Stable peptide compositions for the treatment of moderate / severe dry eye-related ocular symptoms and methods of using the same

Abstract

The present disclosure generally relates to stable peptide compositions and kits containing low concentrations of buffering and chelating agents, and methods of using them for the treatment of patients with moderate / severe or severe dry eye.

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims priority based on U.S. Provisional Application No. 63 / 357,638, filed on July 1, 2022, and U.S. Provisional Application No. 63 / 390,550, filed on July 19, 2023, each of which is incorporated herein by reference in its entirety.

[0002] (Field of the Invention) This application relates to the fields of chemistry, biochemistry, and medicine. More specifically, some embodiments of this application relate to stable diluted peptide compositions for use in the treatment of eye disorders, particularly moderate / severe or severe dry eye, and kits containing such compositions.

[0003] (Description of Related Art) Dry - eye treatments are commercially available, but there is still a need for effective treatments for common dry eye, particularly in patients with moderate / severe or severe dry - eye symptoms.

Summary of the Invention

[0004] There is an unmet need for effective treatment of dry eye, particularly in patients having moderate / severe dry - eye symptoms as disclosed herein (e.g., patients with an ocular dryness score of at least 60). There is also an unmet need for peptide compositions that provide a therapeutic amount of peptide, are stable at room temperature, and contain only a minimal amount or no stabilizer and / or preservative.

[0005] Non - limiting embodiments of the present disclosure include the following numbered embodiments: 1. A method of treating moderate / severe dry - eye - related ocular symptoms, the method comprising: identifying a subject suffering from moderate / severe dry - eye - related ocular symptoms; and administering a liquid composition to the subject wherein the liquid composition comprises the following: A polypeptide or a pharmaceutically acceptable salt thereof at a concentration of 0.0001 to 0.005% (w / v), wherein the polypeptide has a sequence consisting of Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-NH2 (SEQ ID NO: 1), in which "Ac" represents an acetyl group and the C-terminus is amidated; A buffer at a concentration of 0.01 to 0.6% (w / v); Disodium EDTA at a concentration of 0.0005 to 0.01% (w / v); Tyloxapol at a concentration of 0.0001 to 0.01% (w / v) or 0.0001 to 0.05% (w / v); and Sodium chloride at a concentration of 0.1 to 1.0% (w / v) comprising wherein the pH of the composition is from about 6.2 to about 6.8, A method of administering 1 drop of the composition to the eye of a subject up to three times a day. 2. The method according to embodiment 1, wherein the subject satisfies at least one of the criteria selected from the group consisting of: a) an FCS total score of ≧4 and <15 on the NEI / Industry Workshop scale; b) an eye dryness score of ≧60 using VAS instantaneous measurement; c) a score of ≦5 mm wetting / 5 minutes in the Schirmer 1 test under anesthesia; and d) an LGCS total score of ≧5 (0 = no staining) using the NEI / Industry Workshop scale from the group consisting of 3. The method according to embodiment 1 or 2, wherein the subject has an eye dryness score of ≧60 using VAS instantaneous measurement. 4. The method according to embodiment 1 or 2, wherein the subject satisfies all four criteria in one eye. 5. The method according to any one of embodiments 1 to 4, wherein the subject does not have Sjögren's syndrome. 6. The method according to any one of the preceding embodiments, wherein the polypeptide or a pharmaceutically acceptable salt thereof is present in the liquid composition at 0.00025 to 0.005% (w / v) or 0.0001 to 0.001% (w / v). 7. The method according to any one of the preceding embodiments, wherein the polypeptide or a pharmaceutically acceptable salt thereof is present in the liquid composition at 0.00025% or 0.001% (w / v). 8. The method according to any one of the preceding embodiments, wherein tyloxapol is present in the liquid composition at 0.0005 to 0.01% (w / v) or 0.0005 to 0.05% (w / v). 9. The method according to any one of the preceding embodiments, wherein tyloxapol is present in the liquid composition at about 0.001% (w / v) or about 0.01% (w / v) or about 0.05% (w / v). 10. The method according to any one of the preceding embodiments, wherein the pH of the composition is about 6.0 to about 7.0, or about 6.4 to about 6.6. 11. The method according to embodiment 9 or 10, wherein the pH of the composition is about 6.5. 12. The method according to any one of the preceding embodiments, wherein the amount of NaCl is about 0.4% (w / v) to about 0.6% (w / v). 13. The method according to any one of the preceding embodiments, wherein the amount of NaCl is about 0.5% (w / v). 14. The method according to embodiment 11 or 13, wherein the osmolality of the composition is about 190 to 210 mOsm / kg. 15. The method according to any one of the preceding embodiments, wherein the buffer is a citrate buffer. 16. The method according to embodiment 13 or 15, wherein the citrate buffer contains 0.0098% of citric anhydride and 0.279% of sodium citrate dihydrate. 17. The method according to any one of the preceding embodiments, wherein the amount of EDTA is about 0.0005% (w / v) to about 0.005% (w / v). 18. The method according to any one of the preceding embodiments, wherein the amount of EDTA is about 0.001% (w / v). 19. The method according to any one of the preceding embodiments, wherein the composition further comprises 0.04% methylparaben. 20. The method according to any one of the preceding embodiments, wherein the composition is sterile. 21. The method according to any one of the preceding embodiments, further comprising determining that the subject has a history of one or more dry eye-related eye symptoms. 22. The method according to any one of the preceding embodiments, wherein the one or more dry eye-related eye symptoms include the subject's use of eye moisturizing agents within the past six months. 23. The method according to any one of the preceding embodiments, wherein the composition is administered three times a day. 24. The method according to any one of the preceding embodiments, wherein the composition is administered up to three times a day for at least one week. 25. The method according to any one of the preceding embodiments, wherein the composition is administered up to three times a day for 1 to 6 weeks. 26. The method according to any one of the preceding embodiments, wherein the administration improves the total FCS score (NEI / Industry Workshop 0 - 15 scale) in the subject's eye after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or at least 6 weeks after the start of 4 weeks of treatment, as compared to the baseline measurement before the start of treatment. 27. The administration is as follows: Eye dryness after at least 2 weeks of treatment or after at least 4 weeks of treatment, compared to the baseline on the visual analog scale; The total score of SANDE (total score of SANDE 1) after at least 2 weeks of treatment, compared to the baseline measurement before the start of treatment; The average score of the total score of SANDE (total score of SANDE - 1) after at least 2 weeks of treatment, compared to the baseline measurement before the start of treatment; Individual symptom evaluation (instantaneous) after at least 2 weeks of treatment, compared to the baseline measurement before the start of treatment; The average score of individual symptom evaluation (reflex) after at least 2 weeks of treatment, compared to the baseline measurement before the start of treatment; The LGCS of the eye of the subject after at least 2 weeks of treatment, compared to the baseline measurement before the start of treatment; The Schirmer test under anesthesia of the eye of the subject after at least 2 weeks of treatment, compared to the baseline measurement before the start of treatment; The TFBUT of the eye of the subject after at least 2 weeks of treatment, compared to the baseline measurement before the start of treatment; The FCS of the eye of the subject after at least 2 weeks of treatment, compared to the baseline measurement before the start of treatment; The SANDE (total score of SANDE 1) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before the start of treatment; Individual symptoms (instantaneous) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before the start of treatment; The average score of (total score of SANDE-2) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before the start of treatment; The average score of the individual symptom evaluation (reflex) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before the start of treatment; FCS and SANDE 1 and individual symptom evaluation (instantaneous) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, compared to the baseline measurement before the start of treatment; The LGCS after at least 2 weeks of treatment, or after at least 4 weeks of treatment, compared to the baseline measurement before the start of treatment; The result of the Schirmer test under anesthesia after at least 2 weeks of treatment, or after at least 4 weeks of treatment, compared to the baseline measurement before the start of treatment; The TFBUT after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before the start of treatment The method according to any one of the preceding embodiments, which improves one or more of them. 28. The method according to embodiment 27, wherein the improvement is 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100% of the measured or evaluated value, approximately that, at least that, at least approximately that, or within a range defined by any two of the foregoing values. 29. The subject meets the following criteria: 18 years of age or older; A recorded history or current diagnosis of dry eye-related eye symptoms; and A history of dry eye-related eye symptoms and self-reported use of over-the-counter eye lubricants within the past 120 days The method according to any one of the preceding embodiments, further satisfying all of the above. 30. The subject meets the following criteria: Any active infectious eye condition; Monocular or, when evaluated by the Early Treatment Diabetic Retinopathy Study (ETDRS), best corrected visual acuity (BCVA) of 1.0 logMAR or less using corrective lenses as needed; Ocular inflammatory conditions not related to dry eye syndrome (e.g., conjunctivitis, keratitis, anterior blepharitis, etc.); Clinical evidence of cicatricial ocular surface disease, such as cicatricial pemphigoid or Stevens-Johnson syndrome; During the induction and treatment periods, the use of any topical ophthalmic drug other than the test drug (including topical cyclosporine) cannot be interrupted; Restasis® (topical cyclosporine) was used within 60 days before the start of treatment with the composition; Xiidra® (topical lifitegrast) was used within 60 days before the start of treatment with the composition; The subject's eye has a total fluorescein corneal staining (FCS) score = 15 or a score = 3 on the upper region NEI / Industry Workshop scale, or the subject's eye has FCS with diffuse confluent staining, filaments, or epithelial defects; Subjects who have active herpetic keratitis, have developed herpetic keratitis, or are taking chronic oral antiviral drugs for herpes disease within 365 days of treatment initiation; During treatment, the use of contact lenses cannot be interrupted and the use cannot be refrained from; Has used amiodarone or its use is predicted; Within 30 days before treatment initiation, the dosage of tetracycline, omega-3 or omega-6 is changed or a dosage change is predicted; Within 60 days before treatment initiation and / or during the treatment period, the dosage of anticholinergic drugs, antidepressants, oral contraceptives, isotretinoin, oral systemic corticosteroids, oral systemic immunosuppressive drugs is changed or a dosage change is predicted, Within 30 days before treatment initiation and / or during the treatment period, topical ocular antihistamines, ocular, inhaled or intranasal corticosteroids, topical or oral mast cell stabilizers, oral antihistamines, topical or nasal vasoconstrictors, topical ocular NSAIDs, topical ocular antibiotics are used; In the subject's eye, within the past 90 days, before treatment initiation, has undergone punctal cautery or a change (insertion or removal) to punctal plugs; In the subject's eye, has undergone corneal refractive surgery (LASIK, PRK, RK); Has a history of any surgical procedure on the ocular surface or eyelid within 365 days before treatment initiation and has a history of intraocular surgery within 90 days before treatment initiation; Is pregnant or has a suspicion of pregnancy; Is breastfeeding or has an intention to breastfeed; Has any physical or mental disorder that impedes the ability to participate and give informed consent; and Participated in a device or investigational drug trial or clinical trial within 30 days of treatment initiation The method according to any one of the prior embodiments that does not satisfy one or more of the above. 31. The subject has the following criteria in one or both eyes: a) The total FCS score on the NEI / Industry Workshop scale is <4 or ≧15, b) The ocular dryness score using instantaneous VAS is < 60, c) The score of the Schirmer I test under anesthesia is > 5 mm wetting / 5 min, or d) The total LGCS score using the NEI / Industry Workshop scale is < 5 (0 = no staining) The method according to any one of the prior embodiments, which does not satisfy at least one of the above. 32. The method according to any one of the prior embodiments, wherein the subject is identified as suffering from severe dry eye rather than moderate / severe dry eye. 33. The method according to embodiment 32, wherein the subject has an ocular dryness score of ≧ 70 using instantaneous VAS measurement in at least one eye. 34. The method according to any one of the prior embodiments, wherein the composition comprises, consists of, or consists essentially of a composition selected from the compositions of the formulations of Tables A, B, C, 1.1, 1.2, and 1.3. 35. A liquid composition for use in treating moderate / severe dry eye-related ocular symptoms in a subject's eye, the composition comprising: 0.0001 to 0.006% (w / v) of a polypeptide or a pharmaceutically acceptable salt thereof, wherein the polypeptide has a sequence consisting of Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-NH2 (SEQ ID NO: 1), wherein "Ac" represents an acetyl group and the C-terminus is amidated, the polypeptide or a pharmaceutically acceptable salt thereof; 0.01 to 0.6% (w / v) of a buffer; 0.0005 to 0.01% (w / v) of disodium EDTA; 0.0001 to 0.01% (w / v) or 0.0001 to 0.05% (w / v) of tyloxapol; and 0.1 to 1.0% (w / v) of sodium chloride comprising The composition, wherein the pH of the composition is from about 6.2 to about 6.8. 36. The liquid composition according to embodiment 35, wherein the composition comprises, consists of, or consists essentially of a composition selected from the compositions of the formulations of Tables A, B, C, 1.1, 1.2, and 1.3. 37. The subject is as follows: a) The total FCS score is ≧4 and <15 on the NEI / Industry Workshop scale; b) The dry eye score using VAS instantaneous measurement is ≧60; c) The score of the Schirmer I test under anesthesia is ≦5 mm wetting / 5 min; and d) The total LGCS score using the NEI / Industry Workshop scale is ≧5 (0 = no staining) The liquid composition according to any one of embodiments 35 - 36, which satisfies at least one of the criteria selected from the group consisting of: 38. The liquid composition according to any one of embodiments 35 - 37, wherein the subject has a dry eye score of ≧60 using VAS instantaneous measurement. 39. The liquid composition according to any one of embodiments 35 - 38, wherein the subject satisfies all four criteria in one eye. 40. The liquid composition according to any one of embodiments 35 - 39, wherein the subject does not have Sjögren's syndrome. 41. The subject has the following criteria in one or both eyes: a) The total FCS score is <4 or ≧15 on the NEI / Industry Workshop scale, b) The dry eye score using VAS instantaneous is <60, c) The score of the Schirmer I test under anesthesia is >5 mm wetting / 5 min, or d) The total LGCS score using the NEI / Industry Workshop scale is <5 (0 = no staining) The liquid composition according to any one of embodiments 35 - 40, which does not satisfy at least one of the above. 42. The liquid composition according to any one of embodiments 35 - 41, wherein the subject is identified as suffering from severe dry eye rather than moderate / severe dry eye. 43. The liquid composition according to embodiment 42, wherein the subject has an eye dryness score of ≧70 using VAS instantaneous measurement in at least one eye.

Brief Description of the Drawings

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MODE FOR CARRYING OUT THE INVENTION

[0016] (TERMS) The following are exemplary definitions of terms used in this specification. Unless otherwise explicitly stated, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art in light of the entire specification. All patents, applications, published applications, and other publications referenced in this specification are incorporated by reference in their entirety as part of this specification, unless otherwise stated.

[0017] As used herein, the term "about" refers to a quantity, value, number, frequency, percentage, amount, or weight that varies by ±10% from the quantity, value, number, frequency, percentage, amount, or weight being referred to.

[0018] Unless otherwise indicated, when a percentage (%) value is used in the present application, the value refers to a weight / volume percent value.

[0019] As used herein, the term "tonicity agent" is given its ordinary meaning and is intended to include substances whose purpose is to change the osmolality of a composition. Suitable tonicity agents include, but are not limited to, propylene glycol, polyethylene glycol, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, monosaccharides such as dextrose, fructose, galactose, and / or simple polyols such as the sugar alcohols mannitol, sorbitol, xylitol, lactitol, isomaltitol, maltitol, hydrolyzed hydrogenated starch, glycerin, and combinations thereof.

[0020] As used herein, the term "stabilizer" is given its ordinary meaning and is intended to include substances that inhibit chemical reactions with peptides. Stabilizers can include, for example, antioxidants such as sodium pyrosulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole, butylated hydroxytoluene, and combinations thereof.

[0021] As used herein, the term "surfactant" is given its ordinary meaning and is intended to include amphiphilic molecules, meaning that it includes both a hydrophobic group (tail) and a hydrophilic group (head). Thus, a surfactant includes both a water-insoluble (or oil-soluble) component and a water-soluble component. Surfactants used herein can be detergents, wetting agents, emulsifiers, foaming agents, or dispersants. In some embodiments, a polypeptide can act as a surfactant.

[0022] As used herein, the term "chelating agent" is given its ordinary meaning and includes compounds that can form two or more bonds with metal ions, i.e., compounds containing multidentate ligands. Chelating agents include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), ethylenediamine, amino acids such as glutamic acid and histidine, organic diacids such as oxalic acid, malonic acid, succinic acid, etc., and pharmaceutically acceptable salts thereof. In some embodiments, the chelating agent is EDTA or a pharmaceutically acceptable salt thereof. In some embodiments, the polypeptide can act as a chelating agent.

[0023] As used herein, the term "thickening agent" is given its ordinary meaning and includes substances that affect the viscosity (centipoise, or Cp) of a composition. Examples of thickening agents include, but are not limited to, polysaccharides such as hyaluronic acid and its salts, chondroitin sulfate and its salts, dextran, various polymers of the cellulose family (and their derivatives), vinyl polymers, and acrylic acid polymers. Non-limiting examples of thickening agents include polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), and polyacrylic acid (PAA).

[0024] As used herein, the term "ophthalmically acceptable" is given its ordinary meaning and includes substances that are compatible with eye tissues, i.e., do not cause significant or excessive harmful effects when in contact with eye tissues.

[0025] As used herein, the terms "stable", "stability" or "stabilized" are given their ordinary meaning and include products and compositions that improve the primary, secondary and / or tertiary structure of a polypeptide. In some embodiments, a stabilized composition may have an acceptable percentage of peptide degradation products, or aggregates, products after a given time. These peptide degradation products can be, for example, the result of oxidation and / or hydrolysis of the peptide.

[0026] As used herein, the terms "peptide", "polypeptide" and "protein" are used interchangeably and shall be given their ordinary meanings. Unless otherwise clearly indicated from the context, the terms recited shall include polymers having at least two or more amino acids linked by peptide bonds. Thus, the terms include oligopeptides, analogs, derivatives, acetylated derivatives, glycosylated derivatives, PEGylated derivatives, and the like.

[0027] The term "pharmaceutically acceptable salt" shall be given its ordinary meaning and shall include salts of a compound that do not cause significant irritation to the organism to which it is administered and do not inhibit or substantially reduce the biological activity and properties of the compound. In some embodiments, the salt of the compound may improve the biological activity and properties of the compound. In other embodiments, the salt may further improve the structural integrity or chemical stability of the compound. In some embodiments, the salt is an acid addition salt of the compound. Pharmaceutical salts can be obtained by reacting the compound with an inorganic acid, such as a hydrohalic acid (e.g., hydrochloric acid or hydrobromic acid), sulfuric acid, nitric acid, or phosphoric acid. Pharmaceutical salts can also be obtained by reacting the compound with an organic acid, such as an aliphatic or aromatic carboxylic acid or sulfonic acid, such as formic acid, acetic acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, nicotinic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, or naphthalenesulfonic acid. Pharmaceutical salts can also be obtained by reacting the compound with a base that forms salts, such as an ammonium salt, an alkali metal salt, such as a sodium salt or a potassium salt, an alkaline earth metal salt, such as a calcium salt or a magnesium salt, dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, C 1- C7 alkylamine, cyclohexylamine, salts of organic bases such as triethanolamine, ethylenediamine, and salts with amino acids such as arginine and lysine. In some embodiments, the polypeptide is an acetate.

[0028] Provided herein is a method of treating dry eye-related ocular symptoms by administering a therapeutically effective amount of a polypeptide to a subject in need thereof. As used herein, "dry eye-related ocular symptoms" has the conventional and ordinary meaning as understood by one of ordinary skill in the art in view of the present disclosure. The ocular symptoms can include one or more symptoms associated with dry eye disease (or keratoconjunctivitis sicca). Dry eye disease is a multifactorial disease of the ocular surface characterized by the loss of tear film homeostasis. It can be accompanied by ocular symptoms in which tear film instability and hyperosmolarity, ocular surface inflammation and damage, and neurosensory abnormalities play etiological roles. Dry eye-related ocular symptoms can include, but are not limited to, blurred vision, photosensitivity, gritty or sandy sensation, eye irritation, eye pain, burning or discomfort, eye stinging, and eye itching. In some embodiments, the subject does not have Sjögren's syndrome (primary or secondary). As used herein, "moderate / severe" or "moderate to severe" when referring to dry eye disease or symptoms are used interchangeably and have the conventional and ordinary meaning as understood by one of ordinary skill in the art in view of the present disclosure. In some embodiments, when the subject meets the following criteria in at least one eye: a) the total FCS score is ≧4 and <15 on the NEI / Industry Workshop scale, b) the ocular dryness score using VAS instantaneous is ≧60, c) the score of the Schirmer I test under anesthesia is ≦5 mm wetting / 5 minutes, and d) the total LGCS score using the NEI / Industry Workshop scale is ≧5 (0 = no staining) then the subject has moderate / severe dry eye and / or dry eye symptoms. As used herein, "severe" when referring to dry eye disease or symptoms has the conventional and ordinary meaning as understood by one of ordinary skill in the art in view of the present disclosure. In some embodiments, when the subject has an ocular dryness score of ≧70 using VAS instantaneous measurement in at least one eye, the subject has severe dry eye and / or dry eye symptoms.

[0029] The therapeutic polypeptides of the present disclosure include lactoferrin and / or therapeutically active fragments thereof, such as Lacripep described herein TM The present application discloses the surprising finding that the therapeutic effect of Lacripep in alleviating the symptoms of subjects with moderate / severe dry eye disease and, optionally, subjects with severe dry eye is higher when the polypeptide is administered at a concentration of 22 μM or less (corresponding to about 0.005% w / v) in the composition, or more preferably 4 μM (corresponding to about 0.001% w / v) or 1 μM (corresponding to about 0.00025% w / v), than when administered at a higher concentration (e.g., 44 μM, corresponding to about 0.01% w / v). In some embodiments, the subject does not have Sjögren's syndrome (primary or secondary). TM

[0030] Some embodiments of the present application, as described above and herein, provide compositions that are stable at room temperature. In some embodiments, the composition reduces the concentration of stabilizers and other additives that can cause undesirable side effects while still providing the desired stability. In some embodiments, the composition provides stability in the eye, nasal cavity, oral cavity, epithelium, and other tissues for up to 1, 3, 6, 12, 24, and 48 hours. In some embodiments, the composition is formulated such that some or all of the components are not evaporated, absorbed, excreted, or otherwise eliminated after application to the eye or other area, but instead remain stable and active for several hours (e.g., 1 - 3 hours, 3 - 6 hours, 6 - 12 hours, 12 - 24 hours, and ranges therein). In some embodiments, the composition is a peptide, such as Lacripep TM ​or comprises other sequences specified herein, wherein the peptide is applied to the eye, and the peptide is incorporated into the lipid layer of the tear film covering the eye or at the interface between the oily and aqueous components of the tear film, and the peptide stabilizes the tear film for at least 1 to 3 hours, at least 3 to 6 hours, or at least 12 to 24 hours, or for more than 24 hours and remains in the tear fluid. In some embodiments, this feature is particularly advantageous because the active ingredient (e.g., the peptide) can be kept stable and effective for a long time. In some embodiments, the reduced dosing frequency results in a reduction of the overall burden of the component on sensitive areas of the body (e.g., the eye).

[0031] Peptides are provided in some embodiments herein, but other compounds can be used as active ingredients in addition to the peptides.

[0032] Peptides are highly selective and effective, while at the same time being relatively safe and well-tolerated. Because peptides can be chemically and physically unstable compared to certain small-molecule-based therapeutic agents, peptides are particularly well-suited to the compositions described herein. For example, peptides are prone to hydrolysis, oxidation, and aggregation. Polypeptide compositions are typically aqueous solutions containing the active peptide together with a number of stabilizers, preservatives, and other agents that maintain the effectiveness of the peptide. Stabilizers, preservatives, and other agents can maintain the chemical and / or structural integrity of the polypeptide and thus retain its effectiveness. Certain additives, such as stabilizers and preservatives, can cause undesirable side effects, including allergic reactions, itching, and stinging or burning sensations. However, these additives are required in amounts that cause undesirable results in most peptide compositions in order to maximize the shelf life of the peptide and maintain its effectiveness. Even in compositions with all of these additives, peptide therapeutics typically have to be refrigerated, which makes transportation difficult and still results in a short shelf life. Furthermore, because peptides degrade and / or aggregate over time (especially upon warming and cooling when going from refrigerated storage to room temperature for use), the by-products can be not only inactive but also toxic and / or immunogenic. Formulators may attempt to increase the potency of peptide compositions by increasing the amount of active peptide in the composition. However, an increase in peptide concentration also increases the rate of peptide aggregation and inactivation.

[0033] Accordingly, some embodiments herein provide peptide compositions that provide a therapeutic amount of a combination of peptides, are stable at room temperature, and contain reduced amounts (e.g., only trace amounts) or no stabilizers and / or preservatives.

[0034] In some embodiments, the peptide is (a) the amino acid sequence KQFIENGSEFAQKLLKKFS, Ac-KQFIENGSEFAQKLLKKFS-NH2, or Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-NH2 (where "Ac" represents an acetyl group and the C-terminus is amidated (SEQ ID NO: 1), "Lacripep TM ", also referred to herein as); and (b) selected from the group consisting of the amino acid sequences KQFIENGSEFAQKLLKKFSLLKPWA, Ac-KQFIENGSEFAQKLLKKFSLLKPWA-NH2, or Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-Leu-Leu-Lys-Pro-Trp-Ala-NH2 (where "Ac" represents an acetyl group and the C-terminus is amidated (SEQ ID NO: 2)). In other embodiments, the peptide has one of the following amino acid sequences, or a fragment thereof, and optionally, the N-terminus is acetylated and the C-terminus is amidated:

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

[0035] In some embodiments, the peptide is represented by the amino acid sequence Ac-KQFIENGSEFAQKLLKKFS-NH2 or Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-NH2, where "Ac" represents an acetyl group and the C-terminus is amidated (SEQ ID NO: 1). In some embodiments, the peptide is Lacripep TM . That is. In some embodiments, the peptide is any one or more of SEQ ID NOs: 1-9.

[0036] (Buffer and pH) Buffers stabilize the pH of a solution, i.e., resist changes in pH when acidic or alkaline substances are added to the solution. Suitable buffer compositions for use in the compositions of the present application include, but are not limited to, glycine hydrochloride, sodium acetate, phosphate buffered saline (PBS) (containing monohydrogen phosphate and dihydrogen), citrate buffer (citric acid and sodium citrate), phosphate-citrate buffer, tris(hydroxymethyl)aminomethane (Tris), carbonate buffer (sodium carbonate and sodium bicarbonate), borate buffer, and combinations thereof.

[0037] In some embodiments, the buffer solution includes one or more of sodium acetate, phosphate buffered saline (PBS), citrate buffer (citric acid and sodium citrate), and phosphate-citrate buffer. In some embodiments, the buffer solution is selected from the group consisting of sodium acetate, phosphate buffered saline (PBS), citrate buffer (citric acid and sodium citrate), and phosphate-citrate buffer.

[0038] In some embodiments, the amount of buffer is less than 0.1, 0.2, 0.3, or 0.4%, or is limited within the range defined by any two of the foregoing values.

[0039] In one embodiment, the buffer is a citrate buffer (citric acid and sodium citrate). In one embodiment, the only buffer is the citrate buffer and no other buffers are present in the composition.

[0040] In some embodiments, the pH of the composition is 6.0 - 7.4; 6.1 - 7.3; 6.2 - 7.2; 6.3 - 7.1; 6.4 - 7.0; 6.5 - 6.9; 6.6 - 6.8 or any pH in between.

[0041] In some embodiments, the pH of the composition is 6.0; 6.1; 6.2; 6.3; 6.4; 6.5; 6.6; 6.7; 6.8; 6.9; 7; 7.1; 7.2; 7.3; 7.4 or approximately so or is in a range defined by any two of the foregoing values, for example 6.0 - 7.0, 6.2 - 6.5, 6.3 - 6.5, 6.0 - 6.5, 6.3 - 6.7, or 6.3 - 7.0. In some embodiments, the pH of the composition is 6.2, 6.3, 6.4, 6.5, 6.6, 6.7 or 6.8 or approximately so. In some embodiments, the pH of the composition is 6.0, 6.1, 6.2, 6.3, 6.4, 6.5 or approximately so. In some embodiments, the pH of the composition is 6.2, 6.3, 6.4, 6.5, 6.6, 6.7 or 6.8 or approximately so.

[0042] The pH of the composition can be adjusted as needed by the addition of a solution of an acid or a base. Any acid or base that is ophthalmically acceptable for the conjugate can be used. Acids include, for example, hydrochloric acid and bases include, for example, sodium hydroxide and potassium hydroxide.

[0043] In some embodiments, the pH is measured using the method of USP <791>.

[0044] (Chelating agent) In some embodiments, the composition further comprises one or more chelating agents. In some embodiments, the chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid, disodium edetate (EDTA), ethylenediamine, amino acids such as glutamic acid and histidine, organic diacids such as oxalic acid, malonic acid, succinic acid, etc., 3-dimercaptopropanesulfonic acid (DMPS), alpha-lipoic acid (ALA), 2,3-dimercaptopropanesulfonic acid (DMPS), thiamine tetrahydrofurfuryl disulfonic acid (TTFD), penicillamine, dimercaptosuccinic acid (DMSA), combinations thereof, and pharmaceutically acceptable salts thereof.

[0045] In some embodiments, the chelating agent, such as EDTA or a pharmaceutically acceptable salt thereof by way of non-limiting example, is present at 0.0001% to 0.1%; 0.0005% to 0.05%; 0.0006% to 0.04%; 0.0007% to 0.003%; 0.0008% to 0.002%; 0.0009% to 0.001%; or any value or range therebetween. In some embodiments, the chelating agent is present in an amount that is 0.1%; 0.09%; 0.08%; 0.07%; 0.06%; 0.05%; 0.04%; 0.03%; 0.02%; 0.01%; 0.009%; 0.008%; 0.007%; 0.006%; 0.005%; 0.004%; 0.003%; 0.002%; 0.001%; 0.0009%; 0.0008%; 0.0007%; 0.0006%; 0.0005%; 0.0004%; 0.0003%; 0.0002%; or 0.0001% or less, or within a range defined by any two of the foregoing values.

[0046] In some embodiments, the chelating agent, such as EDTA or otherwise, or a pharmaceutically acceptable salt thereof, is present at less than about 0.05% or less than about 0.005% (e.g., about 0.001%).

[0047] (Stabilizer) Buffers and chelating agents can stabilize the peptide component of the composition by maintaining the pH and reducing the degradation of peptides via metal ions. In some embodiments, the composition further comprises one or more peptide stabilizers in addition to the buffer and / or chelating agent. In some embodiments, one or more stabilizers in addition to the buffer and / or chelating agent are selected from the group consisting of disaccharides, polysaccharides (e.g., hyaluronic acid), polyols, sugar alcohols, amino acids, proteins (e.g., serum albumin), and combinations thereof. In some embodiments, non-limiting examples of stabilizers include trehalose, sucrose, mannitol, sorbitol, polysorbate 20, polysorbate 80, histidine, glycine, and arginine, and combinations thereof. In one embodiment, the composition does not contain a stabilizer in addition to the buffer and / or chelating agent.

[0048] (Polypeptide degradation) Polypeptides are prone to physical and chemical degradation, such as aggregation, shear, oxidation, deamidation, and hydrolysis. In fact, liquid peptide compositions are at high risk of becoming physically and chemically unstable during manufacture and storage. Reduction of polypeptide degradation is particularly important for low-concentration peptide formulations that initially contain very small amounts of specific peptides. Even a very small loss of the initial very small amount can significantly affect the effectiveness of the composition.

[0049] In some embodiments, the stability of the composition is determined by high performance liquid chromatography (HPLC). In some embodiments, the stability of the composition is determined by high performance liquid chromatography-mass spectrometry (HPLC-MS).

[0050] In some embodiments, the stability of the composition is determined after being placed in the dark or under light exposure at room temperature for several days, weeks, or months (e.g., 1 to 24 days or 1 to 24 months, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 days or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 months).

[0051] In some embodiments, the stability of the composition is determined after being placed in the dark or under light exposure at 2 to 8 °C, e.g., 5 °C, or any value therebetween, for several days, weeks, or months (e.g., 1 to 24 days or 1 to 24 months, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 days or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 months).

[0052] In some embodiments, the stability of the composition is determined after being placed in the dark or under light exposure at -10 to -30 °C, e.g., -25 °C, or any value therebetween, for several days, weeks, or months (e.g., 1 to 24 days or 1 to 24 months, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 days or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 months).

[0053] In some embodiments, the stability of the composition is determined after being placed in the dark or under light exposure and transferred from 2 to 8 °C (storage), or any value therebetween, to room temperature 1, 2, or 3 times per day for 1 to 60 days and then allowed to stand for 5 minutes.

[0054] In some embodiments, after exposure to one or more of the above or herein-described conditions, the composition provides at least 99.99%, 99.95%, 99.9%, 99%; 98%; 97%; 96%; 95%; 94%; 93%; 92%; 91%; 90%; 89%; 88%; 87%; 86%; 85%; 84%; 83%; 82%; 81%; 80%; 79%; 78%; 77%; 76%; 75%; 74%; 73%; 72%; 71%; 70%; or any value therebetween of the original amount or activity of the polypeptide or a pharmaceutically acceptable salt thereof in an intact, non-degraded or non-aggregated form. In some embodiments, the amount or activity of the intact polypeptide or a pharmaceutically acceptable salt thereof is at least 80%, 85%, 90% or 95% of the original amount. In some embodiments, the amount or activity of the intact polypeptide or a pharmaceutically acceptable salt thereof is at least 97% of the original amount.

[0055] In some embodiments, after exposure to one or more of the above or herein-described conditions, the composition comprises 30%; 29%; 28%; 27%; 26%; 25%; 24%; 23%; 22%; 21%; 20%; 19%; 18%; 17%; 16%; 15%; 14%; 13%; 12%; 11%; 10%; 9%; 8%; 7%; 6%; 5%; 4%; 3%; 2%; 1% or less of peptide aggregation products or peptide degradation products, or is within the range defined by any two of the foregoing values. In some embodiments, the composition comprises about 15% or less or 20% or less of inert peptides.

[0056] In some embodiments, after exposure to one or more of the above or herein-described conditions, the composition comprises 30%; 29%; 28%; 27%; 26%; 25%; 24%; 23%; 22%; 21%; 20%; 19%; 18%; 17%; 16%; 15%; 14%; 13%; 12%; 11%; 10%; 9%; 8%; 7%; 6%; 5%; 4%; 3%; 2%; 1% or less of the total amount of peptide degradation products or peptide aggregation products, or is within the range defined by any two of the foregoing values.

[0057] In some embodiments, the composition comprises a very low concentration of a buffering agent in combination with a very low concentration of a chelating agent. In some embodiments, the buffering agent is a citrate buffer and the chelating agent is EDTA. The combination of a low concentration of citrate buffer (e.g., 0.012% - 0.020%) and EDTA (e.g., 0.0005% - 0.005%) provides a surprising and unexpected benefit for a stabilizing composition containing a low concentration of a peptide (e.g., 0.001 - 0.01%). Such a stabilizing composition reduces peptide aggregation and degradation, thereby maintaining the effectiveness of the peptide composition and reducing the accumulation of undesirable degradation products in the composition, providing advantages in manufacturing, transporting, storing, and using the peptide composition.

[0058] In some embodiments, the stabilizing composition reduces the rate of formation of degradation and / or aggregation products.

[0059] In some embodiments, the peptide is Lacripep TM and is. In some embodiments, the stabilizing composition comprises less than about 5%, 4%, 3%, 2% or about 1% total degradation products. In some embodiments, the stabilizing composition comprises any one degradation product of 0.25%, 0.5%, 0.75%, 1.0%, 1.25%, 1.5%, 1.75% or 2.0% or less. In some embodiments, the stabilizing composition comprises less than about 5%, 4%, 3%, 2% or about 1% total degradation products and any one degradation product of 0.25%, 0.5%, 0.75%, 1.0%, 1.25%, 1.5%, 1.75% or 2.0% or less.

[0060] In some embodiments, the aggregation products include dimers, trimers, tetramers, or higher order peptide aggregates.

[0061] (Preservative) In some embodiments, the composition further comprises one or more preservatives to prevent the growth of microorganisms in the composition. In some embodiments, the composition further comprises one or more preservatives to maintain the sterility of the composition. In some embodiments, the composition further comprises one or more preservatives to prevent the growth of microorganisms and maintain the sterility of the composition. However, in many embodiments, the preservatives are provided in reduced amounts. In some embodiments, the one or more preservatives are selected from the group consisting of benzalkonium chloride, cetylpyridinium chloride, chlorobutanol, benzalkonium bromide, methylparaben, propylparaben, phenylethyl alcohol, sodium perborate, disodium edetate, chlorobutanol, sorbic acid, benzethonium chloride, sodium acetate, polyquaternium-1, phenylmercuric nitrate, phenylmercuric borate, sodium propionate, chlorhexidine, thimerosal, and combinations thereof. In some embodiments, the composition does not contain a preservative. In some embodiments, the composition does not contain a detectable concentration of a preservative. In some embodiments, the polypeptide can be self-preserving, i.e., further preservatives are not necessary to maintain the sterility of the composition.

[0062] In some embodiments, the preservative is present in an amount of 0.0001% to 1%; 0.01% to 0.9%; 0.05% to 0.8%; 0.1% to 0.7%; 0.2% to 0.3%; 0.4% or 0.5%, or any value included therein. In some embodiments, the preservative is present in an amount of 1%; 0.9%; 0.8%; 0.7%; 0.6%; 0.5%; 0.4%; 0.3%; 0.2%; 0.1%; 0.09%; 0.08%; 0.07%; 0.06%; 0.05%; 0.04%; 0.03%; 0.02%; 0.01%; 0.009%; 0.008%; 0.007%; 0.006%; 0.005%; 0.004%; 0.003%; 0.002%; or 0.001% or less, or in a range defined by any two of the foregoing values.

[0063] In some embodiments, the composition is sterile. In some embodiments, the composition is manufactured from sterile components in a sterile environment. In some embodiments, the composition is sterilized prior to packaging. In some embodiments, the composition is sterilized by one or more of (1) addition of one or more quaternary ammonium chlorides to the composition; (2) exposure of the composition to ionizing radiation; (3) filtration of the composition; (4) exposure of the composition to ionizing radiation after packaging; and any combination of the foregoing. In some embodiments, filtration includes passing the composition through a filter (including, but not limited to, a 0.22 μm filter having polyvinylidene fluoride or other suitable membrane (e.g., polyethersulfone)).

[0064] In some embodiments, the peptide is provided in bacteriostatic and / or bactericidal amounts. In some embodiments, the amount of peptide provided in the composition is bacteriostatic and / or bactericidal when 1, 2, or 3 drops of the composition are administered to the surface of the eye. In some embodiments, the peptide is bacteriostatic and / or bactericidal against Gram-positive and / or Gram-negative bacteria, for example when administered to the eye. In some embodiments, the amount of peptide in the composition is sufficient to inhibit bacterial growth by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% compared to a control composition that does not contain the peptide in a standard bacteriological assay. In some embodiments, the bacteria in the bacteriological assay are selected from P. aeruginosa, E. coli, S. epidermidis, S. aureus, or combinations thereof. In some embodiments, the bacteriological assay is selected from a bacterial growth assay, SYTOX Green assay, well diffusion assay, broth or canten dilution assay, time kill test, antimicrobial gradient assay, ATP bioluminescence assay, or propidium-iodide flow cytometry assay. In some embodiments, the peptide provided in bacteriostatic and / or bactericidal amounts is Lacripep TM is

[0065] In some embodiments, the bacteriological assay is an assay of section <51> of the USP or an assay mandated by the FDA. For example, the original product container contains a peptide solution, and each container is seeded with and mixed with one of the prepared and standardized inoculum solutions (e.g., P. aeruginosa, E. coli, S. epidermidis, S. aureus, or a combination thereof). The amount of the suspended inoculum solution is about 0.5% - 1.0% of the amount of the product, and the concentration of the test preparation immediately after seeding is 1x10 5 ~1x10 6 colony forming units (CFU) per mL of the product (e.g., measured by the plate count method or another microbial counting test).

[0066] The seeded containers are incubated at 22.5 ± 2.5 °C in a controlled environment and sampled at specific intervals (e.g., 7, 14, and 28 days). For each sampling, changes in appearance are recorded and the CFU / mL is determined. The change in the log 10 value of the CFU / mL provides the change over time from the point of logarithmic decrease. The product provides a logarithmic decrease of 1.0 or less from the first calculated number on day 7, a logarithmic decrease of 3.0 or less from the first number on day 14, and no increase from the number on day 14 at day 28 for bacteria, and no increase from the first number for yeast and mold. In some embodiments, the peptide provided in bacteriostatic and / or bactericidal amounts is Lacripep TM .

[0067] (Surfactant) In some embodiments, the composition further comprises one or more surfactants. In some embodiments, the one or more surfactants are selected from detergents, wetting agents, emulsifiers, foaming agents, dispersing agents, and combinations thereof.

[0068] In some embodiments, the surfactant is an anionic surfactant. An anionic surfactant contains an anionic functional group, such as sulfuric acid, sulfonic acid, phosphoric acid, and carboxylic acid, at its head. In some embodiments, the surfactant is a sulfate ester, a sulfonate ester, or a phosphate ester, such as a sulfate ester. In some embodiments, the surfactant is selected from the group consisting of or including ammonium lauryl sulfate and sodium lauryl sulfate, such as sodium lauryl sulfate (also called SDS, sodium dodecyl sulfate). In some embodiments, the surfactant is an alkyl-ether sulfate, such as sodium laureth sulfate (also known as sodium lauryl ether sulfate), and sodium myreth sulfate, and is selected from the group consisting of or including them. In some embodiments, the surfactant is doxate, such as sodium dioctyl sulfosuccinate, perfluorooctanesulfonic acid (PFOS), perfluorobutanesulfonic acid, linear alkylbenzene sulfonic acid (LAB). In some embodiments, the surfactant is a carboxylate, such as an alkyl carboxylate (soap), such as sodium stearate; sodium lauroyl sarcosinate and carboxylic acid-based fluorosurfactants, such as perfluorononanoic acid, perfluorooctanoic acid (PFOA or PFO). In some embodiments, the polypeptide contributes to the surfactant properties of the composition.

[0069] In some embodiments, the surfactant is a cationic surfactant, the charge of which can be pH-dependent, such as a primary, secondary or tertiary amine, such as octenidine dihydrochloride; or a permanently charged quaternary ammonium cation, such as an alkyltrimethylammonium salt, such as cetyltrimethylammonium bromide (CTAB) or cetyltrimethylammonium chloride (CTAC); cetylpyridinium chloride (CPC); benzalkonium chloride (BAC); benzethonium bromide (BZT); 5-bromo-5-nitro-1,3-dioxane; dimethyldioctadecylammonium chloride; or dioctadecyldimethylammonium bromide (DODAB). In some embodiments, the surfactant is an amphoteric surfactant (i.e., having both cationic and anionic centers bonded to the same molecule). The cationic moiety can be based on a primary, secondary or tertiary amine or a quaternary ammonium cation. The anionic moiety can be variable and include sulfonic acid, such as CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid). Other anionic groups can be thiones exemplified by cocamidopropyl hydroxysultaine; betaines such as cocamidopropyl betaine; and phosphates such as lecithin. In some embodiments, the surfactant can be a nonionic surfactant (not charged).

[0070] Many long-chain alcohols exhibit some surfactant properties and are provided herein as part of the composition in some embodiments. Among these are the notable ones, which are the fatty alcohols, cetyl alcohol, stearyl alcohol, and cetostearyl alcohol (consisting mainly of cetyl and stearyl alcohol), and oleyl alcohol. Other surfactants include cocamide MEA, cocamide DEA, dodecyl dimethyl amine oxide, and polyethoxylated tallow amine (POEA). Examples of nonionic surfactants include polyoxyethylene glycol alkyl ethers, such as octaethylene glycol monodecyl ether or pentaethylene glycol monododecyl ether; polyoxypropylene glycol alkyl ethers; glucoside alkyl ethers, such as decyl glucoside, lauryl glucoside, or octyl glucoside; polyoxyethylene glycol octylphenol ethers, such as Triton X-100; polyoxyethylene glycol alkylphenol ethers, such as nonoxynol-9; glycerol alkyl esters, such as glycerol laurate; polyoxyethylene glycol sorbitan alkyl esters (polysorbate); sorbitan alkyl esters (span); block copolymers of polyethylene glycol and polypropylene glycol, or poloxamers.

[0071] In some embodiments, the composition may include one or more components found in artificial tears in amounts known in the art, including, but not limited to, carboxymethylcellulose, polyvinyl alcohol, hydroxypropylmethylcellulose (also known as HPMC or hypromellose), hydroxypropylcellulose, hydroxyethylcellulose (HEC), and hyaluronic acid (also known as hyaluronan, HA), and combinations thereof. In some embodiments, the composition does not contain any of the aforementioned artificial tear components.

[0072] In some embodiments, the surfactant is another peptide or protein. In some embodiments, by way of non-limiting example, the surfactant is human serum albumin. In some embodiments, by way of another non-limiting example, the surfactant is Lacripep TM is.

[0073] In some embodiments, the surfactant is tyloxapol (formaldehyde; oxirane; 4-(2,4,4-trimethylpentan-2-yl)phenol). In one embodiment, the only surfactant is tyloxapol and no other surfactants are present in the composition.

[0074] In some embodiments, the surfactant, by way of non-limiting example tyloxapol, is present in an amount of 0.01% to 1%; 0.05% to 0.9%; 0.1% to 0.8%; 0.2% to 0.7%; 0.3% to 0.6%; 0.4% to 0.5%, or any value included therein. In some embodiments, the surfactant is 1%; 0.9%; 0.8%; 0.7%; 0.6%; 0.5%; 0.4%; 0.3%; 0.2%; 0.1%; 0.09%; 0.08%; 0.07%; 0.06%; 0.05%; 0.04%; 0.03%; 0.02%; 0.01%; 0.009%; 0.008%; 0.007%; 0.006%; 0.005%; 0.004%; 0.003%; 0.002%; or 0.001% or less, or is present in an amount within a range defined by any two of the foregoing values.

[0075] In some embodiments, the composition comprises tyloxapol at 0.0001%, 0.0002%, 0.0005%, 0.001%, 0.002%, 0.005%, 0.01%, 0.02% or 0.05% (w / v), or approximately so, or in a range defined by any two of the foregoing values (e.g., 0.0001 - 0.05%, 0.0001 - 0.02%, 0.001 - 0.01%, 0.0001 - 0.005%, 0.0002 - 0.002%, 0.0005 - 0.002%, 0.0002 - 0.001%, 0.001 - 0.01%, 0.001 - 0.05% or 0.001 - 0.005%, etc.). In some embodiments, tyloxapol is present in the liquid composition at 0.0001 - 0.002% (w / v). In some embodiments, tyloxapol is present in the liquid composition at 0.0005 - 0.005% (w / v). In some embodiments, tyloxapol is present in the liquid composition at 0.001 - 0.01% (w / v). In some embodiments, tyloxapol is present in the liquid composition at approximately 0.001% (w / v). In some embodiments, tyloxapol is present in the liquid composition at approximately 0.01% (w / v). In some embodiments, tyloxapol is present in the liquid composition at approximately 0.05% (w / v).

[0076] In some embodiments, the composition does not contain a surfactant. In some embodiments, the composition does not contain a detectable concentration of surfactant.

[0077] (Isotonic Agents and Osmolarity) In some embodiments, the composition further comprises one or more tonicity agents. Such tonicity agents are added to any polypeptide or buffer having an osmotic pressure modifying effect. In some embodiments, the one or more tonicity agents are selected from propylene glycol, polyethylene glycol, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, monosaccharides such as dextrose, fructose, galactose, and / or simple polyols such as the sugar alcohols mannitol, sorbitol, xylitol, lactitol, isomaltitol, maltitol, hydrolyzed hydrogenated starch, glycerin, and combinations thereof.

[0078] In some embodiments, the one or more tonicity agents are selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, dextrose, mannitol, and combinations thereof.

[0079] In some embodiments, the tonicity agent is sodium chloride. In some embodiments, sodium chloride is present at 0.01% - 1%; 0.05% - 0.9%; 0.1% - 0.8%; 0.2% - 0.75%; 0.3% - 0.7%; 0.4% - 0.6%; or any value included therein. In some embodiments, sodium chloride is 1%; 0.95%; 0.9%; 0.85%; 0.8%; 0.75%; 0.7%; 0.65%; 0.6%; 0.55%; 0.5%; 0.45%; 0.4%; 0.35%; 0.3%; 0.25%; 0.2%; 0.15%; 0.1%; 0.09%; 0.08%; 0.07%; 0.06%; 0.05%; 0.04%; 0.03%; 0.02%; or 0.01%, or is present in an amount approximately that, or is in a range defined by any two of the foregoing values.

[0080] In some embodiments, the only tonicity agent is sodium chloride and no other tonicity agents are present in the composition.

[0081] In some embodiments, an isotonic agent, such as sodium chloride by way of non-limiting example, is added to the composition to adjust the osmolality to a desired level. In some embodiments, the osmolality of the composition is from about 150 to about 400 mOsm / kg; from about 170 to about 380 mOsm / kg; from about 190 to about 360 mOsm / kg; from about 210 to about 340 mOsm / kg; from about 230 to about 320 mOsm / kg; from about 250 to about 300 mOsm / kg; from about 270 to about 280 mOsm / kg; or any value therebetween. In some embodiments, the osmolality of the composition is from about 250 to about 350 mOsm / kg; from about 260 to about 340 mOsm / kg; from about 270 to about 330 mOsm / kg; from about 280 to about 320 mOsm / kg; from about 290 to about 310 mOsm / kg; or any value therebetween. In some embodiments, the osmolality of the composition is from about 150 to about 300 mOsm / kg; from about 150 to about 250 mOsm / kg; from about 160 to about 230 mOsm / kg; from about 170 to about 220 mOsm / kg; from about 180 to about 220 mOsm / kg; from about 190 to about 210 mOsm / kg; from about 190 to about 200 mOsm / kg; from about 170 to about 210 mOsm / kg; from about 180 to about 200 mOsm / kg; or any value therebetween.

[0082] In some embodiments, the osmolality of the composition is 150 mOsm / kg; 160 mOsm / kg; 170 mOsm / kg; 180 mOsm / kg; 190 mOsm / kg; 200 mOsm / kg; 210 mOsm / kg; 220 mOsm / kg; 230 mOsm / kg; 240 mOsm / kg; 250 mOsm / kg; 260 mOsm / kg; 270 mOsm / kg; 280 mOsm / kg; 290 mOsm / kg; 300 mOsm / kg; 310 mOsm / kg; 320 mOsm / kg; 330 mOsm / kg; 340 mOsm / kg; or 350 mOsm / kg, or approximately so, or within a range defined by any two of the foregoing values.

[0083] In some embodiments, the osmolality of the composition is from about 280 mOsm / kg to about 320 mOsm / kg. In some embodiments, the osmolality of the composition is about 300 mOsm / kg. In some embodiments, NaCl is used to adjust the osmolality of the solution to the desired level. In one embodiment, the composition is isotonic or approximately isotonic with human tears.

[0084] In some embodiments, the osmolality is measured using the method of USP <785>.

[0085] (Polypeptide and other components) In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof has 10 to 150 amino acids; 10 to 50 amino acids; 100 to 150 amino acids; 30 to 70 amino acids; or any number included therein. In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof has 10 to 30 amino acids; 11 to 29 amino acids; 12 to 28 amino acids; 13 to 27 amino acids; 14 to 26 amino acids; 15 to 25 amino acids; 16 to 24 amino acids; 17 to 23 amino acids; 18 to 22 amino acids; 19 to 21 amino acids; or any number included therein. In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof is, has approximately, or is in a range defined by any two of the foregoing values, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 amino acids in length.

[0086] In some embodiments, the C-terminus of the polypeptide or a pharmaceutically acceptable salt thereof is amidated. In some embodiments, the N-terminus of the polypeptide or a pharmaceutically acceptable salt thereof is acetylated. In some embodiments, one or more side chains of the polypeptide or a pharmaceutically acceptable salt thereof are acetylated. In some embodiments, one or more side chains of the polypeptide or a pharmaceutically acceptable salt thereof are amidated. In some embodiments, the N-terminus of the polypeptide or a pharmaceutically acceptable salt thereof is acetylated and the C-terminus of the polypeptide or a pharmaceutically acceptable salt thereof is amidated.

[0087] In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof comprises, consists of, or consists essentially of the amino acid sequence: Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-Leu-Leu-Lys-Pro-Trp-Ala-NH2 (SEQ ID NO: 2), wherein "Ac" represents an acetyl group and the C-terminus is amidated (indicated by "NH2"). In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof comprises the amino acid sequence: Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-NH2 (SEQ ID NO: 1), wherein "Ac" represents an acetyl group and the C-terminus is amidated (indicated by "NH2"). In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof comprises, consists of, or consists essentially of a sequence selected from the group of SEQ ID NOs: 3-9, or a fragment thereof or a pharmaceutically acceptable salt thereof.

[0088] In some embodiments, the amount of the polypeptide or a pharmaceutically acceptable salt thereof in the composition is 0.0001% to 0.005%; 0.0005% to 0.005%; or 0.001% to 0.005%, or approximately so. In one embodiment, the polypeptide or a pharmaceutically acceptable salt thereof is present in the composition at about 0.003% to 0.09% (e.g., 0.005%, 0.01%, 0.02%, 0.03% and ranges thereof).

[0089] In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof is present in the composition in an amount that is 0.0001, 0.00025, 0.0005, 0.00075, 0.001, 0.002, 0.003, 0.004, 0.005, or 0.006%, approximately that, greater than that, less than that, or within a range defined by any two of the foregoing values. In some embodiments, the composition comprises the polypeptide or a pharmaceutically acceptable salt thereof at 0.0001%, 0.0002%, 0.00025%, 0.0003%, 0.0005%, 0.00075%, 0.001%, 0.002%, 0.005% or 0.006% (w / v), approximately that, or within a range (e.g., 0.00025 - 0.006%, 0.00025 - 0.005%, 0.00025 - 0.001%, 0.0001 - 0.001%, 0.0001 - 0.002%, or 0.001 - 0.005%) defined by any two of the foregoing values. In some embodiments, the composition comprises the polypeptide or a pharmaceutically acceptable salt thereof at 0.00025 - 0.005% (w / v). In some embodiments, the composition comprises the polypeptide or a pharmaceutically acceptable salt thereof at 0.0001 - 0.002% (w / v). In some embodiments, the composition comprises the polypeptide or a pharmaceutically acceptable salt thereof at about 0.00025% (w / v). In some embodiments, the composition comprises the polypeptide or a pharmaceutically acceptable salt thereof at about 0.001% (w / v). In some embodiments, the composition comprises the polypeptide or a pharmaceutically acceptable salt thereof at about 0.005% (w / v).In some embodiments, the amount of the polypeptide or a pharmaceutically acceptable salt thereof is 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 10, 15, 20, 21, 22, 23, 24, or 25 μM, approximately that, at least that, at least approximately that, less than that, approximately less than that, or in a range defined by any two of the foregoing values (e.g., 0.1 - 25, 0.1 - 6.0, 0.1 - 4.0, 1.0 - 5.0, 1.0 - 22, or 1.0 - 4.0 μM). In some embodiments, the composition comprises the polypeptide or a pharmaceutically acceptable salt thereof at about 1.0 μM. In some embodiments, the composition comprises the polypeptide or a pharmaceutically acceptable salt thereof at about 4.0 μM. In some embodiments, the composition comprises the polypeptide or a pharmaceutically acceptable salt thereof at about 22 μM. In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof has a sequence consisting of or consisting essentially of Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-NH2 (SEQ ID NO: 1), where "Ac" represents an acetyl group and the C-terminus is amidated.

[0090] In some embodiments, the composition is a polypeptide or a pharmaceutically acceptable salt thereof at 0.0001 to 0.005% (w / v), wherein the polypeptide consists of or consists essentially of the sequence Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-NH2 (SEQ ID NO: 1), wherein "Ac" represents an acetyl group and the C-terminus is amidated, a polypeptide or a pharmaceutically acceptable salt thereof; a buffer at 0.01 to 0.6% (w / v); disodium EDTA at 0.0005 to 0.01% (w / v); tyloxapol at 0.0001 to 0.02% or 0.0001 to 0.06% (w / v); and about 0.1 to 1.0% NaCl; the pH of the composition is about 6.0 to about 7.0, and the osmolality is 170 to 210 mOsm / kg. In some embodiments, the composition is a sterile aqueous composition.

[0091] In some embodiments, the composition is a sterile aqueous composition, the composition comprising about 0.005% ± 0.0005% of a polypeptide, such as Lacripep TM (SEQ ID NO: 1) or other peptides identified herein, or a pharmaceutically acceptable salt thereof; about 0.0098% ± 0.001% of anhydrous citric acid; about 0.279% ± 0.028% of sodium citrate dihydrate; about 0.001% ± 0.0001% of disodium EDTA; about 0.001% ± 0.0001% of tyloxapol; about 0.50% ± 0.05% of NaCl, comprising, consisting of, or consisting essentially of the foregoing; the pH of the composition is adjusted to about 6.2 to about 6.8 using NaOH or HCl, and the osmolality is 170 to 210 mOsm / kg. In some embodiments, the composition is a sterile aqueous composition, the composition comprising about 0.005% ± 0.0005% of a polypeptide, such as Lacripep TM(SEQ ID NO:1) or another peptide specified herein, or a pharmaceutically acceptable salt thereof; about 0.0098% ± 0.001% anhydrous citric acid; about 0.279% ± 0.028% sodium citrate dihydrate; about 0.001% ± 0.0001% disodium EDTA; about 0.01% ± 0.001% tyloxapol; about 0.50% ± 0.05% NaCl, comprising, consisting of, or essentially consisting of the foregoing; the pH of the composition is adjusted to about 6.2 to about 6.8 using NaOH or HCl, and the osmolality is 170 to 210 mOsm / kg. In some embodiments, the composition is a sterile aqueous composition, the composition comprising about 0.005% ± 0.0005% polypeptide, such as Lacripep TM (SEQ ID NO:1) or another peptide specified herein, or a pharmaceutically acceptable salt thereof; about 0.0098% ± 0.001% anhydrous citric acid; about 0.279% ± 0.028% sodium citrate dihydrate; about 0.001% ± 0.0001% disodium EDTA; about 0.05% ± 0.005% tyloxapol; about 0.50% ± 0.05% NaCl, comprising, consisting of, or essentially consisting of the foregoing; the pH of the composition is adjusted to about 6.2 to about 6.8 using NaOH or HCl, and the osmolality is 170 to 210 mOsm / kg. In one embodiment, the pH is about 6.3 to about 6.5 and the osmolality is 180 to 200 mOsm / kg. In one embodiment, the composition consists only of the listed components and does not contain any additional active ingredients, additives (e.g., thickeners, buffers, chelating agents, stabilizers, preservatives, surfactants, and tonicity agents), carriers, or diluents.

[0092] In some embodiments, the composition is a sterile aqueous composition, the composition comprising about 0.001% ± 0.0001% polypeptide, such as Lacripep TM(SEQ ID NO:1) or other peptides specified herein, or a pharmaceutically acceptable salt thereof; about 0.0098% ± 0.001% anhydrous citric acid; about 0.279% ± 0.028% sodium citrate dihydrate; about 0.001% ± 0.0001% disodium EDTA; about 0.001% ± 0.0001% tyloxapol; about 0.50% ± 0.05% NaCl, comprising, consisting of, or consisting essentially of the foregoing; the pH of the composition is adjusted to about 6.2 to about 6.8 using NaOH or HCl, and the osmolality is 170-210 mOsm / kg. In some embodiments, the composition is a sterile aqueous composition, the composition comprising about 0.001% ± 0.0001% polypeptide, such as Lacripep TM (SEQ ID NO:1) or other peptides specified herein, or a pharmaceutically acceptable salt thereof; about 0.0098% ± 0.001% anhydrous citric acid; about 0.279% ± 0.028% sodium citrate dihydrate; about 0.001% ± 0.0001% disodium EDTA; about 0.01% ± 0.001% tyloxapol; about 0.50% ± 0.05% NaCl, comprising, consisting of, or consisting essentially of the foregoing; the pH of the composition is adjusted to about 6.2 to about 6.8 using NaOH or HCl, and the osmolality is 170-210 mOsm / kg. In some embodiments, the composition is a sterile aqueous composition, the composition comprising about 0.001% ± 0.0001% polypeptide, such as Lacripep TM(SEQ ID NO:1) or another peptide identified herein, or a pharmaceutically acceptable salt thereof; about 0.0098% ± 0.001% anhydrous citric acid; about 0.279% ± 0.028% sodium citrate dihydrate; about 0.001% ± 0.0001% disodium EDTA; about 0.05% ± 0.005% tyloxapol; about 0.50% ± 0.05% NaCl, comprising, consisting of, or essentially consisting of the foregoing; the pH of the composition is adjusted to about 6.2 to about 6.8 using NaOH or HCl, and the osmolality is 170 - 210 mOsm / kg. In one embodiment, the pH is about 6.3 to about 6.5 and the osmolality is 180 - 200 mOsm / kg. In one embodiment, the composition consists only of the listed components and does not contain any additional active ingredients, additives (e.g., thickeners, buffers, chelating agents, stabilizers, preservatives, surfactants, and tonicity agents), carriers, or diluents.

[0093] In some embodiments, the composition is a sterile aqueous composition, the composition comprising about 0.00025% ± 0.000025% polypeptide, such as Lacripep TM (SEQ ID NO:1) or another peptide identified herein, or a pharmaceutically acceptable salt thereof; about 0.0098% ± 0.001% anhydrous citric acid; about 0.279% ± 0.028% sodium citrate dihydrate; about 0.001% ± 0.0001% disodium EDTA; about 0.001% ± 0.0001% tyloxapol; about 0.50% ± 0.05% NaCl, comprising, consisting of, or essentially consisting of the foregoing; the pH of the composition is adjusted to about 6.2 to about 6.8 using NaOH or HCl, and the osmolality is 170 - 210 mOsm / kg. In some embodiments, the composition is a sterile aqueous composition, the composition comprising about 0.00025% ± 0.000025% polypeptide, such as Lacripep TM(SEQ ID NO:1) or another peptide specified herein, or a pharmaceutically acceptable salt thereof; about 0.0098% ± 0.001% anhydrous citric acid; about 0.279% ± 0.028% sodium citrate dihydrate; about 0.001% ± 0.0001% disodium EDTA; about 0.01% ± 0.001% tyloxapol; about 0.50% ± 0.05% NaCl, comprising, consisting of, or consisting essentially of these; the pH of the composition is adjusted to about 6.2 to about 6.8 using NaOH or HCl, and the osmolality is 170 - 210 mOsm / kg. In some embodiments, the composition is a sterile aqueous composition, and the composition contains about 0.00025% ± 0.000025% polypeptide, such as Lacripep TM (SEQ ID NO:1) or another peptide specified herein, or a pharmaceutically acceptable salt thereof; about 0.0098% ± 0.001% anhydrous citric acid; about 0.279% ± 0.028% sodium citrate dihydrate; about 0.001% ± 0.0001% disodium EDTA; about 0.05% ± 0.005% tyloxapol; about 0.50% ± 0.05% NaCl, comprising, consisting of, or consisting essentially of these; the pH of the composition is adjusted to about 6.2 to about 6.8 using NaOH or HCl, and the osmolality is 170 - 210 mOsm / kg. In one embodiment, the pH is about 6.3 to about 6.5 and the osmolality is 180 - 200 mOsm / kg. In one embodiment, the composition consists of only the listed components and does not contain any additional active ingredients, additives (such as thickeners, buffers, chelating agents, stabilizers, preservatives, surfactants, and tonicity agents), carriers, or diluents.

[0094] In some embodiments including, but not limited to, the above sterile composition, the polypeptide is Lacripep having SEQ ID NO:1 TMor a pharmaceutically acceptable salt thereof. In some embodiments, the polypeptide is the polypeptide having SEQ ID NO: 2, or a pharmaceutically acceptable salt thereof. In some embodiments, the polypeptide is a polypeptide having a sequence selected from the group consisting of SEQ ID NOs: 3-9, or a pharmaceutically acceptable salt or one or more fragments thereof.

[0095] In some embodiments including, but not limited to, the above sterile composition, the pH of the composition is from about 6.5 to about 6.6. In some embodiments including, but not limited to, the above sterile composition, the pH of the composition is from about 6.3 to about 6.5. In some embodiments including, but not limited to, the above sterile composition, the pH of the composition is about 6.5.

[0096] In some embodiments including, but not limited to, the above sterile composition, the osmolality of the composition is from about 270 to about 210 mOsm / kg. In some embodiments including, but not limited to, the above sterile composition, the osmolality of the composition is from about 180 to about 200 mOsm / kg. In some embodiments, the osmolality of the composition is about 190 mOsm / kg.

[0097] In one embodiment including, but not limited to, the above sterile composition, the composition consists of only the recited ingredients and does not contain further active ingredients, additives (such as thickeners, buffers, chelating agents, stabilizers, preservatives, surfactants, and tonicity agents), carriers or diluents. In some embodiments, the amount of any one or more of the recited ingredients is provided in an amount of 5% and / or ±1% of the recited amount.

[0098] In some embodiments, the compositions described herein are manufactured as a solution, gel or ointment. The gel or ointment may be advantageous in providing a composition that contacts the eye for a longer time than a solution, or provides other benefits. Thus, in some embodiments, the gel or ointment is useful when applied to a subject when the subject is sleeping or when the subject's eyes are closed for an extended period of time (e.g., 1, 2, 3, 4, 5 or more hours). The gel or ointment may be used at other times based on the preference of the user.

[0099] Non-limiting exemplary compositions that may be used in the methods and kits disclosed herein include the following compositions in Tables A, B and C below, and Tables 1.1, 1.2 and 1.3 of the Examples. [Table 1] [Table 2] [Table 3]

[0100] Embodiments of the compositions of Table A and / or B and / or C (and Tables 1.1 and / or 1.2 and / or 1.3) also include compositions having a pH of 6.3 to 6.5. Embodiments of the compositions of Table A and / or B and / or C (and Tables 1.1 and / or 1.2 and / or 1.3) also include compositions having an amount disclosed within the range of ±1%, ±2%, ±3%, ±4%, or ±5% of the disclosed amount. In some embodiments, the compositions of Table A and / or B and / or C (and Tables 1.1 and / or 1.2 and / or 1.3) are stored at -20 ± 5°C, 5 ± 3°C, or 25 ± 2°C and 25 ± 5% relative humidity for at least 1 or 2 weeks, 1, 2, 3, 4, or 5 months, and after storage of the composition, the initial undegraded form of Lacripep of SEQ ID NO: 1 in the composition TM maintains at least about 99.0%, 99.9%, 99.95%, or 99.99% of the polypeptide. In some embodiments, the compositions of Table A and / or B and / or C (and Tables 1.1 and / or 1.2 and / or 1.3) are stored at -20 ± 5°C or 5 ± 3°C for at least 12 months, and after storage of the composition, the initial undegraded form of Lacripep of SEQ ID NO: 1 in the composition TM maintains at least about 80% or 90% of the polypeptide.

[0101] (Other therapeutic components) In some embodiments, the composition comprises one or more additional therapeutic agents in addition to the polypeptides described herein. These therapeutic agents can include substances known to those of skill in the art for the treatment of dry eye and related syndromes and conditions, including common dry eye diseases. A non-exhaustive list of additional therapeutic agents includes cholinergic agents (e.g., pilocarpine, cevimeline), cyclosporine, lifitegrast, dexamethasone (or other corticosteroids, such as prednisolone), hyaluronic acid (and its derivatives) with or without chondroitin sulfate, Cyclocut, SI-614, skQ1, Cis-UCA, CycloASol, RGN-259, diclofasol, anakinra, tofacitinib, EBI-005, EGP-437, KP-121, MIM-D3, OTX-DP, rebamipide (OPC-12759), and RU-101. In some embodiments, the additional therapeutic agent is Xiidra (lifitegrast, SAR-1118). In some embodiments, one or more additional therapeutic agents are affected as salts of the polypeptide. Artificial tears and other lubricants, including carboxymethylcellulose, polyvinyl alcohol, hydroxypropylmethylcellulose (also known as HPMC or hypromellose), hydroxypropylcellulose, ethylene glycol polymers, and hyaluronic acid (also known as hyaluronan, HA), and tear ointments such as white petrolatum, mineral oil, and similar lubricants, can also be included in the composition. These additional therapeutic agents can be included in known therapeutic amounts or amounts below the therapeutic amount.

[0102] (Containers and Kits) In some embodiments, the composition is provided in a kit that includes one or more multi-use containers. In some embodiments, the multi-use container includes a protective cap and a liquid storage bottle, where the cap is connected to the bottle via a flexible connector. A blocking plug is disposed at the center of the upper surface of the protective cap. A conical or other suitable shaped liquid outlet is disposed at the center of the bottle cover and mates tightly with the blocking plug of the protective cap. Accordingly, the sterile composition can be placed in a container for multiple uses.

[0103] In some embodiments, the amount of the composition in the container is 0.1 - 0.5, 0.5 - 1.0, 1 - 2, 2 - 5, 5 - 10, 10 - 20, 20 - 30, or 30 - 60 mL or any range therebetween or approximately therewith. The container can be a bottle, tube, vial or other suitable container. For reusable containers, instructions may accompany for use up to 12 hours, 24 hours, 2 - 7 day cycles, 1 month cycles or a specified expiration date. Single-use containers may be suitable for use in one or both eyes for a single application cycle.

[0104] In some embodiments, the composition is provided in a kit that includes single-use containers. In some embodiments, the composition is provided in a kit that includes multiple single-use containers. In some embodiments, the single-use container includes a vessel for holding the liquid, a removable seal top for sealing the vessel, and optionally a neck portion for interconnecting the vessel and the seal top. In some embodiments, a kit is provided that includes multiple single-use containers along with instructions for use.

[0105] In some embodiments, the container includes a pharmaceutically inert material. In some embodiments, the container includes glass, polyvinyl chloride, polypropylene, polyethylene terephthalate, polyethylene terephthalate, polyethylene terephthalate G, high density polyethylene, low density polyethylene, polybutylene terephthalate, polyurethane, ethylene vinyl acetate, silicone, acrylonitrile butadiene styrene, polytetrafluoroethylene, polycarbonate, polystyrene, polymethyl methacrylate, polysulfone, polyvinylidene chloride, or combinations thereof.

[0106] In some embodiments, the container includes polyvinyl chloride, polypropylene, low density polyethylene, polyurethane, ethylene vinyl acetate, silicone, or combinations thereof.

[0107] In some embodiments, the amount of the composition in the container is 0.02 mL; 0.05 mL to 1 mL; 0.1 mL to 0.95 mL; 0.15 mL to 0.8 mL; 0.2 mL to 0.85 mL; 0.25 mL to 0.8 mL; 0.3 mL to 0.75 mL; 0.35 mL to 0.7 mL; 0.4 mL to 0.65 mL; 0.45 mL to 0.6 mL; 0.5 mL to 0.55 mL; or any value therebetween, or approximately so.

[0108] In some embodiments, the amount of the composition in the container is 0.02 mL; 0.025 mL; 0.030 mL; 0.035 mL; 0.040 mL; 0.045 mL; 0.050 mL; 0.055 mL; 0.060 mL; 0.065 mL; 0.070 mL; 0.075 mL; 0.1 mL; 0.15 mL; 0.2 mL; 0.25 mL; 0.3 mL; 0.35 mL; 0.4 mL; 0.45 mL; 0.5 mL; 0.55 mL; 0.6 mL; 0.65 mL; 0.7 mL; 0.75 mL; 0.8 mL; 0.85 mL; 0.9 mL; 0.95 mL; or 1 mL of the composition, or approximately so, or within a range defined by any two of the foregoing values.

[0109] (Eye drops and other administrations) Provided is a method for treating moderate / severe dry eye-related eye symptoms by administering the polypeptide described herein to the eye of a subject identified as suffering from moderate / severe dry eye and in need of treatment. In some embodiments, the subject is identified as suffering from severe dry eye. In some embodiments, the subject does not have Sjögren's syndrome (primary or secondary). Referring to FIG. 1, provided is a method for treating moderate / severe dry eye-related eye symptoms in a subject (e.g., a subject having common dry eye). Method 100, at block 100, has the following criteria: a) The total FCS score is ≧4 and <15 on the NEI / Industry Workshop scale, b) The eye dryness score using VAS instantaneous is ≧60, c) the score of the Schirmer I test under anesthesia is ≤ 5 mm wetting / 5 min, and d) the total LGCS score using the NEI / Industry Workshop scale is ≥ 5 (0 = no staining) identifying a subject suffering from one or more moderate / severe dry eye-related eye symptoms (e.g., a subject having common dry eye and optionally not having Sjögren's syndrome (primary or secondary)), where the subject satisfies one or more (e.g., 1, 2, 3, or 4) of the following:

[0110] In some embodiments, a subject suffering from moderate / severe dry eye satisfies all four of the above criteria in at least one eye. In some embodiments, a subject is identified as suffering from severe dry eye by having a SANDE score ≥ 60 (not shown).

[0111] In block 120, the method includes administering to the subject a composition of the present disclosure, e.g., a liquid composition comprising an amount of a polypeptide of the present disclosure (e.g., Lacripep TM (SEQ ID NO: 1) or a pharmaceutically acceptable salt thereof, e.g., a composition of Table A, B, or C or 1.1, 1.2, or 1.3).

[0112] In some embodiments, a subject has, in at least one eye, the following criteria: a) the total FCS score on the NEI / Industry Workshop scale is < 4 or ≥ 15, b) the eye dryness score using VAS instantaneous is < 60, c) the score of the Schirmer I test under anesthesia is > 5 mm wetting / 5 min, or d) the total LGCS score using the NEI / Industry Workshop scale is < 5 (0 = no staining) If the subject satisfies at least one of the above, the subject has no moderate / severe or severe dry eye and / or dry eye symptoms in at least one eye.

[0113] In some embodiments, the subject does not have moderate / severe or severe dry eye and / or dry eye symptoms in at least one eye if the total FCS score on the NEI / Industry Workshop scale is <4 in at least one eye. In some embodiments, the subject does not have moderate / severe or severe dry eye and / or dry eye symptoms in at least one eye if the dry eye score using VAS instantaneous is <60 in at least one eye. In some embodiments, the subject does not have moderate / severe or severe dry eye and / or dry eye symptoms in at least one eye if the score of the Schirmer 1 test under anesthesia is >5 mm wetting / 5 minutes in at least one eye. In some embodiments, the subject does not have moderate / severe or severe dry eye and / or dry eye symptoms in at least one eye if the total LGCS score using the NEI / Industry Workshop scale is <5 (0 = no staining) in at least one eye.

[0114] Subjects suffering from one or more moderate / severe dry eye-related ocular symptoms can be identified using any suitable option. In some embodiments, a subject is identified as suffering from moderate / severe dry eye-related ocular symptoms based at least on an evaluation of the subject for one or more dry eye-related ocular symptoms such as, but not limited to, pain, stinging, burning, or itching sensations. In some embodiments, a subject is identified as suffering from moderate / severe dry eye-related ocular symptoms based at least on an evaluation of the subject for eye stinging and / or burning sensations. In some embodiments, the evaluation of the symptoms is an immediate evaluation of the symptoms by the subject. In some embodiments, the symptoms are evaluated on a visual analog scale (VAS), for example a scale of 0 to 100 mm (0 mm being the minimum intensity and 100 mm being the maximum intensity), to provide an eye dryness score (EDS) (e.g., evaluated on a scale of 0 to 100 mm, 0 mm being the minimum score and 100 mm being the maximum score). The EDS can be obtained, for example, as shown in FIG. 6, by evaluating the symptoms of the subject on a questionnaire. Using the VAS, the evaluation of the symptoms reported by the subject is measured in millimeters on the linear dimension of this scale (e.g., taking the left end of each line as 0) to provide the EDS (see Schaumberg D, et al. Development and Validation of a Short Global Dry Eye Symptom Index. The Ocular Surface. January 2007, Vol 5; 1; 50-57, which is incorporated herein by reference in its entirety by express citation). The EDS score is the value of the "eye dryness" line of the VAS questionnaire, for example the questionnaire of FIG. 6.

[0115] In some embodiments, when at least the subject has an EDS above a predetermined EDS threshold, the subject is identified as suffering from moderate / severe dry eye-related eye symptoms. In some embodiments, the EDS threshold is determined based on the distribution of EDS in a cohort of patients known to have moderate / severe dry eye disease. In some embodiments, the EDS threshold is determined based on the distribution of EDS in a cohort of subjects not known to have moderate / severe dry eye disease. In some embodiments, the EDS threshold is a value within 60, 65, 70, 75, 80, 85, 90 or more, or a range defined by any two of the foregoing values (e.g., 60-90, 60-80, 60-70, or 60-100). In some embodiments, one or more dry eye-related eye symptoms include an EDS of 60 or more. In some embodiments, the subject has a common dry eye disease. In some embodiments, the subject does not have Sjögren's syndrome (primary or secondary).

[0116] In some embodiments, the method includes at least using responses to one or more questionnaires designed to measure, for example, but not limited to, one or more of ocular surface discomfort or visual symptoms associated with dry eye disease, the impact of dry eye disease on daily function, and health-related quality of life, and can be used to identify a subject as suffering from one or more dry eye-related ocular symptoms, instead of or in addition to the instant assessment of the subject on one or more dry eye-related ocular symptoms on the VAS scale described above. In some embodiments, the questionnaires include, but are not limited to: 1) Ocular Surface Disease Index (OSDI); 2) Impact of Dry Eye on Everyday Life (IDEEL); 3) National Eye Institute-Visual Function Questionnaire (NEIVFQ); 4) Symptom Assessment in Dry Eye (SANDE); 5) Dry Eye-Related Quality-of-Life Score Questionnaire (DEQS); 6) McMonnies Dry Eye Questionnaire; 7) Women's Health Study Questionnaire; 8) Dry Eye Questionnaire (DEQ); 9) North Carolina Dry Eye Management Scale (UNC DEMS); 10) Subjective Evaluation of Symptom of Dryness (SESoD); 11) Standard Patient Evaluation of Eye Dryness (SPEED); 12) Dry Eye Epidemiology Project Questionnaire (DEEP);13) Canada Dry Eye Epidemiology Study (CANDEES); 14) Salisbury eye evaluation; 15) Melbourne visual impairment project; 16) Bjerrum questionnaire; 7) one or more of the Japanese dry eye awareness study. Suitable non-decreasing examples of questionnaires are summarized in Stapleton et al. TFOS DEWS II Epidemiology Report. Ocul Surf. 2017 Jul;15(3):334-365, and Wolffsohn et al. TFOS DEWS II Diagnostic Methodology report. Ocul Surf. 2017 Jul;15(3):539-574 (each of which is incorporated herein by reference in its entirety). In some embodiments, the subject has a measured value of symptoms by any suitable questionnaire, such as those described above, that is at least a threshold measurement value corresponding to the EDS threshold for the instant evaluation of one or more dry eye-related eye symptoms in the subject on the VAS scale, and is identified as having moderate / severe dry eye-related eye symptoms. In some embodiments, the subject does not have Sjögren's syndrome (primary or secondary).;

[0117] In some embodiments, a subject suffering from one or more moderate / severe dry eye-related ocular symptoms is at least identified by one or more ocular measurements, such as one or more ocular symptoms not measured based on a report by the subject. Any suitable ocular measurement can be used to identify a subject as suffering from one or more dry eye-related ocular symptoms. In some embodiments, the symptoms of dry eye are determined based at least on one or more measurements of, but not limited to, tear film stability, tear secretion, tear volume, tear film composition, damage to the ocular surface, inflammation of the ocular surface, and eyelid function. In some embodiments, the symptoms of dry eye are determined at least by one or more of, but not limited to, an increase in corneal staining, an increase in conjunctival staining, a decrease in tear breakup time, and a decrease in Schirmer tear test score (with or without anesthesia). Suitable non-limiting examples of ocular measurements are summarized in Stapleton et al. TFOS DEWS II Epidemiology Report. Ocul Surf. 2017 Jul;15(3):334-365, and Wolffsohn et al. TFOS DEWS II Diagnostic Methodology report. Ocul Surf. 2017 Jul;15(3):539-574, each of which is incorporated herein by reference in its entirety. In some embodiments, the subject does not have Sjögren's syndrome (primary or secondary).

[0118] In some embodiments, moderate / severe dry eye symptoms are determined at least by one or more of, but not limited to, the Schirmer test (e.g., Schirmer test under anesthesia), Lissamine Green conjunctival staining (LGCS), and fluorescein corneal staining (FCS). "Fluorescein corneal staining", "FCS", "corneal fluorescein staining" and "CFS" are used interchangeably herein. In some embodiments, the method includes measuring an ocular surface staining, e.g., FCS or LGCS, to determine whether a subject has one or more dry eye-related ocular symptoms. In some embodiments, the ocular surface staining is scored using the National Eye Institute (NEI) / Industry Workshop scale. In some embodiments, FCS in the NEI / Industry Workshop scale is determined by a trained practitioner (e.g., a trained ophthalmologist) scoring the level of CFS within a grid that divides the corneal region into five sections (e.g., as shown in FIG. 7), with each section assigned a score (or grade) of 0-3 depending on the amount and distribution of staining. A score / grade of 0 indicates no staining in the section. The total FCS score can range from 0 / 15 (indicating the absence of corneal epitheliopathy) to 15 / 15 (indicating severe epitheliopathy) in the NEI / Industry Workshop scale. In some embodiments, LGCS in the NEI / Industry Workshop scale is determined by a trained practitioner (e.g., a trained ophthalmologist) scoring the level of LGCS within a grid that divides the conjunctival region into six sections (superior limbus of both nasal and temporal sides, inferior limbus, peripheral region) (e.g., as shown in FIG. 8), with each section assigned a score (or grade) of 0-3 depending on the amount and distribution of staining. A score / grade of 0 indicates no staining in the section. The total LGCS score can range from 0 / 18 to 18 / 18 in the NEI / Industry Workshop scale.

[0119] Ocular surface staining can be scored using any suitable option equivalent to the above NEI / Industry Workshop scale. In some embodiments, ocular surface staining is scored according to one or more of the following grading scales: the van Bijsterveld system, the Collaborative Longitudinal Evaluation of Keratoconus (CLEK) schema, the Oxford schema, the area-density combined index, and the Sjögren's International Collaborative Clinical Alliance ocular staining score. Suitable non-limiting examples of scoring of ocular surface staining are summarized in Wolffsohn et al. TFOS DEWS II Diagnostic Methodology report. Ocul Surf. 2017 Jul;15(3):539-574 (incorporated herein by reference in its entirety).

[0120] In some embodiments, a subject is identified as suffering from moderate / severe dry eye-related eye symptoms if at least the subject's total FCS score exceeds an FCS threshold. In some embodiments, one or more dry eye-related eye symptoms include a total FCS score of ≧ about 4 and < about 15 on the NEI / Industry Workshop scale. In some embodiments, the FCS threshold is a value within the range defined by 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15, or any two of the foregoing values (e.g., 4-14, 6-12, 4-10, 6-8, 6-14, 8-14, 8-12, or 4-6) on the NEI / Industry Workshop scale, where 0 indicates no staining. In some embodiments, a subject is identified as suffering from dry eye-related eye symptoms if at least the subject's total FCS score determined using another scoring option is above a threshold corresponding to the threshold defined using the NEI / Industry Workshop scale.

[0121] In some embodiments, the subject is identified as suffering from moderate / severe dry eye-related eye symptoms if at least the resampling green conjunctival staining (LGCS) score of the subject exceeds the LGCS threshold. In some embodiments, one or more dry eye-related eye symptoms include an LGCS score of ≧5 on the National Eye Institute (NEI) / Industry Workshop scale, where 0 indicates no staining. In some embodiments, the LGCS threshold is a value within at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more, or a range defined by any two of the foregoing values (e.g., 5-14, 5-12, 8-14, 5-8, 10-14, or 5-6) on the NEI / Industry Workshop scale, where 0 indicates no staining. In some embodiments, the subject is identified as suffering from dry eye-related eye symptoms if at least the LGCS score of the subject determined using another scoring option is above a threshold corresponding to the threshold defined using the NEI / Industry Workshop scale.

[0122] In some embodiments, the subject is identified as suffering from moderate / severe dry-eye related eye symptoms based at least on the measurement of tear secretion. In some embodiments, tear secretion is measured using a Schirmer test or a modification thereof. The Schirmer test can be any suitable form of Schirmer test. In some embodiments, one or more dry-eye related eye symptoms include a Schirmer test score of ≤5 mm wetting / 5 minutes under anesthesia. In some embodiments, the Schirmer test is, but not limited to, Schirmer 1 under anesthesia, Schirmer 2, Schirmer 3, the thread method, or a 1-minute Schirmer test. In some embodiments, one or more dry-eye related eye symptoms include a Schirmer test score below the Schirmer test threshold, e.g., the score of the Schirmer 1 test under anesthesia below the Schirmer 1 test threshold under anesthesia. Appropriate Schirmer tests and their modifications, as well as the corresponding thresholds, are summarized in Savini et al. The challenge of dry eye diagnosis. Clin Ophthalmol. 2008 Mar;2(1):31-55 (incorporated herein by reference in its entirety). In some embodiments, the Schirmer test is the Schirmer 1 test under anesthesia. In some embodiments, one or more dry-eye related eye symptoms include a Schirmer test score of ≤5 mm wetting / 5 minutes under anesthesia. In some embodiments, any one of the modifications of the Schirmer test (including Schirmer 1 under anesthesia) can be used to identify the subject as suffering from one or more moderate / severe dry-eye related eye symptoms.

[0123] In some embodiments, the subject is identified as suffering from severe dry eye rather than moderate / severe dry eye by having a SANDE score ≥60.

[0124] In some embodiments, the subject has a history of moderate / severe dry eye-related ocular symptoms. In some embodiments, the method includes determining that the subject has a history of one or more moderate / severe dry eye-related ocular symptoms. In some embodiments, the subject has experienced or been diagnosed with one or more previous moderate / severe dry eye-related ocular symptoms for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 16, 20, 24, 30, 36 months or more, or within a period defined by any two of the foregoing values (e.g., 1-36, 1-24, 1-12, 3-30, 3-24, 6-12, 6-24 months), prior to being identified as having one or more dry eye-related ocular symptoms. In some embodiments, one or more of the subject's moderate / severe dry eye-related ocular symptoms include the subject's use of an eye lubricant (e.g., artificial tears) within the past 6, 5, 4, 3, 2, or 1 month, or within a period defined by any two of the foregoing values (e.g., 1-6, 2-6, 3-5 months).

[0125] In some embodiments, the subject is at least 18 years old. In some embodiments, the subject is 18-25, 25-40, 40-55, 55-60, 60-65, 65-70, 70-75, 75-80, 80-85 years old, or older.

[0126] In some embodiments, subjects meeting one or more of the following criteria are excluded: 1. Subjects having any active infectious ocular condition. 2. Subjects who are monocular or have a BCVA of 1.0 logMAR or less, as evaluated by the Early Treatment Diabetic Retinopathy Study (ETDRS), using corrective lenses as needed. 3. Subjects having an ocular inflammatory condition (e.g., conjunctivitis, keratitis, anterior blepharitis, etc.) not related to dry eye syndrome. 4. Subjects having clinical evidence of a cicatricial ocular surface disease, such as cicatricial pemphigoid or Stevens-Johnson syndrome. 5. Subjects for whom the use of any topical ophthalmic drug (including topical cyclosporine) other than the composition cannot be interrupted during the induction and treatment periods. 6. Subjects who have used Restasis® (topical ophthalmic cyclosporine) within 60 days before Visit 1. 7. Subjects who have used Xiidra® (topical ophthalmic lifitegrast) within 60 days before Visit 1. 8. Subjects with a total fluorescein corneal staining (FCS) score = 15 or an upper region score = 3 on the NEI / Industry Workshop scale in the eye to be treated, or subjects with FCS with diffuse confluent staining, filaments or epithelial defects. 9. Subjects who have had active herpetic keratitis or developed herpetic keratitis within 365 days of identification, or subjects who are taking chronic oral antiviral drugs for herpetic diseases. 10. Subjects who cannot interrupt the use of contact lenses and cannot refrain from using them during administration. 11. Subjects who have used amiodarone or for whom its use is predicted. 12. Subjects who change or for whom a change in the dose of tetracycline, omega 3 or 6 is predicted within 30 days before identification. 13. Subjects who change or for whom a change in the dose of anticholinergic drugs, antidepressants, oral contraceptives, isotretinoin, oral systemic corticosteroids, oral systemic immunosuppressive drugs is predicted within 60 days before identification and during the treatment period. 14. Subjects who use topical ophthalmic antihistamines, ocular, inhaled or intranasal corticosteroids, topical or oral mast cell stabilizers, oral antihistamines, topical or nasal vasoconstrictors, topical ophthalmic NSAIDs, topical ophthalmic antibiotics within 30 days before identification and during the treatment period. 15. Subjects who have undergone punctal cautery or a change (insertion or removal) to punctal plugs in the eye to be treated within the past 90 days before identification. Note: If the punctal plug is randomized / baseline and removed, the plug needs to be replaced as soon as possible. 16. A subject who has undergone corneal refractive surgery (LASIK, PRK, RK) in the eye to be treated. 17. A subject who has a history of any surgical treatment on the eye surface or eyelid within 365 days prior to identification, or a history of intraocular surgery within 90 days prior to identification, in the eye to be treated. 18. A subject who is pregnant or suspected of being pregnant, and a subject who is breastfeeding or intends to breastfeed. Female subjects who may be pregnant are required to have a negative urine pregnancy test at the time of screening and must consent to use an acceptable method of contraception from the time of signing the informed consent until the end of treatment. Acceptable medical methods of contraception include intrauterine contraceptive devices; barrier methods such as diaphragms, condoms, caps or sponges used with spermicides; or hormonal contraception. 19. A subject who has any physical or mental disability that prevents the ability to participate and give informed consent. 20. A subject who has participated in a device or investigational drug trial or clinical trial within 30 days of identification. Participation in another study during treatment is excluded throughout the treatment period. In some embodiments, the subject does not have Sjögren's syndrome (primary or secondary).

[0127] In some embodiments, the composition is administered topically to the eye. In some embodiments, the composition is administered for the treatment thereof to an individual suffering from (e.g., identified as suffering from) any form of moderate / severe dry eye. In some embodiments, the subject does not have Sjogren's syndrome (primary or secondary). The compositions described herein may be provided as a liquid (solution, gel, ointment, etc.) or in other forms, such as a powder or patch, tab, etc. In some embodiments, the compositions described herein are used to achieve one or more of the following: restore basal tearing, restore general mucosal and ocular surface wetness; restore the integrity of the ocular surface and mucosa, rapidly but transiently promote autophagy, and eliminate pressure, stress or degenerative diseases throughout the eye or in other organs; reduce inflammation, promote wound healing (e.g., corneal and oral wound healing after refractive surgery), stabilize the tear film, and suppress bacterial infection.

[0128] In some embodiments, topical administration to the eye comprises administering one or more drops of the composition to the surface of the eye. For example, in some embodiments, the user is instructed to apply to the surface of the eye, rather than wearing contact lenses. In other embodiments, eye drops (or other applications) are suitable for administration while wearing contact lenses. In some embodiments, the composition is administered from a container as a single dose delivered as one drop to each eye. In some embodiments, the eye drops are from about 0.020 mL to about 0.050 mL, or any amount therebetween. In some embodiments, the eye drops are about 0.035 mL.

[0129] The composition can be administered to a subject at any suitable dosing frequency and amount. In some embodiments, the composition is administered to the subject's eye (e.g., the ocular surface) as 1 drop or more (e.g., 1, 2, 3, 4, 5 drops or more). In some embodiments, 1 drop of the composition is administered to the subject's eye per administration. In some embodiments, 1 drop of the composition contains an amount of the composition within about 0.005 mL, about 0.01 mL, about 0.02 mL, about 0.03 mL, 0.04 mL, about 0.05 mL, about 0.06 mL, about 0.07 mL, about 0.08 mL, about 0.09 mL, about 0.1 mL, about 0.2 mL, about 0.5 mL or more, or a range defined by any two of the foregoing values (e.g., 0.005 - 0.5 mL, 0.01 - 0.1 mL, 0.02 - 0.08 mL, 0.03 - 0.07 mL, etc.). In some embodiments, 1 drop or more of the composition is administered to the subject's eye up to 3 times a day, e.g., 1, 2, or 3 times a day. In some embodiments, 1 drop or more of the composition is administered to the subject's eye up to 3 times a day, up to 3 times every other day, up to 3 times every 3 days, up to 3 times every 4 days, up to 3 times every 5 days, up to 3 times a week, or more. In some embodiments, 1 drop or more of the composition is administered to the subject's eye up to 3 times a day for a period of at least 5 days, at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 10 weeks, at least 3 months, at least 4 months, at least 6 months, at least 8 months, at least 12 months, at least 18 months, at least 24 months or more, or a range defined by any two of the foregoing values (e.g., 5 days - 24 months, 1 week - 24 months, 1 - 6 weeks, 1 week - 12 months, 6 - 10 weeks, 3 - 12 months, etc.).

[0130] In some embodiments, the composition is administered to one eye of the subject, e.g., an eye exhibiting one or more moderate / severe dry eye-related ocular symptoms. In some embodiments, the composition is administered to both eyes of the subject. In some embodiments, the composition is administered to one eye of the subject, e.g., an eye exhibiting one or more severe dry eye-related ocular symptoms. In some embodiments, the composition is administered to both eyes of the subject. In some embodiments, the subject does not have Sjögren's syndrome (primary or secondary).

[0131] In some embodiments, the subject is identified as suffering from severe dry eye rather than moderate / severe dry eye. In some embodiments, if the subject has an eye dryness score of ≧70 using VAS instantaneous measurement in at least one eye, the subject has severe dry eye and / or dry eye symptoms.

[0132] In some embodiments, the ocular administration of the composition improves the reported symptoms of one or more patients or the clinical signs of moderate / severe dry eye. In some embodiments, the subject does not have Sjögren's syndrome (primary or secondary). Improvement of dry eye symptoms or signs is as follows: · Fluorescein corneal staining (FCS) (0 - 3 scale for each of 5 regions using the NEI / Industry Workshop scale, total 0 - 15 scale) · Lissamine green conjunctival staining (LGCS) (0 - 3 scale for each region using the NEI / Industry Workshop scale, total 0 - 18 scale) · Schirmer test under anesthesia (mm of wetting in 5 minutes) · Tear film break-up time (seconds) · Eye dryness reported by the patient on a visual analog scale and aggregated as the mean change from baseline ·Dry eye-related eye symptom questionnaire (SANDE: frequency and severity of dry eye symptoms) (Schaumberg D, et al. Development and Validation of a Short Global Dry Eye Symptom Index. The Ocular Surface. January 2007, Vol 5; 1; 50-57 (incorporated herein by reference in its entirety)) can be evaluated by one or more of

[0133] In some embodiments, evaluating improvement of moderate / severe or severe dry eye symptoms or signs includes reflective and / or instantaneous evaluation of the symptoms. In some embodiments, reflective evaluation includes scoring the difference in symptoms experienced by the subject after treatment compared to before treatment. In some embodiments, reflective evaluation of symptoms includes scoring the difference in symptoms currently experienced by the subject compared to the last day of treatment (e.g., the first day of treatment). In some embodiments, reflective evaluation of symptoms includes scoring the difference in symptoms currently experienced by the subject compared to the last day of nitration of dry eye symptoms. In some embodiments, improvement in the measurement or evaluation of dry eye symptoms or signs (e.g., those listed above) is 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% of the value of the measurement or evaluation, approximately that, at least that, at least approximately that, or within a range defined by any two of the foregoing values.

[0134] In some embodiments, the composition administered to a subject suffering from moderate / severe dry eye symptoms is a sterile aqueous composition comprising, consisting of, or consisting essentially of the compositions disclosed herein. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of the formulation of Table A, B, or C. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of Formulation A of Table A. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of Formulation B of Table A. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of Formulation C of Table A. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of Formulation D of Table B. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of Formulation E of Table B. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of Formulation F of Table B. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of Formulation G of Table C. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of Formulation H of Table C. In some embodiments, the composition is a sterile aqueous composition, wherein the composition comprises, consists of, or consists essentially of Formulation I of Table C.

[0135] Some embodiments include methods of treating moderate / severe dry eye, including administering the compositions disclosed herein to the eyes of a subject having moderate / severe dry eye. In some embodiments, the subject does not have Sjogren's syndrome (primary or secondary). In some embodiments, the compositions described herein are used to treat a subject having moderate / severe dry eye symptoms. In some embodiments, the subject meets one, two, three, or four of the following criteria: a) an FCS total score of ≧4 and <15 on the NEI / Industry Workshop scale, b) an ocular dryness score of ≧60 using VAS instantaneous, c) a score of ≦5 mm wetting / 5 minutes on the Schirmer 1 test under anesthesia, and d) an LGCS total score of ≧5 (0 = no staining) using the NEI / Industry Workshop scale and is identified as having moderate / severe dry eye.

[0136] In some embodiments, the compositions described herein are used to treat a subject having all four of the criteria in the above list. In some embodiments, the compositions described herein are used to treat a subject having all four of the criteria in the above list in at least one eye (e.g., in one or both eyes). In some embodiments, the subject does not have Sjogren's syndrome (primary or secondary). In some embodiments, the subject does not meet one, two, three, or four of the following criteria in one or both eyes: a) an FCS total score of <4 or ≧15 on the NEI / Industry Workshop scale; b) an ocular dryness score of <60 using VAS instantaneous; c) a score of >5 mm wetting / 5 minutes on the Schirmer 1 test under anesthesia; and d) an LGCS total score of <5 (0 = no staining) using the NEI / Industry Workshop scale. In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof in the composition is Lacripep in an amount of 0.005%, 0.001%, or 0.00025% (w / v) TM(It has a sequence consisting of Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-NH2, where "Ac" represents an acetyl group and the C-terminus is amidated (SEQ ID NO: 1)). In some embodiments, the ophthalmic solution is a polypeptide, such as Lacripep TM In addition to, it further contains citric acid (about 0.0098% anhydrous), sodium citrate (about 0.279% sodium citrate anhydrate), disodium EDTA (about 0.001%), NaCl (about 0.5%), tyloxapol (about 0.001%, 0.01%, or 0.05%), NaOH or HCl (up to pH about 6.5), purified water, USP. In some embodiments, a subject having moderate / severe dry eye symptoms is administered a sterile aqueous composition comprising, consisting of, or essentially consisting of Formulation A, B, C, D, E, F, G, H, or I in Tables A, B, and C, or Formulation 1, 2, 3, 4, 5, 6, 7, 8, or 9 in Tables 1.1, 1.2, and 1.3. To evaluate efficacy, in some embodiments, a placebo is used that includes an ophthalmic vehicle solution without the polypeptide. In some embodiments, 1 drop of the composition is administered to the subject's eye up to 3 times a day. In some embodiments, the subject is identified as suffering from severe dry eye rather than moderate / severe dry eye. In some embodiments, if the subject has an eye dryness score of ≧70 using VAS instantaneous measurement in at least one eye, the subject has severe dry eye and / or dry eye symptoms.

[0137] In some embodiments, the method includes measuring the change in one or more dry eye-related eye symptoms after administration compared to before administration. In some embodiments, the method includes measuring one or more scores of eye dryness (EDS), SANDE, LGCS, FCS, Schirmer test (e.g., Schirmer test under anesthesia), and TBUT after administration and comparing them to the corresponding scores before administration.

[0138] In some embodiments, administration improves the total FCS score (NEI / Industry Workshop 0 - 15 scale) in the eye of the subject after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or at least 6 weeks after the start of 4 weeks of treatment, as compared to the baseline measurement before treatment initiation. In some embodiments, administration is as follows: Dry eye after at least 2 weeks of treatment or at least 4 weeks of treatment, compared to baseline on a visual analog scale; SANDE (total score of SANDE 1) after at least 2 weeks of treatment, compared to the baseline measurement before treatment initiation; Average score of SANDE (total score of SANDE - 1) after at least 2 weeks of treatment, compared to the baseline measurement before treatment initiation; Individual symptom assessment (instantaneous) after at least 2 weeks of treatment, compared to the baseline measurement before treatment initiation; Average score of individual symptom assessment (reflex) after at least 2 weeks of treatment, compared to the baseline measurement before treatment initiation; LGCS of the subject's eye after at least 2 weeks of treatment, compared to the baseline measurement before treatment initiation; Schirmer test under anesthesia of the subject's eye after at least 2 weeks of treatment, compared to the baseline measurement before treatment initiation; TFBUT of the subject's eye after at least 2 weeks of treatment, compared to the baseline measurement before treatment initiation; FCS of the subject's eye after at least 2 weeks of treatment, compared to the baseline measurement before treatment initiation; SANDE (total score of SANDE 1) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before treatment initiation; Individual symptoms (instantaneous) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before treatment initiation; The mean score of (the overall score of SANDE-2) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before treatment initiation; The mean score of the individual symptom assessment (reflex) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before treatment initiation; FCS and SANDE 1 and the individual symptom assessment (instantaneous) after at least 2 weeks of treatment, or after at least 4 weeks of treatment, compared to the baseline measurement before treatment initiation; LGCS after at least 2 weeks of treatment, or after at least 4 weeks of treatment, compared to the baseline measurement before treatment initiation; The Schirmer test results under anesthesia after at least 2 weeks of treatment, or after at least 4 weeks of treatment, compared to the baseline measurement before treatment initiation; TFBUT after at least 2 weeks of treatment, or after at least 4 weeks of treatment, or 1 week after 4 weeks of treatment, compared to the baseline measurement before treatment initiation Improve one or more of the following.

[0139] In some embodiments, the improvement in the measurement or evaluation of the symptoms or signs of dry eye (such as those listed above) is 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100% of the value of the measurement or evaluation, approximately that, at least that, at least approximately that, or within a range defined by any two of the foregoing values.

[0140] In some embodiments, the comparison includes, alternatively or additionally, a comparison with a vehicle control.

[0141] In some embodiments, one or more of any of the following drugs / treatments are not administered with the composition described herein, and in some embodiments, one or more of any of the following drugs / treatments are administered with the composition described herein: 1. Ophthalmic drugs (any topical ophthalmic drug), including prescription drugs and over-the-counter (OTC) drugs 2. Contact lenses 3. Any surgical procedure on the ocular surface or eyelid within 365 days before the start of treatment, or intraocular surgery within 90 days before the start of treatment 4. Amiodarone 5. Topical ophthalmic antihistamines 6. Ocular, inhaled or intranasal corticosteroids 7. Topical or oral mast cell stabilizers 8. Oral antihistamines 9. Topical or nasal decongestants 10. Topical ophthalmic NSAIDs 11. Topical ophthalmic antibiotics 12. Within 60 days before and / or during treatment: topical cyclosporine, topical lifitegrast 13. Within 90 days before and / or during treatment: punctal cautery or punctal plug replacement (insertion or removal) or nasolacrimal duct surgery 14. Chronic oral antiviral drugs for ocular herpes diseases.

Examples

[0142] The following are some non-limiting examples of the embodiments described herein.

[0143] Example 1: Treatment of moderate / severe dry eye A randomized placebo-controlled double-blind parallel-group trial was conducted to determine the therapeutic effect of 0.005% (22 μM), 0.001% (4 μM) or 0.00025% (1 μM) Lacripep in a cohort of patients with moderate / severe dry eye-related symptoms TM .

[0144] Exclusion and inclusion criteria Select subjects who meet the following criteria: 1. Subjects who are 18 years of age or older at the time of obtaining informed consent. 2. Subjects who have a history of dry eye-related eye symptoms and self-reported the use of over-the-counter eye lubricants within the past 120 days. 3. For both screening and the Visit 2 (randomization / baseline) examination, the following criteria: a) The total FCS score on the NEI / Industry Workshop scale is ≥ 4 and < 15, b) The eye dryness score (EDS) using VAS instantaneous is ≥ 60, c) The score of the Schirmer 1 test under anesthesia is ≤ 5 mm wetting / 5 minutes, d) The total LGCS score using the NEI / Industry Workshop scale is ≥ 5 (0 = no staining) Subjects who meet the criteria.

[0145] Note: Subjects must meet all criteria, and the eligible scores for FCS, EDS, Schirmer, and LGCS must be present in at least one eye and be in the same eye at the time of the visit.

[0146] FCS is scored by a trained ophthalmologist who scores the level of corneal fluorescein staining within a grid that divides the corneal area into 5 sections (Figure 7), and each section is assigned a score (or grade) of 0 - 3 according to the amount and distribution of the staining. A score / grade of 0 indicates no staining in the section. The total FCS score is determined in the range of 0 / 15 (indicating the absence of corneal epitheliopathy) to 15 / 15 (indicating severe epitheliopathy).

[0147] The LCGS is scored by a trained ophthalmologist who scores the level of resazurin conjunctival staining within a grid that divides the conjunctival region into six sections (both nasal and temporal superior limbal, inferior limbal, peripheral regions) (Figure 8), and a score (or grade) of 0 - 3 is assigned to each section according to the amount and distribution of staining. A score / grade of 0 indicates no staining in the section. The total FCS score ranges from 0 / 15 (indicating absence of corneal epitheliopathy) to 15 / 15 (indicating severe epitheliopathy), and for each of these, a score (or grade) of 0 - 3 is assigned according to the amount and distribution of staining. A score / grade of 0 indicates no staining in the section. Determine the total LGCS score in the range of 0 / 18 - 18 / 18.

[0148] The EDS is scored by the subject's immediate assessment of symptoms as reported by the questionnaire shown in Figure 6. In the questionnaire, the subject is asked to evaluate dry eye symptoms by marking an "X" on each line. The EDS is the value of the "Eye Dryness" line of the questionnaire.

[0149] The Schirmer test is performed by folding a Schirmer strip (5×35 mm) at the notch and hooking the folded end over the temporal one-third of the lower eyelid margin. The score is the measured length of wetting from the notch after 5 minutes. For the Schirmer test score under anesthesia, anesthetic is applied to the subject's eye before applying the Schirmer strip.

[0150] Subjects who meet any of the following criteria at Visit 1 (screening) or Visit 2 (randomization / baseline) are excluded: 1. Subjects with any active infectious eye condition. 2. Subjects who are monocular or have a BCVA of 1.0 logMAR or less using corrective lenses as needed when evaluated by the Early Treatment Diabetic Retinopathy Study (ETDRS). 3. Subjects with an ocular inflammatory condition not related to dry eye syndrome (e.g., conjunctivitis, keratitis, anterior blepharitis, etc.). 4. Subjects having clinical evidence of cicatricial ocular surface diseases, such as cicatricial pemphigoid or Stevens-Johnson syndrome. 5. Subjects who cannot discontinue the use of any topical ophthalmic medications (including topical cyclosporine) other than the investigational drug during the lead-in and treatment periods. 6. Subjects who used Restasis® (topical ophthalmic cyclosporine) within 60 days prior to Visit 1. 7. Subjects who used Xiidra® (topical ophthalmic lifitegrast) within 60 days prior to Visit 1. 8. Subjects with a total fluorescein corneal stain (FCS) score = 15 or a score = 3 in the superior region by the NEI / Industry Workshop scale in the test eye, or subjects with FCS with diffuse confluent staining, filaments, or epithelial defects. 9. Subjects who had or developed active herpetic keratitis within 365 days prior to Visit 1, or subjects taking chronic oral antiviral medications for herpetic diseases. 10. Subjects who cannot discontinue the use of contact lenses and cannot refrain from using them from Screening Visit (Visit 1) to the end of the study (Visit 5). 11. Subjects who were using or whose use of amiodarone was predicted. 12. Subjects who changed or whose change in the dose of tetracycline, omega 3 or 6 was predicted within 30 days prior to Visit 1. 13. Subjects who changed or whose change in the dose of anticholinergic drugs, antidepressants, oral contraceptives, isotretinoin, oral systemic corticosteroids, oral systemic immunosuppressive drugs was predicted within 60 days prior to Visit 1 and during the study period. 14. Subjects who used topical ocular antihistamines, ocular, inhaled or intranasal corticosteroids, topical or oral mast cell stabilizers, oral antihistamines, topical or nasal vasoconstrictors, topical ocular NSAIDs, topical ocular antibiotics within 30 days prior to Visit 1 and during the study period. 15. Subjects who, in the test eye, have undergone punctal cauterization or a change (insertion or removal) to punctal plugs within 90 days prior to the first visit. Note: If the punctal plug is present at the second visit (randomization / baseline) and is removed, the plug should be replaced as soon as possible. 16. Subjects who, in the test eye, have undergone corneal refractive surgery (LASIK, PRK, RK). 17. Subjects who, in the test eye, have a history of any surgical procedure on the ocular surface or eyelid within 365 days prior to the first visit, or a history of intraocular surgery within 90 days prior to the first visit. 18. Subjects who are pregnant or suspected of being pregnant, and subjects who are breastfeeding or intend to breastfeed. Female subjects with a possible pregnancy are required to have a negative urine pregnancy test at the time of screening and must agree to use an acceptable method of contraception from the time of signing the informed consent until the end of the test visit. Acceptable methods of medical contraception include intrauterine contraceptive devices; barrier methods such as diaphragms, condoms, caps, or sponges used with spermicides; or hormonal contraception. 19. Subjects who have any physical or mental disability that prevents their ability to participate and give informed consent. 20. Subjects who have participated in a device or investigational drug trial or clinical trial within 30 days of the first visit. Participation in another study during this trial is excluded for the duration of this trial.

[0151] Randomly assign eligible subjects to one of four treatment groups: one of three Lacripep TM ophthalmic solution strengths (0.005% (formulation 1 or 4), 0.001% (formulation 2 or 5) or 0.00025% (formulation 3 or 6) w / v) or placebo (individual vehicle). Each test group contains 65 subjects, for a total of 260 subjects. Lacripep TM formulations are shown in Tables 1.1, 1.2 and 1.3.

[0152] Instill 1 drop of the test drug three times a day (TID) into both eyes for 1 week, 2 weeks, 4 weeks, or 6 weeks. Efficacy and safety are evaluated at predetermined intervals.

Table 4

Table 5

Table 6

[0153] Efficacy: Conduct all efficacy evaluations in both eyes by the same person if possible. Evaluate corneal fluorescein staining (CFS) on a scale of 0 - 3 (total 0 - 15) in five regions (central, inferior, superior, temporal, and nasal). Evaluate lissamine green conjunctival staining (LGCS) on a scale of 0 - 3 (total 0 - 15) in six regions. All facilities receive training and are tested using the NEI scoring method.

[0154] After drying the lower cul-de-sac, instill approximately 50 μL (1 drop) of 0.5% proparacaine, then perform a Schirmer test under anesthesia and record the wetting in mm over 5 minutes. Measure the tear break-up time (TBUT) in seconds. Measure the severity and frequency of symptoms evaluated by patients using the Visual Analogue Scale for the Dry Eye Symptom Assessment (SANDE) Inventory Version 1 and Version 2. Measure the individual symptom evaluations (instantaneous and reflex) of symptoms reported by patients using the Visual Analogue Scale (VAS). For SANDE Version 2 and individual symptom evaluations (reflex), ask patients to evaluate the difference in symptoms compared to the last visit and administer to the subjects on days 14, 28, and 42 of treatment.

[0155] The primary efficacy measure is the mean change from baseline to day 28 in the total CFS score (NEI / Industry Workshop 0 - 15 scale, 0 - 3 scores in each of 5 domains) in the test eye. The key secondary efficacy measure is the mean change from baseline to day 28 in the dry eye score (0 - 100 mm VAS) from individual symptom assessments.

[0156] Changes from baseline in CFS at day 14 and post - treatment follow - up (day 42) are additional secondary measures. Other secondary measures include changes from baseline to days 14 and 28 in LGCS score, TFBUT, and Schirmer test. Secondary measures include changes from baseline to days 14 and 28 in each of SANDE and individual symptom assessments.

[0157] The test results show a significant improvement in one or more symptoms compared to vehicle treatment.

[0158] Example 2: Treatment of Primary Sjögren's Syndrome The primary objective of this study was to evaluate the safety and tolerability of 22 and 44 μM Lacripep ophthalmic solution, administered three times daily for 28 days, compared to placebo (vehicle), in subjects with a history of ocular surface disease associated with primary Sjögren's syndrome, including subjects with mild dry eye symptoms. TM Many dry eye signs and symptoms were also evaluated.

[0159] Lacripep TM is a synthetic 19 - amino acid peptide fragment of lactoferrin (SEQ ID NO: 1). Proteomics studies have revealed that active monomeric form of lactoferrin is decreased in the tears of patients suffering from many forms of dry eye disease, including water - deficiency, evaporation, contact - lens related, but is most prominent in dry eye patients with primary and secondary Sjögren's syndrome.

[0160] Study Design This was a multicenter, randomized, placebo-controlled, double-blind, parallel-group trial conducted at 35 sites in the United States and approved by local institutional review boards. Written informed consent was obtained from all subjects after consideration of the risks and benefits of participation. The study was conducted in accordance with the 1996 Health Insurance Portability and Accountability Act and the 1996 Declaration of Helsinki and was registered at ClinicalTrials.gov: NCT 03226444.

[0161] The study period was 56 days, including a 14-day lead-in period, a 28-day active treatment period, and a 14-day follow-up (washout) period (see Figure 2). Measurements were collected at five scheduled visits.

[0162] Study Procedures Screening and Eligibility: At Visit 1 (screening), informed consent was obtained from subjects, eligibility was determined, and documentation of primary Sjögren's syndrome was required according to the Sjögren's syndrome criteria of the American-European Consensus Group. Subjects were at least 18 years old, had a history of dry eye-related symptoms, and had used an ocular lubricant within the past 120 days. The inclusion criteria at screening and subsequent baseline visit were as follows: 1. Total CFS score on the National Eye Institute Industry Workshop (NEI) scale (Protocol Appendix 6) was ≥4 and <15 2. Symptom severity score using symptom assessment on the Dry Eye (SANDE) questionnaire was ≥40 3. Schirmer test score under anesthesia was ≤5 mm wetting / 5 minutes 4. Total LGCS score using the NEI scale was ≥5 including.

[0163] Subjects with the following were excluded: 1. Active infectious eye conditions 2. Ocular inflammatory conditions not associated with dry eye syndrome 3. Clinical evidence of cicatricial ocular surface disease 4. Use of Restasis® (topical ophthalmic cyclosporine) or Xiidra® (topical ophthalmic lifitegrast) within 14 days prior to Visit 1 5. History of collagen vascular disease, autoimmune disease, or rheumatic disease other than primary Sjögren's syndrome (e.g., lupus, rheumatoid arthritis, etc.) 6. History or current presence of anterior membrane dystrophy, corneal transplantation, corneal refractive surgery, or other recent eye procedures 7. Possibility of pregnancy not desiring contraception, or pregnancy or lactation 8. Any physical or mental disorder that would prevent the ability to participate and give informed consent

[0164] Eligible subjects entered a 14-day lead-in period that included instillation of 1 drop of single-blind placebo (vehicle) 3 times a day in each eye.

[0165] Randomization and treatment: At Visit 2 (baseline / randomization), eligibility was confirmed. All criteria were met in the same study eye. For each subject, the study eye was the one eligible for inclusion in the study group, or if both were eligible, the one with the higher baseline total corneal fluorescein staining score, or the right eye if both eyes had the same baseline score. Eligible subjects were randomly assigned to 1 of 3 treatment groups: 2 Lacripep TM ophthalmic solution strengths (22 μM or 44 μM) or placebo (vehicle).

[0166] One drop of the study drug was administered 3 times a day (TID) to both eyes for 28 days. At Visit 3 (2 weeks) and Visit 4 (4 weeks), assessments of efficacy and safety were performed. Subjects who discontinued prior to Visit 4 received the Visit 4 assessment (early discontinuation).

[0167] 14 - day follow - up period: After discontinuation of the investigational drug, there was a 14 - day follow - up period, and the subjects instilled 1 drop of Refresh Plus® (Allergan, Dublin, Ireland) into each eye TID.

[0168] At Visit 5 (6 - week follow - up), efficacy and safety evaluations were performed.

[0169] Concomitant medications / treatments: Subjects who had records indicating use of drugs (topical, topical ocular, systemic, and / or injectable) prohibited during the appropriate pre - washout period before randomization and / or during the 14 - day vehicle lead - in period were excluded from the efficacy analysis before database lock.

[0170] Subjects did not receive any investigational drug or device, except as per protocol within 30 days of screening and during the trial. Subjects receiving systemic (oral) treatment for the treatment of Sjögren's syndrome had to have received a stable systemic treatment defined as the same treatment for the previous 90 days. Use of cyclosporine (formulated or Restasis® Allergan, Irvine, CA or Cequa, Sun Pharmaceuticals, Mumbai, India) or lifitegrast (Xiidra® Novartis, Basel, CH) within 14 days before the screening examination was prohibited. Subjects did not make any changes (insertion or removal) to punctal plugs in the study eye within 14 days before the screening examination and throughout the trial.

[0171] Topical or systemic drugs known to exacerbate dry eye were prohibited during the trial.

[0172] Trial masking: Subjects were randomly assigned to receive either placebo or the investigational drug. Placebo (vehicle) and Lacripep TMThe containers were identical in appearance during the double-blind treatment phase. The subjects, the investigators, and their staff were masked to the identity of the treatment until the final database was locked.

[0173] Outcome Measures Efficacy: All efficacy evaluations were performed in both eyes by the same person if possible. Corneal fluorescein staining (CFS) was evaluated on a scale of 0 - 3 (total 0 - 15) in five regions (central, inferior, superior, temporal, and nasal). Lissamine green conjunctival staining (LGCS) was evaluated on a scale of 0 - 3 (total 0 - 15) in six regions. All sites were trained and tested using the NEI scoring method.

[0174] After drying the inferior fornix, approximately 50 μL (one drop) of 0.5% proparacaine was instilled, and then the Schirmer test under anesthesia was performed and recorded in millimeters of wetting over 5 minutes. Tear breakup time (TBUT) was measured in seconds. The Dry Eye Symptom Assessment (SANDE) Inventory Version 1 and Version 2 were used to measure the severity and frequency of symptoms evaluated by the patients using a visual analog scale. Individual symptom evaluations (instantaneous and reflex) were measured on a visual analog scale (VAS) for symptoms reported by the patients. SANDE Version 2 and individual symptom evaluations (reflex) were administered to the subjects on days 14, 28, and 42 of treatment and asked to evaluate the difference in symptoms compared to their last visit.

[0175] The primary efficacy measure was the mean change from baseline to day 28 in the total CFS score (NEI / Industry Workshop 0 - 15 scale, 0 - 3 score in each of five regions) in the study eye. The important secondary efficacy measure was the mean change from baseline to day 28 in the dry eye score (0 - 100 mm VAS) from individual symptom evaluations.

[0176] Changes from baseline at day 14 and follow-up after treatment (day 42) in the CFS were secondary measurements. Other secondary measurements included changes from baseline at day 14 and day 28 in the LGCS score, TFBUT, and Schirmer test. Secondary measurements included changes from baseline at day 14 and day 28 in each of the SANDE and individual symptom assessments.

[0177] Safety: Safety measurements included external eye examinations, dilated ophthalmoscopy, intraocular pressure (IOP), slit-lamp biomicroscopy, and best-corrected visual acuity (BCVA). If the subject was diagnosed with or found to have meibomian gland disease (MGD), the severity was evaluated.

[0178] All treatment-emergent adverse events (TEAEs), their severity, and their relationship to the study drug were recorded.

[0179] Statistical analysis: All analyses were performed using SAS (version 9.4, SAS Institute, Cary NC).

[0180] Analysis sets: The intention-to-treat (ITT) set included all subjects who took at least one dose of the study drug, as shown in the dosing records. All safety variables were analyzed using the safety analysis set, including only the observed data (i.e., missing data remained missing in the safety analysis).

[0181] The full analysis set (FAS) was composed of all subjects in the ITT set who met the predefined inclusion criteria. Prior to database lock, several subjects were excluded from the FAS based on expected eligibility exclusions and / or expected defined protocol violations.

[0182] Baseline and Safety Analysis: Demographic and baseline efficacy assessment parameters were summarized for each treatment group. Baseline eye evaluations were summarized for the study eyes. Adverse events were classified according to the Medical Dictionary for Regulatory Activities (MedDRA version 20.0, MedDRA MSSA, McLean, VA). Adverse events and any adverse findings from standard safety tests were counted. The number and percentage of adverse events were summarized by treatment group and by the presence of any serious adverse events. Safety data were examined for trends between treatment groups.

[0183] Pre-specified Efficacy Analysis: Pre-specified primary and secondary assessment item analyses were performed in the FAS.

[0184] The difference between placebo and each of the two active dose groups in the change from baseline to day 28 in the total CFS score, the primary assessment item, was first tested. The Bonferroni correction was used to control the overall type I error (i.e., these were formally tested to a significance level of p < 0.025). The important secondary assessment item (dry eye score) was tested in the same way using the Bonferroni gatekeeping procedure, i.e., if the primary assessment item was met.

[0185] Next, important secondary analyses were performed using statistical inference conditional on primary inference by the Bonferroni gatekeeping procedure. Other secondary efficacy assessment items were examined for any trends between treatment groups. Summaries of all inferences for these analyses were used for illustrative purposes. Point estimates and 95% confidence intervals (CIs) for treatment differences were calculated using the t-test, along with p-values for treatment comparisons.

[0186] All two-sample t-tests performed in the analysis did not assume equal variances and used the Satterthwaite approximation for different sample sizes being compared. Missing values were imputed prior to day 28, except for the reflexive individual symptom evaluations where patients explicitly compared symptoms to previous visits, using last observation carried forward (LOCF).

[0187] Post-hoc efficacy analysis: Post-hoc, all primary and secondary assessment items were evaluated for treatment effects controlling for baseline eye dryness score (EDS) using analysis of covariance (ANCOVA). All ANCOVA models were fitted using the Proc Mixed procedure in SAS. Post-hoc analysis was also performed without imputation using all observed data in the ITT set. Data from the ITT set were examined for trends and significant efficacy signals using the mean ± standard error of the assessment item and the p-value from the non-parametric Wilcoxon rank sum test as descriptors of statistical strength. Subsequently, subgroups were evaluated according to baseline EDS.

[0188] Results Subject disposition: In the trial, 350 subjects were screened, 204 of whom were enrolled in the treatment, with 68 enrolled in each of the three treatment groups (ITT set). FAS excluded 27 subjects based on exclusion criteria or protocol violations. Excluded treated subjects reported use of prohibited drugs such as cyclosporine or lifitegrast during the lead-in or exhibited signs of co-existing autoimmune / connective tissue diseases characteristic of secondary Sjögren's.

[0189] FAS consisted of 177 subjects, 60 in the placebo group, 57 in the 22 μM Lacripep TM group, and 60 in the 44 μM Lacripep TMThey were a group. Five subjects in the efficacy analysis set were unable to complete the test: three were unable to complete the active treatment of the 28-day course, and two were unable to complete the follow-up period. In the pre-specified analysis, missing values from early dropouts were imputed using LOCF.

[0190] Demographics of the subjects: A detailed summary of the demographic data is in Table 2.1. Women constituted 96% of the subjects. The average age of the subjects was 60 years, 87% were identified as white, and the majority were not Hispanic or Latino. There were no significant differences in demographics among the three study groups.

[0191] Summarize the baseline disease characteristics in Table 2.1. The subjects showed a baseline CFS total score of 9.0 ± 2.7 (mean ± SD). The baseline dry eye score was 65.5 ± 25.5 (mean ± SD). All three groups had similar baseline disease characteristics. Similar baseline characteristics were observed for the ITT set and the FAS.

[0192]

Table 7

[0193] Post-hoc efficacy analysis: The secondary evaluation items yielded potential signals of treatment effects in the changes in lower-region CFS and the changes in burning / stinging symptoms. These were repeated in an ANCOVA analysis using the ITT set and showed a highly significant effect of baseline EDS. For illustration, Figure 3 shows the lower-region CFS (ICFS) for the entire ITT set and the subgroup with baseline EDS of ≥40 to ≥80. The subgroup with baseline EDS of ≥60 showed the strongest treatment effect but maintained 74 / 129 subjects (57%) in the sample set. The evaluation items were re-evaluated in a subset of ITT subjects with EDS ≥60 at baseline. A significant treatment effect was seen using the LGCS in total CFS and lower CFS, burning / stinging symptoms, and in sub-regions of the conjunctiva.

[0194] Figure 3: In the post hoc analysis, the inferior corneal fluorescein staining (ICFS) scores were calculated for subsets of the ITT population according to the baseline eye dryness score (EDS). For each subset, the mean and standard error of the results are plotted. The number of patients in each subset is shown above the bar graph. The subgroup with EDS ≥ 60 showed the most statistically significant effect (p = 0.0001 on day 14 and p = 0.026 on day 28). For descriptive values, the p-value levels from the post hoc Wilcoxon test are shown within the bar graph (*p < 0.05, **p < 0.005, ***p < 0.0005).

[0195] Corneal staining: The improvement in the inferior region CFS score from baseline to day 14 was statistically significant in the 22 μM Lacripep TM group compared to the placebo group. The post hoc analysis showed an improvement of 0.4, p = 0.005 (see Figure 4A). In the subset of patients with baseline EDS ≥ 60, the effect was more pronounced (improvement of 0.8, p = 0.0001), and a significant effect was also seen on day 28 (improvement of 0.4, p = 0.026). A positive (but not statistically significant) difference was seen in the high-dose group (see Figure 4B).

[0196] In the group with baseline EDS ≥ 60, the total CFS score for the predefined primary evaluation item was significantly improved on day 14 for the 22 μM dose versus placebo (1.2, p = 0.03).

[0197] Figures 4A, 4B: Post hoc analysis of CFS signs. Figure 4A. The primary evaluation item, the CFS score on day 28, was not met, but there was a significant treatment effect for the 22 μM dose on day 14 in the inferior corneal region. Figure 4B. In the subset of subjects with baseline EDS ≥ 60, significant treatment effects were seen for the 22 μM dose group on days 14 and 28, decreased after the 42-day washout, and a positive but not significant effect was seen in the 44 μM dose group.

[0198] Individual symptom assessment (burning / stinging): Improvement from baseline to day 14 in the burning / stinging symptom (reflex VAS assessment) was 22 μM Lacripep TM in the group compared to the placebo group was statistically significant in the pre-specified analysis. Post hoc analysis of the change in the immediate assessment of burning / stinging showed a 11.6 mm improvement in VAS vs placebo, p = 0.006 (see Figure 5A). In a subset of patients with a baseline EDS of ≥60, a significant effect (14.0 mm improvement, p = 0.027) was seen at the same time point, and a significant effect was also shown in the high-dose group (14.2 mm, p = 0.038, see Figure 5B). Both dose groups trended better than the placebo group at days 28 and 42 (not statistically significant).

[0199] Figure 5A, 5B: Post hoc analysis of the burning / stinging symptom. Figure 5A. There was a significant treatment effect for the 22 μM dose at day 14 in the burning / stinging symptom. Figure 5B. A significant treatment effect was seen at day 14 for both doses in a subset of subjects with a baseline EDS of ≥60. Positive but not significant effects were seen at day 28 for both doses.

[0200] Resamine staining: Improvement in LGCS was not statistically significant in the pre-specified secondary assessment item analysis. However, in a subset of patients with a baseline EDS of ≥60, the change from the baseline LCGS total score was 1.5 improved for 22 μM Lacripep TM in the group compared to placebo at day 14 (p = 0.017). The same dose and time point showed significant improvement in segment 1 (0.4, p = 0.049) and segment 5 (0.4, p = 0.038).

[0201] Furthermore, although the foregoing has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be understood by those of ordinary skill in the art that numerous and various modifications may be made without departing from the spirit of the present disclosure. Accordingly, it should be clearly understood that the forms disclosed herein are merely illustrative and not limiting of the scope of the present disclosure, but rather are intended to encompass all modifications and alternatives that accompany the true scope and spirit of the embodiments of the present invention.

[0202] In this application, particularly in the claims, the terms and phrases used and their variations should be construed as open-ended rather than limiting, unless otherwise expressly stated. By way of example, the term "including" should be interpreted to mean "including without limitation," "including but not limited to," and the like.

[0203] The indefinite articles "a" or "an" do not exclude a plurality. The use of "about" before a number includes the number itself. For example, "about 5" provides explicit support for "5".

Claims

1. A liquid composition for treating moderate / severe dry eye-related ocular symptoms, The composition is administered to subjects identified as suffering from moderate / severe dry eye-related ocular symptoms; The composition is as follows: A polypeptide comprising 0.0001–0.005% (w / v) or a pharmaceutically acceptable salt thereof, wherein the polypeptide is Ac-Lys-Gln-Phe-Ile-Glu-Asn-Gly-Ser-Glu-Phe-Ala-Gln-Lys-Leu-Leu-Lys-Lys-Phe-Ser-NH 2 A polypeptide having the sequence (SEQ ID NO: 1), wherein "Ac" represents an acetyl group and the C-terminus is amidated, or a pharmaceutically acceptable salt thereof; 0.01–0.6% (w / v) of buffering agent; 0.0005–0.01% (w / v) of disodium EDTA; 0.0001–0.01% or 0.0001–0.05% (w / v) of tyroxapole; and 0.1–1.0% (w / v) sodium chloride Includes, The pH of the composition is 6.2 to 6.

8. A liquid composition administered to the eye of a subject up to three times a day, one drop of the composition.

2. The target object is as follows: a) The total FCS score on the NEI / Industry Workshop scale is ≥4 and <15; b) Eye dryness score using instantaneous VAS measurement is ≥60; c) Schirmer 1 test score under anesthesia is ≤5mm infiltration / 5 min; and d) Total LGCS score using the NEI / Industry Workshop scale is ≥ 5 (0 = no staining) The liquid composition according to claim 1, satisfying at least one criterion selected from the group consisting of the following.

3. The liquid composition according to claim 2, wherein the subject satisfies all four criteria in one eye.

4. The liquid composition according to claim 1, wherein a polypeptide or a pharmaceutically acceptable salt thereof is present in the liquid composition at a concentration of 0.00025 to 0.005% (w / v) or 0.0001 to 0.001% (w / v).

5. The liquid composition according to claim 1, wherein a polypeptide or a pharmaceutically acceptable salt thereof is present in the liquid composition at a concentration of 0.00025% or 0.001% (w / v).

6. The liquid composition according to claim 1, wherein tyroxapol is present in the liquid composition at a concentration of 0.0005 to 0.01% (w / v) or 0.0005 to 0.05% (w / v).

7. The liquid composition according to claim 5, wherein tyroxapol is present in the liquid composition at a concentration of 0.05% (w / v).

8. The liquid composition according to claim 7, wherein the pH of the composition is 6.3 to 6.

6.

9. The liquid composition according to claim 1, wherein the amount of NaCl is 0.4% (w / v) to 0.6% (w / v).

10. The liquid composition according to claim 8, wherein the amount of NaCl is 0.5% (w / v).

11. The liquid composition according to claim 10, wherein the osmolality of the composition is 190 to 210 mOsm / kg.

12. The liquid composition according to claim 1, wherein the buffering agent is a citrate buffering agent.

13. The liquid composition according to claim 11, wherein the buffering agent comprises 0.0098% anhydrous citric acid and 0.279% sodium citrate dihydrate.

14. The liquid composition according to claim 1, wherein the amount of EDTA is 0.0005% (w / v) to 0.005% (w / v).

15. The liquid composition according to claim 13, wherein the amount of EDTA is approximately 0.001% (w / v).

16. The liquid composition according to any one of claims 1 to 15, wherein administration improves the total FCS score (NEI / Industry Workshop 0-15 scale) in the eyes of a subject at least two weeks after treatment, or at least four weeks after treatment, or at least six weeks after the start of treatment at four weeks, compared to a baseline measurement before the start of treatment.

17. The administration is as follows: Dry eyes at least two weeks after treatment or at least four weeks after treatment, compared to baseline on a visual analog scale; SANDE (overall score on SANDE 1) at least two weeks after treatment, compared to baseline measurements before treatment initiation; The mean SANDE (overall SANDE-1 score) score at least two weeks after treatment, compared to baseline measurement before treatment initiation; Individual symptom assessments (instantaneous) at least two weeks after treatment, compared to baseline measurements before treatment initiation; The mean score of individual symptom assessments (reflexes) at least two weeks after treatment, compared to baseline measurements before the start of treatment; LGCS of the subject's eye at least two weeks after treatment, compared to baseline measurement before treatment commencement; Schirmer test of the eye under anesthesia in the subject at least two weeks after treatment, compared to baseline measurements before the start of treatment; TFBUT of the subject's eye at least two weeks after treatment, compared to baseline measurements before the start of treatment; FCS of the subject's eye at least two weeks after treatment, compared to baseline measurement before treatment commencement; SANDE (overall SANDE 1 score) at least two weeks after treatment, or at least four weeks after treatment, or one week after four weeks of treatment, compared to baseline measurements before treatment initiation; Individual symptoms (instantaneous), compared to baseline measurements before the start of treatment, at least two weeks after treatment, or at least four weeks after treatment, or one week after four weeks of treatment; The mean score (overall SANDE-2 score) at least two weeks after treatment, or at least four weeks after treatment, or one week after four weeks of treatment, compared to baseline measurement before treatment initiation; The mean score of individual symptom assessments (reflexes) at least two weeks after treatment, or at least four weeks after treatment, or one week after four weeks of treatment, compared to baseline measurements before the start of treatment; FCS and SANDE 1 and individual symptom assessments (instantaneous) at least 2 weeks after treatment, or at least 4 weeks after treatment, compared to baseline measurements before treatment initiation; LGCS at least 2 weeks after treatment, or at least 4 weeks after treatment, compared to baseline measurements before treatment initiation; Schirmer test results under anesthesia, at least two weeks after treatment or at least four weeks after treatment, compared to baseline measurements before the start of treatment; TFBUT at least 2 weeks after treatment, or at least 4 weeks after treatment, or 1 week after 4 weeks of treatment, compared to baseline measurements before treatment initiation. The liquid composition according to claim 1, which improves one or more of the above.

18. The liquid composition according to claim 1, wherein the subject is identified as suffering from severe dry eye rather than moderate / severe dry eye.

19. The liquid composition according to claim 18, wherein the subject has an eye dryness score of ≥70 in at least one eye using VAS instantaneous measurement.

20. The liquid composition according to claim 19, wherein the subject does not have Sjögren's syndrome.

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