptives, antihistamines, nasal decongestants, beta blockers, phenothiazines, atropine and opiate pain relievers such as morphine.

Diagnostic testing for dry eye includes assessment of corneal sensitivity (in severe and chronic dry eye disease, corneal hyperesthesia and/or decreased sensitivity may be present) using, for example, a cotton swab or, with greater accuracy, a Cochet-Bonnet esthesiometer; measurement of tear breakup time using, for example, a fluorescein strip moistened with preservative-free saline or more objective computerized techniques that do not require fluorescein instillation; performing ocular surface stains, for example, with sodium fluorescein, rose bengal, lissamine green; performing the Schirmer test (relatively insensitive in patients with mild dry eye), testing for tear outflow retardation; determining the height of the tear meniscus; MMP-9 measurement (MMP-9 levels have been shown to be elevated in the tears of patients with dry eye disease and to correlate with examination findings in patients with moderate to severe dry eye); tear osmolarity measurement and tear film interferometry; Sjo test (detection of serum autoantibodies to SS-A (anti-Ro antibodies) and SS-B (anti-La antibodies), salivary gland protein 1 (SP-1), carbonic anhydrase 6 (CA6), and parotid secretory protein (PSP) - SP-1, CA6, and PSP).

Artificial tears, lubricating ointments, and corticosteroids (e.g., 0.5% loteprednol eye drops four times daily) are used as initial treatment. Prescription medications include cyclosporine, lifitegrast, diquafosol, rebamipide, and corticosteroids (e.g., 0.5% loteprednol eye drops four times daily).

The term "tear film dysfunction" refers to a condition in which the tear film breaks down in various places on the cornea and conjunctiva, leading not only to symptoms of irritation but also to unstable and intermittent vision. For example, painful dry eye syndrome is characterized by tear film dysfunction. Symptoms of tear film dysfunction include watery eyes, burning, sharp pain, itching, a grainy or gritty sensation, a scratchy or foreign body sensation, discharge, frequent blinking, pus or clumping of eyelashes (usually worse on awakening), redness, blurred or fluctuating vision (worse with reading, computer work, watching television, driving, or playing video games), sensitivity to light, eye pain and/or headache, heaviness in the eyelids, and eye fatigue.

Adenoviral keratoconjunctivitis, also known as epidemic keratoconjunctivitis, is a common and highly contagious viral ocular infection. The clinical course of adenoviral keratoconjunctivitis is divided into an acute phase with varying degrees of conjunctival inflammation, with or without corneal involvement, and a chronic phase with corneal opacity.

Vernal keratoconjunctivitis (VKC) is an atopic condition of the outer surface of the eye characterized by symptoms of severe itching, photophobia, foreign body sensation, mucous discharge (often described as "stringy"), 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 usually bilateral but can be asymmetric in nature. It characteristically affects young males seasonally in dry, hot climates; 23% of patients may have a year-round form (Kumar, S., Acta Ophthalmologica, 2009. 87(2): p. 133-147 ; Bonini, S., et al., Ophthalmology, 2000. 107(6): p. 1157-63).

Signs of VKC can be divided into conjunctival, limbal and corneal signs:

Conjunctival features include diffuse conjunctival injection and discrete giant papillae on the tarsal conjunctiva of the upper eyelid with a diameter > 1 mm;

Limbal features include thickening and opacification of the limbal conjunctiva, as well as a gelatinous appearance and, in places, confluent limbal papillae. Perilimbal Horner-Trantas spots are white, focal limbal spots 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 process and include macroerosions, ulcers, and corneal scars (Buckley, RJ, Int Ophthalmol Clin, 1988. 28(4): p. 303-8).

Patients with active VKC (defined by moderate to severe ocular discomfort, including photophobia, tarsal papillae on the upper tarsal conjunctiva, or Horner-Trantas spots clearly visible at the time of examination) demonstrated a significant increase in the intensity of symptoms and signs of ocular surface disease. Patients with inactive VKC (defined by no symptoms or mild discomfort and no corneal abnormalities at the time of examination) demonstrated an increase in photophobia, lissamine green staining intensity, and Schirmer test values, as well as a decrease in fluorescein tear breakup time (BUT) and corneal sensitivity. This syndrome appears to affect the ocular surface in all phases (active and asymptomatic), causing abnormalities in tear film stability, epithelial cell integrity, and corneal nerve function (Villani E et al., Medicine (Baltimore). 2015 Oct; 94(42): e1648).

The following factors are thought to play a role in VKC: IgE-mediated response via mast cell release; type IV hypersensitivity mediated by activated eosinophils, mononuclear cells, and neutrophils, as well as CD4+ T helper type 2 cells, with immunomodulators such as IL-4, IL-5, and bFGF (Buckley, RJ, Int Ophthalmol Clin, 1988. 28(4): p. 303-8 ; Kumar, S., Acta Ophthalmologica, 2009. 87(2): p. 133-147 ; La Rosa, M., et al., Ital J Pediatr, 2013. 39: p. 18).

Treatment involves the use of cold compresses and eyelid scrubs, saline eye drops, which may help relieve symptoms, along with topical antihistamines, nonsteroidal anti-inflammatory drugs or corticosteroids, such as low-absorption corticosteroids (fluorometholone, loteprednol, rimexolone, etc.), topical mast cell stabilizers (cromolyn sodium, nedocromil sodium, and lodoxamide), topical cyclosporine A, or tacrolimus. See, 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, R. K. et al.J Clin Diagn Res.2016 Jun;10(6):NC05-9; Wan Qet al., Ophthalmic Res.2018; 59(3):126-134.

Atopic keratoconjunctivitis (AKC) typically has an onset at an older age, between the 20s and 50s, compared to VKC, which typically occurs before age 10. Classically, conjunctival involvement occurs in the cartilage of the upper eyelid in VKC and in the cartilage of the lower eyelid in AKC. AKC is typically more chronic in nature and more often leads to corneal scarring and conjunctival scarring.

Sjögren's syndrome (Sjögren's syndrome associated with dry eye) is a chronic inflammatory disorder characterized by dysfunction of the exocrine glands, including the salivary and lacrimal glands, often resulting in severe dry eye. The main symptoms are dry eyes (keratitis sicca or keratoconjunctivitis sicca) and dry mouth (xerostomia). Severe forms of dry eye can cause corneal pain, scarring, ulceration, infection, and even corneal perforation. Differential diagnosis includes conditions such as adult blepharitis, dry eye disease, and uveitis associated with juvenile idiopathic arthritis, as well as forms of keratopathy such as superficial punctate, filamentous, neurotrophic, and exposure keratopathy. Treatment of Sjögren's syndrome is aimed at maintaining the integrity of the tear film by preserving, enhancing, and/or replacing deficient tear production. Treatment of Sjögren's syndrome therefore includes the use of artificial tears and lubricating ointments; autologous serum in the form of eye drops; essential omega-6 fatty acids for oral use; gas-permeable scleral contact lenses with liquid ventilation; topical corticosteroids; punctal occlusion to reduce tear outflow; partial lateral tarsorrhaphy; environmental humidification; hydrophilic bandage contact lenses; bromhexine and 3-isobutyl-1-methylxanthine (IBMX) (to enhance tear production/secretion); muscarinic receptor stimulators (pilocarpine and cevimeline); immunosuppressants such as methotrexate, antimalarial drugs, cyclophosphamide, leflunomide, or tumor necrosis factor (TNF) drugs such as infliximab, a monoclonal antibody to TNF-alpha; cyclosporine A; a bandage contact lens.

Stevens-Johnson syndrome (SJS) is a dermatological emergency or a type of severe skin reaction characterized by bullous lesions of the epidermis and mucous membranes affecting less than 10% of the total body surface area. Early symptoms of SJS include fever and flu-like symptoms, which may precede or occur simultaneously with the development of a macular rash affecting the trunk and face. As the disease progresses, the macular lesions coalesce, blisters develop in the affected areas, and ultimately, the epidermal layer separates. During the acute phase of SJS-TEN, ocular involvement occurs in 80% of patients.

A constellation of high fever (>102.2), malaise, joint pain, macular rash affecting the trunk, neck, and face, and a recent history of exposure to a new drug or a recent increase in the dose of an existing drug are indicators used to diagnose SJS. A skin biopsy from the lesion can be performed to confirm the diagnosis. Granulysin can be used as a marker for diagnosing SJS. The concentration of granulysin in blister fluid correlates with the severity of the acute phase of SJS (Chung WH et al. Nat Med. 2008;14(12):1343-50).

Ocular manifestations of SJS include conjunctivitis, subconjunctival hemorrhage, subconjunctival scarring, conjunctival membranes, conjunctival ulceration, superficial punctate epithelial erosions, epithelial defects, lid margin ulcerations, lid margin keratinization, symblepharon, ankyloblepharon, trichiasis, anterior blepharitis, punctal autoocclusion, meibomian gland disease, corneal opacities, dry eye, districhiasis, limbal stem cell failure, and corneal vascularization. Ocular treatment for SJS involves the use of saline eye drops, preservative-free artificial tears, and ointments to ensure adequate lubrication and reduce the intensity of epithelial damage. Patients with any corneal or conjunctival epithelial defects are treated with prophylactic topical antibiotics, preferably fourth-generation fluoroquinolones. Patients with mild to moderate ocular involvement (less than one-third of the eyelid margin is involved, the largest diameter of the conjunctival defect is less than 1 cm, and no corneal epithelial defects are present) are typically treated with topical moxifloxacin 0.5% four times daily, cyclosporine 0.05% twice daily, and topical steroids (1% prednisolone acetate four to eight times daily or 0.1% dexamethasone twice daily). Patients characterized by severe or extremely severe ocular involvement (more than one-third of the eyelid margin is affected, conjunctival defect size is greater than 1 cm, and there are corneal epithelial defects) undergo amniotic membrane transplantation (AM) in addition to the above treatments.

In some embodiments, the subject to be treated suffers from corneal epitheliopathy. Corneal epitheliopathy is a disease affecting the corneal epithelium, manifested, for example, by altered barrier function of the corneal epithelium.

In some embodiments, the subject to be treated suffers from corneal neuropathy or corneal neuralgia. Corneal neuropathy or corneal neuralgia Corneal neuropathy is a disorder associated with corneal pain caused by damage to the corneal nerve fibers—the sensory nerve fibers. One example of corneal neuropathy is LASIK-induced corneal neuropathy. Corneal neuropathy can typically be identified and diagnosed through dry eye testing. Although the causes and risk factors remain unclear, patients with dry eye-like symptoms, increased corneal sensitivity, and changes in the morphological characteristics of the corneal nerves, but without signs of dryness, may have corneal neuropathy.

In some embodiments, the subject to be treated suffers from an ocular surface disease or disorder. The terms "ocular surface diseases" or "ocular surface disorders" encompass entities and their associated symptoms that are caused by a variety of abnormalities, including abnormal anatomical characteristics or function of the eyelid, abnormal or altered tear production or composition, and associated subclinical features. Many diseases can cause ocular surface disorders. Patients with ocular surface disorders may exhibit clinical features characteristic of multiple diseases and include chronic punctate keratopathy, filiform keratopathy, recurrent corneal erosions, bacterial conjunctivitis, culture-negative conjunctivitis, scarring conjunctivitis, persistent epithelial defect, infectious keratitis, corneal softening, and ocular surface tissue failure. The most common ocular surface disorders result from abnormalities of the tear film and/or dysfunction of the eyelid glands ("blepharitis").

In some embodiments, the subject to be treated suffers from neurotrophic keratitis or neurotrophic keratopathy. Neurotrophic keratitis or neurotrophic keratopathy (NK) is a degenerative corneal disease characterized by decreased or absent corneal sensation. In NK, corneal innervation by the trigeminal nerve is impaired. Because corneal sensory innervation is impaired in NK, patients typically do not complain of ocular surface symptoms. However, blurred vision may be reported due to the presence of irregular epithelium or epithelial defects (PEDs), scarring, or edema. NK is typically classified into three different stages according to the "Mackie classification." Stage II NK is defined by recurrent or persistent epithelial defects, most often in the superior half of the cornea. One treatment option for stage II NK involves topical nerve growth factor. Patients typically experience pain during NGF treatment due to nerve regrowth.

In some embodiments, the subject to be treated suffers from blepharitis. Blepharitis is an inflammatory condition of the eyelid margin that can lead to persistent eyelid margin changes or vision loss due to superficial keratopathy, neovascularization, and corneal ulceration. Based on anatomical location, blepharitis can be classified as anterior or posterior. Anterior blepharitis affects the eyelid skin, eyelash roots, and eyelash follicles and, according to traditional classification, includes staphylococcal and seborrheic blepharitis. Posterior blepharitis affects the meibomian glands and gland orifices, with meibomian gland dysfunction being the primary cause. Symptoms of chronic blepharitis can include redness, burning, irritation, lacrimation, crusting of the eyelids, and adhesion of the eyelids, as well as vision problems such as photophobia and blurred vision. Long-term symptom control may involve daily standard eyelid irrigation procedures and the use of therapeutic agents that reduce the intensity of infection and inflammation. Treatment includes 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, such as loteprednol etabonate, fluorometholone; topical antibiotic-corticosteroid combinations, such as tobramycin/dexamethasone or tobramycin/loteprednol; and topical cyclosporine 0.05%.

In some embodiments, the subject to be treated suffers from meibomian gland dysfunction. The meibomian gland is an exocrine gland of the holocrine type within the tarsal plate at the margin of the eyelid that is responsible for supplying meibomian secretion, an oily substance that prevents evaporation of the ocular tear film. Meibomian gland dysfunction (MGD), also known as meibomitis, posterior blepharitis, or meibomian inflammation, is a chronic, diffuse abnormality of the meibomian glands typically characterized by obstruction of the terminal ducts and/or qualitative/quantitative changes in the secretion of these glands (Nelson JD, et al., Invest Ophthalmol Vis Sci2011;52:1930-7). This can lead to changes in the tear film, symptoms of eye irritation, clinically significant inflammation, and ocular surface disease..MGD It often causes dry eyes and can contribute to blepharitis. In some cases, topical steroids and topical/oral antibiotics are also prescribed to reduce inflammation. Intense pulsed light (IPL) treatments or other forms of mechanical therapy that use heat and pressure to empty the glands (such as LipiFlow) have also been shown to reduce inflammation and improve gland function in patients.

In some embodiments, the subject to be treated suffers from graft-versus-host disease. Graft-versus-host disease (GVHD) is an inflammatory disorder that occurs only in allogeneic transplantation. It involves an attack by transplanted white blood cells on the recipient's tissues, which can occur even when the donor and recipient are HLA-identical. Acute GVHD typically appears within the first 3 months after transplantation and can affect the skin, intestine, or liver. Corticosteroids, such as prednisone, are the standard treatment. Chronic GVHD can also develop after allogeneic transplantation and is a major source of late complications. In addition to inflammation, chronic graft-versus-host disease can lead to the development of fibrosis or scar tissue similar to scleroderma or other autoimmune diseases and can cause functional disability and the need for long-term immunosuppressive therapy.

In some embodiments, the subject to be treated suffers from ocular graft-versus-host disease. GVHD occurs in patients who have undergone allogeneic hematological stem cell transplantation. It can occur in patients who have acute or chronic GVHD, although it is more common in patients with the chronic form. Approximately 40-90% of patients with chronic GVHD develop ocular symptoms. Ocular manifestations may include moderate to severe keratoconjunctivitis sicca, bilateral marginal keratitis, anterior uveitis, corneal ulceration, or corneal neovascularization. Treatment includes topical lubricants, including preservative-free artificial tears, autologous serum drops, and other immunosuppressive therapies for topical and systemic use; systemic steroids; and 0.5% topical cyclosporine.

EXAMPLES

The following examples are included to demonstrate non-limiting embodiments of the present invention.

Example 1. First-in-human study of compound I to evaluate safety and pharmacokinetic parameters

This example describes a first-dose human study of compound I conducted in healthy volunteers. The study drug was administered to a total of 54 subjects. In Part 1 of the first-dose human study, single escalating doses of 0.15%, 1.5%, and 2.5% w/v (1 drop) of compound I were administered as eye drops.

In Part 2, multiple ascending doses (MADs) of Compound I were tested by administering (i) 1 drop of 0.15%, 1.5%, 2.5% concentration 4 times daily (every 6 hours) for 7 days or (ii) 1 drop of 2.5% concentration 8 times daily (every 3 hours) for 7 days (supratherapeutic dose) as eye drops.

Part 3 was an esthesiometric evaluation to determine the anesthetic properties of compound I. Part 3 of the study consisted of four groups receiving compound I, vehicle, tetracaine (0.5% ophthalmic solution, used as a positive control), and diclofenac sodium (0.1% ophthalmic solution, used as an NSAID comparator). The ocular anesthetic was chosen due to its established anesthetic effect; however, it is not a standard of care due to its negative effect on wound healing. NSAIDs are the current standard of care for corneal pain after PRK, with minimal anesthetic but significant analgesic properties. Vehicle eye drops were used as a negative control to better determine the potential anesthetic effect. Twelve healthy subjects who met the study eligibility criteria were randomized to receive one eye drop of each of the four study treatments, each administered on one of four different study days (days 1, 4, 7, and 10). Each subject was randomized to receive the study treatment according to one of the following four sequences:

Sequence 1: anesthetic, NSAID, 2.5% compound I, vehicle

Sequence 2: NSAID, vehicle, anesthetic, 2.5% Compound I

Sequence 3: vehicle, 2.5% compound I, NSAID, anesthetic

Sequence 4: 2.5% Compound I, anesthetic, vehicle, NSAID

Safety assessments. Safety assessments included collecting information on all adverse events (AEs), serious adverse events (SAEs), including systemic and ocular adverse events, their severity, and relationship to the study drug. Systemic safety assessments included regular monitoring of complete blood count, blood chemistry, and urinalysis results performed at the study center, and regular assessments of physical examination data, vital signs (systolic and diastolic blood pressure, pulse, and body temperature), ECG, pregnancy, and fertility assessments, and a 49°C hand immersion test during the MAD portion. Ocular safety assessments included visual acuity assessment according to the Early Treatment Diabetic Retinopathy Study (ETDRS), intraocular pressure assessment, slit-lamp biomicroscopy, corneal staining, and dilated fundus examination.

Subjects were selected based on the following inclusion and exclusion criteria.

Inclusion criteria:

Written informed consent was obtained before any assessments were performed.

Healthy male and female subjects aged 18-50 years (inclusive) in good health as determined by history, physical examination, vital signs, ECG, and laboratory tests at screening.

At screening and baseline, vital signs (systolic and diastolic blood pressure (BP) and pulse) were assessed in the sitting position after the subject had been at rest for at least 3 minutes and again after 3 minutes of standing. Vital signs in the sitting position were required to be within the reference range for the following ranges:

body temperature when measured orally 35.0-37.5°C

systolic blood pressure (SBP) 90-150 mmHg

diastolic blood pressure (DBP) 50-90 mmHg

pulse 40-100 beats/min.

To participate in the study, subjects were required to have a body weight of at least 50 kg and a body mass index (BMI) in the range of 18-29 kg/m ² . BMI = body weight (kg) / [height (m)] ²

Subjects were able to communicate freely with the researcher and understand and comply with the requirements of the study.

For Part 3, subjects' baseline ocular sensitivity levels were required to be in the range of 50 to 60 mm (inclusive) as measured using the Cochet-Bonnet esthesiometer.

Exclusion criteria:

Women of childbearing potential are defined as all women physiologically capable of becoming pregnant if they were not using effective contraception at the time of administration of study treatment doses.

Subjects who demonstrated the presence of any medical condition (systemic or ophthalmological) that, in the opinion of the investigator and based on the contents of the investigator's brochure, precluded safe administration of the test product or safe participation in this study.

Part 3 (esthesiometry): Subjects who used contact lenses during the study or had used them within the last 3 years were excluded to minimize variability in corneal sensitivity due to contact lens use.

A history of any eye surgery or laser procedure within the last 6 months prior to screening. A history of any chronic eye disease other than refractive error, early cataracts, disbinocular amblyopia, or anisometropic amblyopia. Subjects with a history of an acute eye disease (such as infection, corneal abrasion, or allergy) within the last 6 months of screening were eligible for the study if the disease was not active.

Any currently active ocular condition that has required topical eye drops.

Subjects using continuous positive airway pressure or other sleep apnea devices.

Safety results from the first-in-human study

Based on the results of Parts 1, 2, and 3, the maximum tolerated dose (MTD) was identified as the maximum achievable concentration of 2.5% administered 8 times daily for 7 days. No dose-limiting adverse events were identified at this dose level. All adverse events with a presumed causal relationship to Compound I were mild in severity, with the exception of moderate ocular irritation, which led to treatment discontinuation in one patient in the 2.5% 4 times daily cohort. The most common ocular adverse events in patients treated with Compound I were corneal staining, hyperemia, and mild anterior chamber inflammation at levels similar to placebo. A summary of adverse events from the SAD study is shown in Table 1.

Table 1. Overall AE incidence – number of events and number of subjects (Part 1: SAD) (safety analysis set)

0.15% compound I 1.5% compound I 2.5% compound I Carrier medium Total N=6 N=6 N=6 N=6 N=24 nE, nS (%) nE, nS (%) nE, nS (%) nE, nS (%) nE, nS (%) AE, subjects with AE 3, 1 (16.7) 3, 2 (33.3) 0, 0 (0,0) 4.3 (50.0) 10.6 (25.0) Mild AE 3, 1 (16.7) 3, 2 (33.3) 0, 0 (0,0) 3, 2 (33.3) 9.5 (20.8) Moderate AE 0, 0 (0,0) 0, 0 (0,0) 0, 0 (0,0) 1, 1 (16.7) 1, 1 (4.2) AEs related to the study drug 3, 1 (16.7) 3, 2 (33.3) 0, 0 (0,0) 3, 2 (33.3) 9.5 (20.8) Serious AE 0, 0 (0,0) 0, 0 (0,0) 0, 0 (0,0) 0, 0 (0,0) 0, 0 (0,0) AE leading to termination of participation in 0, 0 (0,0) 0, 0 (0,0) 0, 0 (0,0) 1, 1 (16.7) 1, 1 (4.2) research AEs related to the study drug leading to study discontinuation 0, 0 (0,0) 0, 0 (0,0) 0, 0 (0,0) 1, 1 (16.7) 1, 1 (4.2)

N=number of subjects enrolled in the study and receiving study drug; nE=number of treatment-emergent AE events in a given category

nS = number of subjects with at least one AE occurring after the start of treatment in a given category. Percentage is determined by the number of subjects

A summary of adverse events from the SAD study is shown in Table 2.

Table 2. Overall AE incidence – number of events and number of subjects (Part 2: MAD) (safety analysis set)

0.15% compound I
4 times a day 1.5% compound I
4 times a day 2.5% compound I
4 times a day 2.5% compound I
8 times a day Carrier medium Total N=6 N=6 N=6 N=6 N=8 N=32 nE, nS (%) nE, nS (%) nE, nS (%) nE, nS (%) nE, nS (%) nE, nS (%) AE, subjects with AE 6.3 (50.0) 12.5 (83.3) 4, 2 (33.3) 3, 2 (33.3) 13.3 (37.5) 38.15 (46.9) Mild AE 6.3 (50.0) 11.5 (83.3) 3, 2 (33.3) 3, 2 (33.3) 12.3 (37.5) 35.15 (46.9) Moderate AE 0, 0 (0,0) 1, 1 (16.7) 1, 1 (16.7) 0, 0 (0,0) 1, 1 (12.5) 3, 3 (9.4) AEs related to the study drug 5.2 (33.3) 9.3 (50.0) 4, 2 (33.3) 2, 1 (16.7) 10.3 (37.5) 30.11 (34.4) Serious AE (0.0) 0, 0 0, 0 (0,0) 0, 0 (0,0) 0, 0 (0,0) 0, 0 (0,0) 0, 0 (0,0) AEs leading to study discontinuation 0, 0 (0,0) 0, 0 (0,0) 1, 1 (16.7) ^a 0, 0 (0,0) 0, 0 (0,0) 1, 1 (3,1) AEs related to the study drug 0, 0 (0,0) 0, 0 (0,0) 1, 1 (16.7) 0, 0 (0,0) 0, 0 (0,0) 1, 1 (3,1)

N=number of subjects enrolled in the study and receiving study drug; nE=number of treatment-emergent AE events in a given category

nS = number of subjects with at least one AE occurring after the start of treatment in a given category. Percentage is determined by the number of subjects

^a A subject treated with 2.5% Compound I 4 times daily experienced ocular discharge, ocular hyperemia (both mild), and ocular irritation (moderate) in the left eye on Day 6, leading to discontinuation of study drug administration.

In the hand immersion test, all subjects in the treatment cohorts withdrew their hands from water at 49°C within a time interval ranging from 0 to ≤50 s. No subject withstood the test for longer than 22 s, and no significant change in immersion time was observed between subjects treated with compound I and vehicle. Therefore, the results indicate that compound I does not alter temperature sensitivity in subjects.

Esthesiometric study (part 3)

In the first human use study, the potential anesthetic effect of 2.5% Compound I for topical ocular application was further assessed by esthesiometric testing, i.e., measuring the thread length (cm) at which the threshold level of corneal touch perception is achieved.

Analysis of the results demonstrated that 0.5% tetracaine had an anesthetic effect with a duration of action of approximately 10 minutes (positive control). Neither 0.1% diclofenac nor the vehicle had an anesthetic effect on the cornea, as expected given the known clinical practice of using diclofenac. 2.5% Compound I did not demonstrate an anesthetic effect at any time after treatment.

In corneal sensitivity measurements, statistical and clinical significance was observed when comparing the least squares means (test and reference samples) in the following cases:

0.5% tetracaine (N=12) versus vehicle (N=12) at 2.5 minutes, 10 minutes, and 20 minutes post-dose;

0.5% tetracaine (N=12) versus 0.1% diclofenac (N=12) at 2.5 minutes, 10 minutes, and 20 minutes post-dose;

2.5% Compound I (N=11) versus 0.5% tetracaine (N=12) at 2.5 minutes and 10 minutes post-dose

No differences in corneal sensitivity were observed between 2.5% Compound I and vehicle, with all p-values greater than or equal to 0.395 at scheduled time points up to 30 minutes post-dose. This study demonstrates the lack of anesthetic effect of 2.5% Compound I compared to 0.5% tetracaine (a drug with anesthetic effect as a positive control) and vehicle (placebo).

Summary of pharmacokinetic parameters in the first-in-human study

Following both single and multiple doses of 0.15%, 1.5%, and 2.5% Compound I topical ocular suspension, the plasma PK profile demonstrated rapid absorption of Compound I into the systemic circulation and achievement of exposure at low concentrations with moderate intersubject variability. Increasing the dose from 0.15% to 2.5% resulted in a less than dose-proportional increase in systemic exposure. The accumulation of Compound I at steady state was minor (~1.7-fold) following administration of 0.15%, 1.5%, or 2.5% suspension four times daily and also minor (~1.3-fold) following administration of 2.5% eight times daily for 7 days.

Example 2. Clinical study of compound I in the treatment of postoperative pain

This example describes a clinical study of compound I (4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile) in the treatment of postoperative ocular pain in patients undergoing photorefractive keratectomy (PRK) surgery.

Objectives of the study

Primary goal Endpoints associated with the primary goal To assess pain relief in the immediate postoperative period. Visual analogue scale (VAS) pain assessment pre-dose to 6 hours post-operatively
Mean VAS ocular pain score from first postoperative assessment to 12-hour predose assessment Secondary goals Endpoints associated with secondary objectives To evaluate the efficacy of 2.5% Compound I eye drops administered four times daily in reducing the frequency of oral analgesic use after PRK procedure. Frequency of use and amount of oral backup analgesics required at 6 hours, 12 hours, 24 hours, 2 days, and 3 days postoperatively after each PRK surgery. To evaluate the safety and tolerability of 2.5% Compound I as eye drops when administered four times daily. Adverse events (AEs) and serious adverse events (SAEs)
Visual acuity, intraocular pressure (IOP), dilated fundus examination, ocular hyperemia
Size of epithelial defect as assessed by slit lamp examination
Blinking rate, tear production
Vital signs (blood pressure, pulse, and body temperature) Assess the severity of pain after surgery All measurements according to VAS during the first 3 days after surgery To evaluate systemic exposure following ocular administration of 2.5% Compound I eye drops four times daily at various time points in patients undergoing PRK Plasma concentration of compound I Search objectives Endpoints associated with search intents To study the rate of wound healing The size of the epithelial defect according to optical coherence tomography of the anterior segment of the eye (OCT) To study the effect of compound I on pain relief compared to vehicle after instillation of eye drops VAS scores before and after eye drop instillation (at three time points) To study the effect of compound I on ocular pain and quality of life Ocular Pain Assessment Questionnaire (OPAS) Exposure to compound I in a bandage contact lens (BCL) Residual amount of compound I after treatment

Research plan

This study was a randomized, vehicle-controlled, double-masked proof-of-concept study of compound I administered as eye drops in addition to standard treatment in patients undergoing PRK surgery. PRK surgery was performed as an outpatient procedure under local anesthesia with removal of the corneal epithelium to expose the stroma for laser ablation. Doses of compound I were administered as a single eye drop of 2.5% concentration (25 mg/mL, 0.925 mg/drop) administered to one eye four times daily (every six hours), starting from the time immediately postoperatively (time 0) until the last dose 72 hours later.

Vehicle doses were administered as one eye drop injected into one eye four times daily (every six hours), starting from the time immediately after surgery (time 0) until the last dose at 72 hours.

The study consisted of two treatment periods using a crossover design. Patients underwent PRK surgery on two separate occasions (periods) in one eye at a time. Patients were randomized to receive either compound 1 or vehicle after procedure 1 and the alternative after procedure 2.

Forty patients were randomized 1:1 to either Compound I for Period 1 followed by vehicle for Period 2 or vehicle for Period 1 followed by Compound I for Period 2. Each patient received one drop four times daily for 72 hours after PRK surgery in their study eye (Day 1, Periods 1 and 2). The study eye was initially the non-dominant eye, as determined at screening and by agreement between the patient and investigator. Following surgery in the first eye, patients returned for follow-up visits on Days 2, 3, 4, and 8 of Period 1, with optional daily follow-up visits until wound healing was complete. Surgery in the second eye was not performed if any complications were noted.

Patients underwent PRK surgery in their second study eye (dominant eye) on Day 1 of Period 2. PRK surgery was performed after repair of the epithelial defect in the first eye and at the investigator's discretion. Following PRK surgery, patients received the opposite treatment four times daily for 72 hours. Patients returned daily for the first 3 days postoperatively (Period 2, Days 2-4) and one week after the second surgery (Period 2, Day 8), with optional daily follow-up visits until wound healing was complete. The end-of-study visit (EOS) occurred 30 days after the second eye surgery (or after the last dose of study medication if the patient completed treatment early during Period 1).

All patients received standard treatment during Periods 1 and 2 of this study, including wearing a bandage contact lens (Air Optix® Night and Day® Aqua or equivalent), after the PRK procedure and before receiving study drops for pain. After the first dose of study drops, a course of topical ocular antibiotic (moxifloxacin or equivalent, 1 eye drop four times daily) was started and continued for 4-7 days. Ophthalmic prednisolone acetate was administered as one eye drop four times daily for one week after PRK, with a gradual dose reduction thereafter. Preservative-free artificial tears were used in single doses as needed. The first dose of study drops after each PRK procedure was administered by study center personnel. Subsequent doses were self-administered. A 5-minute interval was allowed between eye drop administrations in a sequence. Generally, patients undergoing PRK had a bandage contact lens placed on the cornea, Compound I or vehicle injected, an antibiotic injected approximately 5 minutes later, and prednisolone injected 5 minutes later.

The rescue medication consisted of an oral analgesic (300 mg acetaminophen + 30 mg codeine), administered at a total of up to 10 tablets/day or 1-2 tablets every 4 hours as needed.

Additional investigational treatment

All patients after PRK surgery received standard supportive care, including the following:

Bandage contact lens (Air Optix ^® Night and Day ^® Aqua or equivalent)

A course of topical ocular antibiotic (moxifloxacin or equivalent, 1 eye drop four times daily) was started immediately after PRK surgery and after the first dose of study eye drops and continued for 4-7 days at the discretion of the administering physician.

Ophthalmic prednisolone acetate: One eye drop four times daily was started immediately after PRK surgery and after antibiotic eye drop instillation and administered for 1 week after PRK, followed by gradual dose reduction according to local procedures.

Preservative-free artificial tears were used in single doses as needed. Artificial tears were not allowed to be cooled to achieve an analgesic effect.

The drops were administered sequentially with an interval of at least 5 minutes between eye drop administrations.

Inclusion criteria

The population eligible for inclusion in this study had to meet all of the following criteria:

Male and female patients aged 18-75 years eligible for bilateral PRK surgery.

The results of the vision examination are within normal limits except for the refractive error at the baseline.

For planned correction, myopia should not exceed -4.00 diopters (for the spherical component), and astigmatism should not exceed 3.00 diopters with a spherical equivalent of no more than -4.50, confirmed by manifest refraction at baseline. Treatment with monovisual correction was permitted (such as distance vision correction in one eye and intermediate vision correction in the other eye).

Written informed consent was obtained before any assessments were performed.

Exclusion criteria

A patient with one eye (including amblyopia) or with a best-corrected visual acuity (BCVA) score worse than 20/80 (Snellen chart) or 55 letters [according to the Early Treatment Diabetic Retinopathy Study (EDTRS)] at baseline.

Any systemic or ocular diseases that affect wound healing (such as severe rheumatoid arthritis, diabetes mellitus, or a history of keloid formation), or a history of ocular trauma, uveitis, infection, or inflammation within 6 months prior to baseline. Specifically, with regard to diabetes mellitus, patients with severe diabetes mellitus, uncontrolled diabetes mellitus, diabetic keratopathy, diabetic retinopathy, diabetic macular edema, diabetic nephropathy, diabetic foot ulcers, or other systemic complications of diabetes mellitus were excluded. Patients with mild, well-controlled diabetes mellitus without evidence of ocular or systemic complications of diabetes mellitus were included.

Patients with active inflammatory or infectious ocular conditions, severe or progressive retinal disease, who were using topical or systemic steroids or were using Coumadin or similar medications within the last 6 months prior to baseline.

Patients with any form of corneal dystrophy (epithelial, stromal, or endothelial) or any corneal disease (including significant scarring (at the investigator's discretion), ocular herpes, or pterygium).

Prior refractive surgery or corneal surgery (such as LASIK, PRK, radial keratotomy, pterygium removal, corneal transplant).

History of allergic or hypersensitivity reaction or significant AE to any of the medications used in this study, including tetracaine or similar topical ocular anesthetic, NSAIDs and aspirin, oral analgesics (including acetaminophen and codeine), antibiotics, steroids, and inability to tolerate or wear a bandage contact lens.

Concomitant therapy or long-term therapy or history of abuse with NSAIDs for systemic or ocular use, analgesics, pain medications (including gabapentin or pregabalin and similar), opiates, or marijuana.

Patients who had used any topical ocular medication, except lubricating eye drops, within two weeks prior to surgery in the study eye were excluded. Patients who met any of the following criteria were excluded.

Use of a topical NSAID within 30 days prior to baseline, OR

Systemic/persistent use of a systemic NSAID within 30 days prior to baseline, OR

Occasional use of a systemic NSAID within 3 days prior to baseline, OR

Use of cyclosporine ophthalmic (or similar medication) within 3 months prior to surgery.

Patients with a body weight < 50 kg or a body mass index (BMI) outside the range of 18-35 kg/m ² . BMI = body weight (kg) / [height (m)] ²

Pregnant or lactating women. Women of childbearing potential are defined as all women physiologically capable of becoming pregnant if they are not using primary contraception during study drug administration.

The investigator did not apply additional exclusion criteria to ensure that the study population represented all patients eligible for participation in the study.

Study population

The study population consisted of male and female patients aged 18 to 75 years (inclusive) eligible for PRK surgery. A total of 40 patients were planned for participation. A total of 44 patients were screened, of which 40 were included in the study and randomized.

Patient selection was based on consideration of all study eligibility criteria during screening and baseline determination. A record (e.g., a checklist) of all study eligibility criteria was maintained with the primary documentation at the study center. Deviation from any study inclusion criterion excluded the patient from inclusion in the study.

Patient demographic data are presented in Table 3.

Table 3. Patient demographics by treatment sequence (safety analysis set)

Carrier medium/
2.5% compound I
N=20 2.5% compound I/
Carrier medium
N=20 Total
N=40 Age (years) Mean (SD) 34.4 (10.77) 33.7 (8.94) 34.0 (9.78) Median value 33.5 32.0 33.0 Range 20-54 23-56 20-56 Gender - n (%) Male 11 (55%) 10 (50%) 21 (53%) Female 9 (45%) 10 (50%) 19 (48%) Race - n (%) Caucasoid 18 (90%) 17 (85%) 35 (88%) Mongoloid 2 (10%) 1 (5%) 3 (8%) Negroid or African American 0 1 (5%) 1 (3%) Mixed 0 1 (5%) 1 (3%) Ethnicity (n (%)) Origin other than Spanish or Latin American 19 (95%) 18 (90%) 37 (93%) Spanish or Latin American descent 1 (5%) 2 (10%) 3 (8%) Body weight (kg) Mean (SD) 84.1 (14.86) 78.8 (14.07) 81.5 (14.53) Median value 84.7 76.1 79.2 Range 60-119 57-107 57-119 Height (cm) Mean (SD) 170.4 (7.40) 173.2 (6.96) 171.8 (7.23) Median value 171.3 174.1 172.7 Range 155-184 161-185 155-185 BMI (kg/ ^m2 ) Mean (SD) 28.9 (3.98) 26.2 (3.73) 27.5 (4.04) Median value 29.7 26.0 27.5 Range 21-35 21-35 21-35

BMI = body mass index

Treatment groups

Patients were assigned to receive one of the following 2 treatment sequences, each lasting 3 days, in a 1:1 ratio.

Subsequence Period 1 Period 2 1 2.5% compound I
4 times a day (every 6 hours) for 72 hours (inclusive) Carrier medium as a control
4 times a day (every 6 hours) for 72 hours (inclusive) 2 Carrier medium as a control
4 times a day (every 6 hours) for 72 hours (inclusive) 2.5% compound I
4 times a day (every 6 hours) for 72 hours (inclusive)

Compound I was administered to the patient as eye drops. The drops were administered at the study center by study staff on the day of PRK surgery. During daily postoperative follow-up visits, study staff administered the eye drops while the patient was at the study center at the expected time of eye drop administration. The remaining drops were administered by the patient at home or when the patient was at the study center for the visit (e.g., 24, 48, and 72 hours after surgery).

Visual analogue scale

Patients' subjective experience of pain was recorded using the VAS, a numeric pain rating scale ranging from 0 to 100, where 0 represents no pain and 100 represents the worst pain imaginable. Previous studies of pain in PRK have demonstrated that the most severe pain is experienced within the first 12 hours after surgery, peaking at approximately 4-6 hours after surgery (Sher et al., Refract Corneal Surg . Nov-Dec;9(6):425-36 (1993)), and these time points were chosen for the primary endpoint analysis. Because it was clinically important to reduce both maximum pain and total pain experienced by the patient during the immediate postoperative period, two periods of 6 hours and up to 12 hours after surgery were assessed as primary endpoints. All evaluable VAS data were collected using an ePRO, which was an electronic device (cell phone with a software application) on which patients reported their pain levels at relevant time points.

Oral rescue analgesics . It is unethical to withhold pain medications from patients after surgery in a clinical trial. A review of previous clinical trials indicated that NSAIDs are used for postoperative pain after PRK as oral rescue analgesics, similar to standard treatment after PRK. Because the use of an oral rescue medication may be a potential confounder for VAS pain assessments, three approaches were used to account for the influence of analgesic medications when analyzing VAS pain scores (assuming a 4-hour effect of the rescue medication): (1) any VAS scores recorded within 4 hours of rescue medication administration were considered missing; (2) all recorded VAS scores were used; and (3) any VAS scores recorded within 4 hours of rescue medication administration were replaced with entries made before the rescue medication administration.

Results of the primary analysis of effectiveness

The mean VAS pain severity scores at 6 hours and 0-12 hours postoperatively are presented in Table 4 and Table 5, respectively, and shown in Figure 1. The primary analysis was performed using the primary PD analysis set.

The differences between treatments for Compound I and vehicle in mean VAS pain severity scores at 6 hours and 0-12 hours postoperatively were statistically significant, with p-values less than 0.10.

The model-predicted mean difference between treatments (compound I-vehicle) in VAS pain severity scores at 6 hours postoperatively was -11.1 (90% CI: -17.54, -4.71; p=0.005) and at 0-12 hours postoperatively was -8.56 (90% CI: -14.29, -2.83; p=0.016). Thus, the primary objective of this study to assess efficacy was met.

The differences between treatments for Compound I and vehicle in mean VAS pain severity scores at 6 hours and 0-12 hours postoperatively were statistically significant, with p-values less than 0.10.

Table 4. Mean pain severity scores according to VAS at 6 hours after surgery (primary PD analysis sample)

The mean value estimated using the model ^* (SE) Comparison of mean values determined using the model: compound I and vehicle Compound I (N=30) ^# Carrier medium (N=29) Difference (compound I−carrier medium) % difference 90% CI P-value 34.63 (4.05) 45.76 (4.10) -11.1 -25% (−17.54, −4.71) 0.005

^* To account for the effect of oral rescue medication (ORM) use, any recorded VAS pain scores obtained within 4 hours of ORM administration were replaced with the VAS score recorded immediately before ORM administration, consistent with 2014 FDA guidance.

^# ePRO data for the first 10 patients (out of a total of 40) were not evaluable due to failure of the ePRO device from the first supplier

Table 5. Mean pain severity scores according to VAS for the period 0-12 hours after surgery (sample for primary PD analysis)

The mean value estimated using the model ^* (90% CI) Comparison of mean values determined using the model: compound I and vehicle Compound I (N=30) Carrier medium (N=29) Difference (compound I−carrier medium) % difference 90% CI P-value 30.90
(24.04, 37.76) 39.46
(32.54, 46.39) -8.56 -22% (−14.29, −2.83) 0.016

^* To account for the effect of oral rescue medication (ORM) use, any recorded VAS pain scores obtained within 4 hours of ORM administration were replaced with the VAS score recorded immediately before ORM administration, consistent with 2014 FDA guidance.

Results of the secondary analysis of effectiveness

The number of patients who did not use the oral rescue medication (ORM) was higher in the Compound I group than in the vehicle group during the 0-6, 0-12, and 0-24 hour postoperative periods. At 36 hours postoperatively, the number of patients in the Compound I group and the vehicle group using ORM was similar (Table 6).

Table 6. Summary of incidence of oral rescue medication use (number of patients who did not use oral rescue medication) (secondary PD analysis sample)

Time interval 2.5% compound I
N=40
n (%) Carrier medium
N=40
n (%) p-value 0-6 hours after surgery 23 (57.5%) 19 (47.5%) 0.2891 0-12 hours after surgery 19 (47.5%) 16 (40.0%) 0.5811 0-24 hours after surgery 16 (40.0%) 12 (30.0%) 0.3438 0-2 days after surgery 11 (27.5%) 11 (27.5%) 1,0000 0-3 days after surgery 11 (27.5%) 11 (27.5%) 1,0000

n: number of patients who did not use the oral rescue medication (ORM)

* In both columns of treatment sequence data, n represents the number of patients who did not use the rescue oral medication (ORM) in any treatment group.

The histogram of the incidence of oral rescue medication (ORM) use (number of patients who did not use ORM) is shown in Figure 2.

The summary information on the number of ORM (number of pills per patient) and its analysis are presented in Table 7.

Table 7. Summary of the quantity of oral refill medication (number of pills) (secondary PD analysis sample)

Time interval Statistical indicators Compound I
N=40 Carrier medium
N=40 % change (compound I−carrier medium)/
carrier environment 0-6 hours
after surgery Mean (SD) 0.75 (1.032) 1.00 (1.132) -25% Range 0.0-4.0 0.0-4.0 p-value 0.10* 0-12 hours
after surgery Mean (SD) 1.40 (1.780) 1.65 (1.847) -15% Range 0.0-6.0 0.0-6.0 p-value 0.26 0-24 hours
after surgery Mean (SD) 2.35 (2.751) 2.80 (3.006) -16% Range 0.0-10.0 0.0-12.0 p-value 0.05* 0-48 hours
after surgery Mean (SD) 4.05 (4.466) 4.68 (5.225) -13% Range 0.0-16.0 0.0-20.0 p-value 0.05* 0-72 hours
after surgery Mean (SD) 4.33 (4.896) 5.05 (5.574) -14% Range 0.0-21.0 0.0-22.0 p-value 0.07*

*: obtained based on the Wilcoxon signed-rank test

N=total number of patients in each treatment group

* : statistical significance at p ≤ 0.10 (according to the calculation of the statistical power of the primary endpoint)

As can be seen in Table 7 and Figure 2, at each time point during the study, less ORM was taken during Compound I treatment compared to vehicle treatment.

Summary information on the amount of ORM (mg/kg body weight) and its analysis are presented in Table 8.

At each time point, the amount of ORM (mg/kg body weight) was lower during treatment with compound I compared to treatment with vehicle (Table 8). During the 0-6, 0-24, 0-48, and 0-72 hour postoperative periods, the difference in milligrams per kilogram of body weight was statistically significant (p ≤ 0.10).

Table 8. Summary of the amount of rescue drug for oral administration (mg/kg body weight) (secondary PD analysis sample)

Time interval Statistical indicators Compound I
N=40 Carrier medium
N=40 % change (compound I−carrier medium)/
carrier environment 0-6 hours
after surgery Mean (SD) 3.32 (4.684) 4.41 (5.038) -25% Range 0.0-19.2 0.0-17.1 p-value 0.09* 0-12 hours
after surgery Mean (SD) 6.10 (8.062) 7.39 (8.478) -17% Range 0.0-28.7 0.0-28.7 p-value 0.21 0-24 hours
after surgery Mean (SD) 10.17 (12.335) 12.41 (14.130) -18% Range 0.0-47.9 0.0-57.5 p-value 0.05* 0-48 hours
after surgery Mean (SD) 17.59 (19.850) 20.51 (23.991) -14% Range 0.0-76.6 0.0-95.8 p-value 0.05* 0-72 hours
after surgery Mean (SD) 18.70 (21.509) 22.06 (25.527) -15% Range 0.0-87.4 0.0-105.4 p-value 0.06*

*: obtained based on the Wilcoxon signed-rank test

N=total number of patients in each treatment group

* : statistical significance at p ≤ 0.10 (according to the calculation of the statistical power of the primary endpoint)

Each acetaminophen/codeine tablet is considered equivalent to 330 mg when calculating the amount (mg/kg body weight).

Pain severity according to VAS during the first 3 days after PRK surgery

Following PRK surgery, patients reported statistically significant reductions in VAS pain severity scores after treatment with Compound I compared to vehicle at a p-value cutoff of 0.10 at 6 of 7 time points tested during the first 18 hours after surgery, beginning at 1 hour after surgery. VAS pain severity scores were lower for Compound I than for vehicle at all scheduled time points up to and including 36 hours after surgery. At all time points from 36 hours until the end of the VAS data collection period at 72 hours after PRK surgery, the difference between VAS scores for Compound I and VAS scores for vehicle was very small, and no statistically significant difference between scores was observed at a p-value cutoff of 0.10.

Pain severity scores according to VAS before and after instillation of the eye drops under study

The mean changes in VAS scores at 6.5, 18.5, and 24.5 hours postoperatively compared with the time points half an hour before and immediately before eye drop instillation (namely, at 6, 18, and 24 hours) were -3.1, 2.8, and 1.3, respectively, for eyes treated with compound I. For eyes treated with vehicle, the same differences were -5.6, 2.2, and -0.2, respectively.

Ocular Pain Assessment Questionnaire (OPAS)

OPAS is a validated tool for quantifying and monitoring corneal and ocular surface pain and quality of life, developed in response to a need identified at a 2010 National Eye Institute workshop. See Qazi et al.,OphthalmologyJuly 123(7):1458–1468 (2016). Qazi OPAS pain score for overall pain severityet al.The OPAS questionnaire included ratings from 0 (no pain) to 10 (severe pain) or from 0% (never) to 100% (all the time) for symptom frequency. Patients were asked to complete the OPAS questionnaire on day 2, day 4 (at the end of the study medication treatment period), and day 8. Statistical analysis of the OPAS results was not performed. Of the 27 questions in the OPAS, the results for seven questions are presented below.

For the questions about the intensity of eye pain over the previous 24 hours, for the level of eye pain that was most severe (Question 4), the level of pain that was least severe (Question 5), and the level of eye pain on average (Question 6) on Day 2, all 3 responses numerically indicated a benefit for Compound I compared to vehicle (Figure 3A, Figure 3B, Figure 3C).

For questions about how often eye pain was associated with the following symptoms: redness (question 22), burning (question 23), light sensitivity (question 24), and lacrimation (question 25), all responses favored compound I over vehicle (Figures 4A, 4B, 4C, and 4D). Thus, patients administered compound I exhibited lower levels of eye pain associated with redness, burning, light sensitivity (photophobia), and lacrimation compared to patients administered placebo.

Summary of the results of the eye study

The Ocular Pain Assessment Questionnaire (OPAS) demonstrated better pain relief and quality of life in patients during the treatment period with Compound I compared to vehicle.

For the questions about the intensity of eye pain over the previous 24 hours, for the level of eye pain that was most severe (Question 4), the level of pain that was least severe (Question 5), and the level of eye pain on average (Question 6) on Day 2, all 3 responses indicated a benefit for Compound I compared to vehicle (Figures 3A-3C).

For questions about how often eye pain was associated with the following symptoms: redness (question 22), burning (question 23), sensitivity to light (question 24), and lacrimation (question 25), all responses favored compound I over vehicle (Figures 4A-4D).

The VAS scores at 6.5, 18.5, and 24.5 hours postoperatively compared with the time points half an hour before and immediately before eye drop instillation (namely, 6, 18, and 24 hours) were -3.1, 2.8, and 1.3, respectively, for eyes treated with compound I. For eyes treated with vehicle, the same differences were -5.6, 2.2, and -0.2, respectively.

Pharmacokinetic parameters

Pharmacokinetic parameters assessments

Pharmacokinetic (PK) samples were collected at the time points defined in the visit schedule above, at the PK blood sampling time points shown in Table 9.

Table 9. Acceptable time interval for blood sampling for PK assessment

Before dosing After administration of the dose Within 90 minutes before dosing (samples number 101, 106, 107, 112, 117 and 118) Within ±5 min from the expected time point relative to dose administration, up to 2 hours after dose administration
Within ± 5 min from the time point 72.25-74 hours (0.25-2 hours relative to the time frame of administration of the last dose)

All blood samples (3 ml) were collected from the arm via direct venipuncture or via an indwelling catheter inserted into a forearm vein. After blood collection, each tube was immediately and gently inverted 8-10 times to ensure mixing of the tube contents with the anticoagulant (3 ml K2-EDTA). Prolonged contact of the sample with the rubber stopper was avoided, and the tubes were placed upright in a test tube rack surrounded by liquid ice until centrifugation.

Within 30 minutes, the sample was centrifuged at approximately 5°C for 10 minutes at approximately 2000 g (or the sample was placed on ice and centrifuged at room temperature). Immediately after centrifugation, the entire supernatant (approximately 1.5 ml) was transferred to the first 1.8 ml NUNC cryovial with a 2D barcode. After thorough mixing of the blood plasma, half of the blood plasma was transferred from the first cryovial to the second cryovial, and both were securely capped. Appropriate PK cryofreezing labels were attached to each cryovial, and the labels were securely secured with transparent adhesive tape. Cryovials were immediately frozen over dry ice, then maintained frozen at ≤ -20˚C until shipment to the central laboratory. Vials were shipped in batches once every two weeks.

Compound I was quantified in plasma using a validated LC-MS/MS method; the lower limit of quantification (LLOQ) was 0.05 ng/mL. Concentrations were expressed as ng/mL. When possible, bandage contact lenses (BCL) exposed to study drug were collected and analyzed for residual drug exposure after treatment (LLOQ: 5.00 ng/mL in 0.55 mL extraction fluid or 2.75 ng/BCL). Concentrations below the LLOQ were reported as "zero," and missing data were similarly noted.

The following PK parameters were determined to be relevant using actual recorded sampling time points and the non-compartmental method using Phoenix™ ^WinNonlin® (version 6.4): Cmax, Tmax, AUClast (calculated), Clast, and Tlast from plasma concentration-time data. Predose concentrations were determined by observation at the specified time points of 0, 24, and 72 hours. The linear trapezoidal rule was used to calculate AUClast. PK parameters were not calculated from the concentration data for compound I in bandage contact lenses.

Pharmacokinetic parameters of compound I in blood plasma

The mean arithmetic values for the plasma concentration-time profiles and PK parameters of compound I are presented in Figure 5A and Figure 5B, respectively, and in Table 10.

Following unilateral topical ocular dosing of Compound I, absorption into the systemic circulation was rapid, with median Tmax of 0.459 h and 0.467 h after the first (day 1; range 0.167-2.00 h) and 13th (day 4; range 0.00-2.08) doses, respectively. Cmax was determined and AUClast was calculated for 34/40 patients on day 1 and for 33/40 (Cmax) or 31/40 (AUClast) patients on day 4. All pre-dose day 1 concentrations were below the limit of quantification and were substituted with values of 0.00 ng/mL. The residual (at the end of the 6-hour interdose interval and before the next dose) mean concentration (% CV) before the 5th dose on day 2 (24 hours after the 1st dose; time point 0 hour on day 2) was 1.25 ng/mL (54.9%) and was slightly higher, at 1.99 ng/mL (76.3%), before the 13th dose on day 4 (72 hours after the 1st dose; time point 0 hour on day 4).

Concentrations were generally low, with observed Cmax ranging from 0.195 to 7.56 ng/mL and AUClast ranging from 0.261 to 14.5 ng*hr/mL, across all 4 days of repeated QID dosing. The mean arithmetic mean Cmax (% CV) after the first dose on day 1 was 0.454 ng/mL (49.9%) and 2.40 ng/mL (63.5%), a 5.3-fold increase, after the 13th dose on day 4. The corresponding mean AUClast (% CV) values were 0.638 ng*hr/mL (46.0%) and 4.38 ng*hr/mL (67.0%), representing a 6.9-fold increase over 4 days.

Table 10. Summary statistics of PK parameters of compound I after topical ocular administration of 2.5% compound I (PK analysis sample)

A day in profile Statistical indicators Cmax (ng/ml) Tmax (h) AUClast (ng*h/ml) ¹ 1 N 34 34 34 Mean (SD) 0.454 (0.227) - 0.638 (0.293) % CV 49.9 - 46.0 Median value 0.375 0.459 0.569 Minimum, maximum 0.195, 1.05 0.167, 2.00 0.261, 1.58 4 N 33 33 31 Mean (SD) 2.40 (1.53) - 4.38 (2.94) % CV 63.5 - 67.0 Median value 2.07 0.467 3.42 Minimum, maximum 0.639, 7.56 0.00, 2.08 1.18, 14.5

¹ Median Tlast (range): day 1-2.00 h (1.95-2.05 h); day 4-2.00 h (1.77-2.10 h)

Clast not shown.

Bandage contact lens

Bandage contact lenses (BCL) were collected from 40 patients on day 4 of dosing and analyzed for compound I. Values from three patients who were noncompliant with IMP treatment were excluded from the BCL summary statistics, leaving 37 evaluable values, including one value from a patient for whom plasma PK data were unavailable. The mean compound I concentration (% CV) in 0.55 mL of extraction fluid was 8500 ng/mL (73.1%), with a wide range of 284 to 22600 ng/mL. These concentrations translate to an estimated mean value of 4680 ng/lens (0.55 mL × 8500 ng/mL) and a range of 156–12400 ng/lens. The amount of BCL was very small relative to the administered dose of 0.925 mg of compound I: a mean of 0.51% and a range of 0.017% to 1.3% of the dose. This indicates that BCL absorbs and retains compound I for topical ocular application, but in negligible quantities compared to the topical ocular dose.

Summary results of pharmacokinetic analysis

Absorption of compound I into the systemic circulation was rapid (median plasma Tmax was approximately 0.5 hour) after both single (first dose) and repeated topical ocular administration 4 times daily.

Following both single and multiple topical ocular dosing of Compound I, systemic exposure was low, ranging from 0.195 ng/mL to 7.56 ng/mL across all 4 days of repeated QID dosing, and demonstrated moderate variability. The mean Cmax was 0.454 ng/mL after the first dose on day 1 and 2.40 ng/mL after the ^13th dose on day 4.

The % CV values for Cmax and AUClast after a single dose were 49.9% and 46.0%, respectively. The corresponding values after repeated administration were 63.5% and 67.0%.

The mean Cmax after repeated administration (2.40 ng/mL) was 5.3-fold higher than after a single dose on day 1 (0.454 ng/mL). Similarly, the mean AUClast was 6.9-fold higher after repeated administration compared with a single dose (4.38 vs. 0.638 ng*h/mL). These data indicated accumulation over the 4-day period of dosing 2.5% of compound I four times daily.

Bandage contact lenses (BCL) were collected from 40 patients on day 4 of dosing. The mean concentration (% CV) in 0.55 mL of extraction fluid was 8500 ng/mL (73.1%), which translates to 4680 ng/lens. The amount of BCL was very small relative to the administered dose of 0.925 mg of compound I: 0.51% and a range of 0.017% to 1.3% of the dose. This indicates that BCL absorbs and retains compound I for topical ocular application, but in negligible quantities compared to the topical ocular dose.

Safety

Safety assessments included collecting information on all AEs and SAEs, along with their severity and relationship to the study drug. Table 11 includes a list of signs and symptoms common after PRK surgery. Only signs and symptoms with severity or duration exceeding the listed ranges were reported as AEs.

Table 11. Signs and symptoms after PRK surgery that do not require reporting

Sign/symptom Expected severity Expected survival after PRK surgery Corneal epithelial defect Mild to moderate 1 week Swollen eye Mild to moderate 1 week Eyelid edema Mild to moderate 1 week Punctate keratitis Mild to moderate 1 week Conjunctival hyperemia Mild to moderate 1 week A decrease in BCVA of 10 letters or more from baseline Mild to moderate 1 month Eye pain Mild to moderate 1 week Eye irritation/eye discomfort Mild to moderate 1 month Foreign body sensation Mild to moderate 1 month Eyelid margin irritation/crusting of the eyelid margins Mild to moderate 1 month Increased tear production Mild to moderate 1 month Blurred vision Mild to moderate 1 month Blurred vision Mild to moderate 1 month Blurred vision Mild to moderate 1 month Photophobia Mild to moderate 1 month Dry eyes Mild to moderate 1 month Blindness Mild to moderate 1 month Halos Mild to moderate 1 month

One week indicates resolution by postoperative visit day 8 (visit 6 in treatment period 1 or visit 11 in period 2), including the allowable time interval. The table was compiled prior to the study and incorporated comments and suggestions from all principal investigators and surgeons based on their combined experience with common and expected clinical manifestations after standard PRK surgery.

One month indicates resolution by the EOS visit or within 37 days of surgery in cases where an EOS visit was not performed or where more than one week elapsed between study surgeries.

The assessment of adverse events and serious adverse events included both systemic and ocular assessments. Systemic safety assessments included regular assessments of height and weight, vital signs, medication error reports, including misuse/abuse, pregnancy reports, and early safety monitoring. In this study, misuse/abuse, pregnancy reports, and early safety monitoring were not assessed. Ocular safety assessments included the following:

Best-corrected visual acuity (BCVA) and uncorrected visual acuity (UCVA) were measured at each visit using the ETDRS visual acuity chart at 4 meters (13 feet) or at 1 meter (for patients who could not read the chart at 4 meters). BCVA was scored based on the number of letters read correctly plus 30. If visual acuity was so poor that the patient could not read any of the larger letters at 1 meter, the ability to count fingers, see hand movements, and perceive light were tested.

Intraocular pressure (IOP): IOP measurements were performed using applanation tonometry or Tono-Pen.

Dilated fundus examination: Dilated fundus examination included ophthalmoscopic assessment of the vitreous, retina/macula/choroid, and optic nerve. Retinal tears/detachments, retinal hemorrhages, vitreous hemorrhage density, vitreous opacification stage, and any abnormalities detected were assessed and scored according to the classification criteria.

Ocular hyperemia: Conjunctival redness of the bulbar conjunctiva in each eye was graded using the McMonney Redness Scale using a slit lamp. Hyperemia was assessed in four areas (superior, inferior, temporal, and nasal) of each eye, and severity was rated from 0 to 5 for each area.

Epithelial defect size as assessed by slit lamp examination: Using oblique observation relative to the light source and the light beam from a narrow slit, the cornea was sectioned to visualize the vertical and horizontal boundaries of the corneal wound. By adjusting the height and width of the light beam from a calibrated slit, the maximum horizontal (wound width) and vertical (wound height) dimensions of the epithelial surgical wound were estimated. This assessment was performed before wound closure and reporting of the wound size, corresponding to 0 in the horizontal and 0 in the vertical dimensions.

Slit-lamp biomicroscopy: Ocular features (eyelids/conjunctiva, cornea, lens, and iris/anterior chamber) were assessed in both eyes by slit-lamp biomicroscopy according to the classification criteria.

Blink rate: A blink was defined as a bilateral paroxysmal eyelid closure (lasting <1 second) in the absence of external provoking stimuli. Each blink rate assessment lasted for 2 minutes (or as close to 2 minutes as possible, with a minimum time of 1 minute), and the resulting blink rates were averaged to calculate the blink rate in blinks/min.

Lacrimal fluid production (Schirmer's test without anesthesia): The test was performed without anesthesia on both eyes simultaneously. Lacrimal fluid secretion was measured in millimeters along a tear strip moistened with tear fluid. Measurements were made to the nearest whole number.

Corneal staining: This test was performed by gently touching the moistened end of a sodium fluorescein strip to the inferior conjunctival sac. The strip was moistened with 1 drop of sterile saline and shaken with a gentle flick to remove excess saline. The patient blinked several times to ensure distribution of the dye throughout the tear film before rating on a scale of 0 to 3 (0 = normal, no staining; 1 = mild, superficial granular staining of tiny dots; 2 = moderate, staining of large dots with a few connecting areas; and 3 = severe, numerous connecting areas and/or large dotted spots) for each of 5 zones (central and 4 quadrants).

Safety assessment

There were no deaths or SAEs, no discontinuations, or no study withdrawals due to AEs in this study. A total of 18 AEs were reported in 10 patients (25% of the 40 patients enrolled in the study), of which 12 occurred after treatment with Compound I, five after treatment with vehicle, and one occurred in one patient before study drug administration. All AEs were mild or moderate in severity.

A total of 10 patients (25%) experienced at least one AE that occurred after treatment initiation—five patients experienced one AE, and five patients experienced more than one AE. Five patients experienced AEs only during treatment with compound I, two patients experienced AEs only during treatment with vehicle, and three patients experienced AEs during treatment with both compound I and vehicle. One AE (headache) was observed, which occurred before the first dose of study drug.

Six ocular AEs were observed in four patients (10% of the 40 patients included in the study), all of which were mild in severity (in three eyes, each treated with compound I and vehicle). None of the AEs were suspected to be related to either study drug (compound I or vehicle). The investigator believed that five of the six ocular AEs were related to the PRK procedure (two in eyes treated with compound I and three in eyes treated with vehicle).

Twelve non-ocular AEs were observed in seven patients (17.5%) (six eyes were treated with compound I, two eyes were treated with vehicle, and in one patient, these occurred during the drug-free period). Five AEs in four patients (10%) were moderate in severity (four eyes were treated with compound I and one eye was treated with vehicle). The remaining AEs were mild in severity. In the investigator's opinion, none of the AEs were suspected to be related to compound I.

Table 12 shows the overall frequency of adverse events.

Table 12. Overall AE incidence – number of events and number of patients (safety analysis sample)

2.5% compound I
N=40
n (%) Carrier medium
N=40
n (%) Without taking medication*
N=40
n (%) Total
N=40
n (%) Patients with at least one AE 8 (20) 5 (12.5) 1 (2.5) 10 (25) Number AE 12 5 1 18 Patients with at least one ocular AE 3 (7.5) 3 (7.5) 0 4 (10) Number of eye AEs 3 3 0 6 Patients with at least one non-ocular AE 6 (15) 2 (5) 1 (2.5) 7 (17.5) Number of non-ocular AEs 9 2 1 12 Mild AE number 8 4 1 13 Number of moderate AEs 4 1 0 5 Number of severe AEs 0 0 0 0 Number of AEs associated with the procedure 2 3 0 5 Number of drug-related AEs 0 0 0 0

An AE that started in one period and continued into the next was counted only in the period in which it began to appear.

N=number of patients participating in the study.

*Without drug administration or AEs occurring at baseline: AEs occurring after informed consent was signed and before administration of any study drug (compound I or vehicle).

Eye safety ratings

Best-corrected visual acuity (BCVA) and uncorrected visual acuity (UCVA): Compound I administration did not result in any trend or significant difference between eyes treated with Compound I and vehicle throughout the study. Thus, these data indicate that Compound I reduces pain intensity without adversely affecting BCVA and UCVA.

Intraocular pressure (IOP): No trend was observed across all planned time points or significant differences between eyes treated with Compound I and vehicle throughout the study. At the end of the study, a slight increase (< 5 mmHg) in mean IOP values compared to baseline was observed. The small change in mean scores over the course of the study was not clinically significant.

Dilated fundus examination: At all scheduled time points, the investigator reported no abnormal findings in the eyes treated with compound I and vehicle.

Ocular hyperemia: A bar graph of the intensity of ocular hyperemia over time for eyes treated with Compound I and placebo is shown in Figures 6A and 6B, respectively. Figures 6A and 6B indicate grade 4 and grade 3 ocular hyperemia (measured by the McMonney scale) on days 3 and 4, respectively. On postoperative day 2 (24 hours after PRK surgery), eyes treated with Compound I had less grade 4 hyperemia (all quadrants) compared to eyes treated with vehicle. In the superior quadrant, the p-value for this difference was 0.04. On day 3 (48 hours after surgery), eyes treated with Compound I had less grade 3 hyperemia compared to eyes treated with vehicle. None of these observations were recorded as AEs.

Epithelial defect size as assessed by slit lamp examination: To evaluate the rate of wound healing following compound I administration, epithelial defect size was measured in both the compound I-treated and vehicle-treated populations. The elliptical area for epithelial wound size was calculated as follows: area ( ^mm2 ) = width × height × π, using the width and height of the epithelial defect measured by slit lamp examination. The difference in epithelial wound area between compound I and vehicle was not significant at any time point except day 2 after PRK surgery (all p-values for area differences were > 0.35). On day 2 (24 hours after surgery), the difference in mean epithelial defect area between patients treated with compound I versus vehicle was 11.23 ^mm2 (p-value=0.034). This difference was not clinically significant in the immediate postoperative period. By day 3 (48 hours after surgery), no difference was observed between eyes treated with compound I and those treated with vehicle, and the wound area was very small. On day 4 (48 hours after surgery), almost all eyes had healed, and no difference was observed between eyes treated with compound I and those treated with vehicle. Compound I did not demonstrate a delay in wound healing compared to vehicle.

Figure 7 and Table 13 show a comparison of the epithelial defect size in eyes treated with Compound I and eyes treated with vehicle.

Table 13. Statistical analysis of epithelial defect size (safety analysis sample)

Parameter (unit of measurement): epithelial wound size (mm)
Subcategory: area (mm ² ) Raw mean (SD) Visit Compound I Carrier medium Difference,
I-medium-carrier compound p-value* Day 1 220.15 (32.26) 217.16 (34.72) 2.98 (27.33) 0.494 Day 2 83.02 (32.08) 71.79 (26.96) 11.23 (32.30) 0.034 Day 3 6.75 (12.66) 5.73 (9.73) 1.02 (8.78) 0.467 Day 4 0.39 (2.04) 0.08 (0.50) 0.31 (2.11) 0.352 Day 8 0.00 (0.00) 0.00 (0.00) 0.00 (0.00)

*Based on a paired t-test

The size of the epithelial defect was calculated according to the slit lamp examination as the area of the ellipse calculated by the maximum horizontal and vertical distance from the center of the epithelial defect.

Area ( ^mm2 ) = width*height*π

Slit-lamp biomicroscopy: Slit-lamp biomicroscopy included examination of ocular structures (eyelids/conjunctiva, cornea, iris/anterior chamber, lens, aqueous opacities, and aqueous inflammatory cell abundance). No ocular structural abnormalities were observed in aqueous opacities, aqueous inflammatory cell abundance, iris/anterior chamber, and lens in patients treated with either Compound I or vehicle at all scheduled time points.

Blink rate: Fluctuations in mean blink rate were observed; however, no trend was observed across all planned time points or clear differences between blink rate after treatment with Compound I and vehicle. At the end of the study, there were no clinically significant changes in blink rate compared to baseline.

Corneal staining: Most patients had normal staining (grade 0) measured at baseline, Day 8, and EOS visits. No clinically significant differences in corneal staining were observed between eyes treated with Compound I versus vehicle at any of the baseline, Day 8, or EOS visits.

Vital Signs, Physical Examination Results, and Other Safety Observations: Vital sign parameters (systolic and diastolic blood pressure, pulse, and body temperature) were within the normal range in all patients during the study. No AEs related to vital signs were observed. Electrocardiograms and special safety considerations were not performed or evaluated (as per protocol).

Summary of safety analysis results

There were no deaths or serious or severe AEs, treatment discontinuations, or study withdrawals reported in this study.

No AEs were assessed as related to the study drug (compound I or vehicle). Eight of 40 patients and five of 40 patients developed AEs following treatment with compound I and vehicle, respectively. All ocular AEs were mild and evenly distributed between compound I and vehicle, with the vast majority being well-known AEs associated with the PRK procedure.

There were no clinically significant differences in safety observed between Compound I and vehicle.

Compound I did not exhibit delayed wound healing compared to vehicle.

Fewer eyes treated with Compound I than with vehicle-treated eyes demonstrated severe conjunctival hyperemia on day 2 (24 hours postoperatively).

There were no clinically significant changes observed in BCVA, IOP, slit-lamp biomicroscopy, dilated ophthalmoscopy, blink rate, tear production, corneal staining, or vital signs following Compound I administration compared to vehicle.

The frequency of occurrence of AEs by preferred terms is presented in Table 14.

Table 14. Incidence of AEs by Preferred Terms - n (percentage) of Patients (Safety Analysis Set)

2.5% compound I
N=40
n (%) Carrier medium
N=40
n (%) Without taking the drug (before administration of the drug)*
N=40
n (%) Total
N=40
n (%) AE by preferred terms n (%) Heaviness Related to the procedure n (%) Heaviness Related to the procedure n (%) Heaviness Related to the procedure Patients with at least one AE 8 (20.0) 5 (12.5) 1 (2.5) 10 (25) Eye AE Corneal infiltrate 0 1 (2.5) Easy Yes 0 1 (2.5) Corneal opacity 1 (2.5) Easy Yes 1 (2.5) Easy Yes 0 1 (2.5) Punctate keratitis 1 (2.5) Easy Yes 1 (2.5) Easy Yes 0 1 (2.5) Posterior vitreous detachment 1 (2.5) Easy No 0 0 1 (2.5) Non-ocular AE Headache 1 (2.5) Easy No 1 (2.5) Moderate No 1 (2.5) Easy No 3 (7.5) Arthralgia 1 (2.5) Easy No 0 0 1 (2.5) Nasopharyngitis 2 (5.0) Easy, easy No 0 0 2 (5.0) Pain in the oropharynx 1 (2.5) Easy No 0 0 1 (2.5) Tinnitus 0 1 (2.5) Easy No 0 1 (2.5) Sinus congestion 1 (2.5) Moderate No 0 0 1 (2.5) Sinusitis 1 (2.5) Moderate No 0 0 1 (2.5) Fever 1 (2.5) Moderate No 0 0 1 (2.5) Vomit 1 (2.5) Moderate No 0 0 1 (2.5)

An adverse event that begins in one period and continues into the next is counted only in the period in which it begins.

N=number of patients participating in the study.

n=number of patients with at least one AE in a given category.

*Without drug administration or AEs occurring at baseline: AEs occurring after informed consent was signed and before administration of any study drug (compound I or vehicle).

All publications and patent documents cited in this document are herein incorporated by reference as if each such publication or document were specifically and individually indicated to be incorporated by reference. The present invention and its embodiments have been described in detail. However, specific embodiments of any process, manufacturing procedure, composition, compounds, means, methods and/or steps described in the present specification are not intended to limit the scope of the present invention. Various modifications, substitutions and variations can be made with respect to the disclosed material without departing from the spirit and/or essential characteristics of the present invention. Accordingly, one of ordinary skill in the art, upon familiarization with the present invention, will readily understand that later modifications, substitutions and/or variations that perform substantially the same function or achieve substantially the same result as the embodiments disclosed herein can be used in accordance with such related embodiments of the present invention. Thus, the following claims are intended to cover, within their scope, modifications, substitutions, and alterations of the processes, manufacturing procedures, compositions, compounds, means, methods, and/or steps disclosed herein. The claims should not be construed as limited to the order or elements described unless expressly stated. It should be understood that various changes in form and detail may be made without departing from the scope of the appended claims.

Claims (21)

1. A method of treating ocular surface pain in a subject in need thereof, comprising ocularly administering to the subject an effective amount of 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (formula I) having the structure: Formula I, or its pharmaceutically acceptable salt. 2. The method according to claim 1, comprising administering a compound of formula I at a concentration of from 0.5% w/v to 3.5% w/v, from 0.5% w/v to 2.5% w/v, or from 0.5% w/v to 1.5% w/v, from 0.5% to 3.0% w/v, from 1.0% to 2.5% w/v, from 1.5% to 3.0% w/v, or from 0.5% to 2.5% w/v. 3. The method of claim 1 or 2, wherein the subject suffers from one or more of dry eye disease, Sjogren's syndrome, conjunctivitis, including keratoconjunctivitis, vernal keratoconjunctivitis, allergic conjunctivitis, punctate map-like fingerprint dystrophy, acanthamoebiasis, fibromyalgia, meibomian dysfunction, thyroid ophthalmopathy, acne rosacea, ptosis, keratoconus, ocular pain syndrome, Stevens-Johnson syndrome, forms of corneal epitheliopathy, forms of corneal neuropathy, including forms of LASIK-induced corneal neuropathy, forms of corneal dystrophy, including forms of recurrent corneal dystrophy, epithelial basement membrane dystrophy, forms of erosion corneal abrasions or abrasions, including recurrent forms of corneal abrasions or abrasions, ocular surface diseases, blepharitis, graft-versus-host disease, meibomitis, glaucoma, conjunctivochalasis, forms of keratopathy, including herpetic keratopathy, filiform keratopathy, band or bullous keratopathy, exposure keratopathy, keratitis, including keratitis caused by the herpes simplex virus, iritis, episcleritis, corneal surgery, multiple sclerosis, trichiasis, pterygium, neuralgia, xerophthalmia, refers to patients who are recovering from neurotrophic keratitis or have ocular pain that persists for at least three months after photorefractive surgery keratectomy (PRK) or laser in situ keratomileusis (LASIK) surgery. 4. The method of any one of the preceding claims, wherein the administration results in a reduction in pain score by at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 compared to placebo when measured using a visual analogue scale (VAS). 5. The method of any one of the preceding claims, wherein the administration results in a reduction in the subject's pain score by at least 6, at least 7, at least 8, at least 9, or at least 10 compared to placebo when measured using a VAS. 6. The method of any one of claims 1-3, wherein the administration results in a reduction in pain intensity in the subject by at least 10%, at least 15%, at least 20%, or at least 25% compared to placebo. 7. The method of claim 4 or 5, wherein the reduction in pain score results from a difference in pain scores before and after administration of compound I to the subject. 8. The method according to any one of claims 4-7, wherein the reduction in the VAS score occurs within half an hour after administration of compound I to the subject. 9. The method according to any one of the preceding paragraphs, wherein the administration results in a decrease in the intensity of hyperemia in the subject by at least 1, at least 2, at least 3, at least 4, or at least 5 on the McMonney scale. 10. The method according to any one of the preceding claims, wherein the compound of formula I is administered for at least one, at least two, or at least three months. 11. The method according to any of the preceding claims, wherein the compound of formula I is administered for more than 3 months. 12. The method according to any one of the preceding claims, wherein the compound of formula I is administered from one to four times per day. 13. A method for reducing the intensity of ocular surface pain in a subject in need thereof, comprising ocularly administering to the subject 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (formula I) having the structure: Formula I, or a pharmaceutically acceptable salt thereof; wherein the compound of formula I is administered at a concentration of from 0.5% w/v to 3.5% w/v, from 0.5% w/v to 2.5% w/v, or from 0.5% w/v to 1.5% w/v, from 0.5% to 3.0% w/v, from 1.0% to 2.5% w/v, from 1.5% to 3.0% w/v, or from 0.5% to 2.5% w/v. 14. A method for reducing the intensity of ocular hyperemia in a subject in need thereof, comprising ocularly administering to the subject 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (formula I) having the structure: Formula I, or a pharmaceutically acceptable salt thereof; wherein the compound of formula I is administered at a concentration of from 0.5% w/v to 3.5% w/v, from 0.5% w/v to 2.5% w/v, or from 0.5% w/v to 1.5% w/v, from 0.5% to 3.0% w/v, from 1.0% to 2.5% w/v, from 1.5% to 3.0% w/v, or from 0.5% to 2.5% w/v. 15. The method according to claim 14, wherein the reduction in the intensity of ocular hyperemia is at least 1, at least 2, at least 3, at least 4, or at least 5 on the McMonney scale.