Novel Polypeptides

Abstract

The present disclosure provides novel polypeptides. Also provided are ophthalmic compositions for treating or preventing dry eye (DE) or DE-related disorders. Also provided are methods for treating or preventing DE or DE-related disorders in a subject in need thereof, stimulating tear fluid, stabilizing tear film, or any combination thereof, comprising administering to the subject an effective amount of the novel peptide. Also provided are contact lens care products comprising the novel peptides and methods for preparing the same.

Description

[Technical field]

[0001] cross reference This application claims priority to PCT Application No. PCT / CN2021 / 118499, filed September 15, 2021, which is incorporated by reference in its entirety. [Background technology]

[0002] background The hydrophobic and / or hydrophilic properties of a peptide are essential for peptide stability, e.g., shelf life, and biological activity, including secondary structure configuration, peptide-protein interactions, adsorption and desorption kinetics, cell membrane translocation and permeation, etc. Hydrophobic or hydrophilic characteristics are also referred to as hydropathic character, hydropathicity, or hydropathy. Improving the hydropathic properties of a peptide, e.g., by replacing one or more amino acids with amino acids having the opposite hydropathic character, may sometimes improve the stability or biological activity of the peptide.

[0003] Tears are essential for clear vision and for maintaining eye health. They spread over the front of the eye with each blink, providing moisture, washing away foreign material to keep the ocular surface lubricated and clean, and reducing the risk of infection. Dry eye (DE) is a condition in which the eye does not produce enough tears or the tears evaporate too quickly. Dry eye is one of the most common eye disorders and is often a chronic problem, especially in the elderly. Symptoms associated with DE include dryness, burning, irritation, redness, eye discharge, eye fatigue, and blurred vision. In some patients, DE can lead to further scarring and even eye inflammation if not treated properly. Summary of the Invention [Means for solving the problem]

[0004] overview The present disclosure provides a novel peptide. Also provided is an ophthalmic formulation comprising the novel peptide for treating DE. Also provided is a method for treating or preventing DE or DE-related disorders, stimulating tear fluid, stabilizing tear film, or any combination thereof in a subject in need thereof, comprising administering to the subject an effective amount of the novel peptide. Also provided is a contact lens care product comprising the novel peptide and a method for preparing the same.

[0005] In one embodiment, the compound of formula (I): XY(I) Artificial polypeptides (In the formula, X is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:1, Y is a moiety comprising 10 to 50 amino acids, at least 50% of which are selected from I, V, L, F, C, M, and A; The total number of cysteines (C) contained in Y is less than 5. is provided.

[0006] In some embodiments, X comprises a sequence having at least 90% identity to SEQ ID NO: 1. In some embodiments, X comprises a sequence having at least 95% identity to SEQ ID NO: 1. In some embodiments, X comprises the sequence of SEQ ID NO: 1. In some embodiments, at least 50% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, X comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 81-83.

[0007] In some embodiments, the total number of cysteines (C) contained in Y is less than 4. In some embodiments, the total number of cysteines (C) contained in Y is less than 3. In some embodiments, the total number of cysteines (C) contained in Y is less than 2. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 8. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 12. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 15. In some embodiments, the total number of hydrophilic amino acids in Y is 5 or less. In some embodiments, Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments, Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 3-18, 58-61, and 84-85.

[0008] In some embodiments, the artificial polypeptide comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 29-41, 54-57, and 91-95.

[0009] In another embodiment, the compound of formula (II): XY(II) Artificial polypeptides (In the formula, X is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:1, Y is a moiety comprising 10 to 50 amino acids, at least 50% of which are selected from I, V, L, F, C, M, and A; Y comprises a sequence having up to 90% identity to SEQ ID NO:2. is provided.

[0010] In some embodiments, X comprises a sequence having at least 90% identity to SEQ ID NO: 1. In some embodiments, X comprises a sequence having at least 95% identity to SEQ ID NO: 1. In some embodiments, X comprises the sequence of SEQ ID NO: 1. In some embodiments, at least 50% of the amino acids of X are selected from R, K, N, D, Q, E, and H.

[0011] In some embodiments, Y comprises a sequence having up to 80% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having up to 70% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having up to 50% identity to SEQ ID NO:2. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 8. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 12. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 15. In some embodiments, the total number of hydrophilic amino acids in Y is 5 or less. In some embodiments, Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments, Y comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 16-18.

[0012] In some embodiments, the artificial polypeptide comprises a sequence having at least 90% identity to SEQ ID NOs:42-44.

[0013] In another embodiment, the compound of formula (III): XY(III) Artificial polypeptides (In the formula, X is a moiety comprising 40-65 amino acids, at least 50% of which are selected from H, R, K, D, Q, N and E; Y is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:2, The total number of H, R, K, D, Q, N, and E in X is less than 33. is provided.

[0014] In some embodiments, Y comprises a sequence having at least 90% identity to SEQ ID NO: 2. In some embodiments, Y comprises a sequence having at least 95% identity to SEQ ID NO: 2. In some embodiments, Y comprises the sequence of SEQ ID NO: 2. In some embodiments, at least 50% of the amino acids of Y are selected from I, V, L, F, C, M, and A.

[0015] In some embodiments, the total number of H, R, K, D, Q, N, and E in X is less than 30. In some embodiments, the total number of H, R, K, D, Q, N, and E in X is less than 25. In some embodiments, the total number of H, R, K, D, Q, N, and E in X is less than 20. In some embodiments, the total number of hydrophilic amino acids in X is greater than 10. In some embodiments, the total number of hydrophilic amino acids in X is greater than 15. In some embodiments, the total number of hydrophilic amino acids in X is greater than 20. In some embodiments, the total number of hydrophilic amino acids in X is greater than 25. In some embodiments, the total number of hydrophobic amino acids in X is 15 or less. In some embodiments, the total number of hydrophobic amino acids in X is 10 or less. In some embodiments, X comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 23-27. In some embodiments, X comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 23-27.

[0016] In some embodiments, the artificial polypeptide comprises a sequence having at least 90% identity to SEQ ID NOs:49-53.

[0017] In another embodiment, the compound of formula (IV): XY(IV) Artificial polypeptides (In the formula, X is a moiety comprising 40-65 amino acids, at least 50% of which are selected from H, R, K, D, Q, N and E; Y is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:2, X comprises a sequence having up to 90% identity to SEQ ID NO:1. is provided.

[0018] In some embodiments, X comprises a sequence having up to 80% identity to SEQ ID NO: 1. In some embodiments, X comprises a sequence having up to 70% identity to SEQ ID NO: 1. In some embodiments, X comprises a sequence having up to 50% identity to SEQ ID NO: 1.

[0019] In some embodiments, Y comprises a sequence having at least 90% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at least 95% identity to SEQ ID NO:2. In some embodiments, Y comprises the sequence of SEQ ID NO:2. In some embodiments, at least 50% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the total number of hydrophilic amino acids in X is greater than 10. In some embodiments, the total number of hydrophilic amino acids in X is greater than 15.

[0020] In some embodiments, the total number of hydrophilic amino acids in X is greater than 20. In some embodiments, the total number of hydrophilic amino acids in X is greater than 25. In some embodiments, the total number of hydrophobic amino acids in X is 15 or less. In some embodiments, the total number of hydrophobic amino acids in X is 10 or less. In some embodiments, X comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 19-22 and 76-79. In some embodiments, X comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 19-22 and 76-79.

[0021] In some embodiments, the artificial polypeptide comprises a sequence having at least 90% identity to SEQ ID NOs:45-48 and 86-89.

[0022] In another embodiment, the compound of formula (V): XY(V) Artificial polypeptides (In the formula, X is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:1, Y is a moiety containing 10 to 30 amino acids, and Y contains a sequence having at least 10 consecutive AA of SEQ ID NO: 2. is provided.

[0023] In some embodiments, Y comprises 10 to 25 amino acids. In some embodiments, Y comprises 10 to 20 amino acids. In some embodiments, Y comprises 10 to 15 amino acids.

[0024] In some embodiments, X comprises a sequence having at least 90% identity to SEQ ID NO: 1. In some embodiments, X comprises a sequence having at least 95% identity to SEQ ID NO: 1. In some embodiments, X comprises the sequence of SEQ ID NO: 1.

[0025] In some embodiments, at least 50% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, Y comprises a sequence having up to 80% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having up to 70% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having up to 60% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having up to 50% identity to SEQ ID NO:2.

[0026] In some embodiments, the polypeptide comprises a sequence having at least 90% identity to SEQ ID NOs: 53-56. In some embodiments, the polypeptide comprises a sequence having at least 95% identity to SEQ ID NOs: 53-56. In some embodiments, the polypeptide comprises the sequence of any of SEQ ID NOs: 53-56.

[0027] In another embodiment, the compound of formula (VI): XY(VI) Artificial polypeptides (In the formula, X is a moiety comprising a variant of the sequence SEQ ID NO: 1, characterized in that the variant is one in which at least 1, 2, 3, 4, 5, 6, 7, 8 or 9 of the Ds in SEQ ID NO: 1 are mutated to S and / or at least 1, 2, 3, 4, 5 or 6 of the Es in SEQ ID NO: 1 are mutated to T, Y is a moiety comprising a variant of the sequence of SEQ ID NO:2, characterized in that at least one, two, three, four or five of the C's in SEQ ID NO:2 are mutated to A. is provided.

[0028] In some embodiments, X comprises a sequence having at least 70% identity to SEQ ID NO:1. In some embodiments, X comprises a sequence having at most 95% identity to SEQ ID NO:1. In some embodiments, the total number of R, K, T, A, N, Q, D, E, S, and G in X is greater than 30. In some embodiments, the total number of W, Y, F, M, L, I, and V in X is 20 or less. In some embodiments, X comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs:80-83. In some embodiments, X comprises a sequence that is any one selected from SEQ ID NOs:80-83. In some embodiments, X is a sequence that is any one selected from SEQ ID NOs:80-83.

[0029] In some embodiments, Y comprises a sequence having at least 80% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having up to 95% identity to SEQ ID NO:2. In some embodiments, at least 50% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the total number of R, K, T, A, N, Q, D, E, S, and G in Y is greater than 10. In some embodiments, the total number of W, Y, F, M, L, I, and V in Y is 20 or less. In some embodiments, Y comprises a sequence having at least 80% identity to SEQ ID NO:3. In some embodiments, Y comprises the sequence of SEQ ID NO:3. In some embodiments, Y is the sequence of SEQ ID NO:3.

[0030] In some embodiments, the artificial polypeptide comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide comprises a sequence that is any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide is a sequence that is any one selected from SEQ ID NOs: 90-93.

[0031] In another embodiment, the compound of formula (I) XY(I) Artificial polypeptides (In the formula, X is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:1, Y is a moiety comprising 10 to 50 amino acids, at least 50% of which are selected from I, V, L, F, C, M, and A; The total number of cysteines (C) contained in Y is less than 5. wherein at least one, two, three, four, five, six, seven, eight, or nine of the D's in X are mutated to S; and / or at least one, two, three, four, five, or six of the E's in X are mutated to T.

[0032] In some embodiments, at least 20% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, X comprises a sequence having at least 70% identity to SEQ ID NO:1.

[0033] In some embodiments, the total number of cysteines (C) contained in Y is less than 4. In some embodiments, the total number of cysteines (C) contained in Y is less than 3. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 8. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 12. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 15. In some embodiments, the total number of hydrophilic amino acids in Y is 5 or less. In some embodiments, Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments, Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 3-18, 58-61, and 84-85.

[0034] In another embodiment, the compound of formula (II) XY(II) Artificial polypeptides (In the formula, X is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:1, Y is a moiety comprising 10 to 50 amino acids, at least 50% of which are selected from I, V, L, F, C, M, and A; Y comprises a sequence having up to 90% identity to SEQ ID NO:2. wherein at least one, two, three, four, five, six, seven, eight, or nine of the D's in X are mutated to S; and / or at least one, two, three, four, five, or six of the E's in X are mutated to T.

[0035] In some embodiments, at least 20% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, X comprises a sequence having at least 70% identity to SEQ ID NO:1.

[0036] In some embodiments, Y comprises a sequence having up to 80% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having up to 70% identity to SEQ ID NO:2. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 8. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 12. In some embodiments, the total number of hydrophobic amino acids in Y is greater than 15. In some embodiments, the total number of hydrophilic amino acids in Y is 5 or less. In some embodiments, Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments, Y comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 16-18.

[0037] In another embodiment, the compound of formula (III) XY(III) Artificial polypeptides (In the formula, X is a moiety comprising 40-65 amino acids, at least 50% of which are selected from H, R, K, D, Q, N and E; Y is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:2, The total number of H, R, K, D, Q, N, and E in X is less than 33. wherein at least one, two, three, four, five, six, seven, eight, or nine of the D's in X are mutated to S; and / or at least one, two, three, four, five, or six of the E's in X are mutated to T.

[0038] In some embodiments, at least 20% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, X comprises a sequence having at least 70% identity to SEQ ID NO: 1. In some embodiments, the total number of H, R, K, D, Q, N, and E in X is less than 30. In some embodiments, the total number of H, R, K, D, Q, N, and E in X is less than 25. In some embodiments, the total number of H, R, K, D, Q, N, and E in X is less than 20. In some embodiments, the total number of hydrophilic amino acids in X is greater than 10. In some embodiments, the total number of hydrophilic amino acids in X is greater than 15. In some embodiments, the total number of hydrophilic amino acids in X is greater than 20. In some embodiments, the total number of hydrophilic amino acids in X is greater than 25. In some embodiments, the total number of hydrophobic amino acids in X is 15 or less. In some embodiments, the total number of hydrophobic amino acids in X is 10 or less.

[0039] In some embodiments, Y comprises a sequence having at least 90% identity to SEQ ID NO: 2. In some embodiments, Y comprises the sequence of SEQ ID NO: 2. In some embodiments, at least 50% of the amino acids of Y are selected from I, V, L, F, C, M, and A.

[0040] In another embodiment, the compound of formula (IV) XY(IV) Artificial polypeptides (In the formula, X is a moiety comprising 40-65 amino acids, at least 50% of which are selected from H, R, K, D, Q, N and E; Y is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:2, X comprises a sequence having up to 90% identity to SEQ ID NO:1. wherein at least one, two, three, four, five, six, seven, eight, or nine of the D's in X are mutated to S; and / or at least one, two, three, four, five, or six of the E's in X are mutated to T.

[0041] In some embodiments, at least 20% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, X comprises a sequence having at least 70% identity to SEQ ID NO: 1. In some embodiments, the total number of hydrophilic amino acids in X is greater than 10. In some embodiments, the total number of hydrophilic amino acids in X is greater than 15. In some embodiments, the total number of hydrophilic amino acids in X is greater than 20. In some embodiments, the total number of hydrophilic amino acids in X is greater than 25. In some embodiments, the total number of hydrophobic amino acids in X is 15 or less. In some embodiments, the total number of hydrophobic amino acids in X is 10 or less.

[0042] In some embodiments, Y comprises a sequence having at least 90% identity to SEQ ID NO: 2. In some embodiments, Y comprises the sequence of SEQ ID NO: 2. In some embodiments, at least 50% of the amino acids of Y are selected from I, V, L, F, C, M, and A.

[0043] In another embodiment, the compound of formula (V) XY(V) Artificial polypeptides (In the formula, X is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:1, Y is a moiety containing 10 to 30 amino acids, and Y contains a sequence having at least 10 consecutive AA of SEQ ID NO: 2. wherein at least one, two, three, four, five, six, seven, eight, or nine of the D's in X are mutated to S; and / or at least one, two, three, four, five, or six of the E's in X are mutated to T.

[0044] In some embodiments, at least 20% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, X comprises a sequence having at least 70% identity to SEQ ID NO:1.

[0045] In some embodiments, Y comprises 10-25 amino acids. In some embodiments, Y comprises 10-20 amino acids. In some embodiments, Y comprises 10-15 amino acids. In some embodiments, Y comprises a sequence having up to 80% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having up to 70% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having up to 50% identity to SEQ ID NO:2.

[0046] In another aspect, there is provided an ophthalmic formulation for treating or preventing dry eye (DE) or a DE-related disorder, stimulating tear fluid, stabilizing the tear film, or any combination thereof, comprising an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of the artificial polypeptide of (I), (II), (III), (IV) or (V), or a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof.

[0047] In some embodiments, the DE or DE-related disorder is selected from the group consisting of dry eye syndrome, aqueous tear dysfunction syndrome, keratoconjunctivitis sicca (KCS), lacrimal keratoconjunctivitis, aqueous tear-deficient DE, evaporative DE, DE in Sjogren's syndrome, DE in non-Sjogren's syndrome, conjunctivitis-associated DE, post-viral conjunctivitis DE, post-cataract surgery DE, VDT work-associated DE, contact lens wear-associated DE, environmental DE, corneal neovascularization DE, allergic DE, LASIK-induced neuroepitheliopathy, aqueous tear deficiency (hypolacrimation), xerophthalmia, Stevens-Johnson syndrome, ocular pemphigoid, blepharitis marginalis, eyelid insufficiency, corneal ulcer, blepharitis, and ocular hyperemia.

[0048] In some embodiments, the polypeptide of the present disclosure (e.g., an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of (I), (II), (III), (IV) or (V), or a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof) has a concentration of about 0.1 μM to about 100 μM. In some embodiments, the polypeptide has a concentration of about 0.1 μM to about 50 μM. In some embodiments, the polypeptide has a concentration of about 1 μM to about 5 μM. In some embodiments, the ophthalmic composition further comprises one or more pharma- ceutically acceptable excipients. In some embodiments, the pharma-ceutically acceptable excipients include stabilizers, buffers, preservatives, tonicity agents, antioxidants, emulsifiers, and thickeners. In some embodiments, the ophthalmic composition is formulated as a liquid, gel, ointment, suspension, semi-liquid, semi-solid gel, foam gel, cream, contact lens solution, or eye wash. In some embodiments, the ophthalmic composition is formulated as an eye drop. In some embodiments, the ophthalmic composition is formulated for topical, subconjunctival, retrobulbar, periocular, subretinal, suprachoroidal, or intraocular administration.

[0049] In another aspect, there is provided a method for treating or preventing dry eye (DE) or a DE-related disorder, stimulating tear fluid, stabilizing the tear film, or any combination thereof, in a subject in need thereof, comprising administering to the subject an effective amount of an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of the artificial polypeptide of (I), (II), (III), (IV) or (V), or a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof.

[0050] In some embodiments, the artificial polypeptide is a polypeptide of any of SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments, the artificial polypeptide is a polypeptide of any of SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74. In some embodiments, the artificial polypeptide is a polypeptide of any of SEQ ID NO:28 and SEQ ID NO:62-74. In some embodiments, the artificial polypeptide is a polypeptide of SEQ ID NO:28.

[0051] In some embodiments, the DE or DE-related disorder is selected from the group consisting of dry eye syndrome, aqueous tear dysfunction syndrome, keratoconjunctivitis sicca (KCS), lacrimal keratoconjunctivitis, aqueous tear-deficient DE, evaporative DE, DE in Sjogren's syndrome, DE in non-Sjogren's syndrome, conjunctivitis-associated DE, post-viral conjunctivitis DE, post-cataract surgery DE, VDT work-associated DE, contact lens wear-associated DE, environmental DE, corneal neovascularization DE, allergic DE, LASIK-induced neuroepitheliopathy, aqueous tear deficiency, xerophthalmia, Stevens-Johnson syndrome, ocular pemphigoid, blepharitis marginalis, eyelid insufficiency, corneal ulcer, blepharitis, and ocular hyperemia.

[0052] In another aspect, there is provided an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), or a variant of an artificial polypeptide of (I), (II), (III), (IV) or (V), for use in therapy.

[0053] In another aspect, there is provided an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), or a variant of an artificial polypeptide of (I), (II), (III), (IV) or (V), for use in treating or preventing dry eye (DE) or a DE-related disorder in a subject in need thereof, stimulating tear fluid, stabilizing the tear film, or any combination thereof.

[0054] In another aspect, there is provided a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof, for use in treating or preventing dry eye (DE) or a DE-related disorder in a subject in need thereof, stimulating tear fluid, stabilizing the tear film, or any combination thereof.

[0055] In some embodiments, the DE or DE-related disorder is selected from the group consisting of dry eye syndrome, aqueous tear dysfunction syndrome, keratoconjunctivitis sicca (KCS), lacrimal keratoconjunctivitis, aqueous tear-deficient DE, evaporative DE, DE in Sjogren's syndrome, DE in non-Sjogren's syndrome, conjunctivitis-associated DE, post-viral conjunctivitis DE, post-cataract surgery DE, VDT work-associated DE, contact lens wear-associated DE, environmental DE, corneal neovascularization DE, allergic DE, LASIK-induced neuroepitheliopathy, aqueous tear deficiency, xerophthalmia, Stevens-Johnson syndrome, ocular pemphigoid, blepharitis marginalis, eyelid insufficiency, corneal ulcer, blepharitis, and ocular hyperemia.

[0056] In another aspect, there is provided a use of an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of (I), (II), (III), (IV) or (V), or a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof, for preparing a medicament for treating or preventing dry eye (DE) or a DE-related disorder in a subject in need thereof, for stimulating tear fluid, for stabilizing the tear film, or any combination thereof.

[0057] In some embodiments, the DE or DE-related disorder is selected from the group consisting of dry eye syndrome, aqueous tear dysfunction syndrome, keratoconjunctivitis sicca (KCS), lacrimal keratoconjunctivitis, aqueous tear-deficient DE, evaporative DE, DE in Sjogren's syndrome, DE in non-Sjogren's syndrome, conjunctivitis-associated DE, post-viral conjunctivitis DE, post-cataract surgery DE, VDT work-associated DE, contact lens wear-associated DE, environmental DE, corneal neovascularization DE, allergic DE, LASIK-induced neuroepitheliopathy, aqueous tear deficiency, xerophthalmia, Stevens-Johnson syndrome, ocular pemphigoid, blepharitis marginalis, eyelid insufficiency, corneal ulcer, blepharitis, and ocular hyperemia.

[0058] In another aspect, a contact lens care product is provided comprising an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of the artificial polypeptide of (I), (II), (III), (IV) or (V), or a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof.

[0059] In some embodiments, the contact lens care product further comprises one or more acceptable ingredients for contact lens care products. In some embodiments, the one or more acceptable ingredients for contact lens care products are selected from the group consisting of inorganic salts, boric acid, surfactants, humectants, preservatives, and solvents. In some embodiments, the inorganic salts comprise sodium chloride, potassium chloride, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium carbonate, sodium borate, or any combination thereof. In some embodiments, the humectant comprises hyaluronic acid or a salt thereof. In some embodiments, the solvent comprises water. In some embodiments, the contact lens care product is a solution for storing, protecting, and / or cleaning contact lenses.

[0060] In another aspect, a method for preparing a contact lens care product is provided, comprising combining an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of (I), (II), (III), (IV) or (V), or a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof, with one or more acceptable ingredients for a contact lens care product. In some embodiments, the one or more acceptable ingredients for a contact lens care product are selected from the group consisting of inorganic salts, boric acid, surfactants, humectants, preservatives, and solvents. In some embodiments, the inorganic salts include sodium chloride, potassium chloride, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium carbonate, sodium borate, or any combination thereof. In some embodiments, the solvent includes water. In some embodiments, the contact lens care product is a solution for storing, protecting, and / or cleaning contact lenses.

[0061] In another aspect, there is provided the use of an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of (I), (II), (III), (IV) or (V), or a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof, for the preparation of a contact lens care product.

[0062] In some embodiments, the contact lens care product further comprises one or more acceptable ingredients for contact lens care products. In some embodiments, the one or more acceptable ingredients for contact lens care products are selected from the group consisting of inorganic salts, boric acid, surfactants, humectants, preservatives, and solvents. In some embodiments, the inorganic salts comprise sodium chloride, potassium chloride, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium carbonate, sodium borate, or any combination thereof. In some embodiments, the humectant comprises hyaluronic acid or a salt thereof. In some embodiments, the solvent comprises water. In some embodiments, the contact lens care product is a solution for storing, protecting, and / or cleaning contact lenses.

[0063] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in the art from the following detailed description, in which merely illustrative embodiments of the present disclosure are shown and described. As will be understood, the present disclosure is capable of other and different embodiments, and its several details are capable of modification in various obvious respects, all without departing from the present disclosure. Accordingly, the drawings and description should be regarded as illustrative in nature, and not as restrictive. Incorporation by Reference

[0064] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0065] The various features of the present disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description and accompanying drawings that set forth illustrative embodiments, in which the principles of the invention are utilized. [Brief description of the drawings]

[0066] [Figure 1]FIG. 1 illustrates the effect of a polypeptide of the present disclosure on corneal fluorescein staining (CFS) scores in DE eyes.

[0067] [Diagram 2] FIG. 2 illustrates the effect of a polypeptide of the present disclosure on tear film breakup time (TBUT) in DE eyes.

[0068] [Figure 3A] Figures 3A-3B and Figures 4A-4B illustrate the effect of the polypeptides of the present disclosure on eyes with acute severe DE. Figures 3A-3B illustrate the CFS scores of eyes with severe DE treated with a placebo and with a polypeptide of the present disclosure, and Figures 4A-4B illustrate the TBUT of eyes with acute severe DE treated with a placebo and with a polypeptide of the present disclosure. [Figure 3B] Figures 3A-3B and Figures 4A-4B illustrate the effect of the polypeptides of the present disclosure on eyes with acute severe DE. Figures 3A-3B illustrate the CFS scores of eyes with severe DE treated with a placebo and with a polypeptide of the present disclosure, and Figures 4A-4B illustrate the TBUT of eyes with acute severe DE treated with a placebo and with a polypeptide of the present disclosure. [Figure 4A] Figures 3A-3B and Figures 4A-4B illustrate the effect of the polypeptides of the present disclosure on eyes with acute severe DE. Figures 3A-3B illustrate the CFS scores of eyes with severe DE treated with a placebo and with a polypeptide of the present disclosure, and Figures 4A-4B illustrate the TBUT of eyes with acute severe DE treated with a placebo and with a polypeptide of the present disclosure. [Figure 4B] Figures 3A-3B and Figures 4A-4B illustrate the effect of the polypeptides of the present disclosure on eyes with acute severe DE. Figures 3A-3B illustrate the CFS scores of eyes with severe DE treated with a placebo and with a polypeptide of the present disclosure, and Figures 4A-4B illustrate the TBUT of eyes with acute severe DE treated with a placebo and with a polypeptide of the present disclosure.

[0069] [Diagram 5] FIG. 5 illustrates the effect of polypeptides of the present disclosure on inhibiting the thermal aggregation of BSA.

[0070] [Figure 6] FIG. 6 illustrates the effect of polypeptides of the present disclosure on oxLDL uptake.

[0071] [Figure 7] FIG. 7 illustrates the cell binding ability of the polypeptides of the present disclosure.

[0072] [Figure 8] FIG. 8 illustrates tear production and CFS scores before and after administration of a polypeptide of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0073] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0074] Detailed Description definition As used herein, the following terms have the meanings ascribed to them unless specified otherwise.

[0075] As used in the specification and claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "a cell" includes a plurality of cells, including mixtures thereof.

[0076] As used herein, the terms "comprise", "include", "contain" and variations thereof are intended to mean open-ended transitional phrases that do not exclude the possibility of additional substances or methods. When such terms are used to describe a particular pharmaceutical composition, formulation, kit, use or method of the present disclosure, they also encompass the situation where the pharmaceutical composition, formulation, kit, use or method consists of the described substances or methods. For example, the phrase "the solvent comprises water" also encompasses the situation where the solvent consists of water, i.e., the solvent contains only water. In the context of the present disclosure, the term "consisting of" is intended to mean a closed-ended transitional phrase that excludes the possibility of additional substances or methods.

[0077] As used herein, ranges set forth in this disclosure are intended to specifically disclose each of the endpoints of the range and each integer contained within the range, unless otherwise specified. For example, "X is a moiety containing 40 to 65 amino acids" means that the number of amino acids in the moiety X can be 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, or 65. As another example, "Y is a moiety comprising 10 to 50 amino acids" means that the number of amino acids in the moiety Y can be 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. Additionally, any subrange of these integers is intended to be encompassed within the scope of this disclosure. Thus, "X is a moiety comprising 40-65 amino acids" is considered to clearly disclose subranges such as "X is a moiety comprising 41-64 amino acids," "X is a moiety comprising 42-63 amino acids," "X is a moiety comprising 43-62 amino acids," etc.

[0078] The term "about" or "approximately" as used herein means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, where the acceptable error range depends in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within a range of 1 or more standard deviations as per convention in the art. Alternatively, "about" can mean within a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value. When a particular value is described in this application and claims, the term "about" or "approximately" is intended to mean within an acceptable error range for the particular value, unless otherwise specified.

[0079] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. The polymers may be linear or branched, may contain modified amino acids, and may be interrupted by non-amino acids. The terms also encompass amino acid polymers that have been modified, for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation, such as conjugation with a labeling moiety.

[0080] The term "fragment," as used herein, when applied to a protein, refers to a truncated form of a native biologically active protein, which may or may not retain at least a portion of its therapeutic and / or biological activity.

[0081] The term "variant," as used herein, when applied to a protein, refers to a protein that has sequence homology to a native biologically active protein and retains at least a portion of the therapeutic and / or biological activity of the biologically active protein. For example, a variant protein may share at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% amino acid sequence identity compared to the reference biologically active protein.

[0082] As used herein, the term "amino acid" refers to either natural and / or unnatural or synthetic amino acids, including, but not limited to, glycine and both the D or L optical isomers, as well as amino acid analogs and peptidomimetics. Standard one-letter or three-letter codes are used to designate amino acids.

[0083] The term "naturally occurring L-amino acids" refers to the L optical isomeric forms of glycine (G), proline (P), alanine (A), valine (V), leucine (L), isoleucine (I), methionine (M), cysteine ​​(C), phenylalanine (F), tyrosine (Y), tryptophan (W), histidine (H), lysine (K), arginine (R), glutamine (Q), asparagine (N), glutamic acid (E), aspartic acid (D), serine (S), and threonine (T).

[0084] The terms "hydrophilic" and "hydrophobic" refer to the degree of affinity that a substance has for water. A hydrophilic substance has a strong affinity for water and tends to dissolve, mix with, or be wetted by water, whereas a hydrophobic substance substantially lacks affinity for water and tends to repel, not absorb, and neither dissolve in, mix with, or be wetted by water. Amino acids can be characterized based on their hydrophobicity. Examples of "hydrophilic amino acids" are arginine, lysine, threonine, alanine, asparagine, and glutamine. Aspartic acid, glutamic acid, and serine, and glycine are hydrophilic amino acids of particular interest. Examples of "hydrophobic amino acids" are tryptophan, tyrosine, phenylalanine, methionine, leucine, isoleucine, and valine.

[0085] A "host cell" includes an individual cell or cell culture that can be or has been a recipient of a subject vector. A host cell includes the progeny of a single host cell. Progeny may not necessarily be completely identical (in terms of morphology or complete DNA genome) to the original parent cell, due to nature.

[0086] A "chimeric" protein contains at least one fusion polypeptide that contains regions in a sequence that are in a different position than they would be in nature. These regions may be those that are normally present in separate proteins and are brought together in the fusion polypeptide; or, these regions may normally be present in the same protein but are placed in a new arrangement in the fusion polypeptide. Chimeric proteins can be created, for example, by chemical synthesis, or by creating and translating a polynucleotide in which the peptide regions are encoded in the desired relationship.

[0087] "Conjugated," "linked," "fused," and "fusion" are used interchangeably herein. These terms refer to the joining of two or more chemical elements or components by any means, including chemical conjugation or recombinant means.

[0088] The terms "polynucleotide", "nucleic acid", "nucleotide" and "oligonucleotide" are used interchangeably. These terms refer to polymeric forms of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof. Polynucleotides may have any three-dimensional structure and may perform any function, known or unknown. The following are non-limiting examples of polynucleotides: coding or non-coding regions of a gene or gene fragment, locus(s) defined by linkage analysis, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. Polynucleotides may include modified nucleotides, such as methylated nucleotides, and nucleotide analogs. Modifications to the nucleotide structure, if present, may have been imparted prior to or after assembly of the polymer. The sequence of nucleotides may be interrupted by non-nucleotide components. A polynucleotide may be further modified after polymerization, such as by conjugation with a labeling component.

[0089] The term "complement of a polynucleotide" refers to a polynucleotide molecule that has a complementary base sequence and reverse orientation compared to a reference sequence and is therefore capable of hybridizing with complete fidelity to the reference sequence.

[0090] "Recombinant," when applied to a polynucleotide, means that the polynucleotide is the product of various combinations of in vitro cloning, restriction and / or ligation steps, and other procedures that result in a construct that has the potential to be expressed in a host cell.

[0091] "Homology" or "homologous" refers to the sequence similarity or interchangeability between two or more polynucleotide sequences or between two or more polypeptide sequences. When using a program such as BestFit to determine sequence identity, similarity or homology between two different amino acid sequences, the default settings can be used, or an appropriate scoring matrix such as blosum45 or blosum80 can be selected to optimize the identity, similarity or homology score. Homologous polynucleotides are preferably those that hybridize under stringent conditions as defined herein and have at least 70%, preferably at least 80%, more preferably at least 90%, more preferably 95%, more preferably 97%, more preferably 98%, and even more preferably 99% sequence identity to their sequences.

[0092] The terms "percent identity" and "% identity", when applied to polynucleotide sequences, refer to the percentage of residues that match between at least two polynucleotide sequences aligned using a standardized algorithm. Such algorithms can insert gaps in a standardized and reproducible manner into the sequences being compared to optimize the alignment between the two sequences, thus achieving a more meaningful comparison of the two sequences. Percent identity can be measured over the full length of a defined polynucleotide sequence, for example, as defined by a particular SEQ ID NO, or over a shorter length, for example, over the length of a fragment obtained from a larger defined polynucleotide sequence, for example, a fragment of at least 45, at least 60, at least 90, at least 120, at least 150, at least 210, or at least 450 consecutive residues. It is understood that such lengths are merely exemplary and that any fragment length supported by the sequences shown in the tables, figures, or sequence listings herein can be used to describe the length at which percentage identity can be measured.

[0093] "Percent (%) amino acid sequence identity" is defined in terms of the polypeptide sequence identified herein as the percentage of amino acid residues in a query sequence that are identical to the amino acid residues of a second, reference polypeptide sequence or a portion thereof, after aligning the sequences and introducing gaps if necessary to achieve maximum percent sequence identity, without considering any conservative substitutions as part of sequence identity. Alignment for the purpose of determining percent amino acid sequence identity can be achieved in various ways that are within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine the appropriate parameters for measuring alignment, including any algorithms required to achieve maximum alignment over the entire length of the sequences to be compared. Percent identity can be measured over the entire length of a defined polypeptide sequence, e.g., defined by a particular SEQ ID NO, or over shorter lengths, e.g., fragments taken from a larger defined polypeptide sequence, e.g., fragments of at least 15, at least 20, at least 30, at least 40, at least 50, at least 70 or at least 150 contiguous residues. It is understood that such lengths are merely exemplary and that any fragment length supported by the sequences shown in the tables, figures or sequence listing herein can be used to describe the length over which percentage identity can be measured.

[0094] A "vector" is a nucleic acid molecule that transfers an inserted nucleic acid molecule into and / or between host cells, and is preferably self-replicating in a suitable host. The term encompasses vectors that function primarily for the insertion of DNA or RNA into a cell, replicating vectors that function primarily for the replication of DNA or RNA, and expression vectors that function for the transcription and / or translation of DNA or RNA. Vectors that provide more than one of the above functions are also encompassed. An "expression vector" is a polynucleotide that can be transcribed and translated into a polypeptide(s) when introduced into a suitable host cell. An "expression system" usually implies a suitable host cell containing an expression vector that can function to produce a desired expression product.

[0095] "t 1 / 2 " as used herein means ln(2) / K el The terminal half-life is calculated as K el is the terminal elimination rate constant calculated by linear regression of the final linear portion of the log concentration-time curve. Half-life generally refers to the time required for half of the administered substance loaded in a living organism to be metabolized or excreted by normal biological processes. "t 1 / 2 The terms "terminal half-life," "elimination half-life," and "circulating half-life" are used interchangeably herein.

[0096] "Physiological conditions" refers to a set of conditions in a living host, including temperature, salt concentration, pH, that mimic those in a living subject, as well as conditions in vitro. Many physiologically relevant conditions have been established for use in in vitro assays. Generally, physiological buffers contain physiological concentrations of salt and are adjusted to a neutral pH of about 6.5 to about 7.8, preferably about 7.0 to about 7.5. Various physiological buffers are listed in Sambrook et al. (1989). Physiologically relevant temperatures range from about 25°C to about 38°C, preferably about 35°C to about 37°C.

[0097] The term "antagonist" as used herein includes any molecule that partially or completely blocks, inhibits, or neutralizes the biological activity of a native polypeptide disclosed herein. A method for identifying an antagonist of a polypeptide may include contacting the native polypeptide with a candidate antagonist molecule and measuring a detectable change in one or more biological activities normally associated with the native polypeptide. In the context of the present invention, an antagonist may include a protein, a nucleic acid, a carbohydrate, an antibody, or any other molecule that reduces the effect of a biologically active protein.

[0098] The term "agonist" is used in the broadest sense and includes any molecule that mimics the biological activity of a native polypeptide disclosed herein. Suitable agonist molecules specifically include agonist antibodies or antibody fragments, fragments or amino acid sequence variants of native polypeptides, peptides, small organic molecules, and the like. Methods for identifying agonists of a native polypeptide may include contacting the native polypeptide with a candidate agonist molecule and measuring a detectable change in one or more biological activities normally associated with the native polypeptide.

[0099] "Activity", for purposes of this invention, refers to an action or effect of a component of the fusion protein that corresponds to that of a corresponding native biologically active protein, and "biological activity" refers to an in vitro or in vivo biological function or effect, including, but not limited to, receptor binding, antagonist activity, agonist activity, or cellular or physiological response.

[0100] As used herein, "treatment" or "treating", "palliating" and "ameliorating" are used interchangeably herein. These terms refer to an approach to obtain beneficial or desired results, including, but not limited to, therapeutic benefit and / or prophylactic benefit. "Therapeutic benefit" refers to eradication or amelioration of the underlying disorder being treated. Therapeutic benefit may also be achieved by eradication or amelioration of one or more of the physiological symptoms associated with the underlying disease state, thus observing improvement in the subject even though the subject still suffers from the underlying disorder. With regard to prophylactic benefit, the composition may be administered to a subject at risk of developing a particular disease state or to a subject who has reported one or more of the disease physiological symptoms, even if the disease may not have been diagnosed.

[0101] "Therapeutic effect" as used herein refers to a physiological effect caused by the fusion polypeptide of the present invention, other than the ability to induce the production of antibodies against an antigenic epitope possessed by the biologically active protein, including but not limited to curing, alleviating, reversing, or preventing a disease state in a human or other animal, or otherwise enhancing the physical or mental well-being of a human or animal. Determination of a therapeutically effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0102] The terms "therapeutically effective amount" and "therapeutically effective dose" as used herein refer to an amount of a biologically active protein alone or as part of a fusion protein composition, which, when administered to a subject in one or multiple doses, is capable of producing some detectable beneficial effect on any symptom, aspect, measured parameter or characteristic of a disease state or condition. Such an effect need not be absolute to be beneficial. A disease state may refer to a disorder or disease.

[0103] The term "therapeutically effective dose regimen," as used herein, refers to a schedule for sequentially administered doses of a biologically active protein, either alone or as part of a fusion protein composition, where the doses are provided in a therapeutically effective amount to produce a sustained beneficial effect on any symptom, aspect, measured parameter or characteristic of a disease state or condition.

[0104] The terms "subject," "individual," or "patient," as used herein, refer to any animal that can be used in connection with the present disclosure, including, but not limited to, humans, primates, rodents, dogs, cats, horses, sheep, pigs, etc.

[0105] The term "in vivo," as used herein, refers to events that take place inside a subject's body.

[0106] The term "in vitro" as used herein refers to events that occur outside of a subject. In some embodiments, an in vitro assay encompasses any assay that is performed outside of a subject. An in vitro assay encompasses cell-based assays that use live or dead cells. An in vitro assay also encompasses cell-free assays that do not use intact cells. Polypeptides

[0107] In one aspect, an artificial polypeptide of formula XY(I) is provided.

[0108] In some embodiments, X in formula (I) may be a hydrophilic moiety, at least 50% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (I) may be a hydrophilic moiety, at least 55% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (I) may be a hydrophilic moiety, at least 60% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (I) may be a hydrophilic moiety, at least 65% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (I) may be a hydrophilic moiety, at least 70% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (I) may be a hydrophilic moiety, at least 75% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (I) can be a hydrophilic moiety, at least 80% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (I) can be a hydrophilic moiety, at least 85% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (I) can be a hydrophilic moiety, at least 90% of which are selected from R, K, N, D, Q, E, and H.

[0109] In some embodiments, X of formula (I) may be a moiety that comprises a sequence having at least 80% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 85% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 90% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 95% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises the sequence of SEQ ID NO:1. In some embodiments, X is a moiety that is the sequence of SEQ ID NO:1.

[0110] In some embodiments, Y in formula (I) may be a hydrophobic moiety, at least 50% of which are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (I) may be a hydrophobic moiety, at least 55% of which are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (I) may be a hydrophobic moiety, at least 60% of which are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (I) may be a hydrophobic moiety, at least 65% of which are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (I) may be a hydrophobic moiety, at least 70% of which are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (I) may be a hydrophobic moiety, at least 75% of which are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (I) can be a hydrophobic moiety, at least 80% of which are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (I) can be a hydrophobic moiety, at least 85% of which are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (I) can be a hydrophobic moiety, at least 90% of which are selected from I, V, L, F, C, M, and A.

[0111] In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is 5. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is more than 5. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is 6. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is more than 6. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is 7. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is more than 7. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is 8. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is greater than 8. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is greater than 9. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is greater than 9. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is greater than 10. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is greater than 10. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is greater than 11. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is greater than 11. In some embodiments, Y in formula (I) may be a moiety comprising 10-50 amino acids, and the total number of hydrophobic amino acids in Y is 12. In some embodiments, Y in formula (I) may be a moiety comprising 10-50 amino acids, and the total number of hydrophobic amino acids in Y is greater than 12.In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is 13. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is more than 13. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is 14. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is more than 14. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is 15. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of hydrophobic amino acids in Y is more than 15.

[0112] In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 5. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is more than 5. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 6. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is more than 6. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 7. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is greater than 7. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 8. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is greater than 8. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 9. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is greater than 9. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 10. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is more than 10. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 11. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is more than 11.In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 12. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is more than 12. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 13. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is more than 13. In some embodiments, Y in formula (I) may be a moiety containing 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is 14. In some embodiments, Y in formula (I) may be a moiety comprising 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is greater than 14. In some embodiments, Y in formula (I) may be a moiety comprising 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is greater than 15. In some embodiments, Y in formula (I) may be a moiety comprising 10-50 amino acids, and the total number of I, V, L, F, C, M, and A in Y is greater than 15.

[0113] In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is 5. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is less than 5. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is 4. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is less than 4. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is 3. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is less than 3. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is 2. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is less than 2. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is 1. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is less than 1. In some embodiments, Y in formula (I) may be a moiety containing 10 to 50 amino acids, and the total number of cysteines (C) in Y is 0.

[0114] In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 5. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 5 or less. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 4. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 4 or less. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 3. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 3 or less. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 2. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 2 or less. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 1. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 1 or less.

[0115] In some embodiments, the total number of R, K, N, D, Q, E, and H in Y of formula (I) is 5. In some embodiments, the total number of R, K, N, D, Q, E, and H in Y of formula (I) is 5 or less. In some embodiments, the total number of hydrophilic amino acids in Y of formula (I) is 4. In some embodiments, the total number of R, K, N, D, Q, E, and H in Y of formula (I) is 4 or less. In some embodiments, the total number of R, K, N, D, Q, E, and H in Y of formula (I) is 3. In some embodiments, the total number of R, K, N, D, Q, E, and H in Y of formula (I) is 3 or less. In some embodiments, the total number of R, K, N, D, Q, E, and H in Y of formula (I) is 2. In some embodiments, the total number of R, K, N, D, Q, E, and H in Y of formula (I) is 2 or less. In some embodiments, the total number of R, K, N, D, Q, E, and H of Y of formula (I) is 1. In some embodiments, the total number of R, K, N, D, Q, E, and H of Y of formula (I) is 1 or less.

[0116] In some embodiments of the artificial polypeptide, X in formula (I) comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 81-83. In some embodiments of the artificial polypeptide, X in formula (I) comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 81-83. In some embodiments of the artificial polypeptide, X in formula (I) comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 81-83. In some embodiments of the artificial polypeptide, X in formula (I) comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 81-83. In some embodiments of the artificial polypeptide, X in formula (I) comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 81-83. In some embodiments of the artificial polypeptide, X in formula (I) comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 81-83. In some embodiments of the artificial polypeptide, X of formula (I) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 81-83. In some embodiments of the artificial polypeptide, X of formula (I) comprises any one sequence selected from SEQ ID NOs: 81-83. In some embodiments of the artificial polypeptide, X of formula (I) is any one sequence selected from SEQ ID NOs: 81-83.

[0117] In some embodiments of the artificial polypeptide, Y in formula (I) comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments of the artificial polypeptide, Y in formula (I) comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments of the artificial polypeptide, Y in formula (I) comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments of the artificial polypeptide, Y in formula (I) comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments of the artificial polypeptide, Y in formula (I) comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments of the artificial polypeptide, Y in formula (I) comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments of the artificial polypeptide, Y in formula (I) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 9 ... one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments of the artificial polypeptide, Y of formula (I) is any one sequence selected from SEQ ID NOs: 3-15, 58-61, and 84-85.

[0118] In some embodiments of an artificial polypeptide of formula (I), the polypeptide comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 29-41, 54-57, and 91-95. In some embodiments of an artificial polypeptide of formula (I), the polypeptide comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 29-41, 54-57, and 91-95. In some embodiments of an artificial polypeptide of formula (I), the polypeptide comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 29-41, 54-57, and 91-95. In some embodiments of an artificial polypeptide of formula (I), the polypeptide comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 29-41, 54-57, and 91-95. In some embodiments of an artificial polypeptide of formula (I), the polypeptide comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 29-41, 54-57, and 91-95. In some embodiments of an artificial polypeptide of formula (I), the polypeptide comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 29-41, 54-57, and 91-95. In some embodiments of an artificial polypeptide of formula (I), the polypeptide comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 29-41, 54-57, and 91-95. In some embodiments of an artificial polypeptide of formula (I), the polypeptide comprises a sequence having at least one of SEQ ID NOs: 29-41, 54-57, and 91-95. In some embodiments of the artificial polypeptide of Formula (I), the polypeptide is any one of SEQ ID NOs: 29-41, 54-57, and 91-95.

[0119] In another aspect, an artificial polypeptide of formula XY(II) is provided.

[0120] In some embodiments, X in formula (II) may be a hydrophilic moiety, at least 50% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (II) may be a hydrophilic moiety, at least 55% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (I) may be a hydrophilic moiety, at least 60% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (II) may be a hydrophilic moiety, at least 65% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (II) may be a hydrophilic moiety, at least 70% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in (II) may be a hydrophilic moiety, at least 75% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (II) can be a hydrophilic moiety, at least 80% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (II) can be a hydrophilic moiety, at least 85% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (II) can be a hydrophilic moiety, at least 90% of which are selected from R, K, N, D, Q, E, and H.

[0121] In some embodiments, X in formula (II) may be a hydrophilic moiety that includes one or more arginines (R). For example, X in formula (II) may be a hydrophilic moiety that includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 arginines (R). In some embodiments, X in formula (II) may be a hydrophilic moiety that includes one or more lysines (K). For example, X in formula (II) may be a hydrophilic moiety that includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 lysines (K). In some embodiments, X in formula (II) may be a hydrophilic moiety that includes one or more asparagines (N). For example, X in formula (II) may be a hydrophilic moiety that includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 asparagines (N). In some embodiments, X in formula (II) may be a hydrophilic moiety that includes one or more aspartic acids (D). For example, X in formula (II) may be a hydrophilic moiety that includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 aspartic acids (D). In some embodiments, X in formula (II) may be a hydrophilic moiety that includes one or more glutamines (Q). For example, X in formula (II) may be a hydrophilic moiety that includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 glutamines (Q). In some embodiments, X in formula (II) can be a hydrophilic moiety that includes one or more glutamic acids (E).For example, X in formula (II) may be a hydrophilic moiety that includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 glutamic acids (E). In some embodiments, X in formula (II) may be a hydrophilic moiety that includes one or more histidines (H). For example, X in formula (II) may be a hydrophilic moiety that includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 histidines (H).

[0122] In some embodiments, X in formula (II) may be a moiety that comprises a sequence having at least 80% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 85% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 90% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 95% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises the sequence of SEQ ID NO:1. In some embodiments, X is a moiety that is the sequence of SEQ ID NO:1.

[0123] In some embodiments of the artificial polypeptide of Formula (II), Y comprises a sequence having at most 50% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 55% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 60% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 65% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 70% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 75% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 80% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 85% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 90% identity to SEQ ID NO:2.

[0124] In some embodiments of an artificial polypeptide of formula (II), Y comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments of an artificial polypeptide, Y comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments of an artificial polypeptide, Y comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments of an artificial polypeptide, Y comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments of an artificial polypeptide, Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments of an artificial polypeptide, Y comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments of the artificial polypeptide, Y comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments of the artificial polypeptide, Y comprises any one sequence selected from SEQ ID NOs: 16-18. In some embodiments of the artificial polypeptide, Y is any one sequence selected from SEQ ID NOs: 16-18.

[0125] In some embodiments of an artificial polypeptide of formula (II), the artificial polypeptide comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 42-44. In some embodiments of an artificial polypeptide, the artificial polypeptide comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 42-44. In some embodiments of an artificial polypeptide, the artificial polypeptide comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 42-44. In some embodiments of an artificial polypeptide, the artificial polypeptide comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 42-44. In some embodiments of an artificial polypeptide, the artificial polypeptide comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 42-44. In some embodiments of an artificial polypeptide, the artificial polypeptide comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 42-44. In some embodiments of the artificial polypeptide, the artificial polypeptide comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 42-44. In some embodiments of the artificial polypeptide, the artificial polypeptide comprises any one sequence selected from SEQ ID NOs: 42-44. In some embodiments of the artificial polypeptide, the artificial polypeptide is any one sequence selected from SEQ ID NOs: 42-44.

[0126] In another aspect, an artificial polypeptide of formula XY(III) is provided.

[0127] In some embodiments, X in formula (III) can be a hydrophilic moiety, at least 50% of which are selected from H, R, K, D, Q, N, and E. In some embodiments, X in formula (III) can be a hydrophilic moiety, at least 55% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (III) can be a hydrophilic moiety, at least 60% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (III) can be a hydrophilic moiety, at least 65% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (III) can be a hydrophilic moiety, at least 70% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (III) can be a hydrophilic moiety, at least 75% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (III) can be a hydrophilic moiety, at least 80% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (III) can be a hydrophilic moiety, at least 85% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (III) can be a hydrophilic moiety, at least 90% of which are selected from R, K, N, D, Q, E, and H.

[0128] In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 33. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 32. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 31. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 30. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 29. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 28. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 27. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 26. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 25. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 24. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 23. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 22. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 21. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 20.

[0129] In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 10. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 11. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 12. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 13. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 14. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 15. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 16. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 17. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 18. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 19. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 20. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 21. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 22. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophilic amino acids in X is greater than 23. In some embodiments, X of formula (III) is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 24. In some embodiments, X of formula (III) is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 25.

[0130] In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 15 or less. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 14 or less. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 13 or less. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 12 or less. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 11 or less. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 10 or less. In some embodiments, X in formula (III) is a moiety containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 5 or less.

[0131] In some embodiments, X in formula (III) comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 23-27. In some embodiments, X in formula (III) comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 23-27. In some embodiments, X in formula (III) comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 23-27. In some embodiments, X in formula (III) comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 23-27. In some embodiments, X in formula (III) comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 23-27. In some embodiments, X in formula (III) comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 23-27. In some embodiments, X in formula (III) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 23 to 27. In some embodiments, X in formula (III) comprises any one sequence selected from SEQ ID NOs: 23 to 27. In some embodiments, X in formula (III) is any one sequence selected from SEQ ID NOs: 23 to 27.

[0132] In some embodiments, Y in formula (III) is a moiety that comprises a sequence having at least 70% identity to SEQ ID NO:2. In some embodiments, Y in formula (III) is a moiety that comprises a sequence having at least 75% identity to SEQ ID NO:2. In some embodiments, Y in formula (III) is a moiety that comprises a sequence having at least 80% identity to SEQ ID NO:2. In some embodiments, Y in formula (III) is a moiety that comprises a sequence having at least 85% identity to SEQ ID NO:2. In some embodiments, Y in formula (III) is a moiety that comprises a sequence having at least 90% identity to SEQ ID NO:2. In some embodiments, Y in formula (III) is a moiety that comprises a sequence having at least 95% identity to SEQ ID NO:2. In some embodiments, Y in formula (III) is a moiety that comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO:2. In some embodiments, Y in formula (III) is a moiety that comprises the sequence of SEQ ID NO:2. In some embodiments, Y in formula (III) is a moiety that is the sequence of SEQ ID NO:2.

[0133] In some embodiments, Y in formula (III) is a hydrophobic moiety, and at least 50% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (III) is a hydrophobic moiety, and at least 55% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (III) is a hydrophobic moiety, and at least 60% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (III) is a hydrophobic moiety, and at least 65% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (III) is a hydrophobic moiety, and at least 70% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (III) is a hydrophobic moiety, and at least 75% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (III) is a hydrophobic moiety, and at least 80% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (III) is a hydrophobic moiety, and at least 85% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (III) is a hydrophobic moiety, and at least 90% of the amino acids of Y are selected from I, V, L, F, C, M, and A.

[0134] In some embodiments, the artificial polypeptide of formula (III) comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 49-53. In some embodiments, the artificial polypeptide of formula (III) comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 49-53. In some embodiments, the artificial polypeptide of formula (III) comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 49-53. In some embodiments, the artificial polypeptide of formula (III) comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 49-53. In some embodiments, the artificial polypeptide of formula (III) comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 49-53. In some embodiments, the artificial polypeptide of formula (III) comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 49-53. In some embodiments, the artificial polypeptide of formula (III) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 49-53. In some embodiments, the artificial polypeptide of formula (III) comprises any one sequence selected from SEQ ID NOs: 49-53. In some embodiments, the artificial polypeptide of formula (III) is any one sequence selected from SEQ ID NOs: 49-53.

[0135] In another aspect, an artificial polypeptide of formula XY(IV) is provided.

[0136] In some embodiments, X in formula (IV) can be a hydrophilic moiety, at least 50% of which are selected from H, R, K, D, Q, N, and E. In some embodiments, X in formula (IV) can be a hydrophilic moiety, at least 55% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (IV) can be a hydrophilic moiety, at least 60% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (IV) can be a hydrophilic moiety, at least 65% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (IV) can be a hydrophilic moiety, at least 70% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (IV) can be a hydrophilic moiety, at least 75% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (IV) can be a hydrophilic moiety, at least 80% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (IV) can be a hydrophilic moiety, at least 85% of which are selected from R, K, N, D, Q, E, and H. In some embodiments, X in formula (IV) can be a hydrophilic moiety, at least 90% of which are selected from R, K, N, D, Q, E, and H.

[0137] In some embodiments, X in formula (IV) comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 10. In some embodiments, X in formula (IV) comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 15. In some embodiments, X in formula (IV) comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 20. In some embodiments, X in formula (IV) comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 25. In some embodiments, X in formula (IV) comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 30. In some embodiments, X in formula (IV) comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 35.

[0138] In some embodiments, X in formula (IV) comprises a sequence having up to 50% identity to SEQ ID NO:1. In some embodiments, X in formula (IV) comprises a sequence having up to 55% identity to SEQ ID NO:1. In some embodiments, X in formula (IV) comprises a sequence having up to 60% identity to SEQ ID NO:1. In some embodiments, X in formula (IV) comprises a sequence having up to 65% identity to SEQ ID NO:1. In some embodiments, X in formula (IV) comprises a sequence having up to 70% identity to SEQ ID NO:1. In some embodiments, X in formula (IV) comprises a sequence having up to 75% identity to SEQ ID NO:1. In some embodiments, X in formula (IV) comprises a sequence having up to 80% identity to SEQ ID NO:1. In some embodiments, X in formula (IV) comprises a sequence having up to 85% identity to SEQ ID NO:1. In some embodiments, X in formula (IV) comprises a sequence having up to 90% identity to SEQ ID NO:1.

[0139] In some embodiments, Y in formula (IV) is a moiety that comprises a sequence having at least 70% identity to SEQ ID NO:2. In some embodiments, Y in formula (IV) is a moiety that comprises a sequence having at least 75% identity to SEQ ID NO:2. In some embodiments, Y in formula (IV) is a moiety that comprises a sequence having at least 80% identity to SEQ ID NO:2. In some embodiments, Y in formula (IV) is a moiety that comprises a sequence having at least 85% identity to SEQ ID NO:2. In some embodiments, Y in formula (IV) is a moiety that comprises a sequence having at least 90% identity to SEQ ID NO:2. In some embodiments, Y in formula (IV) is a moiety that comprises a sequence having at least 95% identity to SEQ ID NO:2. In some embodiments, Y in formula (IV) is a moiety that comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO:2. In some embodiments, Y in formula (IV) is a moiety that comprises the sequence of SEQ ID NO:2. In some embodiments, Y in formula (IV) is a moiety that is the sequence of SEQ ID NO:2.

[0140] In some embodiments, Y in formula (IV) is a hydrophobic moiety, and at least 50% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (IV) is a hydrophobic moiety, and at least 55% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (IV) is a hydrophobic moiety, and at least 60% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (IV) is a hydrophobic moiety, and at least 65% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (IV) is a hydrophobic moiety, and at least 70% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (IV) is a hydrophobic moiety, and at least 75% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (IV) is a hydrophobic moiety, and at least 80% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (IV) is a hydrophobic moiety, and at least 85% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (IV) is a hydrophobic moiety, and at least 90% of the amino acids of Y are selected from I, V, L, F, C, M, and A.

[0141] In some embodiments, X in formula (IV) comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 19-22 and 76-79. In some embodiments, X in formula (IV) comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 19-22 and 76-79. In some embodiments, X in formula (IV) comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 19-22 and 76-79. In some embodiments, X in formula (IV) comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 19-22 and 76-79. In some embodiments, X in formula (IV) comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 19-22 and 76-79. In some embodiments, X in formula (IV) comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 19-22 and 76-79. In some embodiments, X in formula (IV) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 19-22 and 76-79. In some embodiments, X in formula (IV) comprises any one sequence selected from SEQ ID NOs: 19-22 and 76-79. In some embodiments, X in formula (IV) is any one sequence selected from SEQ ID NOs: 19-22 and 76-79.

[0142] In some embodiments, the artificial polypeptide of formula (IV) comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 45-48 and 86-89. In some embodiments, the artificial polypeptide of formula (IV) comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 45-48 and 86-89. In some embodiments, the artificial polypeptide of formula (IV) comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 45-48 and 86-89. In some embodiments, the artificial polypeptide of formula (IV) comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 45-48 and 86-89. In some embodiments, the artificial polypeptide of formula (IV) comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 45-48 and 86-89. In some embodiments, the artificial polypeptide of formula (IV) comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 45-48 and 86-89. In some embodiments, the artificial polypeptide of formula (IV) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 45-48 and 86-89. In some embodiments, the artificial polypeptide of formula (IV) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 45-48 and 86-89. In some embodiments, the artificial polypeptide of formula (IV) is a sequence having at least 95% identity to any one selected from SEQ ID NOs: 45-48 and 86-89.

[0143] In another aspect, an artificial polypeptide of formula (V): XY(V), where X is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:1, and Y is a moiety comprising 10-30 amino acids, and Y comprises a sequence having at least 10 consecutive AA of SEQ ID NO:2, is provided.

[0144] In some embodiments, Y of formula (V) comprises 10-25 amino acids. In some embodiments, Y comprises 10-20 amino acids. In some embodiments, Y comprises 10-15 amino acids. In some embodiments, Y comprises 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 amino acids.

[0145] In some embodiments, X of formula (V) may be a moiety that comprises a sequence having at least 80% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 85% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 90% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 95% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO:1. In some embodiments, X is a moiety that comprises the sequence of SEQ ID NO:1. In some embodiments, X is a moiety that is the sequence of SEQ ID NO:1.

[0146] In some embodiments of an artificial polypeptide of Formula (V), at least 50% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 55% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 60% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 65% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 70% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 75% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 80% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 85% of the amino acids of X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 90% of the amino acids of X are selected from R, K, N, D, Q, E, and H.

[0147] In some embodiments of the artificial polypeptide of Formula (V), Y comprises a sequence having at most 50% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 55% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 60% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 65% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 70% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 75% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 80% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 85% identity to SEQ ID NO:2. In some embodiments, Y comprises a sequence having at most 90% identity to SEQ ID NO:2.

[0148] In some embodiments, an artificial polypeptide of formula (V) comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 53-56. In some embodiments, an artificial polypeptide of formula (V) comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 53-56. In some embodiments, an artificial polypeptide of formula (V) comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 53-56. In some embodiments, an artificial polypeptide of formula (V) comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 53-56. In some embodiments, an artificial polypeptide of formula (V) comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 53-56. In some embodiments, an artificial polypeptide of formula (V) comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 53-56. In some embodiments, the artificial polypeptide of formula (V) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 53-56. In some embodiments, the artificial polypeptide of formula (V) comprises any one sequence selected from SEQ ID NOs: 53-56. In some embodiments, the artificial polypeptide of formula (V) is any one sequence selected from SEQ ID NOs: 53-56.

[0149] In another aspect, an artificial polypeptide of formula XY(VI) is provided.

[0150] In some embodiments, X in formula (VI) comprises a sequence having at least 70% identity to SEQ ID NO:1. In some embodiments, X in formula (VI) comprises a sequence having at least 75% identity to SEQ ID NO:1. In some embodiments, X in formula (VI) comprises a sequence having at least 80% identity to SEQ ID NO:1. In some embodiments, X in formula (VI) comprises a sequence having at least 85% identity to SEQ ID NO:1. In some embodiments, X in formula (VI) comprises a sequence having at least 90% identity to SEQ ID NO:1. In some embodiments, X in formula (VI) comprises a sequence having at least 95% identity to SEQ ID NO:1. In some embodiments, X in formula (VI) comprises a sequence having up to 95% identity to SEQ ID NO:1. In some embodiments, X in formula (VI) comprises a sequence having up to 90% identity to SEQ ID NO:1. In some embodiments, X in formula (VI) comprises a sequence having up to 85% identity to SEQ ID NO:1. In some embodiments, X of formula (VI) comprises a sequence having up to 80% identity to SEQ ID NO: 1. In some embodiments, X of formula (VI) comprises a sequence having up to 75% identity to SEQ ID NO: 1.

[0151] In some embodiments, Y in formula (VI) is a moiety comprising a sequence having at least 70% identity to SEQ ID NO:2. In some embodiments, Y in formula (VI) is a moiety comprising a sequence having at least 75% identity to SEQ ID NO:2. In some embodiments, Y in formula (VI) is a moiety comprising a sequence having at least 80% identity to SEQ ID NO:2. In some embodiments, Y in formula (VI) is a moiety comprising a sequence having at least 85% identity to SEQ ID NO:2. In some embodiments, Y in formula (VI) is a moiety comprising a sequence having at least 90% identity to SEQ ID NO:2. In some embodiments, Y in formula (VI) is a moiety comprising a sequence having at least 95% identity to SEQ ID NO:2. In some embodiments, Y in formula (VI) is a moiety comprising a sequence having at most 95% identity to SEQ ID NO:2. In some embodiments, Y in formula (VI) is a moiety comprising a sequence having at most 90% identity to SEQ ID NO:2. In some embodiments, Y in formula (VI) is a moiety comprising a sequence having at most 85% identity to SEQ ID NO:2. In some embodiments, Y of formula (VI) comprises a sequence having up to 80% identity to SEQ ID NO: 2. In some embodiments, Y of formula (VI) comprises a sequence having up to 75% identity to SEQ ID NO: 2.

[0152] In some embodiments, Y in formula (VI) is a hydrophobic moiety, and at least 50% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (VI) is a hydrophobic moiety, and at least 55% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (VI) is a hydrophobic moiety, and at least 60% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (VI) is a hydrophobic moiety, and at least 65% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y in formula (VI) is a hydrophobic moiety, and at least 70% of the amino acids in Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (VI) is a hydrophobic moiety and at least 75% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (VI) is a hydrophobic moiety and at least 80% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (VI) is a hydrophobic moiety and at least 85% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, Y of formula (VI) is a hydrophobic moiety and at least 90% of the amino acids of Y are selected from I, V, L, F, C, M, and A.

[0153] In some embodiments, X in formula (VI) is a moiety containing 40-65 amino acids, and the total number of R, K, T, A, N, Q, D, E, S, and G in X is greater than 30. In some embodiments, X in formula (VI) is a moiety containing 40-65 amino acids, and the total number of R, K, T, A, N, Q, D, E, S, and G in X is 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40. In some embodiments, X in formula (VI) is a moiety containing 40-65 amino acids, and the total number of W, Y, F, M, L, I, and V in X is 20 or less. In some embodiments, X of formula (VI) is a moiety comprising 40-65 amino acids, and the total number of W, Y, F, M, L, I, and V in X is 19, 18, 17, 16, 15, or 14.

[0154] In some embodiments, Y in formula (VI) is a moiety containing 10-50 amino acids, and the total number of R, K, T, A, N, Q, D, E, S, and G in Y is more than 10. In some embodiments, Y in formula (VI) is a moiety containing 10-50 amino acids, and the total number of R, K, T, A, N, Q, D, E, S, and G in Y is 11, 12, 13, 14, or 15. In some embodiments, Y in formula (VI) is a moiety containing 10-50 amino acids, and the total number of W, Y, F, M, L, I, and V in X is 20 or less. In some embodiments, Y in formula (VI) is a moiety containing 10-50 amino acids, and the total number of W, Y, F, M, L, I, and V in Y is 19, 18, 17, 16, 15, or 14.

[0155] In some embodiments, X in formula (VI) comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 80 to 83. In some embodiments, X in formula (VI) comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 80 to 83. In some embodiments, X in formula (VI) comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 80 to 83. In some embodiments, X in formula (VI) comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 80 to 83. In some embodiments, X in formula (VI) comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 80 to 83. In some embodiments, X in formula (VI) comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 80 to 83. In some embodiments, X in formula (VI) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 80 to 83. In some embodiments, X in formula (VI) comprises any one sequence selected from SEQ ID NOs: 80 to 83. In some embodiments, X in formula (VI) is any one sequence selected from SEQ ID NOs: 80 to 83.

[0156] In some embodiments, Y of formula (VI) comprises a sequence having at least 70% identity to SEQ ID NO:3. In some embodiments, Y of formula (VI) comprises a sequence having at least 75% identity to SEQ ID NO:3. In some embodiments, Y of formula (VI) comprises a sequence having at least 80% identity to SEQ ID NO:3. In some embodiments, Y of formula (VI) comprises a sequence having at least 85% identity to SEQ ID NO:3. In some embodiments, Y of formula (VI) comprises a sequence having at least 90% identity to SEQ ID NO:3. In some embodiments, Y of formula (VI) comprises a sequence having at least 95% identity to SEQ ID NO:3. In some embodiments, Y of formula (VI) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO:3. In some embodiments, Y of formula (VI) comprises the sequence of SEQ ID NO:3. In some embodiments, Y of formula (VI) is the sequence of SEQ ID NO:3.

[0157] In some embodiments, the artificial polypeptide of formula (VI) comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide of formula (VI) comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide of formula (VI) comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide of formula (VI) comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide of formula (VI) comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide of formula (VI) comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide of Formula (VI) comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide of Formula (VI) comprises a sequence that is any one selected from SEQ ID NOs: 90-93. In some embodiments, the artificial polypeptide of Formula (VI) is a sequence that is any one selected from SEQ ID NOs: 90-93.

[0158] In another aspect, variants of the artificial polypeptides of Formula (I) are provided.

[0159] In some embodiments, at least 20% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 25% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 35% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 40% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 45% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 50% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 55% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 60% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 65% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 70% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 75% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 85% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X in the variant are selected from R, K, N, D, Q, E, and H.

[0160] In some embodiments, the variant X comprises a sequence having at least 70% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 75% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 80% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 85% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 90% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 95% identity to SEQ ID NO:1.

[0161] In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is 5. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is less than 5. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is 4. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is less than 4. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is 3. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is less than 3. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is 2. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is less than 2. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is 1. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is less than 1. In some embodiments, the mutant Y may be a moiety comprising 10-50 amino acids, and the total number of cysteines (C) in Y is 0.

[0162] In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 8. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 9. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 10. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 11. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 12. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 13. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 14. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 15.

[0163] In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 5. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 5 or less. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 4. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 4 or less. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 3. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 3 or less. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 2. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 2 or less. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 1. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 1 or less.

[0164] In some embodiments, the mutant Y comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 9 ... is a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 3-15, 58-61, and 84-85.

[0165] In some embodiments, the mutant Y comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 3-18, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 3-18, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 3-18, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 3-18, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 3-18, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 3-18, 58-61, and 84-85. In some embodiments, the mutant Y comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 9 ... is a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 3-18, 58-61, and 84-85.

[0166] In another aspect, variants of the artificial polypeptides of formula (II) are provided.

[0167] In some embodiments, at least 20% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 25% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 35% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 40% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 45% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 50% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 55% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 60% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 65% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 70% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 75% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 85% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X in the variant are selected from R, K, N, D, Q, E, and H.

[0168] In some embodiments, the variant X comprises a sequence having at least 70% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 75% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 80% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 85% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 90% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 95% identity to SEQ ID NO:1.

[0169] In some embodiments, the variant Y comprises a sequence having up to 50% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 55% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 60% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 65% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 70% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 75% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 80% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 85% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 90% identity to SEQ ID NO:2.

[0170] In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 8. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 9. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 10. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 11. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 12. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 13. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 14. In some embodiments, the total number of hydrophobic amino acids in the variant, Y, is greater than 15.

[0171] In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 5. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 5 or less. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 4. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 4 or less. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 3. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 3 or less. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 2. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 2 or less. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 1. In some embodiments, the total number of hydrophilic amino acids in the variant's Y is 1 or less.

[0172] In some embodiments, the variant Y comprises a sequence having at least 70% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments, the variant Y comprises a sequence having at least 75% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments, the variant Y comprises a sequence having at least 80% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments, the variant Y comprises a sequence having at least 85% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments, the variant Y comprises a sequence having at least 90% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments, the variant Y comprises a sequence having at least 95% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments, the variant Y comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NOs: 16-18. In some embodiments, the Y of the mutant comprises any one of the sequences selected from SEQ ID NOs: 16 to 18. In some embodiments, the Y of the mutant is any one of the sequences selected from SEQ ID NOs: 16 to 18.

[0173] In another aspect, variants of the artificial polypeptide of formula (III) are provided.

[0174] In some embodiments, at least 20% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 25% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 35% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 40% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 45% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 50% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 55% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 60% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 65% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 70% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 75% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 85% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X in the variant are selected from R, K, N, D, Q, E, and H.

[0175] In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 33. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 32. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 31. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 30. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 29. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 28. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 27. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 26. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 25. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 24. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 23. In some embodiments, the X of the mutant is a moiety containing 40 to 65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 22. In some embodiments, the X of the variant is a moiety that comprises 40-65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 21. In some embodiments, the X of the variant is a moiety that comprises 40-65 amino acids, and the total number of H, R, K, D, Q, N, and E in X is less than 20.

[0176] In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 10. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 11. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 12. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 13. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 14. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 15. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 16. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 17. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 18. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 19. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 20. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 21. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 22. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 23. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 24. In some embodiments, the X of the variant is a moiety comprising 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 25.

[0177] In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 15 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 14 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 13 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 12 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 11 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 10 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 5 or less.

[0178] In some embodiments, the variant X comprises a sequence having at least 70% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 75% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 80% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 85% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 90% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 95% identity to SEQ ID NO:1.

[0179] In some embodiments, the variant Y is a moiety that comprises a sequence having at least 70% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 75% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 80% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 85% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 90% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 95% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises the sequence of SEQ ID NO:2. In some embodiments, the variant Y is a moiety that is the sequence of SEQ ID NO:2.

[0180] In some embodiments, the variant Y is a hydrophobic portion, and at least 50% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 55% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 60% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 65% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 70% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 75% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic moiety and at least 80% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic moiety and at least 85% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic moiety and at least 90% of the amino acids of Y are selected from I, V, L, F, C, M, and A.

[0181] In another aspect, variants of the artificial polypeptide of formula (IV) are provided.

[0182] In some embodiments, at least 20% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 25% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 35% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 40% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 45% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 50% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 55% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 60% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 65% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 70% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 75% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 85% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X in the variant are selected from R, K, N, D, Q, E, and H.

[0183] In some embodiments, the variant X comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 10. In some embodiments, the variant X comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 15. In some embodiments, the variant X comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 20. In some embodiments, the variant X comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 25. In some embodiments, the variant X comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 30. In some embodiments, the variant X comprises 40-65 amino acids, and the total number of hydrophilic amino acids in X is greater than 35.

[0184] In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 15 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 14 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 13 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 12 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 11 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 10 or less. In some embodiments, the X of the mutant is a portion containing 40 to 65 amino acids, and the total number of hydrophobic amino acids in X is 5 or less.

[0185] In some embodiments, the variant X comprises a sequence having at least 70% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 75% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 80% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 85% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 90% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 95% identity to SEQ ID NO:1.

[0186] In some embodiments, the variant Y is a moiety that comprises a sequence having at least 70% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 75% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 80% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 85% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 90% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 95% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises a sequence having at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO:2. In some embodiments, the variant Y is a moiety that comprises the sequence of SEQ ID NO:2. In some embodiments, the variant Y is a moiety that is the sequence of SEQ ID NO:2.

[0187] In some embodiments, the variant Y is a hydrophobic portion, and at least 50% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 55% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 60% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 65% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 70% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic portion, and at least 75% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic moiety and at least 80% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic moiety and at least 85% of the amino acids of Y are selected from I, V, L, F, C, M, and A. In some embodiments, the variant Y is a hydrophobic moiety and at least 90% of the amino acids of Y are selected from I, V, L, F, C, M, and A.

[0188] In another aspect, variants of the artificial polypeptide of formula (V) are provided.

[0189] In some embodiments, at least 20% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 25% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 30% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 35% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 40% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 45% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, at least 50% of the amino acids of the variant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 55% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 60% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 65% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 70% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 75% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 80% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 85% of the amino acids of the mutant X are selected from R, K, N, D, Q, E, and H. In some embodiments, up to 90% of the amino acids of X in the variant are selected from R, K, N, D, Q, E, and H.

[0190] In some embodiments, the variant Y comprises 10-25 amino acids. In some embodiments, the variant Y comprises 10-20 amino acids. In some embodiments, the variant Y comprises 10-15 amino acids. In some embodiments, the variant Y comprises 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 amino acids.

[0191] In some embodiments, the variant X comprises a sequence having at least 70% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 75% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 80% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 85% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 90% identity to SEQ ID NO:1. In some embodiments, the variant X comprises a sequence having at least 95% identity to SEQ ID NO:1.

[0192] In some embodiments, the variant Y comprises a sequence having up to 50% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 55% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 60% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 65% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 70% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 75% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 80% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 85% identity to SEQ ID NO:2. In some embodiments, the variant Y comprises a sequence having up to 90% identity to SEQ ID NO:2.

[0193] In some embodiments, an artificial polypeptide or variant described herein has enhanced stability compared to a reference polypeptide. In some embodiments, an artificial polypeptide or variant described herein has enhanced chemical stability compared to a reference polypeptide. In some embodiments, the reference polypeptide can be any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0194] In some embodiments, the artificial polypeptides or variants described herein have enhanced stability compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71. In some embodiments, the stability of the artificial polypeptides or variants described herein is enhanced by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 100%, at least 200%, at least 300%, at least 400%, or at least 500% compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71. In some embodiments, the artificial polypeptides or variants described herein have enhanced chemical stability compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71. In some embodiments, the chemical stability of an artificial polypeptide or variant described herein is enhanced by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 100%, at least 200%, at least 300%, at least 400%, or at least 500% compared to any polypeptide selected from SEQ ID NOs:28 and 62-71.

[0195] In some embodiments, the engineered polypeptide or variant described herein has a longer shelf life compared to a reference polypeptide. In some embodiments, the reference polypeptide can be any polypeptide selected from SEQ ID NOs: 28 and 62-71. In some embodiments, the shelf life of the engineered polypeptide or variant is at least 1 day, at least 3 days, at least 5 days, at least 1 week, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 1 year, at least 2 years, at least 3 years longer than the reference polypeptide. In some embodiments, the shelf life of the engineered polypeptide or variant is at least 1 day, at least 3 days, at least 5 days, at least 1 week, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 1 year, at least 2 years, at least 3 years longer than any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0196] In some embodiments, the artificial polypeptides or variants described herein have comparable or superior activity compared to a reference polypeptide. In some embodiments, the artificial polypeptides or variants described herein have comparable or superior activity compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0197] In some embodiments, an engineered polypeptide or variant described herein has equivalent or superior cellular activity compared to a reference polypeptide. In some embodiments, an engineered polypeptide or variant described herein has equivalent or superior cellular activity compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0198] In some embodiments, an engineered polypeptide or variant described herein has comparable or superior in vivo activity compared to a reference polypeptide. In some embodiments, an engineered polypeptide or variant described herein has comparable or superior in vivo activity compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0199] In some embodiments, an engineered polypeptide or variant described herein has comparable or superior in vitro activity compared to a reference polypeptide. In some embodiments, an engineered polypeptide or variant described herein has comparable or superior in vitro activity compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0200] In some embodiments, the artificial polypeptides or variants described herein have comparable or superior activity in inhibiting / blocking DAMPs (damage associated molecular patterns) compared to a reference polypeptide. In some embodiments, the artificial polypeptides or variants described herein have comparable or superior activity in inhibiting / blocking DAMPs (damage associated molecular patterns) compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0201] In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity in inhibiting / blocking ROS compared to a reference polypeptide. In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity in inhibiting / blocking ROS compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0202] In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity in inhibiting thermal aggregation of BSA compared to a reference polypeptide. In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity in inhibiting thermal aggregation of BSA compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0203] In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity in inhibiting intracellular ROS levels compared to a reference polypeptide. In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity in inhibiting intracellular ROS levels compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0204] In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity in inhibiting oxLDL uptake compared to a reference polypeptide. In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity in inhibiting oxLDL uptake compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0205] In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity with respect to binding to oxidized proteins compared to a reference polypeptide. In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity with respect to binding to oxidized proteins compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0206] In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity with respect to binding to cells compared to a reference polypeptide. In some embodiments, an artificial polypeptide or variant described herein has comparable or superior activity with respect to binding to cells compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71.

[0207] In some embodiments, the artificial polypeptides or variants described herein have comparable or superior activity in facilitating cell membrane penetration compared to a reference polypeptide. In some embodiments, the artificial polypeptides or variants described herein have comparable or superior activity in facilitating cell membrane penetration compared to any polypeptide selected from SEQ ID NOs: 28 and 62-71. Preparation of Polypeptides

[0208] Polypeptides of the present disclosure, e.g., artificial polypeptides of Formula (I), (II), (III), (IV), (V) or (VI), variants of artificial polypeptides of Formula (I), (II), (III), (IV) or (V), or polypeptides having at least 70% identity (e.g., at least 80% identity) to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or fragments or variants thereof, can be prepared by any suitable method, including, but not limited to, molecular cloning techniques and synthetic procedures. Standard molecular cloning techniques are well known in the art and are described by Sambrook, J., Fritsch, EF and Maniatis, T. Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, (1989) (Maniatis), and by TJ Silhavy, ML Bennan. and LW Enquist, Experiments with Gene Fusions, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1984), and by Ausubel, FM et al., Current Protocols in Molecular Biology, pub. by Greene Publishing Assoc. and Wiley-Interscience (1987).

[0209] In some embodiments, the polypeptide of the present disclosure is prepared by referring to the fermentation-based production method disclosed in Chinese Patent Application No. 201711320516.4 (Publication No. CN109913483A), the entirety of which is incorporated herein by reference. In some embodiments, the method for preparing the polypeptide of the present disclosure includes the steps of incorporating a target gene fragment into an expression plasmid by genetic engineering, where the incorporated target gene fragment includes at least one purification tag; transforming the expression plasmid into a corresponding expression host to construct a recombinant engineered cell that highly expresses the target polypeptide; subjecting the recombinant engineered cell to fermentation to induce expression, and then performing crude purification to obtain a crude polypeptide; and subjecting the crude polypeptide to stringent purification to obtain a highly purified polypeptide.

[0210] In some embodiments, the target gene fragment is any one selected from the group consisting of a gene fragment capable of encoding a polypeptide of the present disclosure, such as an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of formula (I), (II), (III), (IV) or (V), or a polypeptide having at least 70% identity (e.g., at least 80% identity) to any one selected from SEQ ID NO: 28, SEQ ID NO: 57, and SEQ ID NO: 62-74, or a fragment or variant thereof. In some embodiments, the target gene fragment is a gene fragment capable of encoding an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI). In some embodiments, the target gene fragment is a gene fragment capable of encoding a variant of an artificial polypeptide of formula (I), (II), (III), (IV) or (V). In some embodiments, the target gene fragment is a gene fragment capable of encoding a polypeptide having at least 70% identity (e.g., at least 80% identity) to any one selected from SEQ ID NO: 28, SEQ ID NO: 57, and SEQ ID NO: 62-74, or a fragment or variant thereof. The target gene fragment may be prepared by any suitable method, including, but not limited to, enzymatic synthesis, i.e., synthesizing cDNA by reverse transcription using RNA as a template, and chemical synthesis, i.e., synthesizing the target gene using chemical methods or a combination of chemical and enzymatic methods. If the sequence of the target gene fragment is provided, the preparation of the target gene fragment may be performed commercially by a contract research organization (CRO).

[0211] In some embodiments, the purification tag is an ST sequence tag (an amino acid sequence that assists the polypeptides of the disclosure in forming inclusion bodies) or a His tag.

[0212] In some embodiments, the expression host is a host cell. A host cell includes, but is not limited to, an individual cell, a cell culture, or a cell line. In some embodiments, a host cell includes the progeny of a single host cell. In some embodiments, a host cell can be transfected with a heterologous sequence, including a vector encoding a polypeptide of the present disclosure. In some embodiments, the host cell can be a prokaryotic cell, such as a bacterial cell. In some embodiments, the host cell can be a eukaryotic cell, such as a yeast cell, an animal cell, an insect cell, or a plant cell.

[0213] Examples of bacterial host cells that can be used to produce the polypeptides of the present disclosure include microorganisms belonging to the genera Escherichia, Serratia, Bacillus, Brevibacterium, Corynebacterium, Microbacterium, Pseudomonas, etc. For example, the bacterial host cell can include, but is not limited to, Escherichia coli XL1-Blue, XL2-Blue, DH1, MC1000, KY3276, W1485, JM109, HB101, No. 49, i W3110, NY49, G1698, BL21, or TB1. Other bacterial host cells include, but are not limited to, Serratia ficaria, Serratia fonticola, Serratia liquefaciens, Serratia marcescens, Bacillus subtilis, Bacillus amyloliquefaciens, Brevibacterium ammoniagenes, Brevibacterium immariophilum ATCC 14068, Brevibacterium saccharolyticum ATCC 14066, Brevibacterium flavum ATCC 14067, Brevibacterium lactofermentum ATCC 13869, Corynebacterium glutamicum ATCC 13032, Corynebacterium glutamicum ATCC 13869, Corynebacterium acetoacidophilum ATCC 13870, Microbacterium ammoniaphilum ATCC 15354, Pseudomonas putida, Pseudomonas sp. D-0110, etc.

[0214] Examples of yeast cells that can be used to produce the polypeptides of the present disclosure include microorganisms belonging to the genera Kluyveromyces, Trichosporon, Saccharomyces, Schizosaccharomyces, Schwanniomyces, Pichia, Candida, etc., such as Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces lactis, Trichosporon pullulans, Schwanniomyces alluvius, Candida utilis, etc.

[0215] Examples of animal cells that can be used to produce the polypeptides of the present disclosure include mammalian cells, such as Chinese hamster ovary cells (CHO) or monkey cells, such as COS cells, HepG2 cells, A549 cells, and any other cells available through the ATCC or other depository organizations.

[0216] In some embodiments, the expression host is an Escherichia coli host cell. In some embodiments, the fermentation medium used in the fermentation process has the following formulation: yeast extract powder 10-50 g / L, peptone 10-30 g / L, ammonium sulfate 2-10 g / L, sodium chloride 2-10 g / L, potassium dihydrogen phosphate 0-10 g / L, dipotassium hydrogen phosphate 2-15 g / L, antifoam 0.01-0.1% (v / v), FeSO4·7H2O 0-0.1 g / L, ZnSO4·7H2O 0-0.02 g / L, CuSO4·5H2O 0-0.1 g / L, MnSO4·5H2O 0-0.05 g / L, CaCl2·7H2O 0-0.01 g / L, CoCl2·6H2O 0-0.01 g / L, Na2MoO4·2H2O 0-0.01 g / L, HBO3 0-0.0005 g / L, and biotin 0-0.005 g / L. In some embodiments, the fermentation process is carried out at 37° C.

[0217] In some embodiments, induction of expression is achieved by adding isopropyl-beta-D-thiogalactopyranoside (IPTG) during the fermentation process. In some embodiments, IPTG is added 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, or 5 hours after the start of the fermentation process. In some embodiments, IPTG is added to a final concentration of 0.25 mM, 0.5 mM, 1 mM, 2 mM, or 4 mM. In some embodiments, induction of expression is performed at 37°C.

[0218] The crude purification of the present disclosure is a process of pre-treating the culture produced by the fermentation process. In some embodiments, the crude purification includes the steps of collecting cells, isolating inclusion body proteins and / or cytoplasmic proteins after cell lysis, and subjecting the inclusion body proteins and / or cytoplasmic proteins to denaturation, renaturation and enzymatic digestion to obtain a crude product containing a crude polypeptide. In some embodiments, the crude purification includes the steps of collecting the culture medium, removing cells and impurities, and obtaining a supernatant, i.e., a crude product containing a crude polypeptide.

[0219] The strict purification of the present disclosure is the process of purifying the crude product containing crude polypeptide by using chromatography.The examples of the chromatography methods that can be used to purify the polypeptide of the present disclosure include ion exchange chromatography using strong base anion exchange resin, weak base anion exchange resin or multimodal anion exchange resin; affinity chromatography; reversed phase chromatography using reversed phase packing material; molecular sieve chromatography using size exclusion packing material; and hydrophobic chromatography using hydrophobic packing material.

[0220] More detailed information regarding the preparation of the polypeptides of the present disclosure can be found throughout the CN109913483A disclosure, for example, in Examples 1-4. formulation

[0221] In another aspect, a formulation is provided that includes a polypeptide of the disclosure, e.g., an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of formula (I), (II), (III), (IV) or (V), or a polypeptide having at least 70% identity (e.g., at least 80% identity) to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof, and a pharma- ceutically acceptable excipient. In some embodiments of the formulation, the polypeptide has at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the formulation, the polypeptide has at least 85% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the formulation, the polypeptide has at least 90% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the formulation, the polypeptide has at least 95% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the formulation, the polypeptide has at least 96%, at least 97%, at least 98%, at least 99% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the formulation, the polypeptide comprises or is the sequence of any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof.

[0222] In some embodiments of the formulation, pharma- ceutically acceptable excipients include, but are not limited to, inert solid diluents and fillers, diluents, sterile aqueous solutions and various organic solvents, permeation enhancers, solubilizing agents, and adjuvants.

[0223] In some embodiments of the formulation, the formulation may be in a form suitable for oral administration, for example, as a tablet, capsule, pill, powder, sustained release formulation, liquid, suspension, as a sterile solution, suspension or emulsion, as a form suitable for parenteral injection, as a spray, ointment or cream, as a form suitable for topical administration. The formulation may be in a unit dosage form suitable for single administration. In some embodiments, the pharmaceutical composition may include other medicinal or pharmaceutical agents, carriers, adjuvants, etc.

[0224] In some embodiments, the preparations may be formulated as aqueous or oily suspensions or emulsions using sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or sterile aqueous solutions, and similar pharmaceutical vehicles.

[0225] In some embodiments, the formulations can be prepared as discrete dosage forms such as capsules, cachets, or tablets, or as a liquid or aerosol spray, each containing a predetermined amount of the active ingredient as a powder or in granules, as a solution or suspension in an aqueous or non-aqueous liquid, an oil-in-water emulsion, or a water-in-oil liquid emulsion.

[0226] In some embodiments, binders suitable for use in the formulation include, but are not limited to, corn starch, potato starch, or other starches, gelatin, natural and synthetic gums, such as gum arabic, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropyl methylcellulose, microcrystalline cellulose, and mixtures thereof.

[0227] In some embodiments, fillers suitable for use in the formulation include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.

[0228] In some embodiments, disintegrants that can be used in the formulation include, but are not limited to, agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato starch or tapioca starch, other starches, pregelatinized starch, other starches, clays, other algins, other celluloses, gums, or mixtures thereof.

[0229] In some embodiments, lubricants that can be used in the formulation include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, or mixtures thereof.Additional lubricants include, for example, syloid silica gel, aerosol of synthetic silica, or mixtures thereof.Lubricants can be added in an amount of less than about 1 weight percent of the formulation if necessary.

[0230] In some embodiments, surfactants that can be used in the formulation include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, a mixture of hydrophilic surfactants can be used, a mixture of lipophilic surfactants can be used, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant can be used. A surfactant with a lower HLB value is more lipophilic or hydrophobic and more soluble in oil. On the other hand, a surfactant with a higher HLB value is more hydrophilic and more soluble in aqueous solution. Hydrophilic surfactants are generally considered to be compounds with an HLB value greater than about 10, as well as anionic, cationic, or zwitterionic compounds for which the HLB scale is not generally applicable. Similarly, lipophilic (i.e., hydrophobic) surfactants are compounds with an HLB value equal to or less than about 10. However, the HLB value of surfactants is merely a rough guideline that is generally used to enable the formulation of industrial, pharmaceutical, and cosmetic emulsions.

[0231] Hydrophilic surfactants can be either ionic or nonionic.Suitable ionic surfactants include, but are not limited to, alkyl ammonium salts; fusidate salts; fatty acid derivatives of amino acids, oligopeptides, and polypeptides; glyceride derivatives of amino acids, oligopeptides, and polypeptides; lecithin and hydrogenated lecithin; lysolecithin and hydrogenated lysolecithin; phospholipids and their derivatives; lysophospholipids and their derivatives; carnitine fatty acid ester salts; alkyl sulfate salts; fatty acid salts; docusate sodium; acyl lactate salts; mono- and diacetylated tartaric acid esters of monoglycerides and diglycerides; succinylated monoglycerides and succinylated diglycerides; citric acid esters of monoglycerides and diglycerides; and mixtures thereof.

[0232] Within the above mentioned groups, examples of ionic surfactants include lecithin, lysolecithin, phospholipids, lysophospholipids and derivatives thereof; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; docusate sodium; acyl acrylates; mono- and diacetylated tartaric acid esters of mono- and diglycerides; succinylated mono- and diglycerides; citrate esters of mono- and diglycerides; and mixtures thereof.

[0233] Ionic surfactants include lecithin, lysolecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylserine, PEG-phosphatidylethanolamine, PVP-phosphatidylethanolamine, lactylic acid esters of fatty acids, stearoyl-2-lactylate ... The ionic forms of the fatty acids may be glyceryl esters, oleyl esters, oleic acid, ricinoleic acid, linoleic acid, linolenic acid, stearic acid, lauryl sulfate, tert-butyl sulfate, docusate, lauroyl carnitine, palmitoyl carnitine, myristoyl carnitine, and salts and mixtures thereof.

[0234] Hydrophilic nonionic surfactants may include, but are not limited to, alkyl glucosides; alkyl maltosides; alkyl thioglucosides; lauryl macrogolglycerides; polyoxyalkylene alkyl ethers, such as polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols, such as polyethylene glycol alkylphenols; polyoxyalkylene alkylphenol fatty acid esters, such as polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters; polyethylene glycol glycerol fatty acid esters; polyglycerin fatty acid esters; polyoxyalkylene sorbitan fatty acid esters, such as polyethylene glycol sorbitan fatty acid esters; hydrophilic transesterification products of polyols with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols; polyoxyethylene sterols, derivatives and analogs thereof; polyoxyethylated vitamins and derivatives thereof; polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof; polyethylene glycol sorbitan fatty acid esters, and hydrophilic transesterification products of polyols with at least one member of the group consisting of triglycerides, vegetable oils, and hydrogenated vegetable oils. The polyol can be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol, or a sugar.

[0235] Other hydrophilic non-ionic surfactants include, without limitation, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, PEG-1 stearate, PEG-20 stearate, PEG-10 stearate, PEG-12 stearate, PEG-15 stearate, PEG-20 stearate, PEG-1 ... 00, PEG-20 Dilaurate, PEG-25 Glyceryl Trioleate, PEG-32 Dioleate, PEG-20 Glyceryl Laurate, PEG-30 Glyceryl Laurate, PEG-20 Glyceryl Stearate, PEG-20 Glyceryl Oleate, PEG-30 Glyceryl Oleate, PEG-30 Glyceryl Laurate, PEG-40 Glyceryl Laurate, PEG-40 Palm Kernel Oil, PEG-50 Hydrogenated Castor Oil, PEG-40 Castor Oil, PEG-35 Castor Oil, PEG-60 Castor Oil, PE G-40 Hydrogenated Castor Oil, PEG-60 Hydrogenated Castor Oil, PEG-60 Corn Oil, PEG-6 Caprylic / Caprylic Glycerides, PEG-8 Caprylic / Caprylic Glycerides, Polyglyceryl-10 Laurate, PEG-30 Cholesterol, PEG-25 Phytosterols, PEG-30 Soy Sterols, PEG-20 Trioleate, PEG-40 Sorbitan Oleate, PEG-80 Sorbitan Laurate, Polysorbate 20, Polysorbate 80, POE-9 Lauryl Ether, POE- 23 lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearyl ether, tocopheryl PEG-100 succinate, PEG-24 cholesterol, polyglyceryl-10 oleate, Tween® 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG 10-100 nonylphenol series, PEG 15-100 octylphenol series, and poloxamer.

[0236] Suitable lipophilic surface-active substances include, by way of example only, fatty alcohols, glycerol fatty acid esters, acetylated glycerol fatty acid esters, lower alcohol fatty acid esters, propylene glycol fatty acid esters, sorbitan fatty acid esters, polyethylene glycol sorbitan fatty acid esters, sterols and sterol derivatives, polyoxyethylated sterols and sterol derivatives, polyethylene glycol alkyl ethers, sugar esters, sugar ethers, lactic acid derivatives of monoglycerides and lactic acid derivatives of diglycerides, hydrophobic transesterification products of polyols with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols, oil-soluble vitamins / vitamin derivatives, and mixtures thereof.Preferred lipophilic surface-active substances in this group include glycerol fatty acid esters, propylene glycol fatty acid esters, and mixtures thereof, or preferred lipophilic surface-active substances are hydrophobic transesterification products of polyols with at least one member of the group consisting of vegetable oils, hydrogenated vegetable oils and triglycerides.

[0237] In one embodiment, a solubilizing agent can be included in the formulation to ensure good solubilization and / or dissolution of the artificial polypeptide or variant of the invention, as well as to minimize precipitation of the artificial polypeptide or variant of the invention. This can be particularly important for compositions for parenteral use, e.g., compositions for injection. Examples of suitable solubilizing agents include, but are not limited to, the following: alcohols and polyols, e.g., ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediol and its isomers, glycerol, pentaerythritol, sorbitol, mannitol, transcutol, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclodextrins and cyclodextrin derivatives; ethers of polyethylene glycol with an average molecular weight of about 200 to about 6000, e.g., tetrahydrofurfuryl alcohol PEG ether (glycofurol) or methoxy PEG; amides and other nitrogen-containing compounds, e.g., 2-pyrrolidone, 2-piperidone, ε-caprolactam, N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone; esters such as ethyl propionate, tributyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene glycol monoacetate, propylene glycol diacetate, ε-caprolactone and its isomers, δ-valerolactone and its isomers, β-butyrolactone and its isomers; and other solubilizing agents known in the art such as dimethylacetamide, dimethylisosorbide, N-methylpyrrolidone, monooctanoin, diethylene glycol monoethyl ether, and water.

[0238] Mixtures of solubilizers can also be used. Examples include, but are not limited to, triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethylacetamide, n-methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropylmethylcellulose, hydroxypropylcyclodextrin, ethanol, polyethylene glycol 200-100, glycofurol, transcutol, propylene glycol, and dimethylisosorbide. Particularly preferred solubilizers include sorbitol, glycerol, triacetin, ethyl alcohol, PEG-400, glycofurol, and propylene glycol.

[0239] In addition, acids or bases can be incorporated into the formulation to facilitate processing, enhance stability, or for other reasons. Examples of pharma-ceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium bicarbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminum silicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine, triethylamine, triisopropanolamine, trimethylamine, tris(hydroxymethyl)aminomethane (TRIS), and the like. Also suitable are bases that are salts of pharma- ceutically acceptable acids, such as acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid, and the like. Salts of polybasic acids, such as sodium phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate, can also be used. When the base is a salt, the cation can be any convenient pharma-ceutically acceptable cation, such as ammonium, an alkali metal, an alkaline earth metal, and the like. Examples include, but are not limited to, sodium, potassium, lithium, magnesium, calcium, and ammonium.

[0240] Suitable acids are pharma- ceutically acceptable organic or inorganic acids. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid, etc. Examples of suitable organic acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinonesulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid, etc. Ophthalmic preparations

[0241] Dry eye (DE) is a multifactorial disorder of the tears and ocular surface that results in symptoms such as ocular discomfort, visual disturbance, tear film instability, and ocular surface damage.

[0242] The ocular surface system includes the cornea, conjunctiva, lacrimal gland, meibomian gland, nasolacrimal duct, and their associated connective tissue matrix, as well as the eyelids and eyelashes. In DE, the ocular surface epithelium may undergo squamous metaplasia, which is manifested by goblet cell loss, mucin deficiency, and keratinization, leading to further tear film instability. "Corneal fluorescein staining," which detects ocular surface damage, has been used to assess DE. Fluorescein does not stain intact corneal epithelium, but it does stain the corneal stroma, and therefore can be used to identify areas of epithelial loss. For the test, fluorescein dye is instilled into the eye as a drop or via a paper strip. Damage is then assessed by a fluorescein staining score. Scores range from 0 to 15, with a score of 5 or more being considered DE.

[0243] The tear film is a three-layer structure that includes a mucoid basal layer, an aqueous component, and a superficial lipid layer. The tear film is formed and maintained by blinking, and the composition of the tear film can be influenced by systemic conditions or conditions affecting the eye. Tear film stability can be evaluated by various assessments, such as the "tear breakup time" or "TBUT." TBUT is a clinical test that measures the interval between an individual's last complete blink and the breakup of the tear film. This test can be used to evaluate DE or DE-related disorders. To measure TBUT, fluorescein is dropped onto the subject's tear film, and the subject is then asked not to blink while the tear film is observed under a broad beam of cobalt blue illumination. TBUT is recorded as the number of seconds that elapsed from the last blink until the appearance of the first dry spot on the tear film. A TBUT of less than 10 seconds is considered an abnormality in the tear film, and less than 5 seconds indicates DE.

[0244] "DE or DE-related disorder," as used herein, refers to any form of DE and associated symptoms, including, but not limited to, dry eye syndrome, tear dysfunction syndrome, keratoconjunctivitis sicca (KCS), lacrimal keratoconjunctivitis, aqueous tear-deficient DE, evaporative DE, DE in Sjogren's syndrome, DE in non-Sjogren's syndrome, conjunctivitis-associated DE, post-viral conjunctivitis DE, post-cataract surgery DE, VDT work-associated DE, contact lens wear-associated DE, environmental DE, corneal neovascularization DE, allergic DE, LASIK-induced neuroepitheliopathy, aqueous tear deficiency, xerophthalmia, Stevens-Johnson syndrome, ocular pemphigoid, blepharitis marginalis, eyelid insufficiency, corneal ulcer, blepharitis, and ocular redness.

[0245] In another aspect, the disclosure provides an ophthalmic formulation for treating or preventing dry eye (DE) or a DE-related disorder, the ophthalmic formulation comprising a polypeptide of the disclosure, e.g., an artificial polypeptide of Formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of Formula (I), (II), (III), (IV) or (V), or a polypeptide having at least 70% identity (e.g., at least 80% identity) to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof.

[0246] In another aspect, the disclosure provides an ophthalmic formulation for treating or preventing dry eye (DE) or a DE-related disorder, the ophthalmic formulation comprising a polypeptide having at least 70% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the ophthalmic formulation, the polypeptide comprises a sequence having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the ophthalmic formulation, the polypeptide comprises a sequence having at least 85% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the ophthalmic formulation, the polypeptide comprises a sequence having at least 90% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the ophthalmic formulation, the polypeptide comprises a sequence having at least 95% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the ophthalmic formulation, the polypeptide comprises a sequence having at least 96%, at least 97%, at least 98%, at least 99% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments of the ophthalmic formulation, the polypeptide comprises a sequence of any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof.

[0247] In some embodiments, the ophthalmic formulations of the present disclosure are formulated as liquids. In some embodiments, the ophthalmic formulations of the present disclosure are formulated as eye drops. In some embodiments, the ophthalmic formulations of the present disclosure are formulated as gels. In some embodiments, the ophthalmic formulations of the present disclosure are formulated as ointments. In some embodiments, the ophthalmic formulations of the present disclosure are formulated as suspensions, semi-liquids, semi-solid gels, creams, foam gels, contact lens solutions, eye washes, etc.

[0248] In some embodiments, the ophthalmic formulation is prepared by dissolving the polypeptide of the present disclosure in an aqueous solution. Aqueous solutions and diluents that can be used to prepare the ophthalmic formulation include, but are not limited to, distilled water, physiological saline, and the like.

[0249] In some embodiments, ophthalmic formulations are prepared by dissolving the polypeptides of the present disclosure in non-aqueous solutions or diluents, including but not limited to edible (e.g., vegetable) oils, liquid paraffin, mineral oil, propylene glycol, p-octyldodecanol, polysorbates, macrogols, aluminum monostearate, and the like.

[0250] In some embodiments, ophthalmic formulations can be formulated by mixing, diluting or dissolving the polypeptides of the present disclosure with suitable pharmaceutical excipients, such as disintegrants, binders, lubricants, diluents, buffers, preservatives, wetting agents, emulsifiers, dispersants, stabilizers and solubilizing agents, according to conventional methods and in a conventional manner depending on the dosage form.

[0251] In some embodiments, a buffer is added to keep the pH constant. The buffer includes pharma- ceutically acceptable buffers such as borate buffer, citrate buffer, tartrate buffer, phosphate buffer, acetate buffer and Tris-HCl buffer (containing tris(hydroxymethyl)aminomethane and HCl). For example, a Tris-HCl buffer with a pH of 7.4 contains 3 g / l tris-(hydroxymethyl)aminomethane and 0.76 g / l HCl. In some embodiments, the buffer is a 10x phosphate buffered saline ("PBS") or 5x PBS solution. The buffer is added to the ophthalmic formulation in an amount that provides sufficient buffer capacity under expected physiological conditions.

[0252] Other buffers that can be used in the ophthalmic formulations of the present disclosure include, but are not limited to, buffers based on HEPES (N-{2-hydroxyethyl}piperazine-N'-{2-ethanesulfonic acid}), which has a pKa of 7.5 at 25°C and a pH range of about 6.8-8.2; buffers based on BES (N,N-bis{2-hydroxyethyl}2-aminoethanesulfonic acid), which has a pKa of 7.1 at 25°C and a pH range of about 6.4-7.8; a buffer based on MOPS (3-{N-morpholino}propanesulfonic acid); which has a pKa of 7.4 at 25°C and a pH range of about 6.8-8.2; a buffer based on TES (N-tris{hydroxymethyl}-methyl-2-aminoethanesulfonic acid); which has a pKa of 7.6 at 25°C and a pH range of about 6.9-8.3; a buffer based on MOBS (4-{N-morpholino}butanesulfonic acid); which has a pKa of 7.52 at 25°C and a pH range of about 7-8.2; a buffer based on TAPS ({(2-hydroxy-3{tris(hydroxymethyl)methylamino}-1-propanesulfonic acid)) having a pKa of 7.61 at 25°C and a pH in the range of about 7 to 8.2; a buffer based on TAPS ({(2-hydroxy-1,1-bis(hydroxymethyl)ethyl)aminol-1-propanesulfonic acid)) having a pKa of 8.4 at 25°C and a pH in the range of about 7.7 to 9.1; a buffer based on TAPS ({(2-hydroxy-1,1-bis(hydroxymethyl)ethyl)aminol-1-propanesulfonic acid)) having a pKa of 8.9 at 25°C and a pH in the range of about 8.2 to 9.6; buffers based on TABS (N-tris(hydroxymethyl)methyl-4-aminobutanesulfonic acid) with a pKa of 9.0 at 25°C and a pH range of about 8.3-9.7; buffers based on AMPSO (N-(1,1-dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid) with a pKa of 9.5 at 25°C and a pH range of about 8.6-10.0; buffers based on CHES (2-cyclohexylamino)ethanesulfonic acid) with a pKa of 9.6 at 25°C and a pH range of about 8.9-10.3; and CAPS (3-(cyclohexylamino)-1-propanesulfonic acid) based buffers, which have a pKa of 10.4 at 25°C and a pH range of about 9.7 to 11.1.

[0253] In some embodiments, isotonicity agent can be added to make the formulation isotonic with tear fluid.Isotonicity agent includes, but is not limited to, sugar, such as dextrose, glucose, sucrose and fructose; sugar alcohol, such as mannitol and sorbitol; polyhydric alcohol, such as glycerol, polyethylene glycol and propylene glycol; and salt, such as sodium chloride, sodium citrate, benzalkonium chloride, phedrine chloride, potassium chloride, procaine chloride, chloramphenicol and sodium succinate.Isotonicity agent is added in an amount that makes the osmotic pressure of ophthalmic formulation equal to the osmotic pressure of tear fluid.

[0254] In some embodiments, the ophthalmic formulation comprises an osmolality adjuster. Suitable osmolality adjusters for the ophthalmic formulations of the present disclosure include, but are not limited to, sodium chloride, sodium nitrate, sodium sulfate, sodium hydrogen sulfate, potassium chloride, calcium chloride, magnesium chloride, zinc chloride, potassium acetate, sodium acetate, sodium bicarbonate, sodium carbonate, sodium thiosulfate, magnesium sulfate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, dextrose, mannitol, sorbitol, dextrose, sucrose, urea, propylene glycol, glycerin, and combinations thereof.

[0255] In some embodiments, the ophthalmic formulation comprises a thickening agent.The thickening agent suitable for the ophthalmic formulation of the present disclosure includes, but is not limited to, monomeric polyols, such as tyloxapol, glycerol, propylene glycol, ethylene glycol; polymeric polyols, such as polyethylene glycol (e.g., PEG300, PEG400); cellulose-based polymers, such as cellulose gum, alkyl cellulose, hydroxyl-alkyl cellulose, hydroxyl-alkyl alkyl cellulose, carboxy-alkyl cellulose, hydroxyethyl cellulose, hypromellose, hydroxypropyl methylcellulose, methyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose; dextran, such as dextran 70; water-soluble proteins, such as gelatin; vinyl polymers, such as polyvinyl alcohol, polyvinylpyrrolidine; other polyols, such as polysorbate 80, povidone; polysaccharides and glycosaminoglycans, such as hyaluronan, chondroitin sulfate; and combinations thereof.

[0256] In some embodiments, preservatives can be added to maintain the integrity of the ophthalmic formulation, including but not limited to sorbic acid, benzalkonium chloride, benzododecinium bromide, parabens, chlorobutanol, benzyl alcohol, phenylethyl alcohol, edentate disodium, sorbic acid, polyquaternium-1, or other agents known to those skilled in the art.

[0257] In addition to the above, in some embodiments, it may be desirable to use additional agents suitable for the ophthalmic formulations of the present disclosure, including, but not limited to, stabilizers such as sodium sulfite, sodium carbonate, and propylene glycol; antioxidants such as ascorbic acid, sodium ascorbate, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), tocopherol, sodium thiosulfate; and / or chelating agents such as ethylene-diamine-tetra-acetic acid (EDTA), ethylene glycol-bis-(2-aminoethyl)-N,N,N,N-tetraacetic acid (EGTA) and sodium citrate.

[0258] Eye drops, ophthalmic gels and / or ophthalmic ointments can be prepared by aseptic techniques. Alternatively, sterilization of the composition can be performed at the appropriate preparation stage. In some embodiments, sterile compositions can be prepared by aseptically mixing sterile components. In some embodiments, sterile compositions can be prepared by first mixing the components and then sterilizing the final preparation. Sterilization methods can include, but are not limited to, heat sterilization, irradiation and filtration.

[0259] In some embodiments, ophthalmic ointments (eye ointments) can be prepared aseptically by mixing the polypeptide of the present disclosure into a base used for preparing eye ointments, and then formulating it into a pharmaceutical preparation using any method known in the art. Typical bases for eye ointments are exemplified by petrolatum, jelene 50, plastibase and macrogol. In addition, surfactants can be added to increase water absorption.

[0260] In some embodiments, additives can be added to ophthalmic formulations, such as eye drops, ophthalmic gels, and / or ophthalmic ointments, as needed. In some embodiments, additives include, but are not limited to, additional ingredients, additives, carriers suitable for use in contact with the eye or around the eye without undue toxicity, incompatibility, instability, irritation, allergic response, etc.

[0261] In some embodiments, the ophthalmic formulations of the present disclosure are formulated for topical administration. In some embodiments, the ophthalmic formulations of the present disclosure can be administered locally to the eye, for example, subconjunctivally, retrobulbarly, periocularly, subretinally, suprachoroidally, or intraocularly.

[0262] In some embodiments, the ophthalmic formulations can be delivered to the ocular surface, interconnected innervated areas, the conjunctiva, the lacrimal gland, or the meibomian gland. It is envisioned that effective treatments may include administering the therapeutic agents of the present invention orally, topically, by injection, intranasally, rectally, transdermally, by impregnated or coated devices such as ophthalmic inserts or implants, or by iontophoresis, among other routes of administration.

[0263] In some embodiments, the ophthalmic formulation of the present disclosure is formulated for injection.For administration by injection, the ophthalmic formulation can be injected intramuscularly, intraarterially, subcutaneously, or intravenously.A pump mechanism can be used to administer the pharmaceutical composition over a preselected period of time.For some embodiments of the present invention, it is desirable to deliver the drug locally, and therefore it can be injected periocularly, intraocularly, subconjunctivally, retrobulbarly, or intercamerally.

[0264] In some embodiments, the ophthalmic formulations of the present invention can be administered to the ocular surface by a pump-catheter system or released from a continuous or selective release device, such as, but not limited to, a membrane, such as that used in the Ocusert™ System (Alza Corp, Palo Alto, Calif.). The pharmaceutical composition can be incorporated into, retained on, or attached to a contact lens, which can then be worn by a subject. The pharmaceutical composition can be sprayed onto the ocular surface.

[0265] In some embodiments, the ophthalmic formulation of the present disclosure is formulated for systemic delivery.For systemic administration, the ophthalmic formulation can be formulated for oral administration and administered orally.For administration that can result in either regional or systemic distribution of therapeutic agent, the formulation of the present invention can be administered intranasally, transdermally, or by some form of oral administration, for example, using mouthwash or lozenges that incorporate the polypeptide of the present disclosure that is poorly absorbed in the gastrointestinal tract.For administration that can result in local or topical delivery of the formulation of the present invention, iontophoresis or topical administration can be used.

[0266] In some embodiments, the ophthalmic formulation comprises from about 0.001 μM to about 100 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 0.01 μM to about 20 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 0.1 μM to about 5 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 0.2 μM to about 3 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 0.1 μM to about 10 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 1 μM to about 5 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 1 μM to about 10 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 5 μM to about 10 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 10 μM to about 50 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 20 μM to about 50 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 5 μM to about 50 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 1 μM to about 50 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 1 μM to about 20 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 5 μM to about 20 μM of the polypeptide of the present disclosure. In some embodiments, the ophthalmic formulation comprises from about 10 μM to about 20 μM of the polypeptide of the present disclosure.In some embodiments, the ophthalmic formulation comprises a polypeptide of the present disclosure at greater than about 0.001 μM, greater than 0.05 μM, greater than 0.1 μM, greater than 0.2 μM, greater than 0.3 μM, greater than 0.4 μM, greater than 0.5 μM, greater than 0.6 μM, greater than 0.7 μM, greater than 0.8 μM, greater than 0.9 μM, greater than 1 μM, greater than 1.1 μM, greater than 1.2 μM, greater than 1.3 μM. For example, greater than 1.4 μM, greater than 1.5 μM, greater than 1.6 μM, greater than 1.7 μM, greater than 1.8 μM, greater than 1.9 μM, greater than 2.0 μM, greater than 2.1 μM, greater than 2.2 μM, greater than 2.3 μM, greater than 2.4 μM, greater than 2.5 μM, greater than 2.6 μM, greater than 2.7 μM, greater than 2.8 μM, greater than 2.9 μM, or greater than 3.0 μM. In some embodiments, the ophthalmic formulation comprises a polypeptide of the present disclosure at less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, less than 25 μM, less than 20 μM, less than 19 μM, less than 18 μM, less than 17 μM, less than 16 μM, less than 15 μM, less than 14 μM, less than 13 μM, less than 12 μM, less than 11 μM, less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, or less than 4 μM.

[0267] In some embodiments, ophthalmic formulations can be formulated into dosage forms of about 0.01ml to about 10ml for single or multiple use.In some embodiments, ophthalmic formulations are formulated into unit dosage forms that provide a total daily dosage of about 0.01ml to about 2ml.In some embodiments, ophthalmic formulations can be formulated into unit dosage forms that provide a total weekly dosage of about 1ml to about 5ml.In some embodiments, ophthalmic formulations can be formulated into unit dosage forms that provide a total monthly dosage of about 1ml to about 20ml. DE treatment

[0268] Also provided herein are methods for treating or preventing DE or a DE-related disorder, stimulating tear fluid, stabilizing the tear film, or any combination thereof in a subject in need thereof, comprising administering to a subject in need thereof a therapeutically effective amount of a polypeptide of the disclosure, such as an artificial polypeptide of Formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of Formula (I), (II), (III), (IV) or (V), or a polypeptide having at least 70% identity (e.g., at least 80% identity) to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments, the polypeptide has at least 70% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments, the polypeptide has at least 75% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments, the polypeptide has at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments, the polypeptide has at least 85% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments, the polypeptide has at least 90% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof. In some embodiments, the polypeptide has at least 95% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof.In some embodiments, the polypeptide has at least 96%, at least 97%, at least 98%, or at least 99% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof. In some embodiments, the polypeptide comprises or is the amino acid sequence of any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof.

[0269] In some embodiments, a method is provided for treating or preventing DE or a DE-related disorder, stimulating tears, stabilizing the tear film, or any combination thereof in a subject in need thereof, comprising administering to a subject in need thereof a therapeutically effective amount of an ophthalmic formulation of the present disclosure.

[0270] In some embodiments, the DE or DE-related disorder is selected from the group consisting of dry eye syndrome, tear dysfunction syndrome, keratoconjunctivitis sicca (KCS), lacrimal keratoconjunctivitis, aqueous tear-deficient DE, evaporative DE, DE in Sjogren's syndrome, DE in non-Sjogren's syndrome, conjunctivitis-associated DE, post-viral conjunctivitis DE, post-cataract surgery DE, VDT work-associated DE, contact lens wear-associated DE, environmental DE, corneal neovascularization DE, allergic DE, LASIK-induced neuroepitheliopathy, aqueous tear deficiency, xerophthalmia, Stevens-Johnson syndrome, ocular pemphigoid, blepharitis marginalis, eyelid insufficiency, corneal ulcer, blepharitis, and ocular congestion.

[0271] Age is another factor that may affect tear composition and / or tear film stability. In some embodiments, the subject of the present disclosure is older than 40 years old. In some embodiments, the subject of the present disclosure is older than 50 years old. In some embodiments, the subject of the present disclosure is older than 60 years old. In some embodiments, the subject of the present disclosure is older than 70 years old, older than 75 years old, older than 80 years old, older than 85 years old, or older than 90 years old.

[0272] In some embodiments, the treatments of the present disclosure can be used to treat or prevent aqueous tear-deficient DE, which is the inability of the lacrimal gland to produce sufficient water-rich components of tears to maintain a healthy ocular surface. In some embodiments, the treatments of the present disclosure can be used to stimulate tears, increase tear volume, regulate tear composition, improve tear clearance and / or osmolality, or any combination thereof. In some embodiments, the methods of the present disclosure can be used to improve tear film stability or tear film composition. In some embodiments, the treatments of the present disclosure can be used to improve ocular surface damage.

[0273] In some embodiments, treatment of the present disclosure results in partial or complete elimination of DE or a DE-related disorder, tear stimulation, tear film stabilization, or any combination thereof in a subject. In some embodiments, after administration of a therapeutically effective amount of a polypeptide of the present disclosure to a subject, the severity or duration of one or more signs or symptoms of DE or a DE-related disorder is reduced by about 2%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%. In some embodiments, after administration of a therapeutically effective amount of a polypeptide of the present disclosure to a subject, the severity or duration of one or more signs or symptoms of DE or a DE-associated disorder is reduced by about 2-fold, about 5-fold, about 10-fold, about 15-fold, about 20-fold, about 25-fold, about 30-fold, about 35-fold, about 40-fold, about 45-fold, about 50-fold, about 55-fold, about 60-fold, about 65-fold, about 70-fold, about 75-fold, about 80-fold, about 85-fold, about 90-fold, about 95-fold, about 100-fold, or more.

[0274] In some embodiments, the TBUT before treatment of one or both eyes of the subject of the present disclosure is equal to or less than 10 seconds. In some embodiments, the TBUT before treatment of one or both eyes of the subject of the present disclosure is equal to or less than 9 seconds. In some embodiments, the TBUT before treatment of one or both eyes of the subject of the present disclosure is equal to or less than 8 seconds. In some embodiments, the TBUT before treatment of one or both eyes of the subject of the present disclosure is equal to or less than 7 seconds. In some embodiments, the TBUT before treatment of one or both eyes of the subject of the present disclosure is equal to or less than 6 seconds. In some embodiments, the TBUT before treatment of one or both eyes of the subject of the present disclosure is equal to or less than 5 seconds. In some embodiments, the TBUT before treatment of one or both eyes of the subject of the present disclosure is equal to or less than 4 seconds. In some embodiments, the TBUT before treatment of one or both eyes of the subject of the present disclosure is equal to or less than 3 seconds. In some embodiments, the subject of the present disclosure has a pre-treatment TBUT of one or both eyes of less than 2 seconds. In some embodiments, the subject of the present disclosure has a pre-treatment TBUT of one or both eyes of less than 1 second.

[0275] In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the TBUT of one eye of the subject is prolonged. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the TBUT of both eyes of the subject is prolonged. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the TBUT of one or both eyes of the subject is prolonged by at least 0.5 seconds. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the TBUT of one or both eyes of the subject is prolonged by at least 1 second. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the TBUT of one or both eyes of the subject is prolonged by at least 2 seconds. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the TBUT of one or both eyes of the subject is prolonged by at least 3 seconds. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the TBUT of one or both eyes of the subject is prolonged by at least 4 seconds. In some embodiments, after administering a therapeutically effective amount of a polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 5 seconds. In some embodiments, after administering a therapeutically effective amount of a polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 6 seconds. In some embodiments, after administering a therapeutically effective amount of a polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 7 seconds. In some embodiments, after administering a therapeutically effective amount of a polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 8 seconds. In some embodiments, after administering a therapeutically effective amount of a polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 9 seconds. In some embodiments, after administering a therapeutically effective amount of a polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 10 seconds. In some embodiments, after administering a therapeutically effective amount of a polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 15 seconds.In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 20 seconds.In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 25 seconds.In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to a subject, the TBUT of one or both eyes of the subject is prolonged by at least 30 seconds.

[0276] In some embodiments, a subject of the present disclosure has a pre-treatment corneal fluorescein staining (CFS) score of one or both eyes equal to or higher than 5. In some embodiments, a subject of the present disclosure has a pre-treatment CFS score of one or both eyes equal to or higher than 6. In some embodiments, a subject of the present disclosure has a pre-treatment CFS score of one or both eyes equal to or higher than 7. In some embodiments, a subject of the present disclosure has a pre-treatment CFS score of one or both eyes equal to or higher than 8. In some embodiments, a subject of the present disclosure has a pre-treatment CFS score of one or both eyes equal to or higher than 9. In some embodiments, a subject of the present disclosure has a pre-treatment CFS score of one or both eyes equal to or higher than 10.

[0277] In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the CFS score of one eye of the subject is reduced. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the CFS score of both eyes of the subject is reduced. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the CFS score of one or both eyes of the subject is reduced by at least 0.5. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the CFS score of one or both eyes of the subject is reduced by at least 1. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the CFS score of one or both eyes of the subject is reduced by at least 2. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the CFS score of one or both eyes of the subject is reduced by at least 3. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to the subject, the CFS score of one or both eyes of the subject is reduced by at least 4. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to a subject, the CFS score of one or both eyes of the subject is reduced by at least 5. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to a subject, the CFS score of one or both eyes of the subject is reduced by at least 6. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to a subject, the CFS score of one or both eyes of the subject is reduced by at least 7. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to a subject, the CFS score of one or both eyes of the subject is reduced by at least 8. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to a subject, the CFS score of one or both eyes of the subject is reduced by at least 9. In some embodiments, after administering a therapeutically effective amount of the polypeptide of the present disclosure to a subject, the CFS score of one or both eyes of the subject is reduced by at least 10.

[0278] In some embodiments, the treatment comprises administering a therapeutically effective amount of the polypeptide of the present disclosure to one affected eye or ocular tissue of the subject.In some embodiments, the method comprises administering a therapeutically effective amount of the polypeptide of the present disclosure to both eyes or ocular tissue of the subject.In some embodiments, the therapeutically effective amount of the polypeptide of the present disclosure is about 0.1 μg to about 100 μg per eye, or about 0.1 μg to about 50 μg per eye, or about 0.1 μg to about 20 μg per eye, or about 0.1 μg to about 10 μg per eye, or about 0.5 μg to about 50 μg per eye, or about 0.5 μg to about 20 μg per eye, or about 0.5 μg to about 10 μg per eye, or about 1 μg to about 10 μg per eye.

[0279] In some embodiments, the dosage for each eye of the subject may be about 1 to about 5 drops of the ophthalmic formulation of the present disclosure. In some embodiments, the dosage for each eye of the subject may be 1 drop of the ophthalmic formulation of the present disclosure. In some embodiments, the dosage for each eye of the subject may be 2 drops of the ophthalmic formulation of the present disclosure. In some embodiments, the dosage for each eye of the subject may be 3 drops of the ophthalmic formulation of the present disclosure. In some embodiments, the dosage for each eye of the subject may be 4 drops of the ophthalmic formulation of the present disclosure. In some embodiments, the dosage for each eye of the subject may be 5 drops of the ophthalmic formulation of the present disclosure. In some embodiments, each drop corresponds to about 10 μL to about 150 μL. In some embodiments, each drop corresponds to about 20 μL to about 70 μL.

[0280] In some embodiments, the method comprises administering a therapeutically effective amount of the ophthalmic formulation to each eye of the subject once a day. In some embodiments, the method comprises administering a therapeutically effective amount of the ophthalmic formulation to each eye of the subject twice a day. In some embodiments, the method comprises administering a therapeutically effective amount of the ophthalmic formulation to each eye of the subject three or more times a day. In some embodiments, the method comprises administering a therapeutically effective amount of the ophthalmic formulation to each eye of the subject once every two days. In some embodiments, the method comprises administering a therapeutically effective amount of the ophthalmic formulation to each eye of the subject once every three days. In some embodiments, the method comprises administering a therapeutically effective amount of the ophthalmic formulation to each eye of the subject once a week.

[0281] In some embodiments, the method comprises administering one or more drops of the ophthalmic formulation of the present disclosure to each eye of the subject daily. In some embodiments, the method comprises administering one to multiple drops of the ophthalmic formulation of the present disclosure to each eye of the subject 2, 3, 4, 8, 12, 18, or 24 times daily. In some embodiments, the method comprises administering one or multiple drops of the ophthalmic formulation of the present disclosure to each eye of the subject once every two days.

[0282] In some embodiments, the polypeptide of the present disclosure is administered in combination with one or more additional therapeutic agents. In some embodiments, a therapeutically effective amount of the polypeptide of the present disclosure can be administered simultaneously with the additional therapeutic agent. In some embodiments, a therapeutically effective amount of the polypeptide of the present disclosure can be administered prior to administration of the additional therapeutic agent. In some embodiments, a therapeutically effective amount of the polypeptide of the present disclosure can be administered after administration of the additional therapeutic agent. In some embodiments, the additional therapeutic agent includes, but is not limited to, artificial tears, antibiotics, antiviral agents, antifungal agents, antiprotozoal agents, anti-inflammatory agents, antiallergic agents, anesthetics, analgesics, intraocular pressure reducing agents, immunomodulators, antioxidants, enzyme inhibitors, proteases, peptidases, cytokine inhibitors, vitamins and minerals.

[0283] In some embodiments, a therapeutically effective amount of the polypeptide of the present disclosure is administered simultaneously or separately during a treatment period in which the subject is receiving immunosuppressive therapy, such as azathioprine, cyclophosphoramide, methotrexate, antimalarial drugs, mycophenolan mofetile, daclizumab, intravenous immunoglobulin therapy, etc. In some embodiments, a therapeutically effective amount of the polypeptide of the present disclosure is administered simultaneously or separately during a treatment period in which the subject is receiving other anti-inflammatory treatment, such as cyclosporine A, corticosteroids, NSAIDS1 aspirin, doxycycline, etc. In some embodiments, a therapeutically effective amount of the polypeptide of the present disclosure is administered simultaneously or separately during a treatment period in which the subject is receiving hormone therapy, etc. In some embodiments, a therapeutically effective amount of the polypeptide of the present disclosure is administered simultaneously or separately during a treatment period in which the subject is receiving anti-allergy therapy, palliative care for dry eye, such as artificial tears or artificial saliva, muscarinic M3 receptor agonists to increase water secretion, autologous serum, sodium hyaluronate eye drops, etc.

[0284] The present disclosure also contemplates an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), or a variant of an artificial polypeptide of (I), (II), (III), (IV) or (V), for use in therapy. In some embodiments, the present disclosure also contemplates an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), or a variant of an artificial polypeptide of (I), (II), (III), (IV) or (V), for use in treating or preventing dry eye (DE) or a DE-related disorder in a subject in need thereof, stimulating tear fluid, stabilizing the tear film, or any combination thereof. In some embodiments, the disclosure also contemplates a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NO:62-74, or a fragment or variant thereof, for use in treating or preventing dry eye (DE) or a DE-related disorder in a subject in need thereof, stimulating tear fluid, stabilizing the tear film, or any combination thereof. In some embodiments, the DE or DE-related disorder is selected from the group consisting of dry eye syndrome, tear dysfunction syndrome, keratoconjunctivitis sicca (KCS), lacrimal keratoconjunctivitis, aqueous tear-deficient DE, evaporative DE, DE in Sjogren's syndrome, DE in non-Sjogren's syndrome, conjunctivitis-associated DE, post-viral conjunctivitis DE, post-cataract surgery DE, VDT work-associated DE, contact lens wear-associated DE, environmental DE, corneal neovascularization DE, allergic DE, LASIK-induced neuroepitheliopathy, aqueous tear deficiency, xerophthalmia, Stevens-Johnson syndrome, ocular pemphigoid, blepharitis marginalis, eyelid insufficiency, corneal ulcer, blepharitis, and ocular congestion.

[0285] The disclosure also contemplates the use of an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of (I), (II), (III), (IV) or (V), or a polypeptide having at least 80% identity to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof, for preparing a medicament for treating or preventing dry eye (DE) or a DE-related disorder in a subject in need thereof, for stimulating tear fluid, for stabilizing the tear film, or any combination thereof. In some embodiments, the DE or DE-related disorder is selected from the group consisting of dry eye syndrome, tear dysfunction syndrome, keratoconjunctivitis sicca (KCS), lacrimal keratoconjunctivitis, aqueous tear-deficient DE, evaporative DE, DE in Sjogren's syndrome, DE in non-Sjogren's syndrome, conjunctivitis-associated DE, post-viral conjunctivitis DE, post-cataract surgery DE, VDT work-associated DE, contact lens wear-associated DE, environmental DE, corneal neovascularization DE, allergic DE, LASIK-induced neuroepitheliopathy, aqueous tear deficiency, xerophthalmia, Stevens-Johnson syndrome, ocular pemphigoid, blepharitis marginalis, eyelid insufficiency, corneal ulcer, blepharitis, and ocular congestion. kit

[0286] The present disclosure also provides kits for preventing or treating DE or DE-related disorders, stimulating tears, stabilizing the tear film, or any combination thereof. In some embodiments, the kits include one or more polypeptides of the present disclosure and instructions for use of the kit. In some embodiments, the kits include one or more ophthalmic formulations described herein and instructions for use of the kit.

[0287] The kit may include one or more containers containing one or more ophthalmic formulations or polypeptides of the present disclosure.The polypeptides of the present disclosure may be present in a container as a prepared pharmaceutical composition, or the polypeptides of the present disclosure may not be formulated.In some embodiments, the kit may include the polypeptides of the present disclosure that are not formulated in a container separate from pharmaceutically acceptable excipients.Before use, the polypeptides of the present disclosure are diluted or otherwise mixed with pharmaceutically acceptable excipients.

[0288] In some embodiments, the kits provided herein also include instructions describing how to administer the ophthalmic formulation to treat or prevent one or more symptoms associated with DE or a DE-related disorder, In some embodiments, the instructions also describe how to mix the polypeptide of the present disclosure contained in the kit with a pharma- ceutically acceptable excipient.

[0289] In some embodiments, the container is configured to deliver the polypeptide of the present disclosure or the ophthalmic formulation of the present disclosure. In some embodiments, the container includes a vial, an eye dropper, a bottle, a tube, and a syringe. In certain embodiments, the container is an eye dropper for administering eye drops. In other embodiments, the container is a tube for administering an ophthalmic gel or an ophthalmic ointment. Contact Lens Care Products

[0290] According to the pathogenesis of dry eye (DE), wearing contact lenses can be considered as one of the causes of dry eye. Specifically, wearing contact lenses (contact lens wear, CL wear) is one of the factors that cause instability of the tear film. Other factors that cause instability of the tear film include vitamin A deficiency, ocular allergy, topical use of preservatives, and deficiency or instability of the tear film lipid layer. Wearing contact lenses can also cause a decrease in tear secretion by blocking the ocular reflex, resulting in hypertonicity of the tears.

[0291] Thus, in another aspect, also provided herein are contact lens care products that may be useful in preventing or reducing contact lens wear-related DE. In some embodiments, the contact lens care products comprise a polypeptide of the present disclosure, such as an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of formula (I), (II), (III), (IV) or (V), or a polypeptide having at least 70% identity (e.g., at least 80% identity) to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof. In some embodiments, the contact lens care product is a solution for storing, protecting, and / or cleaning contact lenses.

[0292] In some embodiments, the contact lens care product further comprises one or more acceptable ingredients for contact lens care products. In some embodiments, the one or more acceptable ingredients for contact lens care products are selected from the group consisting of inorganic salts, boric acid, surfactants, humectants, preservatives, and solvents. In some embodiments, the contact lens care product further comprises inorganic salts. In some embodiments, the contact lens care product further comprises boric acid. In some embodiments, the contact lens care product further comprises a surfactant. In some embodiments, the contact lens care product further comprises a humectant. In some embodiments, the contact lens care product further comprises a preservative. In some embodiments, the contact lens care product further comprises a solvent. In some embodiments, the contact lens care product further comprises inorganic salts and a solvent. In some embodiments, the contact lens care product further comprises inorganic salts, boric acid, and a solvent. In some embodiments, the contact lens care product further comprises inorganic salts, boric acid, surfactants, and solvents. In some embodiments, the contact lens care product further comprises inorganic salts, boric acid, humectants, and solvents. In some embodiments, the contact lens care product further comprises inorganic salts, boric acid, surfactants, and solvents. In some embodiments, the contact lens care product further comprises inorganic salts, boric acid, humectants, and solvents. In some embodiments, the contact lens care product further comprises inorganic salts, boric acid, preservatives, and solvents. In some embodiments, the contact lens care product further comprises inorganic salts, boric acid, surfactants, moisturizers and solvents. In some embodiments, the contact lens care product further comprises inorganic salts, boric acid, surfactants, preservatives and solvents. In some embodiments, the contact lens care product further comprises inorganic salts, boric acid, surfactants, moisturizers, preservatives and solvents. In some embodiments, the contact lens care product consists of the polypeptide of the present disclosure, inorganic salts, boric acid, surfactants, moisturizers, preservatives and solvents.

[0293] In some embodiments, the inorganic salt comprises sodium chloride, potassium chloride, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium carbonate, sodium borate, or any combination thereof. In some embodiments, the inorganic salt comprises sodium chloride. In some embodiments, the inorganic salt comprises potassium chloride. In some embodiments, the inorganic salt comprises sodium dihydrogen phosphate, disodium hydrogen phosphate, or a combination thereof. In some embodiments, the inorganic salt comprises sodium carbonate. In some embodiments, the inorganic salt comprises sodium borate. In some embodiments, the inorganic salt comprises sodium chloride, sodium borate, or both. In some embodiments, the inorganic salt comprises sodium chloride and sodium borate. In some embodiments, the inorganic salt consists of sodium chloride and sodium borate.

[0294] In some embodiments, the surfactant comprises a zwitterionic surfactant, a non-ionic surfactant, or both. In some embodiments, the surfactant comprises a zwitterionic surfactant. In some embodiments, the surfactant comprises a non-ionic surfactant. In some embodiments, the surfactant comprises a zwitterionic surfactant and a non-ionic surfactant. Examples of zwitterionic surfactants include, but are not limited to, lecithin. Examples of non-ionic surfactants include, but are not limited to, sorbitan fatty acid esters (Spans), polyoxyethylene sorbitan fatty acid esters (Tweens), polyoxyethylene fatty acid esters (Myri), polyoxyethylene fatty alcohol ethers (Brij®), and polyethylene-polypropylene glycols (e.g., poloxamers).

[0295] In some embodiments, the moisturizer comprises hyaluronic acid or a salt thereof. In some embodiments, the moisturizer comprises hyaluronic acid. In some embodiments, the moisturizer comprises a salt of hyaluronic acid, such as sodium hyaluronate. In some embodiments, the moisturizer comprises hyaluronic acid and a salt of hyaluronic acid, such as sodium hyaluronate. In some embodiments, the moisturizer consists of hyaluronic acid and a salt of hyaluronic acid, such as sodium hyaluronate.

[0296] In some embodiments, the preservative comprises an organic mercury compound, a quaternary ammonium salt, an alcohol, an ester, an acid, or any combination thereof. In some embodiments, the preservative comprises an organic mercury compound. In some embodiments, the preservative comprises a quaternary ammonium salt. In some embodiments, the preservative comprises an alcohol. In some embodiments, the preservative comprises an ester. In some embodiments, the preservative comprises an acid. In some embodiments, the preservative comprises at least two of an organic mercury compound, a quaternary ammonium salt, an alcohol, an ester, and an acid. Examples of organic mercury compounds include, but are not limited to, phenylmercuric nitrate, phenylmercuric acetate, thimerosal, mercuric oxycyanide, and the like. Examples of quaternary ammonium salts include, but are not limited to, benzalkonium chloride, benzalkonium bromide, chlorhexidine, and the like. Examples of alcohols include, but are not limited to, trichlorobutanol, phenethyl alcohol, and the like. Examples of esters include, but are not limited to, methylparaben, ethylparaben, propylparaben, and the like. Examples of acids include, but are not limited to, sorbic acid.

[0297] In some embodiments, the solvent comprises water, hi some embodiments, the solvent consists of water.

[0298] In another aspect, also provided herein is a method for preparing a contact lens care product, the method comprising combining a polypeptide of the present disclosure, e.g., an artificial polypeptide of Formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of Formula (I), (II), (III), (IV) or (V), or a polypeptide having at least 70% identity (e.g., at least 80% identity) to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof, with one or more acceptable ingredients for a contact lens care product.

[0299] In another aspect, also provided herein is the use of a polypeptide of the present disclosure, e.g., an artificial polypeptide of formula (I), (II), (III), (IV), (V) or (VI), a variant of an artificial polypeptide of formula (I), (II), (III), (IV) or (V), or a polypeptide having at least 70% identity (e.g., at least 80% identity) to any one selected from SEQ ID NO:28, SEQ ID NO:57, and SEQ ID NOs:62-74, or a fragment or variant thereof, for preparing a contact lens care product.

[0300] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. The present invention is not limited by the specific examples provided herein. Although the present invention has been described in conjunction with the above specification, the description and illustration of the embodiments herein are not intended to be construed in a limiting sense. Numerous modifications, changes, and substitutions will readily occur to those skilled in the art without departing from the present invention. Furthermore, it should be understood that all aspects of the present invention are not limited to the specific depictions, configurations, or relative proportions set forth herein, which depend upon a variety of conditions and variables. It should be understood that various alternatives to the embodiments of the invention described herein can be used in the practice of the present invention. It is therefore intended that the present invention encompass any such alternatives, modifications, variations, or equivalents. It is intended that the scope of the present invention be defined by the following claims, and that methods and structures falling within the scope of these claims, and their equivalents, are therefore covered. EXAMPLES

[0301] Example 1 Efficacy of the polypeptides of the present disclosure in treating mild DE The efficacy of an exemplary polypeptide of the present disclosure (SEQ ID NO:29, hereinafter also referred to as "CF-04") for the treatment of DE was evaluated in Rhesus macaques as follows: Briefly, six rhesus monkeys (five females and one male, weighing 9-13 kg), aged 12-23 years (equivalent to a human age of 40-70 years), that met the following criteria for mild DE in either of two eyes within six months were enrolled in the study: Corneal Fluorescein Staining (CFS) score: 5–10; or · Tear film breakup time (TBUT): <5 seconds.

[0302] After enrollment, the monkeys were divided into two groups, two in the placebo group and four in the treatment group. Prior to treatment, the animals underwent baseline examinations on days -13 and -5. The monkeys then received placebo (saline) or CF-04 (10 μM, dissolved in saline) eye drops (2 drops per eye, twice per day) from day 0 to day 13 for 14 consecutive days, followed by an 18-day washout period. The monkeys' clinical observations and food intake were also observed from day -13 to day 14.

[0303] 1. Corneal fluorescein staining score

[0304] Corneal fluorescein staining (CFS) scores are indicative of ocular surface damage and are used to assess DE. CFS scores were assessed for each eye of monkeys on days -5 and 14 by using 2% sodium fluorescein solution. Each of the corneal C, S, T, I, and N regions was scored, and the CFS score of the eye was then calculated as the sum of the scores of the five regions. Table 1 shows the CFS scores for both placebo and treatment monkeys. [Table 1]

[0305] From Table 1, it can be seen that in the CF-04 group, the administration of the polypeptide of the present disclosure reduces the CFS score of eyes by at least 3 points (OD of No.3, OD and OS of No.4, OS of No.5 and OD of No.6).After the washout period, the CFS score of eyes of the CF-04 group increases to the level before treatment, which shows the effectiveness of CF-04 throughout the treatment period.However, the CFS score of eyes of DE of the placebo group does not show a significant reduction (OD of No.1 and OD of No.2).

[0306] FIG. 1 also illustrates the effect of the polypeptide of the present disclosure on CFS scores according to the data in Table 1. As shown in FIG. 1, the average CFS scores for all tested eyes of the placebo group at day 14 (red dots) showed no significant change compared to baseline at day -5 (blue dots) (p=0.4287). However, the average CFS scores of the CF-04 group were significantly decreased at day 14 (red dots) compared to day -5 (blue dots) (5.00 to 2.14, p=0.0016), indicating that administration of the polypeptide of the present disclosure improved ocular surface damage and reduced symptoms of DE. After the washout period, the CFS scores of the eyes of the CF-04 group increased to pre-treatment levels, further suggesting the efficacy of CF-04 throughout the treatment period.

[0307] In contrast, administration of CF-04 did not affect food intake and other clinical findings of the animals (data not shown), demonstrating the safety of the polypeptides of the present disclosure.

[0308] 2. Tear film breakup time (TBUT)

[0309] Tear breakup time (TBUT) is an indicator of tear film stability and is widely used to evaluate DE. In this study, TBUT was evaluated on days -13, -5, and 14, respectively (three tests each day). The results are shown in Table 2. [Table 2]

[0310] As can be seen from Table 2, in the CF-04 group, administration of the polypeptide of the present disclosure prolonged the TBUT of the DE eyes (OD of No. 3, OD and OS of No. 5, and OS of No. 6) to more than 5 seconds. However, in the control group, placebo could not prolong the TBUT of the DE eyes (OS of No. 1, OD and OS of No. 2).

[0311] FIG. 2 also illustrates the effect of the polypeptide of the present disclosure on TBUT according to the data in Table 2. As shown in FIG. 2, in the placebo group, TBUT in all tested eyes was not significantly different from baseline 2 (day -5) on day 14 (p=0.8804). For the CF-04 group, TBUT in all tested eyes was not significantly different compared to baseline 2 on day 14 (p=0.2596), but the mean TBUT was prolonged by more than 1 second (5.143±2.183 vs. 6.671±2.630), suggesting that administration of the polypeptide of the present disclosure has the potential to stabilize the tear film and reduce symptoms of DE. After the washout period, the CFS scores of the eyes in the CF-04 group increased to pre-treatment levels, indicating the efficacy of CF-04 throughout the treatment period. Example 2 Effect of the polypeptides of the present disclosure on the treatment of DE

[0312] The efficacy of an exemplary polypeptide of the present disclosure (SEQ ID NO:29, hereinafter also referred to as CF-04) for the treatment of DE was evaluated in rhesus monkeys as follows: Briefly, 22 monkeys aged 12-24 years (equivalent to a human age of 40-70 years) that met the following criteria for DE in either of two eyes within the last six months were enrolled in the study:

[0313] Corneal fluorescein staining (CFS) score: 5–10; and

[0314] Tear film breakup time (TBUT): <5 seconds.

[0315] The monkeys were divided into six groups, including placebo (saline, 2 monkeys), positive control (Restasis, 4 monkeys), CF-04 0.46 μg / ml (4 monkeys), CF-04 2.3 μg / ml (4 monkeys), CF-04 4.6 μg / ml (4 monkeys), and CF-04 50 μg / ml (4 monkeys). Briefly, animals were treated with placebo or CF-04, 2 drops per eye, twice daily, and for the positive control group, animals were treated with Restasis, 1 drop per eye, twice daily, for 28 or 30 consecutive days. Clinical observations and food intake of the monkeys were also observed at baseline, day 14, and at the end of treatment (days 28 or 30).

[0316] 1. Corneal fluorescein staining score

[0317] CFS scores were assessed for each eye of the monkeys at baseline, day 14, and at the end of treatment (day 28 or 30) using a 2% sodium fluorescein solution. The cornea was scored in the C, S, T, I, and N regions, and the CFS score for the eye was then calculated as the sum of the scores for the five regions. Table 3 shows the CFS scores for both the placebo and treatment monkeys. [Table 3-1] [Table 3-2] [Table 3-3]

[0318] Scores were collected and analyzed by ANOVA. Figures 3A-3B illustrate the effect of the polypeptides of the present disclosure on CFS scores according to the data in Table 3. As shown in Figure 3A, the mean CFS scores of the placebo group showed no significant changes compared to baseline on days 14 and 28. The CFS scores of animals treated with 0.46 μg / ml CF-04 showed no significant changes compared to baseline on days 14 (Δ=3.214, p=0.0987) or 28 (Δ=2.893, p=0.1566). Treatment with 2.3 μg / ml CF-04 significantly reduced CFS scores on day 14 (Δ=4.000, p=0.0226) and further reduced scores on day 28 (Δ=4.750, p=0.0049) compared to baseline. Treatment with CF-04 at 4.6 μg / ml showed no significant effect on CFS scores compared to baseline at day 14 (Δ=3.125, p=0.1012), but significantly reduced CFS scores at day 28 (Δ=4.000, p=0.0226). Treatment with CF-04 at 50 μg / ml significantly reduced CFS scores compared to baseline at day 14, but no further reduction in CFS scores was observed at day 28 (Δ=4.000, p=0.0226).

[0319] Figure 3B further illustrates the CFS scores collected from animals in different groups. As can be seen from Figure 3B, on days 14 and 28, treatment with 2.3 μg / ml CF-04 significantly reduced the CFS scores of the animals' eyes compared to the CFS scores of the placebo group's eyes.

[0320] 2. Tear film breakup time (TBUT)

[0321] TBUT for animals in each group was assessed at baseline, day 14, and day 28, as shown in Table 4. [Table 4-1] [Table 4-2] [Table 4-3]

[0322] The data in Table 4 was further analyzed by ANOVA, as shown in Figures 4A-4B. As can be seen from Figures 4A-4B, animals treated with CF-04 at 2.3 μg / ml and CF-04 at 4.6 μg / ml had significantly longer TBUT compared to placebo and comparable to the positive control Restasis, demonstrating the efficacy of CF-04 at 2.3 μg / ml and CF-04 at 4.6 μg / ml in treating DE. Example 3 Efficacy of exemplary polypeptides of the present disclosure in treating acute DE

[0323] The effect of an exemplary polypeptide of the present disclosure (SEQ ID NO:29, CF-04) on the treatment of DE was evaluated in rhesus monkeys as follows. Briefly, healthy adult monkeys were placed in a dry eye chamber for 3 weeks to induce acute severe dry eye symptoms by a dry environment. CFS and TUBT were examined on day 21, and tears were assayed for the inflammatory factor IL-17a on day 23. Animals with a single eye CFS score of 8-13 were then enrolled in the study.

[0324] The enrolled animals were randomly divided into a placebo group (saline, 3 monkeys, 6 eyes) and a CF-04 group (CF-04 at 5 μg / mL, 4 animals, 8 eyes). Both eyes of the animals were treated with 2 drops per eye, twice daily. CFS scores and TUBT were examined on days 7 and 14 after treatment. IL-17a was assayed on day 16 after treatment.

[0325] 1. Corneal fluorescein staining score

[0326] CFS scores were first assessed at baseline (day -2 of treatment). The mean CFS scores in the control and CF-04 groups were 9.0±0.6 and 9.6±0.9, respectively. There was no significant difference in the mean CFS scores between the two groups (p=0.4719). After 7 days of treatment, the mean CFS scores of animals in the control and CF-04 groups were 8.7±1.4 and 5.9±1.8, respectively. The mean CFS of the CF-04 group was significantly lower than the control group (p=0.0194). After 14 days of treatment, the mean CFS scores of animals in the control and CF-04 groups were 9.0±1.3 and 7.0±2.2, respectively, with no significant difference. The CFS scores of each animal are shown in Table 5 below. [Table 5]

[0327] 2. Tear film breakup time (TBUT)

[0328] TUBT was first assessed at baseline (day -2 of treatment). The mean TUBT in the control and CF-04 groups was 4.0±0.8 and 3.8±0.6 seconds, respectively. There was no significant difference in the mean TUBT between the two groups (p>0.9999). After 7 days of treatment, the mean TUBT of animals in the control and CF-04 groups was 3.5±0.6 and 4.9±0.6 seconds, respectively. The mean TUBT of the CF-04 group was significantly longer than the control group (p=0.0042). After 14 days of treatment, the mean TUBT of animals in the control and CF-04 groups was 3.6±0.5 and 4.6±0.9 seconds, respectively, with no significant difference. The TUBT of each animal is shown in Table 6 below. [Table 6] Example 4 Effect of exemplary polypeptides of the present disclosure on inhibiting thermal aggregation of BSA.

[0329] Exemplary polypeptides of the present application shown in Table 7 below were dissolved in PBS and serially diluted three times. 150 uL of the polypeptide was then mixed with 150 μL of 200 uM BSA, and the mixture was incubated in a 98° C. water bath for 30 minutes. 200 μL of the mixture was added to a detection plate and assayed for absorbance at a wavelength of 600 nM. The inhibition rate was calculated as follows: 100-[(OD 試料 -OD PBS ) / (OD BSA -OD PBS )]*100 In the formula, O.D. BSA is the absorbance of the sample containing only BSA, and OD PBS is the absorbance of the sample containing only PBS. IC 50 was calculated and analyzed by Graphpad using log-log concentration. The results are shown in Figure 5 and Table 7 below. [Table 7] Example 5 Effect of the polypeptides of the present disclosure on intracellular ROS levels induced by DAMPs

[0330] 90μL of HUVEC cells were seeded at 10000 cells / well in a 96-well plate and left overnight. One hour before detection, ROS reagent (Thermo, Cat#88-5930-74) was added to the cell culture plate at 90μl / well as a working solution. Meanwhile, the exemplary polypeptides of the present application shown in Table 8 below were serially diluted in three times with cell culture medium. 60μL of polypeptides at various concentrations were mixed with 60μL of 400ug / ml oxHSA. Then, 20μl of the mixture of polypeptides and oxHSA was transferred to the cell culture and incubated with the cells for 40 minutes. Fluorescence was detected at excitation / emission wavelengths of 490nm / 525nm. The inhibition rate was calculated as follows: 100-[(L 試料 -L PBS / 培地 ) / (L oxHSA -L PBS / 培地 )]*100 The results are shown in Table 8 below. [Table 8] Example 6 Effect of polypeptides of the present disclosure on inhibiting oxLDL uptake.

[0331] THP-1 cells (ATCC, 30-2001) were resuspended in RPMI-1640 complete medium (10% fetal bovine serum, 1% bispecific antibody, 0.05 mM β-mercaptoethanol) containing 100 nM PMA (Sigma-Aldrich, P1585) and counted. Then, 30,000 cells were seeded into each well of a 96-well plate and cultured at 37°C for 72 hours. Then, the medium was replaced with RPMI-1640 complete medium without PMA, and the cells were cultured for another 48 hours to obtain THP-1-induced differentiated macrophages. The exemplary polypeptides of the present application were serially diluted in RPMI-1640 complete medium. Then, each polypeptide was mixed with oxLDL-Dylight488 solution (Cayman Chemical, 601180). The cell culture medium was replaced with the mixture of polypeptide and oxLDL-Dylight488, and the cells were cultured at 37°C for 5-6 h. After incubation, the cells in each well were rinsed and washed with PBS, resuspended in 200 μL of PBS supplemented with 7-AAD, collected through a filter into a FACS tube, and analyzed by flow cytometry (BD, FACSCelesta). 7-AAD in the Dylight488 channel - The geometric mean fluorescence intensity (gMFI) of the cells was analyzed. The oxLDL uptake rate was calculated as follows: oxLDL uptake rate = (gMFI of experimental group - gMFI of negative control group) / (gMFI of positive control group - MeanFI of negative control group) × 100% Here, the positive group contains only oxLDL, and the negative group contains neither oxLDL nor the polypeptide of the present application. The results are shown in FIG. Example 7 Binding ability of exemplary polypeptides of the present disclosure to oxidized proteins.

[0332] 1mg / ml HSA protein was oxidized with 8mM NaClO for 30min in the dark. The reactants were added to a 3KD dialysis bag and dialyzed in PBS at 4℃ for 24hrs, with the fluid replaced with fresh dialysate 2-3 times during dialysis. After filter sterilization, the proteins were quantified using the BCA method. The oxidized proteins were mixed with the target protein in a mass ratio of 1:2 and incubated at 4℃ for 24hrs. After the reaction, the samples were subjected to SDS-Page electrophoresis and photographed after Coomassie brilliant blue staining. [Table 9] Example 8 Cell binding ability of the polypeptides of the present disclosure.

[0333] Peripheral blood mononuclear cells (PBMCs, SAILY BIO) were collected by centrifugation at 400 g at room temperature, resuspended in RPMI-1640 complete medium (10% fetal bovine serum, 1% bispecific antibody, 0.05 mM β-mercaptoethanol) and counted. 6 Cells were seeded into each well of a 96-well plate, and each was supplemented with the CF polypeptide of the present application labeled with FITC (Sigma-Aldrich, FITC1). The concentrations of CF polypeptide labeled with FITC in the cell culture were 0 nM, 30 nM, 100 nM, 300 nM, and 1000 nM, respectively. The cells were then cultured at 37°C for 30 minutes. After incubation, the cells in each well were washed with PBS and resuspended in 100 μL of PBS supplemented with BV421-CD16 fluorescent antibody (BD, 3G8) and BV605-CD14 (BD, M5E2) fluorescent antibodies. The cells were then incubated in the dark at 4°C for 30 minutes. After incubation, the cells were washed with PBS, resuspended in 200 μL of PBS, collected through a filter into a FACS tube, and analyzed by flow cytometry (BD, FACSCelesta). CD16 in each PBMC sample in the FITC channel - CD14 + The fluorescent signal of monocytes was analyzed. As shown in Figure 7, the CD16 -CD14 + CF + The monocyte ratio was calculated. Example 9 Pharmacodynamic study of polypeptides against dry eye in mice induced by low humidity environment and scopolamine hydrobromide.

[0334] Thirty-two mice (C57BL / 6JShjh) with similar tear production and corneal sodium fluorescein scores were divided into four groups (n=8): negative control, vehicle control, treatment group 1, and treatment group 2. All animals were subjected to 10%-30% humidity and 0.3 mL of scopolamine hydrobromide (Sigma-Aldrich, 0.75 mg / mL) was administered subcutaneously twice a day from day 1 (D1) to day 15 (D15) to induce dry eye. Animals in the negative group were not treated, whereas the other groups were treated twice a day (approximately 8 hours apart) from D6 to D15 with vehicle only, the polypeptide of SEQ ID NO:29, and the artificial polypeptide of SEQ ID NO:75, respectively, at a dose of 3 mL / eye. Tear secretion was measured by phenol red thread test in all animals on D-1, D5, D10 and D15, and tear secretion was measured as wet thread length (mm) (Figure 8, top panel).

[0335] Corneal sodium fluorescein scores were assessed for all animals on D-1, D5, D10 and D15 according to the following protocol: Sodium fluorescein solution (1.5 μL, 0.5%) was instilled into the superior conjunctival sac of the animals, followed by irrigation of the animals' conjunctival sac three consecutive times, 10 seconds each, with 1.25 mL of sterile saline. After approximately 5 minutes of staining, the ocular surface was observed and photographed using a slit lamp (+ cobalt blue filter), and scoring was performed in situ. The cornea of ​​each animal was scored using the improved NEI scale for grading fluorescein staining. The cornea was divided into five regions (1-central, 2-superior, 3-temporal, 4-nasal, 5-inferior) and each region was given a maximum score of 8 and a minimum score of 0 for no staining at all (grade 1: stained area is 1%-25% of the corresponding area; grade 2: stained area is 26%-50% of the corresponding area; grade 3: stained area is 51%-75% of the corresponding area; grade 4: stained area is 76%-100% of the corresponding area). For dense / confluent staining, additional points were given according to the area of ​​such dense / confluently stained area (1: 1%-25%; 2: 26%-50%; 3: 51%-75% and 4: 76%-100%). The maximum total score for each eye was 40 points. The total score of sodium fluorescein staining was calculated for each eye of all animals (Figure 8, lower panel).

[0336] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. The present invention is not limited by the specific examples provided herein. Although the present invention has been described in conjunction with the above specification, the description and illustration of the embodiments herein are not intended to be construed in a limiting sense. Numerous modifications, changes, and substitutions will readily occur to those skilled in the art without departing from the present invention. Furthermore, it should be understood that all aspects of the present invention are not limited to the specific depictions, configurations, or relative proportions set forth herein, which depend upon a variety of conditions and variables. It should be understood that various alternatives to the embodiments of the invention described herein can be used in the practice of the present invention. It is therefore intended that the present invention encompass any such alternatives, modifications, variations, or equivalents. It is intended that the scope of the present invention be defined by the following claims, and that methods and structures falling within the scope of these claims and their equivalents are therefore covered. [Table 10-1] [Table 10-2] [Table 10-3]

Claims

1. A pharmaceutical composition for treating the eye of a subject, said pharmaceutical composition comprising a compound of formula (I): X-Y (I) Artificial polypeptides (In the formula, X is a moiety comprising an amino acid sequence having at least 80% identity to SEQ ID NO: 1; Y is a moiety comprising 10 to 50 amino acids, at least 50% of which are selected from the group consisting of isoleucine, valine, leucine, phenylalanine, cysteine, methionine, and alanine; The total number of cysteines (C) contained in Y is less than 5.

2. Use of an artificial polypeptide in the manufacture of a medicament for treating an eye of a subject, wherein the artificial polypeptide has formula (I): X-Y (I) (In the formula, X is a moiety comprising an amino acid sequence having at least 80% identity to SEQ ID NO: 1; Y is a moiety comprising 10 to 50 amino acids, at least 50% of which are selected from the group consisting of isoleucine, valine, leucine, phenylalanine, cysteine, methionine, and alanine; The total number of cysteines (C) contained in Y is less than 5.

3. 3. The pharmaceutical composition of claim 1, or the use of claim 2, wherein X comprises a sequence having at least 90% identity to SEQ ID NO:

1.

4. 3. The pharmaceutical composition of claim 1, or the use of claim 2, wherein X comprises a sequence having at least 95% identity to SEQ ID NO:

1.

5. 3. The pharmaceutical composition of claim 1, or the use of claim 2, wherein X comprises the sequence of SEQ ID NO:

1.

6. 3. The pharmaceutical composition of claim 1, or the use of claim 2, wherein at least 50% of the amino acids of X are selected from the group consisting of arginine (R), lysine (K), asparagine (N), aspartic acid (D), glutamine (Q), glutamic acid (E), and histidine (H).

7. The pharmaceutical composition according to claim 1 or the use according to claim 2, wherein the total number of cysteines (C) contained in Y is less than 4.

8. 3. The pharmaceutical composition of claim 1, or the use of claim 2, wherein the total number of hydrophobic amino acids in Y is greater than 8.

9. 3. The pharmaceutical composition of claim 1, or the use of claim 2, wherein the total number of hydrophilic amino acids in Y is 5 or less.

10. The pharmaceutical composition of claim 1, or the use of claim 2, wherein X comprises an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 3-15, 29-41, 54-61, 84, 85, or 91-95.

11. The pharmaceutical composition of claim 1 or the use of claim 2, characterized in that the artificial polypeptide is administered to the eye at a concentration of about 0.1 μM to about 50 μM.

12. A pharmaceutical composition as described in claim 1 or a use as described in claim 2, wherein the artificial polypeptide stimulates tear fluid produced by the eye, stabilizes the tear film of the eye, or both.

13. The pharmaceutical composition of claim 1 or the use of claim 2, wherein the subject has an eye disease, disorder or condition.

14. The pharmaceutical composition of claim 13, or the use of claim 13, wherein the ocular disease, disorder or condition includes ocular discomfort, visual impairment, tear film instability, ocular surface damage, or a combination thereof.

15. The pharmaceutical composition of claim 1 or the use of claim 2, wherein the eye disease, disorder or condition comprises dry eye (DE) or a DE-related disorder.

16. The pharmaceutical composition of claim 15, or the use of claim 15, wherein the DE-related disorder is selected from the group consisting of dry eye syndrome, tear dysfunction syndrome, keratoconjunctivitis sicca (KCS), lacrimal keratoconjunctivitis, aqueous tear-deficient DE, evaporative DE, DE in Sjogren's syndrome, DE in non-Sjogren's syndrome, conjunctivitis-related DE, post-viral conjunctivitis DE, post-cataract surgery DE, VDT work-related DE, contact lens wear-related DE, environmental DE, corneal neovascularization DE, allergic DE, LASIK-induced neuroepitheliopathy, aqueous lacrimation, xerophthalmia, Stevens-Johnson syndrome, ocular pemphigoid, blepharoma, eyelid closure failure, corneal ulcer, blepharitis, and ocular hyperemia.

17. A pharmaceutical composition as described in claim 1 or a use as described in claim 2, characterized in that the artificial polypeptide is administered to the eye in an ophthalmic formulation.

18. The pharmaceutical composition of claim 17, or the use of claim 17, wherein the ophthalmic preparation is selected from the group consisting of eye drops, gels, ointments, suspensions, semi-liquids, semi-solid gels, creams, foaming gels, contact lens solutions, and eyewashes.

19. The pharmaceutical composition of claim 1 or the use of claim 2, wherein the administration comprises topical administration to the eye.

20. The pharmaceutical composition of claim 19, or the use of claim 19, wherein the local administration comprises topical administration, subconjunctival administration, retrobulbar administration, periocular administration, subretinal administration, suprachoroidal administration, or intraocular administration.

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