Compositions, methods, and preparations for ophthalmic applications

Abstract

This document relates to compositions, methods, and preparations assessing and / or treating subjects (e.g., humans) with ophthalmic disease, disorders, and conditions. For example, methods and compositions disclosed herein include an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine that is suitable for eyes.

Description

[0001] COMPOSITIONS, METHODS, AND PREPARATIONS FOR OPHTHALMIC APPLICATIONS

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to: U.S. Patent Application Serial No. 63 / 604,809, filed November 30, 2023; U.S. Patent Application Serial No. 63 / 604,817, filed November 30, 2023; and U.S. Patent Application Serial No. 63 / 604,819, filed November 30, 2023; each of which is herein incorporated by reference in its entirety.

[0004] TECHNICAL FIELD

[0005] The present disclosure belongs to the technical field of ophthalmic drugs, and specifically relates to a peptide and a pharmaceutical composition containing the same, and its ophthalmic application in preparing ophthalmic drugs for alleviating, improving and / or treating ophthalmic diseases and symptoms associated with ophthalmic diseases.

[0006] BACKGROUND

[0007] Ophthalmic diseases, disorders, and conditions can include symptoms or discomfort related to the cornea, conjunctiva, and tear film. Ophthalmic diseases, disorders, and conditions include but are not limited to infectious ocular diseases caused by bacteria (e.g., chlamydia sp ), viruses, fungus and other pathogens, non-infectious ocular diseases include ocular surface diseases dry eye, allergic keratoconjunctivitis, autoimmune or inflammatory ocular diseases, ocular surgery and trauma, and ocular surface (e.g., anterior and posterior) injuries.

[0008] SUMMARY

[0009] Treatment of ophthalmic diseases, disorders, and conditions can include lifestyle management and medicaments. Disclosed herein are ophthalmic pharmaceutical compositions comprising a peptide (e.g., a dipeptide) L-Alanyl-L-Glutamine, its methods of preparation, methods of treatment, and methods of use for treatment, mitigation, ameliorating the symptoms of ophthalmic diseases, disorders, and conditions, including ocular surface symptoms.

[0010] A dipeptide, like L-Alanyl-L-Glutamine, is a compound formed by two amino acids: alanine and glutamine connected by a peptide bond. This structure is fundamentally different from that of a single amino acid. When applied to complex structures such as the eye, these differences are impactful. The ocular environment is a complex system with specific structural considerations, such as the blood-eye barrier, which affect how compounds are delivered and interact with eye tissues. A dipeptide and an individual amino acid will interact differently with these structures due to their distinct properties.

[0011] For instance, L-Alanyl-L-Glutamine has different solubility, polarity, charge distribution, and metabolic pathways compared to either alanine or glutamine alone. Its larger size and greater molecular weight influence how it is absorbed, distributed, metabolized, and excreted by the body. These factors collectively affect the dipeptide's bioavailability and how it distributes within ocular tissues.

[0012] In contrast, individual amino acids, due to their smaller size and different chemical properties, will interact differently with ocular tissues and are likely not produce the same pharmacological effects as a dipeptide. When studying the pharmaceutical activity of peptides like L-Alanyl-L-Glutamine, especially in complex environments like the eye, their behavior cannot be assumed to be the same as that of individual amino acids. Each has unique characteristics and interacts differently with biological systems, making a separate and specific approach necessary for their study.

[0013] One aspect of this document features compositions, methods, and preparations of ophthalmic pharmaceutical compositions comprising a peptide (e.g., a dipeptide) L-Alanyl-L- Glutamine. In some example embodiments, compositions can include, or consist essentially of, an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine that is suitable for eyes. In some embodiments, the ophthalmic composition is in the form of an eye drop, suspension, ointment, emulsion, eye patch, eye mask, eye film, eye cream, spray, gel, injection, or implant. In some embodiments, the concentration of L-Alanyl-L-Glutamine is about 0.2% to about 0.8% (w / v) of the composition. In some embodiments, the concentration of L-Alanyl-L-Glutamine is about 1.8% to about 2.2% (w / v) of the composition. In some embodiments, the concentration of L-Alanyl-L-Glutamine is about 0.5% (w / v) in the composition. In some embodiments, the concentration of L-Alanyl-L-Glutamine is about 1.0% (w / v) of the composition. In some embodiments, the concentration of L-AlanyLL- Glutamine is about 2.0% (w / v) of the composition.

[0014] In some embodiments, the pharmaceutical composition further comprises a bacteriostatic or antimicrobial agent. For example, the bacteriostatic or antimicrobial agent includes one or more of benzalkonium chloride, benzalkonium bromide, chlorhexidine acetate, chlorhexidine gluconate, chlorobutanol, phenoxyethyl alcohol, methyl hydroxybenzoate, ethyl hydroxybenzoate, propyl hydroxybenzoate, and combinations thereof. For example, the concentration of the bacteriostatic or antimicrobial agent is about 0.003 to about 0.5% (w / v) of the composition. In some embodiments, the pharmaceutical composition further comprises a viscosity modifying agent. For example, the viscosity modifying agent includes one or more of sodium hyaluronate, sodium carboxymethyl cellulose, methyl cellulose, polyethylene glycol, polyvinyl alcohol, povidone, and combinations thereof. For example, the concentration of the viscosity modifying agent is about 0.01% to about 0.5% (w / v) of the composition.

[0015] In some embodiments, the pharmaceutical composition further comprises one or more pH regulators. For example, the pH regulator includes one or more of sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, boric acid, borax, acetic acid, sodium acetate, citric acid, sodium citrate, tartaric acid, sodium tartrate, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, sodium hydroxide, potassium hydroxide, hydrochloric acid, phosphoric acid, and combinations thereof. For example, the pH of the pharmaceutical composition is about 5.0 to about 9.0.

[0016] In some example embodiments, methods provided herein can include, or consist essentially of, preparing the pharmaceutical compositions disclosed herein, wherein the method comprising suspending or dissolving L-Alanyl-L-Glutamine in water to form an aqueous solution; adjusting the pH to about 5.0 to about 9.0; and filtering to sterilize the resulting solution with a microporous filter membrane. In another aspect, methods provided herein can include, or consist essentially of, decreasing, alleviating, mitigating, treating ocular surface symptoms and discomforts in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0017] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by ocular surface diseases in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0018] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by non-ocular surface diseases in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0019] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by dry eye in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one any one of the pharmaceutical compositions disclosed herein.

[0020] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by allergies in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of any one of the pharmaceutical compositions disclosed herein.

[0021] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by infections in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of any one of the pharmaceutical compositions disclosed herein.

[0022] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by neuropathic conditions in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0023] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by injury or surgery in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one any one of the pharmaceutical compositions disclosed herein.

[0024] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by foreign body wear in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one any one of the pharmaceutical compositions disclosed herein.

[0025] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by immune mediated diseases or inflammatory conditions in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0026] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by diabetes in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of any one of the pharmaceutical compositions disclosed herein.

[0027] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by surgical-related factors in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0028] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating ocular surface symptoms caused by meibomian glands disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions described herein. For example, the ocular surface symptoms described herein can include one or more of feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the corneal epithelium, density reduction of conjunctival goblet cells, squamous metaplasia of the ocular surface epithelium, and / or ocular surface inflammation.

[0029] In some aspects, methods provided herein can include, or consist essentially of, relieving dry eye symptoms in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0030] In some aspects, methods provided herein can include, or consist essentially of, improving dry eye symptoms in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0031] In some aspects, methods provided herein can include, or consist essentially of, treating dry eye symptoms in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0032] In some aspects, methods provided herein can include, or consist essentially of, decreasing ocular surface damage in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0033] In some aspects, methods provided herein can include, or consist essentially of, decreasing cornea epithelial damage in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0034] In some aspects, methods provided herein can include, or consist essentially of, restoring ocular surface health in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0035] In some aspects, methods provided herein can include, or consist essentially of, restoring conjunctival epithelial condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0036] In some aspects, methods provided herein can include, or consist essentially of, decreasing conjunctival hyperemia in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0037] In some aspects, methods provided herein can include, or consist essentially of, restoring tear film stability in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0038] In some aspects, methods provided herein can include, or consist essentially of, increasing tear breakup time (BUT) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0039] In some aspects, methods provided herein can include, or consist essentially of, increasing goblet cells in the eye of a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0040] In some aspects, methods provided herein can include, or consist essentially of, increasing mucin production in the eye of a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0041] In some aspects, methods provided herein can include, or consist essentially of, restoring or increasing level of Lymphotoxin-alpha in tear film in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0042] In some aspects, methods provided herein can include, or consist essentially of, restoring ocular surface immune homeostasis in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0043] In some aspects, methods provided herein can include, or consist essentially of, increasing tear secretion in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0044] In some aspects, methods provided herein can include, or consist essentially of, increasing goblet cells in the eye of a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions disclosed herein.

[0045] In some aspects, methods provided herein can include, or consist essentially of, relieving, improving, treating dry eye symptoms that include one or more of feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the comeal epithelium, density reduction of conjunctival goblet cells, squamous metaplasia of the ocular surface epithelium, and / or ocular surface inflammation.

[0046] As used herein, “a”, “an”, “the”, “at least one”, and “one or more” are used interchangeably.

[0047] As used herein, the term “about”, when used herein in reference to a value, refers to a value that is similar, in context to the referenced value. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variance encompassed by “about” in that context. For example, in some embodiments, the term “about” may encompass a range of values that are within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.

[0048] The terms “comprises”, and variations thereof do not have a limiting meaning where these terms appear in the description and claims.

[0049] The terms “treat(ment)” or “treating” are used herein to denote delaying the onset of, preventing, inhibiting, alleviating the effects of, or regressing a disease or a symptom thereof in a subject.

[0050] The terms “therapeutically effective amount” and “effective amount” as used herein, refer to an amount or concentration of a composition or treatment described herein, utilized for a period of time (including acute or chronic administration and periodic or continuous administration) that is effective within the context of its administration for causing an intended effect or physiological outcome.

[0051] The term “subject” is used throughout the specification to describe an animal, human or non-human, to whom treatment according to the methods of the present disclosure is provided. Human and vetennary applications are anticipated by the present disclosure. The term includes but is not limited to birds, reptiles, amphibians, and mammals, e.g., humans, other primates, pigs, rodents, such as mice and rats, rabbits, guinea pigs, hamsters, horses, cows, cats, dogs, sheep, chickens and goats. In some embodiments, the subjects are humans, chickens, or mice. In some embodiments, the subject is a human. Both pediatric and adult subjects are included. For example, in any of the methods described herein, the subject can be at least 6 months old (e.g., 6 months or older, 12 months or older, 18 months or older, 2 years or older, 4 years or older, 6 years or older, 10 years or older, 13 years or older, 16 years or older, 18 years or older, 21 years or older, 25 years or older, 30 years or older, 35 years or older, 40 years or older, 45 years or older, 50 years or older, 60 years or older, 65 years or older, 70 years or older, 75 years or older, 80 years or older, 85 years or older, 90 years or older, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16 ,18, 20, 21, 24, 25, 27, 28, 30, 33, 35, 37, 39, 40, 42, 44, 45, 48, 50, 52, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, or more years old).

[0052] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0053] BRIEF DESCRIPTION OF DRAWINGS

[0054] FIG. 1 shows an exemplary schematic of the phase 2, multi-center, double-masked, randomized, vehicle-controlled, dose-response, parallel-group study designed to evaluate the ocular and systemic safety and ocular efficacy of L-Alanyl-L-Glutamine Ophthalmic Solution over a 60-day treatment period in subjects with moderate to severe DED.

[0055] FIG. 2 shows change from baseline in total comeal fluorescein staining (tCFS), wherein the change from baseline was calculated at each visit days (Days 7, 14, and 28).

[0056] FIG. 3 shows change from baseline in eye dryness (VAS), wherein the change from baseline was calculated at each visit days (Days 7, 14, and 28).

[0057] FIG. 4 shows change from baseline in subjects with DED symptoms severity score of SANDE in subgroup (Baseline Tear MMP-9>31.6 ng / mL (1.5 in Log 10 scale).

[0058] FIG. 5 shows change from baseline in subjects with eye dryness (VAS) in subgroup (Baseline Tear MMP-9>31.6 ng / mL (1.5 in Log 10 scale)).

[0059] FIG. 6 shows change from baseline in subjects with central CFS in subgroup (Baseline Tear MMP-9>31.6 ng / mL (1.5 in Log 10 scale)).

[0060] FIG. 7 shows change from baseline in subjects with 0.5% L-Alanyl-L-Glutamine Ophthalmic Solution Tear MMP-9>31.6 ng / mL (1.5 in Log 10 scale) .

[0061] FIG. 8 shows change from baseline in subjects with DED symptoms severity score of SANDE in subgroup with baseline Schirmer test score < 10 at baseline.

[0062] FIG. 9 shows change from baseline in subjects with eye dryness (VAS) in subgroup with Schirmer’s Test Score < 10 at baseline.

[0063] FIG. 10 shows change from baseline in subjects with central CFS in subgroup with Schirmer’s Test Score < 10 at baseline.

[0064] FIG. 11 shows change from baseline in subjects with 0.5% SY-201 Ophthalmic Solution Schirmer’s Test Score < 10 at baseline.

[0065] DETAILED DESCRIPTION

[0066] Ophthalmic disease, disorders, and conditions encompass a range of conditions that affect the eye. For example, ophthalmic diseases, disorders, and / or conditions can include but not limited to neuropathic conditions (e.g., diabetic neuropathy, trigeminal neuralgia, and peripheral neuropathy); diabetes (e.g., diabetic retinopathy, diabetic macular edema); ocular surface disease, disorders, or conditions (dry eye disease (DED), dry eye symptoms, trauma / injury, surgery, allergy related conditions, infections, immune related conditions, contact lens wear, foreign body interaction, neuropathic conditions, foreign object, immune mediated diseases or inflammatory conditions, diabetes, meibomian glands disease); surgery related disease, disorders, and / or conditions; injury; contact lens wear; chemical induced ophthalmic disease, disorders, or conditions; drug induced ophthalmic disease, disorders, or conditions; comeal conditions (e.g., keratoconus, Fuchs' dystrophy, comeal dystrophies, pterygium), meibomian glands disease, inflammatory conditions (e.g., anterior uveitis, posterior uveitis, scleritis, episcleritis, orbital cellulitis, sarcoidosis), orbital disorders (e.g., thyroid eye disease (Graves' Ophthalmopathy), orbital tumor, orbital inflammation), and / or neuro-ophthalmic conditions (e.g., vascular disease, demyelinating diseases (e.g., multiple sclerosis), tumor, trauma).

[0067] Ophthalmic diseases, disorders, and / or conditions can affect the ocular surface of the eye. Many of the ophthalmic diseases, disorders, and conditions described above can cause ocular surface symptoms. For example, non-limiting examples of ophthalmic diseases, disorders, and conditions described above that can cause ocular surface symptoms include dry eye disease DED, dry eye symptoms, trauma / injury, surgery, allergy related conditions, infections, immune related conditions, contact lens wear, foreign body interaction, neuropathic conditions, foreign object, immune mediated diseases or inflammatory conditions, diabetes, and / or meibomian glands disease.

[0068] DED is characterized by inadequate tear production or poor tear quality , leading to ocular surface discomfort and potential vision impairment. Conjunctivitis, known commonly as pink eye, involves inflammation of the conjunctiva and can be triggered by infections, allergies, or irritants. Comeal dystrophies, a group of genetic disorders, affect the clarity and health of the cornea, often resulting in visual distortion. Blephantis, involving inflammation of the eyelid margins, can cause irritation and contribute to ocular surface symptoms. Pterygium, characterized by the growth of a fleshy tissue on the white of the eye, can potentially encroach on the cornea, affect vision, and cause ocular surface symptoms.

[0069] Additionally, conditions like contact lens-related disorders and various forms of trauma can disrupt the delicate balance of the ocular surface, leading to discomfort and, in some cases, more serious vision issues. Infections and inflammations like keratitis, which involves the cornea, often caused by various microorganisms, to autoimmune disorders like Sjogren's Syndrome that lead to ocular surface symptoms. Some individuals may experience ocular herpes, a viral condition that can lead to recurring inflammation and potential scarring of the cornea. In cases of chemical exposure, the eyes can suffer from chemical bums, resulting in acute damage and discomfort. Additionally, severe skin reactions such as Stevens-Johnson Syndrome can extend their effects to the eyes, leading to ocular surface symptoms.

[0070] Ophthalmic diseases, disorders, and / or conditions can include symptoms and discomfort to the ocular surface of the eye. Symptoms and / or discomfort affecting the ocular surface can include but are not limited to feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the comeal epithelium, density reduction of conjunctival goblet cells, squamous metaplasia of the ocular surface epithelium, and / or ocular surface inflammation.

[0071] Symptoms affecting the ocular surface can affect daily activities and tasks, such as reading, using computers, using contact lenses, watching television, driving, and cause longterm ocular discomfort, which may seriously impair quality of life. Serious dry eye can cause keratitis, comeal neovascularization, comeal ulcer and vision impairment which can lead to blindness. The occurrence and development of dry eye are influenced by a plurality of factors. Some examples of risk factors include age, gender, meibomian gland dysfunction, race, use of contact lens, hematopoietic stem cell transplantation, air pollution, digital screen usage, vitamin A deficiency, diet, refractive surgery, diabetes, mental factors, genetic factors, systemic diseases, and medications.

[0072] Dry eye disorders (e.g., DED) can develop as a result of one or more ophthalmic disease, disorders, and conditions through different mechanisms involving inflammation, physical changes to the eye, or autoimmune responses, which can lead to the development or exacerbation of dry eye disease. Symptoms of dry eye can include but are not limited to feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the corneal epithelium, density reduction of conjunctival goblet cells, squamous metaplasia of the ocular surface epithelium, and / or ocular surface inflammation.

[0073] As such, disclosed herein are ophthalmic pharmaceutical compositions comprising a dipeptide (e g., L-Alanyl-L-Glutamine), its methods of preparation, methods of treatment, and methods of use for treatment, mitigation, ameliorating the symptoms of ophthalmic disease, disorders, and conditions.

[0074] Compositions

[0075] Disclosed herein are ophthalmic pharmaceutical compositions comprising L-Alanyl- L-Glutamine that is suitable for eyes. Such solutions suitable for eyes can include one or more of an osmotic agent, a bacteriostatic or antimicrobial agent, a viscosity modifying agent, one or more pH regulators, preservation agent, and water.

[0076] In some embodiments, an ophthalmic pharmaceutical composition includes L-Alanyl- L-Glutarmne at a concentration of about 0.1% to about 10.0% (w / v) of the composition (e.g., about 0.1% to about 9.0% (w / v), about 0.1% to about 8.0% (w / v), about 0.1% to about 7.0% (w / v), about 0. 1% to about 6.0% (w / v), about 0.1% to about 5.0% (w / v), about 0.1% to about 4.0% (w / v), about 0.1% to about 3.0% (w / v), about 0.1% to about 2.5% (w / v), about 0.1% to about 2.0% (w / v), about 0.1% to about 1.5% (w / v), about 0.1% to about 1.0% (w / v), about 0.1% to about 0.5% (w / v), about 0.5% to about 10.0% (w / v), about 0.5% to about 9.0% (w / v), about 0.5% to about 8.0% (w / v), about 0.5% to about 7.0% (w / v), about 0.5% to about 6.0% (w / v), about 0.5% to about 5.0% (w / v), about 0.5% to about 4.0% (w / v), about 0.5% to about 3.0% (w / v), about 0.5% to about 2.5% (w / v), about 0.5% to about 2.0% (w / v), about 0.5% to about 1.5% (w / v), about 0.5% to about 1.0% (w / v), about 1.0% to about 10.0% (w / v), about 1.0% to about 9.0% (w / v), about 1.0% to about 8.0% (w / v), about 1.0% to about 7.0% (w / v), about 1.0% to about 6.0% (w / v), about 1.0% to about 5.0% (w / v), about 1.0% to about 4.0% (w / v), about 1.0% to about 3.0% (w / v), about 1.0% to about 2.5% (w / v), about 1.0% to about 2.0% (w / v), about 1.0% to about 1.5% (w / v), about 1.5% to about 10.0% (w / v), about 1.5% to about 9.0% (w / v), about 1.5% to about 8.0% (w / v), about 1.5% to about 7.0% (w / v), about 1.5% to about 6.0% (w / v), about 1.5% to about 5.0% (w / v), about 1.5% to about 4.0% (w / v), about 1.5% to about 3.0% (w / v), about 1.5% to about 2.5% (w / v), about 1.5% to about 2.0% (w / v), about 2.0% to about 10.0% (w / v), about 2.0% to about 9.0% (w / v), about 2.0% to about 8.0% (w / v), about 2.0% to about 7.0% (w / v), about 2.0% to about 6.0% (w / v), about 2.0% to about 5.0% (w / v), about 2.0% to about 4.0% (w / v), about 2.0% to about 3.0% (w / v), about 2.0% to about 2.5% (w / v), about 2.5% to about 10.0% (w / v), about 2.5% to about 9.0% (w / v), about 2.5% to about 8.0% (w / v), about 2.5% to about 7.0% (w / v), about 2.5% to about 6.0% (w / v), about 2.5% to about 5.0% (w / v), about 2.5% to about 4.0% (w / v), about 2.5% to about 3.0% (w / v), about 3.0% to about 10.0% (w / v), about 3.0% to about 9.0% (w / v), about 3.0% to about 8.0% (w / v), about 3.0% to about 7.0% (w / v), about 3.0% to about 6.0% (w / v), about 3.0% to about 5.0% (w / v), about 3.0% to about 4.0% (w / v), about 4.0% to about 10.0% (w / v), about 4.0% to about 9.0% (w / v), about 4.0% to about 8.0% (w / v), about 4.0% to about 7.0% (w / v), about 4.0% to about 6.0% (w / v), about 4.0% to about 5.0% (w / v), about 5.0% to about 10.0% (w / v), about 5.0% to about 9.0% (w / v), about 5.0% to about 8.0% (w / v), about 5.0% to about 7.0% (w / v), about 5.0% to about 6.0% (w / v), about 6.0% to about 10.0% (w / v), about 6.0% to about 9.0% (w / v), about 6.0% to about 8.0% (w / v), about 6.0% to about 7.0% (w / v), about 7.0% to about 10.0% (w / v), about 7.0% to about 9.0% (w / v), about 7.0% to about 8.0% (w / v), about 8.0% to about 10.0% (w / v), about 8.0% to about 9.0% (w / v), and about 9.0% to about 10.0% (w / v) of the composition). In some embodiments, an ophthalmic pharmaceutical composition includes L-Alanyl-L-Glutamine at a concentration of about 0.2% to about 0.8% (w / v) of the composition. In some embodiments, an ophthalmic pharmaceutical composition includes L-Alanyl-L-Glutamine at a concentration of about 1.8% to about 2.2% (w / v) of the composition. In some embodiments, an ophthalmic pharmaceutical composition includes L-Alanyl-L-Glutamine at a concentration of about 0.5% (w / v) of the composition. In some embodiments, an ophthalmic pharmaceutical composition includes L-Alanyl-L-Glutamine at a concentration of about 1.0% (w / v) of the composition. In some embodiments, an ophthalmic pharmaceutical composition includes L-Alanyl-L- Glutamine at a concentration of about 2.0% (w / v) of the composition.

[0077] In some embodiments, the osmotic agent includes one or more of sodium chloride, potassium chloride, boric acid, borax, sodium sulfate, potassium sulfate, sodium nitrate, potassium nitrate, sodium acetate, mannitol, glycerin, propylene glycol, 2-(4- octylphenylethyl)-2-amino-propylene glycol hydrochloride, glucose, or a combination thereof. In some embodiments, the osmotic agent is about 0.01% to about 3% (w / v) of the composition (e.g., about 0.01% to about 0.05%, about 0.05% to about 0.1%, about 0.1% to about 0.5%, about 0.5% to about 1%, about 1.5% to about 2%, about 2.5% to about 3%, about 0.01%, about 0.1%, about 0.4%, about 0.8%, or about 0.5% (w / v)). In some embodiments, the bacteriostatic or antimicrobial agent includes one or more of benzalkonium chloride, benzalkonium bromide, chlorhexidine acetate, chi orhexi dine gluconate, chlorobutanol, phenoxyethyl alcohol, methyl hydroxybenzoate, ethyl hydroxybenzoate, propyl hydroxybenzoate, or a combination thereof. In some embodiments, the bacteriostatic or antimicrobial agent is about 0.003% to about 0.5% (w / v) of the composition (e.g., about 0.003% to about 0.01%, about 0.01% to about 0.05%, about 0.05% to about 0.1%, about 0.1% to about 0.5%, about 0.003% to about 0.05%, about 0.05% to about 0.5%, about 0.01% to about 0.1%, about 0.003%, about 0.03%, about 0.05%, about 0.1%, or about 0.5% (w / v)).

[0078] In some embodiments, the viscosity modifying agent includes one or more of sodium hyaluronate, sodium carboxymethyl cellulose, methyl cellulose, polyethylene glycol, polyvinyl alcohol, povidone, or a combination thereof. In some embodiments, viscosity modifying agent is about 0.01% to about 0.5% (w / v) of the composition (e.g., about 0.01% to about 0.05%, about 0.05% to about 0.1%, about 0.1% to about 0.5%, about 0.05% to about 0.5%, about 0.01% to about 0.1%, about 0.2%, about 0.15%, or about 0.1% (w / v)).

[0079] In some embodiments, the one or more pH regulators include one or more of sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, boric acid, borax, acetic acid, sodium acetate, citric acid, sodium citrate, tartaric acid, sodium tartrate, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, sodium hydroxide, potassium hydroxide, hydrochloric acid, phosphoric acid, or a combination thereof. In some embodiments, the pH modifying agent adjusts the pH to about 5 to about 9 (e.g., about 6.0 to about 8.0, about 6.5 to about 7.5, or about 7.0.)

[0080] In some embodiments, a preservative includes one or more of benzalkonium chloride and edetate disodium. A preservative can be used in any appropriate concentration. For example, a preservative can be used in a concentration of about 0.005% to about 0.5% (w / v) (e.g., about 0.005% to about 0.01%, about 0.01% to about 0.05%, about 0.05% to about 0.1%, about 0.1% to about 0.5%, about 0.005% to about 0.05%, about 0.05% to about 0.5%, about 0.01% to about 0.1%, about 0.005%, about 0.01%, about 0.05%, about 0.1%, or about 0.5% (w / v)).

[0081] Methods and Uses

[0082] Provided herein are methods comprising administering to an eye of a subject, during a treatment period, an ophthalmic pharmaceutical composition comprising L-Alanyl-L- Glutamine.

[0083] In some embodiments, provided herein are methods of treating ophthalmic disease, disorders, or conditions in a subject, wherein the methods comprise administering to an affected eye of the subject a therapeutically effective amount of the ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0084] In some embodiments, an ophthalmic disease, disorder, or condition can include, but are not limited to, neuropathic conditions (e.g., diabetic neuropathy, trigeminal neuralgia, and peripheral neuropathy); diabetes (e.g., diabetic retinopathy, diabetic macular edema); ocular surface disease, disorders, or conditions (dry eye disease (DED), dry eye symptoms, trauma / injury, surgery, allergy related conditions, infections, immune related conditions, contact lens wear, foreign body interaction, neuropathic conditions, foreign object, immune mediated diseases or inflammatory conditions, diabetes, meibomian glands disease); surgery related disease, disorders, and / or conditions; injury; contact lens wear; chemical induced ophthalmic disease, disorders, or conditions; drug induced ophthalmic disease, disorders, or conditions; comeal conditions (e.g., keratoconus, Fuchs' dystrophy, comeal dystrophies, pterygium), conjunctivitis, blepharitis, meibomian glands disease, inflammatory conditions (e.g., anterior uveitis, posterior uveitis, scleritis, episcleritis, orbital cellulitis, sarcoidosis), orbital disorders (e.g., thyroid eye disease (Graves' Ophthalmopathy), orbital tumor, orbital inflammation), and / or neuro-ophthalmic conditions (e.g., vascular disease, demyelinating diseases (e.g., multiple sclerosis), tumor, trauma).

[0085] In some embodiments, an ophthalmic disease, disorder, and / or condition can affect the ocular surface of the eye. In some embodiments, an ophthalmic disease, disorder, and condition that can cause ocular surface symptoms include dry eye disease DED, dry eye symptoms, trauma / injury, surgery, allergy' related conditions, infections, immune related conditions, contact lens wear, foreign body interaction, neuropathic conditions, foreign object, immune mediated diseases or inflammatory conditions, diabetes, and / or meibomian glands disease.

[0086] In some embodiments, a dry eye disorder (e.g., DED) can develop as a result of one or more ophthalmic disease, disorders, and conditions through different mechanisms involving inflammation, physical changes to the eye, or autoimmune responses, which can lead to the development or exacerbation of dry eye disease. Symptoms of dry eye can include but are not limited to feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the corneal epithelium, density reduction of conjunctival goblet cells, squamous metaplasia of the ocular surface epithelium, and / or ocular surface inflammation.

[0087] In some embodiments, an ophthalmic disease, disorder, and / or condition can also include conditions like contact lens-related disorders and various forms of trauma, wherein the trauma can disrupt the delicate balance of the ocular surface, leading to discomfort and, in some cases, more serious vision issues; infections and inflammations like keratitis, often caused by various microorganisms; autoimmune disorders like Sjogren's Syndrome that lead to ocular surface symptoms; ocular herpes, a viral condition that can lead to recurring inflammation and potential scarring of the cornea; chemical bums, resulting in acute damage and discomfort; and / or severe skin reactions such as Stevens- Johnson Syndrome that can extend their effects to the eyes, leading to ocular surface symptoms. In some embodiments, an ophthalmic disease, disorder, and / or condition can include symptoms and discomfort to the ocular surface of the eye, wherein symptoms and / or discomfort affecting the ocular surface can include, but are not limited to, feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the comeal epithelium, density reduction of conjunctival goblet cells, squamous metaplasia of the ocular surface epithelium, and / or ocular surface inflammation.

[0088] In some embodiments, an ophthalmic disease, disorder, and / or condition can include an treatment-emergent ocular adverse event (TEAE). In some embodiments, an ophthalmic disease, disorder, and / or condition can include an eye disorder, wherein the eye disorder includes conjunctival hyperaemia, eye irritation, blurred vision, blepharitis, abnormal sensation in the eye, and / or periorbital swelling. In some embodiments, an ophthalmic disease, disorder, and / or condition can include bacterial conjunctivitis and / or hordeolum.

[0089] In some embodiments, compositions and methods provided herein can be useful preventing, inhibiting, slowing, or regressing the progression of any of the ophthalmic disease, disorders, or conditions in an eye described herein by administering the ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0090] In some embodiments, provided herein are uses of ophthalmic pharmaceutical compositions comprising L-Alanyl-L-Glutamine for preventing, inhibiting, slowing, or regressing the progression of any of the ophthalmic disease, disorders, or conditions described herein.

[0091] In some embodiments, provided herein are methods of preparation of an ophthalmic medicament for alleviating, ameliorating and / or treating any of the ophthalmic disease, disorder, or conditions described herein comprising suspending or dissolving L-Alanyl-L- Glutamine in an isosmotic solution; adjusting the pH to about 5.0 to about 9.0; and filtering to sterilize the resulting solution with a microporous filter membrane.

[0092] In some embodiments, methods provided herein include relieving symptoms of any of the ophthalmic disease, disorders, or conditions described herein in a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0093] In some embodiments, methods provided herein include improving symptoms of any of the ophthalmic disease, disorders, or conditions described herein in a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0094] In some embodiments, the ophthalmic disease, disorders, or condition includes ocular surface symptoms and / or is an ocular surface disease, disorders, or condition.

[0095] In some embodiments, methods provided herein of treating ocular surface symptoms in a subject comprise administering to an affected eye of the subject a therapeutically effective amount of the ophthalmic pharmaceutical composition comprising L-Alanyl-L- Glutamine.

[0096] In some embodiments, compositions and methods provided herein can be useful preventing, inhibiting, slowing, or regressing the progression of an ocular surface symptoms, or conditions in an eye by administering the ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0097] In some embodiments, provided herein are uses of ophthalmic pharmaceutical compositions comprising L-Alanyl-L-Glutamine for preventing, inhibiting, slowing, or regressing the progression of ocular surface symptoms.

[0098] In some embodiments, provided herein are methods of preparation of an ophthalmic medicament for alleviating, ameliorating and / or treating ocular surface symptoms comprising suspending or dissolving L-Alanyl-L-Glutamine in an isosmotic solution; adjusting the pH to about 5.0 to about 9.0; and filtering to sterilize the resulting solution with a microporous filter membrane. In some embodiments, provided herein are methods of preparation of an ophthalmic medicament for alleviating, ameliorating and / or treating ocular surface symptoms comprising suspending or dissolving L-Alanyl-L-Glutamine at a concentration of about 0.5% (w / v). In some embodiments, provided herein are methods of preparation of an ophthalmic medicament for alleviating, ameliorating and / or treating ocular surface symptoms comprising suspending or dissolving L-Alanyl-L-Glutamine at a concentration of about 1.0% (w / v). In some embodiments, provided herein are methods of preparation of an ophthalmic medicament for alleviating, ameliorating and / or treating ocular surface symptoms comprising suspending or dissolving L-Alanyl-L-Glutamine at a concentration of about 2.0% (w / v).

[0099] In some embodiments, methods provided herein include relieving ocular surface symptoms, or conditions in a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0100] In some embodiments, methods provided herein include improving ocular surface symptoms in a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0101] In some embodiments, the ophthalmic disease, disorders, or condition is DED.

[0102] In some embodiments, methods provided herein of treating in a subject comprise administering to an affected eye of the subject a therapeutically effective amount of the ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0103] In some embodiments, compositions and methods provided herein can be useful preventing, inhibiting, slowing, or regressing the progression of DED in an eye by administering the ophthalmic pharmaceutical composition comprising L-Alanyl-L- Glutamine.

[0104] In some embodiments, provided herein are uses of ophthalmic pharmaceutical compositions comprising L-Alanyl-L-Glutamine for preventing, inhibiting, slowing, or regressing the progression of DED. In some embodiments, provided herein are uses of ophthalmic pharmaceutical compositions comprising L-Alanyl-L-Glutamine at a concentration of about 0.5% (w / v) for preventing, inhibiting, slowing, or regressing the progression of any of the ophthalmic disease, disorders, or conditions in an eye described herein. In some embodiments, provided herein are uses of ophthalmic pharmaceutical compositions comprising L-Alanyl-L-Glutamine at a concentration of about 0.5% (w / v) for preventing, inhibiting, slowing, or regressing the progression of DED. In some embodiments, provided herein are uses of ophthalmic pharmaceutical compositions comprising L-Alanyl-L- Glutamine at a concentration of about 1.0% (w / v) for preventing, inhibiting, slowing, or regressing the progression of any of the ophthalmic disease, disorders, or conditions in an eye described herein. In some embodiments, provided herein are uses of ophthalmic pharmaceutical compositions comprising L-Alanyl-L-Glutamine at a concentration of about 1.0% (w / v) for preventing, inhibiting, slowing, or regressing the progression of DED. In some embodiments, provided herein are uses of ophthalmic pharmaceutical compositions comprising L-Alanyl-L-Glutamine at a concentration of about 2.0% (w / v) for preventing, inhibiting, slowing, or regressing the progression of any of the ophthalmic disease, disorders, or conditions in an eye described herein. In some embodiments, provided herein are uses of ophthalmic pharmaceutical compositions comprising L-Alanyl-L-Glutamine at a concentration of about 2.0% (w / v) for preventing, inhibiting, slowing, or regressing the progression of DED.

[0105] In some embodiments, provided herein are methods of preparation of an ophthalmic medicament for alleviating, ameliorating and / or treating DED comprising suspending or dissolving L-Alanyl-L-Glutamine in an isosmotic solution; adjusting the pH to about 5.0 to about9.0; and filtering to sterilize the resulting solution with a microporous filter membrane.

[0106] In some embodiments, methods provided herein include relieving symptoms of DED in a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0107] In some embodiments, methods provided herein include improving symptoms of DED in a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine. In some embodiments, methods provided herein include increasing tear secretion in a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0108] In some embodiments, methods provided herein include increasing goblet cells in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0109] In some embodiments, methods provided herein include decreasing ocular surface damage in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine. In some embodiments, methods provided herein include decreasing cornea epithelial damage in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0110] In some embodiments, methods provided herein include decreasing ocular surface damage in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0111] In some embodiments, methods provided herein include restoring ocular surface health in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0112] In some embodiments, methods provided herein include restoring conjunctival epithelial condition in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl- L-Glutamme.

[0113] In some embodiments, methods provided herein include restoring tear film stability in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0114] In some embodiments, methods provided herein include increasing goblet cells in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0115] In some embodiments, methods provided herein include increasing mucin production in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine.

[0116] In some embodiments, methods provided herein include restoring or increasing level of lymphotoxin-alpha in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl- L-Glutamme.

[0117] In some embodiments, methods provided herein include restoring ocular surface immune homeostasis in the eye of a subject in need thereof, the method comprising administering to the subject an ophthalmic pharmaceutical composition comprising L-Alanyl- L-Glutamine.

[0118] In some embodiments, ophthalmic pharmaceutical compositions comprising L- Alanyl-L-Glutamine as disclosed herein can be administered to a subject in need thereof to prevent, mitigate, improve, and / or relieve symptoms of ophthalmic disease, disorders, and conditions. For example, ophthalmic pharmaceutical compositions comprising L-Alanyl-L- Glutamine as disclosed herein can be administered to a subject in need thereof to prevent, mitigate, improve, and / or relieve ocular surface symptoms. In some embodiments, the ocular surface symptoms are caused by ocular surface disease, disorders, or condition.

[0119] In some embodiments, diagnostic tests (qualitative and quantitative) can be utilized to diagnose, monitor, or identify, disease, disorder, or conditions. For example, diagnostic test can be performed to establish that ocular surface symptoms of have improved after administration of ophthalmic pharmaceutical compositions disclosed herein. In some embodiments, a subject can report improvement of symptoms. For example, a subject can self-report improvement of symptoms (e.g., foreign body sensation or pain) after administration of an ophthalmic pharmaceutical composition disclosed herein.

[0120] In some embodiments, the diagnostic test used is a cell staining procedure. For example, the diagnostic test used is can be total comeal fluorescein staining. Total comeal fluorescein staining can be used to detect and assess the extent of damage or abnormalities on the surface of the cornea. This procedure involves applying a fluorescein dye to the eye, which is a fluorescent compound that temporarily stains the tear film on the comeal surface. In some cases, the scale used to determine severity is as measured by the National Eye Institute (e.g., 0 (none) to 20 (severe)). In some embodiments, subjective tools can be used. For example, a subject can self-report pain, foreign body sensation, feelings of dryness, photophobia, etc. For example, a subject can record their symptoms in a diary. In some cases, a subject might report a symptom as a score (e.g., an eye dryness score). In some embodiments, the score might be measured by a visual analog scale (0 (none) to 100 (severe). In some cases, the time frame in which such diagnostic procedures are performed / observed is about 30-90 days (e.g., about 30 days, about 60 days, about 90 days, about 30-60 days, or about 60-90 days).

[0121] For example, an ophthalmic pharmaceutical compositions comprising L-Alanyl-L- Glutamine disclosed herein can prevent, mitigate, improve, and / or relieve one or more of feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the corneal epithelium, density reduction of conjunctival goblet cells, squamous metaplasia of the ocular surface epithelium, and / or ocular surface inflammation.

[0122] In some embodiments, an ophthalmic pharmaceutical composition comprising L- Alanyl-L-Glutamine disclosed herein is administered to a subject in need thereof in therapeutically effective amounts to prevent, mitigate, improve, and / or relieve ocular surface symptoms. As described above, a therapeutic effective amount includes, an amount or concentration of a composition or treatment described herein, utilized for a period of time (including acute or chronic administration and periodic or continuous administration) that is effective within the context of its administration for causing an intended effect or physiological outcome. For example, diagnostic test can be performed to establish that symptoms and / or discomfort of the ocular surface have improved after administration of an ophthalmic pharmaceutical composition disclosed herein in a therapeutically effective amount. In some embodiments, a subject can report improvement of symptoms. For example, a subject can self-report improvement of symptoms (e.g., foreign body sensation or pain) after administration of an ophthalmic pharmaceutical composition disclosed herein in a therapeutically effective amount.

[0123] In some embodiments, a therapeutically effective amount of L-Alanyl-L-Glutamine is added to the ophthalmic preparations of the present disclosure in concentrations of about 0.1% to about 10.0% (w / v) of the composition (e.g., about 0.1% to about 9.0% (w / v), about 0.1% to about 8.0% (w / v), about 0.1% to about 7.0% (w / v), about 0.1% to about 6.0% (w / v), about 0.1% to about 5.0% (w / v), about 0.1% to about 4.0% (w / v), about 0.1% to about 3.0% (w / v), about 0. 1% to about 2.5% (w / v), about 0.1% to about 2.0% (w / v), about 0.1% to about 1.5% (w / v), about 0.1% to about 1.0% (w / v), about 0.1% to about 0.5% (w / v), about 0.5% to about 10.0% (w / v), about 0.5% to about 9.0% (w / v), about 0.5% to about 8.0% (w / v), about 0.5% to about 7.0% (w / v), about 0.5% to about 6.0% (w / v), about 0.5% to about 5.0% (w / v), about 0.5% to about 4.0% (w / v), about 0.5% to about 3.0% (w / v), about 0.5% to about 2.5% (w / v), about 0.5% to about 2.0% (w / v), about 0.5% to about 1.5% (w / v), about 0.5% to about 1.0% (w / v), about 1.0% to about 10.0% (w / v), about 1.0% to about 9.0% (w / v), about 1.0% to about 8.0% (w / v), about 1.0% to about 7.0% (w / v), about 1.0% to about 6.0% (w / v), about 1.0% to about 5.0% (w / v), about 1.0% to about 4.0% (w / v), about 1.0% to about 3.0% (w / v), about 1.0% to about 2.5% (w / v), about 1.0% to about 2.0% (w / v), about 1.0% to about 1.5% (w / v), about 1.5% to about 10.0% (w / v), about 1.5% to about 9.0% (w / v), about 1.5% to about 8.0% (w / v), about 1.5% to about 7.0% (w / v), about 1.5% to about 6.0% (w / v), about 1.5% to about 5.0% (w / v), about 1.5% to about 4.0% (w / v), about 1.5% to about 3.0% (w / v), about 1.5% to about 2.5% (w / v), about 1.5% to about 2.0% (w / v), about 2.0% to about 10.0% (w / v), about 2.0% to about 9.0% (w / v), about 2.0% to about 8.0% (w / v), about 2.0% to about 7.0% (w / v), about 2.0% to about 6.0% (w / v), about 2.0% to about 5.0% (w / v), about 2.0% to about 4.0% (w / v), about 2.0% to about 3.0% (w / v), about 2.0% to about 2.5% (w / v), about 2.5% to about 10.0% (w / v), about 2.5% to about 9.0% (w / v), about 2.5% to about 8.0% (w / v), about 2.5% to about 7.0% (w / v), about 2.5% to about 6.0% (w / v), about 2.5% to about 5.0% (w / v), about 2.5% to about 4.0% (w / v), about 2.5% to about 3.0% (w / v), about 3.0% to about 10.0% (w / v), about 3.0% to about 9.0% (w / v), about 3.0% to about 8.0% (w / v), about 3.0% to about 7.0% (w / v), about 3.0% to about 6.0% (w / v), about 3.0% to about 5.0% (w / v), about 3.0% to about 4.0% (w / v), about 4.0% to about 10.0% (w / v), about 4.0% to about 9.0% (w / v), about 4.0% to about 8.0% (w / v), about 4.0% to about 7.0% (w / v), about 4.0% to about 6.0% (w / v), about 4.0% to about 5.0% (w / v), about 5.0% to about 10.0% (w / v), about 5.0% to about 9.0% (w / v), about 5.0% to about 8.0% (w / v), about 5.0% to about 7.0% (w / v), about 5.0% to about 6.0% (w / v), about 6.0% to about 10.0% (w / v), about 6.0% to about 9.0% (w / v), about 6.0% to about 8.0% (w / v), about 6.0% to about 7.0% (w / v), about 7.0% to about 10.0% (w / v), about 7.0% to about 9.0% (w / v), about 7.0% to about 8.0% (w / v), about 8.0% to about 10.0% (w / v), about 8.0% to about 9.0% (w / v), and about 9.0% to about 10.0% (w / v)). In some embodiments, a therapeutically effective amount of the L-Alanyl-L- Glutamine is added to the ophthalmic preparations of the present disclosure in concentrations of about 0.2% to about 0.8% (w / v) in the composition (e.g., about 0.2% to about 0.8%, about 0.3% to about 0.8%, about 0.4% to about 0.8%, about 0.5% to about 0.8%, about 0.6% to about 0.8%, about 0.7% to about 0.8%, about 0.2% to about 0.7%, about 0.2% to about 0.6%, about 0.2% to about 0.5%, about 0.2% to about 0.4%, about 0.2% to about 0.3%, or about 0.4% to about 0.6% (w / v)). In some embodiments, a therapeutically effective amount of the L-Alanyl-L-Glutamine is added to the ophthalmic preparations of the present disclosure in concentrations of about 1.8% to about 2.2% (w / v) of the composition (e g., about 1.8% to about 2.1% (w / v), about 1.8% to about 2.0% (w / v), about 1.8% to about 1.9% (w / v), about 1.9% to about 2.2% (w / v), about 1.9% to about 2.1% (w / v), about 1.9% to about 2.0% (w / v), about 2.0% to about 2.2% (w / v), about 2.0% to about 2.1% (w / v), and about 2. 1% to about 2.2% (w / v)). In some embodiments, a therapeutically effective amount of the L-Alanyl-L- Glutamine is added to the ophthalmic preparations of the present disclosure in concentrations of about 0.5% (w / v) in the composition. In some embodiments, a therapeutically effective amount of the L-Alanyl-L-Glutamine is added to the ophthalmic preparations of the present disclosure in concentrations of about 1.0% (w / v) of the composition. In some embodiments, a therapeutically effective amount of the L-Alanyl-L-Glutamine is added to the ophthalmic preparations of the present disclosure in concentrations of about 2.0% (w / v) of the composition.

[0124] For example, an ophthalmic pharmaceutical comprising L-Alanyl-L-Glutamine disclosed herein can prevent, mitigate, improve, and / or relieve one or more symptoms and / or discomfort of the ocular surface such as feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the corneal epithelium, density reduction of conjunctival goblet cells, squamous metaplasia of the ocular surface epithelium, and / or ocular surface inflammation.

[0125] In some embodiments, a pharmaceutical composition provided is formulated to be compatible with its intended route of administration. Examples of routes of administration include topical ocular administration. The form of such topical ocular administration can include one or more of eye drops, suspensions, ointments, creams, patches, eye masks, eye patches, eye films, eye creams, sprays, gels, injections, or implants. In some embodiments, the pharmaceutical composition can be used in combination with an ocular medical device such as a lens. Pharmaceutically acceptable carriers can include solvents, dispersion media, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Supplementary active compounds can also be incorporated into the compositions.

[0126] In some embodiments, topical preparations have liquid carriers and can be aqueous solutions or suspensions, or emulsions. In some embodiments, topical preparations can include a solution, a suspension, an emulsion, a gel, or a sustained release formulation, including, e g., an implants or an ocular device such as a lens.

[0127] In some embodiments, a composition or formulation (e.g., an ophthalmic preparation) can be applied in a liquid earner. In some embodiments, the earner is an aqueous carrier. In some embodiments, the carrier is water. In some embodiments, quick dissolving forms of the medicaments may be administered in powder form or rubbed into the eye from applicators of various types. Spraying of the eye, eye drops, and other methods of application can be used. In some embodiments, the preparations are packaged as sterile solutions in dropper bottles, as are standard in ophthalmic preparations. Other containers, including eye cups, can also be used. The preparation can, in some embodiments, be packaged with instructions for using the preparation in treating presbyopia, in some embodiments, directing the use of preparation to administer 1 to 2 drops of the solution to each eye.

[0128] In some embodiments, a composition described herein can be administered in a pharmaceutically acceptable formulation (e.g., an ophthalmic preparation), such as topically by application of the formulation to the eye in a non-irritating sterile solution or suspension. In some embodiments, the formulation is preferably at a pH compatible with the eye (e.g., about 6.5 to about 7.5).

[0129] Dosage levels can vary depending upon the individual to be treated, the progression of the disorder, and / or the specific medicament(s) used. In some embodiments, the methods described herein include 1-2 drops per application. In some embodiments, the methods described herein include 1 drop per application. In some embodiments, drop sizes range from about 10 pL to about 80 pL (e.g., about 20 pL to about 80 pL, about 30 pL to about 80 pL, about 40 pL to about 80 pL, about 50 pL to about 80 pL, about 60 pL to about 80 pL, about 70 pL to about 80 pL, about 10 pL to about 70 pL, about 20 pL to about 70 pL, about 30 pL to about 70 pL, about 40 pL to about 70 pL, about 50 pL to about 70 pL, about 60 pL to about 70 pL, about 10 pL to about 60 pL, about 20 pL to about 60 pL, about 30 pL to about 60 pL, about 40 pL to about 60 pL, about 50 pL to about 60 pL, about 10 pL to about 50 pL, about 20 pL to about 50 pL, about 30 pL to about 50 pL, about 40 pL to about 50 pL, about 10 pL to about 40 pL, about 20 pL to about 40 pL, about 30 pL to about 40 pL, about 10 pL to about 30 pL, about 20 pL to about 30 pL, and about 10 pL to about 20 pL). In some embodiments, drop sizes can be about 35 pL. In some embodiments, exemplary dosage amounts can range from about 10 pL to about 480 pL per application. Exemplary dosage regimens useful in some embodiments of the methods described herein include 1 application per day, two applications per day, three applications per day, four applications per day, five applications per day, six applications per day, one application every other day, one application per week, two applications per week, or three applications per week. In some embodiments, the methods described herein include one application per day. In some embodiments, the methods described herein include one application every week (e.g., seven days). In some embodiments, the methods described herein include one application every week for a 60-day period. In some embodiments, ophthalmic pharmaceutical composition comprising L-Alanyl- L-Glutamme can be administered during a treatment period. In some embodiments, exemplary treatment periods include 1 day, up to about 5 days, up to about 10 days, up to about 30 days, up to about 1 week, up to about 2 weeks, up to about 3 weeks, up to about 4 weeks, up to about 5 weeks, up to about 1 month, up to about 2 months, up to about 3 months, up to about 4 months, up to about 5 months, up to about 6 months, up to about 7 months, up to about 8 months, up to about 9 months, up to about 10 months, up to about 11 months, up to about 1 year, up to about 2 years, up to about 3 years, up to about 4 years, up to about 5 years, or up to about 10 years, from about 1 day to about 10 years, from about 1 month to about 10 years, from about 2 months to about 10 years, from about 3 months to about 10 years, from about 4 months to about 10 years, from about 5 months to about 10 years, from about 6 months to about 10 years, from about 6 months to about 9 years, from about 6 months to about 8 years, from about 6 months to about 7 years, from about 6 months to about 6 years, from about 6 months to about 5 years, from about 1 day to about 60 months, from about 6 months to about 4 years, from about 6 months to about 3 years, from about 6 months to about 2 years, from about 6 months to about 1 year, and the like. In some embodiments, ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine can be administered during a treatment period of 2 months (e.g., 60 days). In some embodiments of the methods described herein, treatment regimens may be periodically stopped and restarted according to the subject’s needs.

[0130] In some embodiments, ophthalmic pharmaceutical composition comprising L-Alanyl- L-Glutamme can be administered from 1 to 6 times per day, from 1 to 5 times per day, from 1 to 4 times per day, from 1 to 3 times per day, or from 1 to 2 times per day during the treatment period. In some embodiments, ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine can be administered 1 time per day during the treatment period. In some embodiments, compositions or formulations described herein (e.g., compositions or formulations including the cholinesterase antagonist, the miotic agent, or a combination thereof) can be administered to a subject within, e.g., 2 hours, 1 hour, 45 minutes, 30 minutes, 15 minutes, 10 minutes, or 5 minutes prior to a period of sleep for the subject.

[0131] EXAMPLES

[0132] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims. Example 1 - Multi-Center, Double-Masked, Randomized, Vehicle-Controlled, Dose- Response, Parallel-Group Study of L-alanyl-L-glutamine Ophthalmic Solution versus Vehicle Control in Subjects with Dry Eye Disease

[0133] Overall Study Design and Plan - Description

[0134] A phase 2. multi-center, double-masked, randomized, vehicle-controlled, doseresponse, parallel-group study was designed to evaluate the ocular and systemic safety and ocular efficacy of L-Alanyl-L-Glutamine Ophthalmic Solution over a 60-day treatment period in subjects with moderate to severe Dry Eye Disease (DED).

[0135] During the 14-day single-masked run-in period, approximately 200 subjects instilled vehicle as 1 drop BID in both eyes (OU). At Visit 2, subjects were randomized in a 1 : 1 : 1 : 1 ratio to 4 treatment groups: Vehicle (n=50) and L-Alanyl-L-Glutamine Ophthalmic Solution 2.0% (n=50), 1.0% (n=50), and 0.5% (n=50). Double-masked IP was instilled as 1 drop OU BID for 60 days. The study consisted of 6 clinic visits: Visit 1 (-14 Days, Screening), Visit 2 (Day 1, Randomization), Visit 3 (Day 7 ± 2 days), Visit 4 (Day 14 ± 2 days), Visit 5 (Day 28 ± 2 days), and Visit 6 (Day 60 ± 3 days, End of Study / Early Termination) (Error! Reference source not found.).

[0136] Discussion of Study Design. Including the Choice of Control Groups, and Selection of Study Population

[0137] This study examined the ocular and systemic safety and ocular efficacy of L-Alanyl- L-Glutamme Ophthalmic Solution versus vehicle dosed OU BID for 60 days in subjects with moderate to severe DED.

[0138] L-Alanyl-L-Glutamine Ophthalmic Solution and vehicle were administered as a topical ophthalmic solution. In the single-masked run-in period, vehicle was dosed OU BID by all subjects to normalize baseline conditions between subjects. Subjects self-administered the first dose in-clinic at Visit 1 and all remaining doses at home. In the double-masked treatment period, subjects self-administered 1 dose of IP in-clinic at Visit 2 (Day 1) and Visit 6 (Day 60) and all remaining doses at home, OU BID. Direct instillation is the most efficient method for delivery to the ocular surface and is an accepted and widely used method for topical application to the eye. Each dose was delivered by administering 1 drop (approximately 35 pL of L-Alanyl-L-Glutamine Ophthalmic Solution) in each eye from a single-use vial.

[0139] The study population consisted of 201 adult subjects with moderate to severe DED. The study eye (SE) was defined as the eye meeting all inclusion criteria and no exclusion criteria, and with the highest tCFS scoring at randomization (Visit 2, Day 1). When both eyes met the inclusion criteria and no exclusion criteria and had the same tCFS score, the right eye would be used as the SE.

[0140] Treatments

[0141] Four (4) IPs were administered during this study:

[0142] • L-Alanyl-L-Glutamine Ophthalmic Solution 2.0%

[0143] • L-Alanyl-L-Glutamine Ophthalmic Solution 1.0%

[0144] • L-Alanyl-L-Glutamine Ophthalmic Solution 0.5%

[0145] • Vehicle solution

[0146] Two primary ocular efficacy endpoints (one sign and one symptom) were tested sequentially at Day 60:

[0147] • Mean change from baseline (CFB) in total corneal fluorescein staining (tCFS; modified National Eye Institute [mNEI] scale, 0-20)

[0148] • Mean CFB in eye dryness score on the visual analog scale (VAS, 0-100 points)

[0149] Key secondary endpoints include sequential testing of the primary endpoints at Day 28. The following secondary ocular efficacy endpoints were tested:

[0150] • Mean change from baseline (CFB) in central corneal fluorescein staining (cCFS; modified National Eye Institute [mNEI] scale, 0-4)

[0151] • Mean CFB in bulbar conjunctival hyperemia using the Cornea and Contact Lens Research Unit (CCLRU, 0-4) grading scale

[0152] • Mean CFB in MMP-9 score (study eye [SE])

[0153] • Mean CFB in Symptom Assessment Questionnaire in Dry Eye (SANDE)

[0154] • Mean CFB in unanesthetized Schirmer test score

[0155] • Use of rescue preservative-free artificial tears (AT) after the baseline visit (Visit 2)

[0156] All efficacy assessments were conducted at the time points show n on the Schedule of Visits and Examinations (Table 1).

[0157] Table 1. Schedule of Procedures and Assessments

[0158] Abbreviations: AE = adverse event; AT = artificial tears; BCVA = best corrected visual acuity; CCLRU = Cornea and Contact Lens Research Unit; CFS = corneal fluorescein staining; ET = early termination; IOP = intraocular pressure; IP = investigational product; MMP-9 = matrix metalloproteinase-9; POC: point-of-care; SANDE = Symptom Assessment Questionnaire in Dry Eye; SE = study eye; UPT = urine pregnancy test; WOCBP = women of childbearing potentialaMMP-9 was assessed OU at Visits 1 and 2 and SE thereafter. Wait at least 15 minutes after grading CFS before conducting Schirmer test.

[0159] Statistical Methods Planned in the Protocol and Determination of Sample Size

[0160] Summary statistics for the data collected during this study were presented to give a general description of the subjects studied. Data from all sites were combined in the computation of these descriptive summaries.

[0161] Number of subjects, minimums, and maximums were calculated to the same number of decimal places as the source data. Means, medians, standard deviations, and quartiles were calculated to one more decimal place than the source data. Percentages were calculated to the nearest one decimal place. Zero count cells were displayed as “0” with percentage of (0%). Unless otherwise noted, summaries were performed by the treatment group and presented in the order of: L-Alanyl-L-Glutamine 2.0% or 1.0% or 0.5% (efficacy analyses only), L- Alanyl-L-Glutamine, 2.0%; L-Alanyl-L-Glutamine, 1.0%; L-Alanyl-L-Glutamine, 0.5%; Vehicle.

[0162] Statistical tests were presented as two-sided p-values rounded to four decimal places. Unless otherwise indicated, statistical testing was carried out at the a = 0.05 significance level.

[0163] Baseline values were defined as the last measurement prior to dosing of doublemasked study medication. Ocular measurements will use the most recent measurement for each eye.

[0164] The detection limits for tear biomarker MMP-9 test were: 5 ng / mL and 1000 ng / mL. When MMP-9 test result was below 5, test data would be recorded as < 5 ng / mL, when it was above 1000 ng / mL, it would be recorded as > 1000 ng / mL from the Analyzer. When summarizing the data, the original values were converted to either 4 ng / mL or 1010 ng / mL, respectively. In the data listings, the actual recorded values from the MMP-9 test Analyzer were presented. MMP-9 test values were first log 10 transformed before subjected to statistical analysis.

[0165] All data collected in this study were presented in individual subject data listings for all subjects. Computations for all results were performed using SAS (Version 9.4, SAS / STAT 15.2) computer software package (SAS Institute, Inc, 2013, 2020), unless otherwise specified.

[0166] Subgroup analyses will include testing of the mean CFB in tCFS, eye dryness score, and secondary endpoints including central CFS and SANDE severity score for subgroups based on baseline Schirmer I test score <10 mm / 5 min. Subgroup analyses will include testing of the mean CFB in tCFS, eye dryness score, and secondary endpoints including central CFS and SANDE severity score for subgroups based on baseline tear biomarker MMP-9 test score < 32 ng / mL. Subgroup analyses by sex and age group (<65 years, >65 years) may also be performed. Disposition of Subjects

[0167] A total of 284 subjects were screened and 201 were randomized into four groups: 50 in 2% L-Alanyl-L-Glutamine Group, 51 in 1% L-Alanyl-L-Glutamine Group, 51 in 0.5% L- Alanyl-L-Glutamine Group, and 49 in Vehicle Group. Three subjects were withdrawal before treatment started, 1 in 2% L-Alanyl-L-Glutamine (2.0%), 1 in 0.5% L-Alanyl-L-Glutamine (2.0%) and 1 in Vehicle Group (2.0%) respectively. Of these 201 subjects, 198 (98.5%) received IP treatment and 196 (97.5%) completed dosing. Overall, 195 (97.0%) subjects completed the study and 2 discontinued due to adverse event (AE) (1 (2.0%) in 2% L-Alanyl- L-Glutamine group (acute bacterial conjunctivitis) and 1 (2.0%) in Vehicle Group (worsening of osteoarthritis), and 1 (2.0%) early termination in Vehicle Group due to subject withdrawal (Error! Reference source not found.).

[0168] Table 2. Analysis Sets - All Randomized Subjects

[0169] SY-201 SY-201 SY-201

[0170] Per Protocol Analysis Set (PPAS) 47 (94.0) 48 (94.1) 49 (96.1) 45 (91.8) 189 (94.0)

[0171] Did not complete Study 2 (4.0) 0 1 (2.0) 3 (6.1) 6 (3.0)

[0172] Inclusion and exclusion criteria 1 0 1 2 (1.0)

[0173] Poor Compliance 0 3 0 1 4 (2.0) Demographic and Other Baseline Characteristics

[0174] The demographic characteristics for the 189 subjects in PPAS were those for the subjects in the SAS (Error! Reference source not found.).

[0175] Table 3. Demographics - Per Protocol Analysis Set (PPAS)

[0176] SY-201, SY-201, SY-201,

[0177] 20% 1.0% 0.5% Vehicle Overall

[0178] Mean (SD) 65.6 (12.6) 64.8 (10 9) 65 3 (90) 64.4 (13.0) 65.0 (11.3) SY-201, SY-201, SY-201,

[0179] 20% 1.0% 0.5% Vehicle Overall

[0180] (N=47) (N=48) (N=49) (N=45) (N=189)

[0181] Median 67.0 670 66.0 68.0 67.0

[0182] OD 22 (46.8) 28 (58.3) 28 (57.1) 30 (667) 108 (57.1)

[0183] OS 25 (53.2) 20 (41.7) 21 (42.9) 15 (33 3) 81 (42.9)

[0184] Efficacy Results and Tabulations of Individual Subject Data

[0185] In addition to the primary outcome measures at Day 60, the change from baseline was calculated at each visit (Days 7, 14, and 28). In both the primary sign and primary symptom, there was an onset of activity in all treatment groups starting at the first visit, Day 7. This effect increased over the 60 days of treatment (FIGs. 3 and 4).

[0186] Secondary efficacy measures presented here included central cornea fluorescein staining (cCFS), Bulbar Conjunctiva Hyperemia (CCLRU), dry eye symptoms severity score of SANDE, SANDE global score and dry eye symptom frequency score of SANDE with the PPAS population (Table 4).

[0187] Table 4. Secondary Efficacy Measures: Change from Baseline at Day 60 - PPAS

[0188] SY-201, 2.0% SY-201, 1% SY-201, 0.5% Vehicle

[0189] CFB Statistic(N=47)(N=48) (N=49) (N=45) „ . . SY-201, 2.0% SY-201, 1% SY-201, 0.5% Vehicle

[0190] CFB Statistic(N=47. (N=48) (N=49) (N=45)

[0191] Baseline (Observed) Mean (SD) 1.06 (0.639) 1.19 (0.823) 1.10 (0.661) 0.99 (0.787)

[0192] CFB V6 (Day 60) Mean (SD) -0.33 (0.653) -0.36 (0.933) -0.44 (0.517) -0.18 (0.641) Median -0.50 -0.50 -0.50 0.00 LS Mean -0.4 -0.3 -0.4 -0.2

[0193] L-Alanyl-L- Difference -0.1 (-0.37, -0.1 (-0.34, -0.2 (-0.46,

[0194] Glutamine - Vehicle (95% CI) 0.14) 0.17) 0.04) p-value 0.3689 0.5202 0.1024

[0195] Bulbar Conjunctiva pcrcinia RU) (0-4)

[0196] Baseline (Observed) Mean (SD) 1.8 (0.67) 1.7 (0.76) 1.7 (0.67) 1.7 (0.56)

[0197] CFB V6 (Day 60) Mean (SD) -0.3 (0.61) -0.2 (0.68) -0.1 (0.63) -0.1 (0.63) LS Mean -0.3 -0.3 -0.1 -0.2 , , Glutamine - Vehicle (95% CI) 4.78) 5.85) 0.85) SY-201, 2.0% SY-201, 1% SY-201, 0.5% Vehicle

[0198] CFB Statistic(N 7. _ (N=48) (N=49) _ (N=45) p-value 0.4010 0.5620 0.0783 /

[0199] Sa fety Results

[0200] Overall, 19 / 198 (9.6%) of subjects had at least 1 ocular TEAE (either eye) during the study (8 / 49, (16.3%), 3 / 41 (5.9%), 4 / 50 (8.0%) and 4 / 48 (8.3%), in the L-Alanyl-L-Glutamine 2%, 1%, 0.5% and vehicle groups, respectively. Most ocular TEAEs were mild in severity (95%, 18 / 19), and were judged related to treatment (79%, 15 / 19). There were no ocular SAEs. Two subjects discontinued due to adverse event (AE) (1 (2.0%) in 2% L-Alanyl-L-Glutamine group (acute bacterial conjunctivitis) and 1 (2.0%) in Vehicle Group, worsening of osteoarthritis).

[0201] No deaths occurred in this study. Two serious adverse events were reported in 2 subjects (one in the 0.5% L-Alanyl-L-Glutamine treatment group (Sepsis secondary to urinary tract infection) and one in the 2.0% L-Alanyl-L-Glutamine treatment group (Deep vein thrombosis). Both were judged not related to the investigational drug. Overall, 19 / 198 (9.6%) of subjects had at least 1 ocular TEAE (either eye) during the study (8 / 49, (16.3%), 3 / 41 (5.9%), 4 / 50 (8.0%) and 4 / 48 (8.3%), in the L-Alanyl-L-Glutamine 2%, 1%, 0.5% and vehicle groups, respectively (Table 5).

[0202] Instillation site pain, while relatively infrequent, appeared to have a dose-related incidence: 0, 3 / 50 (6%), 2 / 51 (3.9%) and 6 (10.2%) in the vehicle, 0.5%, 1.0% and 2.0% L-Alanyl-L- Glutamine treatment groups, respectively. Other adverse events were relatively infrequent with no clear dose-response.

[0203] Table 5. Treatment-Emergent Ocular Adverse Events by System Organ Class and Preferred Term - SAS (Either Eye)

[0204] MedDRA v25.0 SY-201, 2.0% SY-201, 1.0% SY-201, 0.5% Vehicle

[0205] System Organ Class (N=49) (N=51) (N=50) (N=48)

[0206] Preferred Term n (%) n (%) n (%) n (%)

[0207] Subjects with Any Ocular TEAE 8 (16.3) 3 (5.9) 4 (8.0) 4 (8.3)

[0208] Eye disorders 3 (6.1) 1 (2.0) 0 4 (8.3)

[0209] Conjunctival hyperaemia 1 (2.0) 0 0 2 (4.2)

[0210] Eye irritation 1 (2.0) 0 0 0

[0211] Vision blurred 1 (2.0) 0 0 0

[0212] Blepharitis 0 1 (2.0) 0 0

[0213] Abnormal sensation in eye 0 0 0 1 (2.1)

[0214] Periorbital swelling 0 0 0 1 (2.1)

[0215] General disorders and administration 5 (10.2) 2 (3.9) 3 (6.0) 0 site conditions MedDRA v25.0 SY-201, 2.0% SY-201, 1.0% SY-201, 0.5% Vehicle

[0216] System Organ Class (N=49) (N=51) (N=50) (N=48)

[0217] Preferred Term n (%) n (%) n (%) n (%)

[0218] Instillation site pain 5 (10.2) 2 (3.9) 3 (6.0) 0

[0219] Infections and infestations 1 (2.0) 0 1 (2.0) 0

[0220] Conjunctivitis bacterial 1 (2.0) 0 0 0

[0221] Hordeolum 0 0 1 (2.0) 0

[0222] Abbreviations: SAS =Safety Analysis Set; AE=adverse event; MedDRA=Medical Dictionary for Regulatory Activities; TEAE=treatment-emergent adverse event.

[0223] Note: A TEAE is defined as an AE that starts on or after the date of the first dose of double-masked study medication up to and including the last dose of double-masked study medication plus 30 days. Note: Subjects with one or more adverse events within a level of MedDRA are counted only once in that level.

[0224] Note: Percentages are based on the number of subjects in each treatment

[0225] OTHER EMBODIMENTS

[0226] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other embodiments, advantages, and modifications are within the scope of the following claims.

Claims

WHAT IS CLAIMED IS1. An ophthalmic pharmaceutical composition comprising L-Alanyl-L-Glutamine that is suitable for eyes.

2. The ophthalmic pharmaceutical composition of claim 1, wherein the ophthalmic composition is in the form of an eye drop, suspension, ointment, emulsion, eye patch, eye mask, eye film, eye cream, spray, gel, injection, or implant.

3. The pharmaceutical composition according to claim 1 or 2, wherein the concentration of L-Alanyl-L-Glutamine is about 0.2% to about 0.8% (w / v) of the composition.

4. The pharmaceutical composition according to claim 3, wherein the concentration of L-Alanyl-L-Glutamine is about 0.5% (w / v) of the composition.

5. The pharmaceutical composition according to claim 1 or 2, wherein the concentration of L-Alanyl-L-Glutamine is about 1.8% to about 2.2% (w / v) of the composition.

6. The pharmaceutical composition according to claim 5, wherein the concentration of L-Alanyl-L-Glutamine is about 2.0% (w / v) of the composition.

7. The pharmaceutical composition according to claim 1 or 2, wherein the concentration of L-Alanyl-L-Glutamine is about 1.0% (w / v) of the composition.

8. The pharmaceutical composition of any one of claims 1-7, wherein the pharmaceutical composition further comprises a bacteriostatic or antimicrobial agent.

9. The pharmaceutical composition of claim 8, wherein the bacteriostatic or antimicrobial agent includes one or more of benzalkonium chloride, benzalkonium bromide, chlorhexidine acetate, chlorhexidine gluconate, chlorobutanol, phenoxyethyl alcohol, methyl hydroxybenzoate, ethyl hy droxy benzoate, propyl hy droxybenzoate, and combinations thereof.

10. The pharmaceutical composition of any one of claims 8-9, wherein the concentration of the bacteriostatic or antimicrobial agent is about 0.003 to about 0.5% (w / v) of the composition.

11. The pharmaceutical composition of any one of claims 1-10, wherein the pharmaceutical composition further comprises a viscosity modifying agent.

12. The pharmaceutical composition of claim 11, wherein the viscosity7modifying agent includes one or more of sodium hyaluronate, sodium carboxymethyl cellulose, methyl cellulose, polyethylene glycol, polyvinyl alcohol, povidone, and combinations thereof.

13. The pharmaceutical composition of any one of claims 11-12, wherein the concentration of the viscosity modifying agent is about 0.01% to about 0.5% (w / v) of the composition.

14. The pharmaceutical composition of any one of claims 1-13. wherein the pharmaceutical composition further comprises one or more pH regulators.

15. The pharmaceutical composition of claim 14, wherein the one or more pH regulators includes one or more of sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, boric acid, borax, acetic acid, sodium acetate, citric acid, sodium citrate, tartaric acid, sodium tartrate, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, sodium hydroxide, potassium hydroxide, hydrochloric acid, phosphoric acid, and combinations thereof.

16. The pharmaceutical composition of any one of claims 14-15, wherein the pH of the pharmaceutical composition is about 5.0 to about 9.0.

17. A method of preparing the pharmaceutical composition of any one of claims 1-16, wherein the method comprising suspending or dissolving L-Alanyl-L-Glutamine in water to form an aqueous solution; adjusting the pH to about 5.0 to about 9.0; and filtering to sterilize the resulting solution with a microporous filter membrane.

18. A method of relieving, improving, treating ocular surface symptoms and discomfort in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-19. A method of relieving, improving, or treating ocular surface symptoms caused by ocular surface diseases in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

20. A method of relieving, improving, or treating ocular surface symptoms caused by non-ocular surface diseases in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

21. A method of relieving, improving, treating ocular surface symptoms caused by dry eye in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1- 16.

22. A method of relieving, improving, treating ocular surface symptoms caused byallergies in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1- 16.

23. A method of relieving, improving, treating ocular surface symptoms caused by infections in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1- 16.

24. A method of relieving, improving, treating ocular surface symptoms caused by neuropathic conditions in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1 -16.

25. A method of relieving, improving, treating ocular surface symptoms caused by injury or surgery in a subject in need thereof, the method comprising administering to the subject atherapeutically effective amount of any one of the pharmaceutical compositions of claims 1- 16.

26. A method of relieving, improving, treating ocular surface symptoms caused by foreign body wear in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

27. A method of relieving, improving, treating ocular surface symptoms caused by immune mediated diseases or inflammatory conditions in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

28. A method of relieving, improving, treating ocular surface symptoms caused by diabetes in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1- 16.

29. A method of relieving, improving, treating ocular surface symptoms caused by surgical-related factors in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

30. A method of relieving, improving, treating ocular surface symptoms caused by meibomian glands disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

31. The method of any one of claims 17-30, wherein the ocular surface symptoms include one or more of feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the comeal epithelium, density' reduction of conjunctival goblet cells, squamous metaplasia of theocular surface epithelium, and / or ocular surface inflammation.

32. A method of relieving dry eye symptoms in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

33. A method of improving dry eye symptoms in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

34. A method of treating dry eye symptoms in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

35. A method of decreasing ocular surface damage in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

36. A method of decreasing cornea epithelial damage in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

37. A method of restoring ocular surface health in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

38. A method of restoring conjunctival epithelial condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

39. A method of decreasing conjunctival hyperemia in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

40. A method of restoring tear film stability in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

41. A method of increasing tear breakup time (BUT) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

42. A method of increasing goblet cells in the eye of a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

43. A method of increasing mucin production in the eye of a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

44. A method of restoring or increasing level of Lymphotoxin-alpha in tear film in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

45. A method of restoring ocular surface immune homeostasis in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the pharmaceutical compositions of claims 1-16.

46. The method of any one of claims 32-34, wherein the dry’ eye symptoms include one or more of feelings of dryness, burning sensation, gritty feeling, eye twitching, soreness, abnormal frequent blinking, eye fatigue, blurred vision, decreased tear production, pain, foreign body sensation, redness, itching, photophobia (sensitivity7to light), lacrimation, changes in goblet cell confluence, delay in epithelial tissue healing, reduction in tear film stability, damage to the ocular surface epithelium, damage to the barrier function of the comeal epithelium, density7reduction of conjunctival goblet cells, squamous metaplasia of the ocular surface epithelium, and / or ocular surface inflammation.

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