Collagen composition and method of using the same

Abstract

The present disclosure relates to a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid. The present disclosure also relates to a composition comprising collagen, hyaluronic acid, and an additional polymer, wherein the collagen is completely miscible with the hyaluronic acid and the polymer. Methods of using the compositions described herein for medical and cosmetic treatments are also provided.

Description

Technical Field

[0001] (Reference to electronically submitted sequence listing) The content of the sequence listing XML (name: 4431_090PC01_SequenceListing_ST26.xml; size: 1,309,312 bytes, and creation date: May 30, 2023) submitted electronically together with this application is hereby incorporated by reference in its entirety into this specification.

Background Art

[0002] Collagen is one of the most important proteins in the human body, present in connective tissues such as cartilage, bone, tendon, ligament, and skin, and is the major protein in the extracellular matrix of human cells. "Recombinant collagen" refers to a family of at least 28 different naturally occurring collagen types prepared using recombinant technology.

[0003] Collagen has many known uses. For example, in the cosmetics and skincare industries, skincare compositions containing collagen can be used to counter the effects of aging and environmental stress on the appearance, elasticity, and thickness of the skin. For example, aging and environmental factors can result in skin conditions such as fine lines, wrinkles, dry skin, enlarged pore size, skin pigmentation disorders, loss of elasticity, vellus hair, thinning of the skin, purpura, actinic keratosis, itching, eczema, acne, rosacea, erythema, telangiectasia, actinic telangiectasia, skin cancer, and nasal polyps, but are not limited to these.

[0004] Separately from this, dermal hyaluronic acid (HA) fillers are common for soft tissue augmentation. Additional age-related conditions that can benefit from collagen and hyaluronic acid treatments include, but are not limited to, arthritis, osteoarthritis, eye problems, cell adhesion, and similar conditions. Numerous skin care products and skin fillers are commercially available for improving the appearance of the skin, but many consumers are hesitant to use chemically synthesized products that they perceive to be environmentally unfriendly or otherwise unsafe. Miscible products containing HA and collagen that retain the advantages of both components are highly desirable, but native collagen is not compatible with HA and tends to separate during storage and use. Similarly, in the art, there is a need for a miscible collagen blend of polymers for use in the delivery of therapeutic agents.

Summary of the Invention

[0005] In some embodiments, the present disclosure provides a composition comprising collagen and hyaluronic acid, wherein the collagen is fully miscible with the hyaluronic acid. In some embodiments, the collagen is recombinantly produced collagen. In some embodiments, the collagen is a recombinantly produced collagen fragment.

[0006] In some embodiments, the composition has an isoelectric point (pI) of from about 4.0 to about 12.0. In some embodiments, the composition has a pI of from about 4.0 to about 5.0. In some embodiments, the composition has a pI of from about 6.0 to about 7. In some embodiments, the composition has a pI of from about 9.0 to about 10.0. In some embodiments, the composition has a pI of from about 9.0 to about 9.7.

[0007] In some embodiments, the molecular weight of the collagen is from about 5 kDa to about 150 kDa.

[0008] In some embodiments, hyaluronic acid has a molecular weight in the range of about 3 kDa to about 10,000 kDa. In some embodiments, hyaluronic acid has a molecular weight in the range of about 20 kDa to about 2,000 kDa. In some embodiments, hyaluronic acid has a molecular weight in the range of about 1,000 kDa to at least about 5,000 kDa. In some embodiments, hyaluronic acid has a molecular weight in the range of about 2,000 kDa to at least about 3,000 kDa. In some embodiments, hyaluronic acid has a molecular weight of about 1,000 kDa. In some embodiments, hyaluronic acid has a molecular weight of about 50 kDa.

[0009] In some embodiments, collagen does not contain hydroxyproline (Hyp) residues. In some embodiments, collagen contains about 1% to about 20% Hyp residues, based on the total number of prolines in the collagen. In some embodiments, collagen does not contain hydroxyl groups. In some embodiments, collagen contains about 1 to about 120 hydroxyl groups. In some embodiments, collagen does not contain amine groups. In some embodiments, collagen contains about 1 to about 100 amine groups. In some embodiments, collagen does not contain carboxylic acid groups. In some embodiments, collagen contains about 1 to about 80 carboxylic acid groups.

[0010] In some embodiments, hyaluronic acid is cross-linked. In some embodiments, hyaluronic acid is not cross-linked. In some embodiments, hyaluronic acid is biphasic cross-linked hyaluronic acid. In some embodiments, hyaluronic acid is monophasic cross-linked hyaluronic acid.

[0011] In some embodiments, the hyaluronic acid solution has a pH in the range of about 5.0 to about 12. In some embodiments, hyaluronic acid has a pH in the range of about 9.0 to about 12. In some embodiments, hyaluronic acid has a pH in the range of about 5.2 to about 7.2.

[0012] In some embodiments, the hyaluronic acid to collagen (w / w) ratio is in the range of about 1:9 to 9:1.

[0013] In some embodiments, the collagen is crosslinked. In some embodiments, the collagen is not crosslinked.

[0014] In some embodiments, the hyaluronic acid is crosslinked by a chemical crosslinking agent, ultraviolet (UV) irradiation, microwave irradiation, or heat. In some embodiments, the collagen is crosslinked by a chemical crosslinking agent, ultraviolet (UV) irradiation, microwave irradiation, or heat.

[0015] In some embodiments, the composition has a viscosity of about 1 cps to about 100,000 cps. In some embodiments, the composition has a viscosity of about 10,000 cps to about 80,000 cps. In some embodiments, the composition has a viscosity of about 50,000 cps to about 70,000 cps.

[0016] In some embodiments, the composition is a solution. In some embodiments, the composition is a gel. In some embodiments, the composition is a foam.

[0017] In some embodiments, the present disclosure provides a film comprising the composition disclosed herein.

[0018] In some embodiments, the present disclosure provides one or more fibers comprising the composition disclosed herein.

[0019] In some embodiments, the present disclosure provides a coating prepared from the composition disclosed herein.

[0020] In some embodiments, the present disclosure provides a powder prepared from the composition disclosed herein.

[0021] In some embodiments, the present disclosure provides a slurry prepared from the compositions disclosed herein.

[0022] In some embodiments, the present disclosure provides one or more particles prepared from the compositions disclosed herein.

[0023] In some embodiments, the collagen is not hydroxylated. In some embodiments, the recombinant collagen fragment is not hydroxylated.

[0024] In some embodiments, the collagen is hydroxylated. In some embodiments, the recombinant collagen fragment is hydroxylated. In some embodiments, the recombinant collagen fragment is about 1% to about 20% hydroxylated. In some embodiments, the recombinant collagen fragment is about 12% hydroxylated. In some embodiments, the recombinant collagen fragment is 5% hydroxylated.

[0025] In some embodiments, the composition further comprises a pharmaceutically active agent. In some embodiments, the composition further comprises a cosmetically active agent.

[0026] In some embodiments, the collagen or recombinant collagen fragment has at least about 85% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the collagen or recombinant collagen fragment has at least about 85% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 2 to 1002.

[0027] In some embodiments, the collagen or recombinant collagen is fermented or chemically synthesized.

[0028] In some embodiments, the present disclosure provides a method of increasing collagen production in cells, comprising administering or applying an effective amount of the compositions disclosed herein.

[0029] In some embodiments, the present disclosure provides a method of medical or cosmetic treatment comprising administering or applying an effective amount of the compositions disclosed herein.

[0030] In some embodiments, the treatment comprises filling wrinkles, increasing viscoelasticity, or increasing the volume of the face or body. In some embodiments, the treatment comprises forming a cosmetic implant in a patient. In some embodiments, the treatment comprises tissue enhancement.

[0031] In some embodiments, the composition is administered to the subject's face, eyes, nose, nasolabial folds, neck, décolletage, arms, abdomen, blood vessels, bladder, urethra, nerves, cartilage, bone, breast, vocal cords, vocal cord folds, tympanic membrane, joints, intervertebral discs, buttocks, hip joints, gastrocnemius, thighs, flanks, or hands.

[0032] In some embodiments, the treatment comprises treating an inflammatory skin disease or mucosal disease. In some embodiments, the treatment comprises treating lipodystrophy.

[0033] In some embodiments, the present disclosure provides a method of treating a wound in a human subject in need thereof, the method comprising applying a composition according to any one of claims 1 to 59 to the subject's wound. In some embodiments, the wound is a chronic wound, a surgical wound, a superficial wound, an abrasion, or the wound is the result of a thermal injury.

[0034] In some embodiments, the present disclosure provides a skin care product comprising a composition disclosed herein for use in reducing the appearance of wrinkles, evening skin tone, providing moisture, reducing the appearance of dark circles under the eyes, increasing the collagen content of the skin, increasing skin density, improving skin firmness and elasticity, improving the appearance of fine lines and wrinkles, smoothing the texture of the skin, increasing skin glow and brightness, improving the appearance of sagging skin, skin whitening, sun care, color cosmetics, hair care, fragrances, oral care, or any combination thereof.

[0035] In some embodiments, the present disclosure provides a dermal filler comprising the compositions disclosed herein.

[0036] In some embodiments, the dermal filler is a heat-stable, sterilizable soft tissue filler. In some embodiments, the dermal filler is heat-stable up to at least about 125°C. In some embodiments, the dermal filler is heat-stable at temperatures in the range up to at least about 135°C. In some embodiments, the dermal filler is ultimately sterilized. In some embodiments, the dermal filler is injectable.

[0037] In some embodiments, the dermal filler further comprises carboxymethylcellulose. In some embodiments, the dermal filler further comprises calcium hydroxyapatite. In some embodiments, the dermal filler further comprises a solid homopolymer of polylactic acid, an aliphatic polyester, a polyorthoester, a polyoxaester, a polyanhydride, a polycarbonate, a polyurethane, a polyamide, a polyesteramide, a hydrolyzable amine, a polyalkylene oxide, a poloxamer, a functionalized isoflavonoid, an amino acid, a solid homopolymer of poly(e-caprolactone), a solid homopolymer of poly(p-dioxanone), a solid homopolymer of poly(trimethylene carbonate), a solid copolymer of a plurality of e-caprolactone repeating units and a third lactone repeating unit, or a solid copolymer of a plurality of trimethylene carbonate repeating units and a second lactone repeating unit, wherein the third lactone repeating unit is selected from the group consisting of a glycolide repeating unit, a lactide repeating unit, a trimethylene carbonate repeating unit, a p-dioxanone repeating unit, a 1,4-dioxepan-2-one repeating unit, a 1,5-dioxepan-2-one repeating unit, or any combination thereof.

[0038] In some embodiments, the dermal filler further comprises a pharmaceutically active agent. In some embodiments, the pharmaceutically active agent comprises an anesthetic.

[0039] In some embodiments, the present disclosure provides for the use of the compositions, skin care products, or dermal fillers disclosed herein for medical or cosmetic treatment of a subject in need thereof.

[0040] In some embodiments, the present disclosure provides a method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid, and comprising dissolving the hyaluronic acid in an NaOH solution; adding the collagen; and adjusting the pH by adding HCl.

[0041] In some embodiments, the present disclosure provides a method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid, and comprising dissolving the hyaluronic acid in an NaOH solution; adjusting the pH by adding HCl; and adding the collagen.

[0042] In some embodiments, the present disclosure provides a method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid, and comprising dissolving the hyaluronic acid in water to form a hyaluronic acid solution; dissolving the collagen in water to form a collagen solution; and mixing the hyaluronic acid solution and the collagen solution.

[0043] In some embodiments, the method further comprises optionally crosslinking the hyaluronic acid and collagen in the solution by adding a coupling agent.

[0044] In some embodiments, the method further comprises optionally crosslinking the hyaluronic acid and collagen in the solution by adjusting the pH with HCl and adding a coupling agent prior to adding the collagen.

[0045] In some embodiments, the method further includes adding a sodium chloride salt to form a gel and solidifying the gel. In some embodiments, the method further includes sterilizing the gel.

[0046] In some embodiments, the solution contains NaCl.

[0047] In some embodiments, the method further includes adding calcium hydroxyapatite, carboxymethyl cellulose, polylactic acid, polylactic acid, aliphatic polyester, polyorthoester, polyoxaester, polyanhydride, polycarbonate, polyurethane, polyamide, polyesteramide, hydrolyzable amine, polyalkylene oxide, poloxamer, functionalized isoflavonoid, amino acid, a solid homopolymer of poly(e-caprolactone), a solid homopolymer of poly(p-dioxanone), a solid homopolymer of poly(trimethylene carbonate), a solid copolymer of a plurality of e-caprolactone repeating units and a third lactone repeating unit, and a solid copolymer of a plurality of trimethylene carbonate repeating units and a second lactone repeating unit; the third lactone repeating unit is selected from the group consisting of glycolide repeating units, lactide repeating units, trimethylene carbonate repeating units, p-dioxanone repeating units, 1,4-dioxepan-2-one repeating units, 1,5-dioxepan-2-one repeating units, and / or combinations thereof, or any combination of these.

[0048] In some embodiments, the method further includes adding an anesthetic. In some embodiments, the method further includes dispensing the gel into a syringe under aseptic conditions.

[0049] In some embodiments, the present disclosure provides a composition comprising collagen and polyvinylpyrrolidone, wherein the collagen is completely miscible with the polyvinylpyrrolidone. In some embodiments, the polyvinylpyrrolidone has a molecular weight of from about 10 kDa to about 400 kDa. In some embodiments, the polyvinylpyrrolidone has a molecular weight of about 50 kDa.

[0050] In some embodiments, the collagen is completely miscible with polyacrylamide. In some embodiments, the polyacrylamide has a molecular weight of from about 5,000 kDa to about 6,000 kDa. In some embodiments, the polyacrylamide has a molecular weight of from about 30 kDa to about 50 kDa.

[0051] In some embodiments, the present disclosure provides a composition comprising collagen and poly(ethylene oxide), wherein the collagen is completely miscible with the poly(ethylene oxide). In some embodiments, the collagen is completely miscible with poly(2-oxazoline). In some embodiments, the collagen is completely miscible with polyethyleneimine.

[0052] In some embodiments, the present disclosure provides a composition comprising collagen and sodium carboxymethylcellulose, wherein the collagen is completely miscible with the carboxymethylcellulose. In some embodiments, the carboxymethylcellulose has a molecular weight of about 250 kDa and a degree of substitution of from about 1.00 to about 1.25. In some embodiments, the carboxymethylcellulose has a molecular weight of about 250 kDa and a degree of substitution of from about 0.70 to about 0.90. In some embodiments, the carboxymethylcellulose has a molecular weight of about 250 kDa and a degree of substitution of about 0.79. In some embodiments, the carboxymethylcellulose has a degree of substitution of about 0.87.

[0053] In some embodiments, the carboxymethylcellulose has a high viscosity.

[0054] In some embodiments, the present disclosure provides a composition comprising collagen and chondroitin sulfate, wherein the collagen is completely miscible with the chondroitin sulfate. In some embodiments, the chondroitin sulfate has a molecular weight of about 50 kDa.

[0055] In some embodiments, the present disclosure provides a composition comprising collagen and acetylated hyaluronate. In some embodiments, the collagen is completely miscible with the acetylated hyaluronate.

[0056] In some embodiments, the present disclosure provides a composition comprising collagen and zinc hyaluronate. In some embodiments, the collagen has at least about 85%, at least about 87%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the collagen has at least about 98% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1.

[0057] In some embodiments, the present disclosure provides a composition comprising a recombinant collagen fragment having at least about 85% amino acid sequence identity to SEQ ID NO: 1, and / or one or more collagen fragment sequence variants having at least about 85% amino acid sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 975 - 1002, and hyaluronic acid, wherein the recombinant collagen fragment is completely miscible with the hyaluronic acid. In some embodiments, the hyaluronic acid is a biphasic cross-linked hyaluronic acid comprising cross-linked HA and non-cross-linked HA.

[0058] In some embodiments, the present disclosure provides a composition comprising a recombinant collagen fragment having at least about 85% amino acid sequence identity to SEQ ID NO: 1 and / or one or more collagen fragment sequence variants having at least about 85% amino acid sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 975-1002, polyvinylpyrrolidone, and polyvinylpyrrolidone, wherein the recombinant collagen fragment is completely miscible with polyvinylpyrrolidone.

[0059] In some embodiments, the present disclosure provides a composition comprising one or more recombinant collagen fragments having at least about 85% amino acid sequence identity to SEQ ID NO: 1 and / or one or more collagen fragment sequence variants having at least about 85% amino acid sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 975-1002, and sodium carboxymethyl cellulose, wherein the recombinant collagen fragment is completely miscible with sodium carboxymethyl cellulose.

[0060] In some embodiments, the present disclosure provides a film comprising a recombinantly produced collagen fragment and hyaluronic acid. In some embodiments, the film is visually transparent. In some embodiments, the film is cloudy. In some embodiments, the film exhibits phase separation.

[0061] In some embodiments, the present disclosure provides a gel comprising a recombinantly produced collagen fragment and hyaluronic acid.

[0062] In some embodiments, the present disclosure provides one or more fibers comprising a recombinantly produced collagen fragment and hyaluronic acid.

[0063] In some embodiments, the present disclosure provides a coating comprising a recombinantly produced collagen fragment and hyaluronic acid.

[0064] In some embodiments, the present disclosure provides a slurry comprising recombinantly produced collagen fragments and hyaluronic acid.

[0065] In some embodiments, the present disclosure provides one or more particles comprising recombinantly produced collagen fragments and hyaluronic acid.

[0066] In some embodiments, the present disclosure provides a foam comprising recombinantly produced collagen fragments and hyaluronic acid.

[0067] In some embodiments, the present disclosure provides a composition comprising collagen and sodium alginate salt, wherein the collagen is completely miscible with the sodium alginate salt. In some embodiments, the sodium alginate salt has a viscosity of about 4 - 12 cps with respect to a 1% aqueous solution at 25°C.

[0068] In some embodiments, the present disclosure provides a composition comprising collagen and sodium alginate salt, wherein the collagen is completely miscible with the sodium alginate salt. In some embodiments, the sodium alginate salt has a viscosity of 2,000 cps or more with respect to a 2% aqueous solution at 25°C.

Brief Description of the Drawings

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Mode for Carrying Out the Invention

[0070] Definition The indefinite articles "a" and "an" used to describe an element or component mean that one or at least one of these elements or components exists. These articles, when used in this specification, have been conventionally used to indicate that the modified noun is a singular noun, but unless otherwise specified in a particular example, the articles "a" and "an" also include the plural form. Similarly, when used in this specification, the definite article "the" also means that the modified noun can be in the singular or plural form, again unless otherwise specified in a particular example.

[0071] As used herein, the term "about" when used with a numerical value means "within 10% of the recited value" unless otherwise specified. For example, "about 5 wt%" means 4.5 wt% to 5.5 wt%.

[0072] The term "modified" as applied to collagen and collagen fragments refers to collagen fragments that contain amino acid sequences that are at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical or similar to the amino acid sequence of a biologically active molecule. In some embodiments, the modified collagen fragment contains an amino acid sequence that is at least 40%, 45%, 50%, 55%, 60%, 65%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of a native or previously engineered sequence. The modified sequence can include additions, deletions, substitutions, or combinations thereof to the amino acid sequence of a native or previously engineered molecule. For example, a modified collagen fragment can incorporate or delete 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid residues compared to a native collagen sequence. Such selections can be made to modify the looseness or tension of recombinant collagen. The degree of hydroxylation of collagen correlates with the looseness or tension of the collagen triple helix. Modified collagen fragments can also include chemical modifications to the polypeptide, such as cross-linking between cysteine residues, or cross-linking between hydroxylated or glycosylated residues. As used herein, the terms "variant" and "sequence variant" refer to polypeptide sequences that are about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 1.

[0073] The terms "pharmaceutically acceptable" and "cosmetically acceptable" when applied to carriers, excipients, or stabilizers that can be used in the compositions described herein refer to carriers, excipients, or stabilizers that are non-toxic to the recipient at the dosages and concentrations employed.

[0074] As used herein, the term "tissue repair" refers to the restoration of tissue structure and function after injury in the context of the healing of damaged tissue. Tissue regeneration refers to a type of healing in which new growth restores damaged tissue parts to a normal state.

[0075] As used herein, the terms "hyaluronic acid" and "HA" refer to acidic polysaccharides having different molecular weights composed of residues of D-glucuronic acid and N-acetyl-D-glucosamine, and their corresponding salts including but not limited to sodium hyaluronate. Compositions described herein as containing HA should be understood to contain a combination of hyaluronic acid and sodium hyaluronate unless otherwise specified. Hyaluronic acid is naturally present on the cell surface, in the basic extracellular substance of vertebrate connective tissue, in the synovial fluid of joints, in the vitreous humor of the eye, and in human umbilical cord tissue. "Hyaluronic acid" can mean the entire series of polysaccharides having alternating residues of D-glucuronic acid and N-acetyl-D-glucosamine or its degradation fractions having various molecular weights. Hyaluronic acid also refers to a non-sulfated glycosaminoglycan composed of repeating disaccharide units of N-acetylglucosamine (GlcNAc) and glucuronic acid (GlcUA) linked together by alternating beta-1,4 and beta-1,3 glycosidic bonds. Hyaluronic acid is also known as hyaluronan, hyaluronate, or HA.

[0076] As used herein, the terms "sodium carboxymethyl cellulose", "carboxymethyl cellulose" and "CMC" can be used interchangeably and refer to polymers having different ranges of molecular weights as derivatives of cellulose having carboxymethyl groups attached to some of the monomers that make up the cellulose backbone. Compositions described herein as containing carboxymethyl cellulose should be understood to contain a combination of carboxymethyl cellulose and sodium carboxymethyl cellulose unless otherwise specified.

[0077] As used herein, the term "polyvinylpyrrolidone" refers to polymers having different ranges of molecular weights made from the monomer N-vinylpyrrolidone. Polyvinylpyrrolidone (PVP) is also known as polyvidone and povidone.

[0078] As used herein, the term "polyacrylamide" refers to polymers having different ranges of molecular weights made from the monomer acrylamide. Polyacrylamide (PAM) is also known as poly(2-propenamide).

[0079] As used herein, the term "poly(ethylene oxide)" refers to polymers having different ranges of molecular weights made from the monomer ethylene oxide. Poly(ethylene oxide) (PEO) is also known as poly(ethylene glycol) (PEG) or poly(oxyethylene).

[0080] As used herein, the term "poly(2-oxazoline)" refers to polymers having different ranges of molecular weights made from the monomer 2-oxazoline and its derivatives such as 2-methyl-2-oxazoline and 2-ethyl-2-oxazoline.

[0081] As used herein, the term "polyethyleneimine" refers to polymers having different ranges of molecular weights made from the monomer aziridine. Polyethyleneimine (PEI) is also known as polyaziridine.

[0082] As used herein, the term "miscibility" means that one substance is soluble with another such that a molecularly homogeneous mixture is formed when the two components are mixed in water or in a buffer system. An exemplary buffer system is phosphate buffered saline (PBS). For example, collagen described as miscible with HA means that an aqueous mixture of a collagen solution and an HA solution (e.g., a 1% - 10% HA solution), where the collagen solution is dissolved in water or the buffer system, forms a homogeneous mixture at room temperature. The miscibility of collagen with hyaluronic acid in a composition may depend on the ratio of hyaluronic acid to collagen, the molecular weights of the collagen and hyaluronic acid used in the composition, and the presence of other additional components in the composition such as salts that affect the compatibility of collagen and hyaluronic acid. Temperature can also affect the separation of hyaluronic acid and collagen in the composition. Collagen described as immiscible with HA means that a mixture of a collagen solution and an HA solution does not form a stable, clear, homogeneous mixture at room temperature. Instead, the solution may exhibit turbidity or phase separation. A solid film prepared from a completely miscible solution of HA and collagen may remain completely miscible as a solid film, as demonstrated by being transparent and homogeneous, or may exhibit small-scale phase separation indicated by cloudiness. Transparency can be readily determined by one of ordinary skill in the art, for example, by measuring using a haze meter.

[0083] "Fully miscible" collagen can be applied to the ability of collagen to form a homogeneous aqueous mixture with any substance or polymer (e.g., polyvinylpyrrolidone (PVP), polyacrylamide (PAM), poly(ethylene oxide) (PEO), poly(2-oxazoline), polyethyleneimine (PEI), carboxymethylcellulose, chondroitin sulfate, acetylated hyaluronate and / or zinc hyaluronate), and is not limited to HA.

[0084] "Viscosity" refers to a measure of the resistance to flow of a liquid at a given temperature. Viscosity can be determined using a variety of methods and instruments known in the art, including but not limited to measurements with rheometers and viscometers. Viscosity can also represent the rigidity of a liquid at a given temperature.

[0085] The compositions disclosed herein can have a viscosity of at least about 1 centipoise (cps), at least about 10 cps, at least about 100 cps, at least about 1000 cps, at least about 10,000 cps, at least about 20,000 cps, at least about 50,000 cps, at least about 100,000 cps or more at a shear rate of 0.1 / second at 25°C. In some embodiments, the viscous carrier in the compositions of the present disclosure can have a viscosity of about 1 cps, about 10 cps, about 100 cps, about 1000 cps, about 10,000 cps, about 20,000 cps, about 50,000 cps, about 100,000 cps or more at a shear rate of 0.1 / second at 25°C.

[0086] As used herein, the term "isoelectric point" or "pI" refers to the pH value at which the overall net charge of a macromolecule such as a protein is zero. In a protein, many charged groups can be present, and at the isoelectric point, the sum of all these charges is zero, i.e., the number of negative charges balances the number of positive charges. At a pH above the isoelectric point, the overall net charge of the protein is negative, while at a pH value below the isoelectric point, the overall net charge of the protein is positive. The isoelectric point of a molecule can be determined by using isoelectric focusing, a well-known technique for separating molecules based on differences in their isoelectric points. The isoelectric point of a protein can also be determined by electrophoresis. The isoelectric point can also be calculated (see, for example, Levene and Simms, "Calculation of isoelectric point" J. Biol. Chem., 1923, pp. 801-813).

[0087] The compositions described herein can have an isoelectric point in the range of from about 4.0 to about 12.0, from about 4.5 to about 11.5, from about 5.0 to about 11.0, from about 5.5 to about 10.5, from about 6.0 to about 10.0, from about 6.5 to about 9.5, from about 7.0 to 9.0, from about 7.5 to 8.5, or about 8.0. In certain embodiments, the compositions described herein can have an isoelectric point of at least about 4.0, about 4.5, about 5.0, about 5.5, about 6.0, at least about 6.5, at least about 7.0, at least about 7.5, at least about 8.0, at least about 8.5, at least about 9, at least about 9.5, at least about 10, at least about 10.5, at least about 11, at least about 11.5, or at least about 12. In some embodiments, the compositions described herein have an isoelectric point of about 4.0, about 4.5, about 5.0, about 5.5, about 6.0, about 6.5, about 7.0, about 7.5, about 8.0, about 8.5, about 9, about 9.5, about 10, about 10.5, about 11, about 11.5, or about 12.

[0088] As used herein, the term "crosslinked" refers to a polymer having at least one covalent bond found either within the repeating units of the polymer or between the repeating units of the polymer. Crosslinking bonds occur between individual chains or molecules of the polymer; intramolecular reactions can occur within the polymer chains and can form macrocyclic structures. The latter are not considered crosslinking in the present disclosure. Crosslinking is formed between any two functional groups of the polymer (e.g., at the ends, on the side chains, etc.). In certain embodiments, the crosslinking is formed between the terminal units of the polymer. In certain embodiments, the crosslinking is formed between the side chain units of the polymer. Any kind of covalent bond can form a crosslink (e.g., carbon-carbon, carbon-oxygen, carbon-nitrogen, oxygen-nitrogen, sulfur-sulfur, oxygen-phosphorus, etc.). The resulting crosslinked material can be branched, linear, dendritic, etc. In certain embodiments, the crosslinking forms a 3-D network of crosslinks. Crosslinking can be formed by any chemical crosslinking agent in a chemical reaction that results in a covalent bond. Crosslinking can be formed by applying ultraviolet (UV) irradiation or microwave irradiation to the polymer. Crosslinking can be formed, for example, by a free radical initiation reaction using a photoinitiator or a thermal initiator, or by applying heat.

[0089] Typical chemical crosslinking agents generally are various kinds of difunctional or polyfunctional crosslinking agents, and can be selected, for example, from DVS (divinyl sulfone) in an alkaline medium (see U.S. Patent No. 4,582,865), difunctional or polyfunctional epoxy compounds (see U.S. Patent No. 4,716,154), carbodiimide, or formaldehyde (see British Patent No. 2 151 244). Other suitable crosslinking agents include epoxides or their derivatives, particularly 1,4-butanediol diglycidyl ether (BDDE), diepoxyoctane, or 1,2-bis(2,3-epoxypropyl)-2,3-ethylene. Known crosslinking agents also include divinyl sulfone, glycerol diglycidyl ether, and 1,4-butanediyl diglycidyl ether.

[0090] In certain embodiments, the compositions according to the present disclosure can react to form a crosslinked polymer matrix. Since the crosslinking reaction occurs in an aqueous solution, the crosslinked polymer matrix can be dispersed in an aqueous liquid in the form of a hydrogel when formed by the crosslinking reaction. As used herein, the term "hydrogel" refers to a three-dimensional network comprising a hydrophilic polymer containing a large amount of water. The hydrogel can contain, for example, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or even more water on a w / w basis. A "hydrogel precursor" is a polymer that is at least partially soluble in an aqueous medium and can crosslink to form a hydrogel.

[0091] In some embodiments, the compositions of the present disclosure can exhibit a physiologically acceptable osmotic pressure. As used herein, the term "osmotic pressure" refers to the concentration of osmotically active solutes in a solution. As used herein, the term "physiologically acceptable osmolality" refers to an osmolality that conforms to or is characteristic of the normal functioning of a living organism. Thus, the administration of the hydrogels disclosed herein exhibits an osmotic pressure that has substantially no long-term or permanent harmful effects when administered to a mammal. Osmolality is expressed as the number of osmoles of osmotically active solutes per liter of solvent (Osmol / L or Osm / L).

[0092] As used herein, "solubility" refers to the amount of a substance that dissolves in a given volume of solvent at a specific temperature and pH to form, for example, a saturated solution. Solubility can be determined, for example, using the shake flask solubility method (ASTM: E 1148-02, Standard Test Method for Measurements of Aqueous Solubility, Book of Standards Volume 11.05). Solubility can be determined at any pH. Solubility can be tested at any temperature from 20°C to 40°C. For example, solubility can be determined at approximately pH 7.0 to 7.4 and at approximately 37°C.

[0093] A. Collagen The term "collagen" refers to any one of the known collagen types, including type I - type XXVIII collagen, and any other collagen that is natural, synthetic, semi - synthetic, or recombinant. The term "collagen" includes collagen, collagen fragments, collagen - like proteins, triple - helix collagen, alpha chains, monomers, gelatin, trimers, and combinations thereof. It includes all of the collagens, modified collagens, and collagen - like proteins described herein. In certain embodiments, the term "collagen" refers to any known type of collagen that is 1) soluble in HA; 2) induces type I collagen production, type III collagen production, ECM production, or combinations thereof. This term also encompasses procollagens and collagen - like proteins or collagenous proteins that contain the motif (Gly - X - Y)n, where n is an integer. It includes molecules of collagen and collagen - like proteins, trimers of collagen molecules, fibrils of collagen, and fibers of collagen fibrils. It also refers to chemically, enzymatically, or recombinantly modified collagens or collagen - like molecules that can be fibrillated, and fragments of collagen, collagen - like molecules, and collagenous molecules that can aggregate into nanofibers. Recombinant collagen molecules generally contain the repeating - (Gly - X - Y)n - sequence, whether natural or engineered. As used herein, collagen is a general term for a family of at least 28 distinct collagen types. In various species, including bovine, ovine, porcine, avian, marine, plant, and human, various distinct collagen types have been identified.

[0094] Collagen has a unique amino acid composition; approximately one-third of its amino acid residues are glycine, and another 15% - 30% are proline or 4-hydroxyproline (Hyp) residues. Hydroxylysine is also characteristic of collagen. These hydroxylated amino acid residues, formed as a result of post-translational modification, confer stability to collagen through intramolecular hydrogen bonds that include bridging water molecules. The collagen content in tissues can be determined by measuring its hydroxyproline content. The hydroxyproline content can be measured by methods known in the art, for example, by liquid chromatography - mass spectrometry (LC-MS). Collagen contains many repeats of the sequence Gly-X-Y, where X is often proline and Y is often 4-hydroxyproline.

[0095] Collagen variants containing such amino acid residue substitutions can be produced by various known methods including genetic engineering techniques and in vitro peptide synthesis. Collagen variants can also be produced by changing the amount of hydroxylysine and / or hydroxyproline present in a given molecule, by changing the expression of lysine hydroxylase and / or proline hydroxylase, where the hydroxylase gene (recombinant or otherwise) is expressed in a host cell for the expression of recombinant collagen or its derivatives. The hydroxyproline content can be measured by methods known in the art, for example, by liquid chromatography - mass spectrometry (LC-MS).

[0096] Collagen is the most abundant protein in vertebrates. Vertebrate collagen is a family of proteins produced by several cell types. Within this protein family, collagen types are distinguishable by their chemical composition, different morphological and pathological characteristics, distribution within tissues, and their functions. Although many types of collagen have been described, five major types are recognized. Mammalian skin is typically type I collagen, but other types of collagen, including type III collagen, can be used in leather formation. The term "collagen" encompasses unprocessed (e.g., procollagen) as well as post-translationally modified and proteolytically degraded collagen having a triple helix structure.

[0097] Any type of collagen, including types I - XXVIII of collagen, is suitable for use in the compositions described herein.

[0098] B. Recombinant Collagen and Recombinant Collagen Fragments As used herein, the term "recombinant collagen" refers to a family of at least 28 distinct naturally occurring collagen types, including but not limited to types I - XXVIII of collagen, prepared using recombinant techniques.

[0099] In some embodiments, the recombinant collagen described herein is a recombinant collagen fragment. The recombinant collagen fragment may be a fragment of the complete amino acid sequence of a native collagen molecule capable of forming tropocollagen (trimeric collagen), or the fragment may be a modified collagen molecule or a fragment of a cleaved collagen molecule having an amino acid sequence that is at least 70, 80, 90, 95, 96, 97, 98, or 99% identical or similar to the native collagen amino acid sequence (or to its fibril-forming region, or to a segment substantially containing [Gly-X-Y]n). In certain embodiments, the term recombinant collagen refers to any known type of recombinant collagen that is (1) soluble in HA and (2) induces the production of type I collagen and / or type III collagen. The production of collagen type I and / or collagen type III can be determined by assays known in the art, including those described in PCT / US2022 / 027016, which is incorporated herein by reference in its entirety.

[0100] Exemplary collagen sequences from which the fragment may be derived include the amino acid sequences of Col1A1, Col1A2, and Col3A1, such as those described in accession numbers P02461.4 (SEQ ID NO: 1003; human Col3A1) (www.ncbi.nlm.nih.gov / protein / 124056490), NP_001029211.1 (SEQ ID NO: 1004; bovine Col1A1) (www.ncbi.nlm.nih.gov / protein / 77404252), NP_776945.1 (SEQ ID NO: 1005 bovine Col1A2) (www.ncbi.nlm.nih.gov / protein / 27806257) and NP_001070299.1 (SEQ ID NO: 1006; bovine (Col3A1) (www.ncbi.nlm.nih.gov / protein / 116003881), which are incorporated herein by reference.

[0101] Genes encoding collagen can be cleaved or otherwise modified to add or remove sequences, for example, to encode collagen fragments disclosed herein. Further, gene modification can be made to customize the size of the polynucleotide or vector, target the expressed protein to the endoplasmic reticulum or other cellular or extracellular compartments, or control the length of the encoded protein. Modifications can be made to the polynucleotide encoding the collagen fragment. For example, the polynucleotide coding sequence of collagen or a collagen fragment can be modified to encode a protein that is at least 70, 80, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 100% identical or similar to a known amino acid sequence. Such modifications can include codon modification or codon optimization of the polynucleotide encoding the collagen fragment.

[0102] In some embodiments, the collagen fragments disclosed herein can have a molecular weight of from about 1 kDa to about 150 kDa. In some embodiments, the collagen fragment can have a molecular weight of about 1 kDa, 2 kDa, 3 kDa, 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa, 9 kDa, 10 kDa, 11 kDa, 12 kDa, 13 kDa, 14 kDa, 15 kDa, 16 kDa, 17 kDa, 18 kDa, 19 kDa, 20 kDa, 21 kDa, 22 kDa, 23 kDa, 24 kDa, 25 kDa, 26 kDa, 27 kDa, 28 kDa, 29 kDa, 30 kDa, 31 kDa, 32 kDa, 33 kDa, 34 kDa, 35 kDa, 36 kDa, 37 kDa, 38 kDa, 39 kDa, 40 kDa, about 41 kDa, about 42 kDa, about 43 kDa, about 44 kDa, about 45 kDa, about 46 kDa, about 47 kDa, about 48 kDa, about 49 kDa, about 50 kDa, about 51 kDa, about 52 kDa, about 53 kDa, about 54 kDa, about 55 kDa, about 56 kDa, about 57 kDa, about 58 kDa, about 59 kDa, about 60 kDa, about 70 kDa, about 80 kDa, about 90 kDa, about 100 kDa, about 110 kDa, about 120 kDa, about 130 kDa, about 140 kDa or about 150 kDa. In certain embodiments, the collagen fragment can have a molecular weight of about 1 kDa. In certain embodiments, the collagen fragment can have a molecular weight of about 10 kDa. In certain embodiments, the collagen fragment can have a molecular weight of about 20 kDa. In certain embodiments, the collagen fragment can have a molecular weight of about 30 kDa. In certain embodiments, the collagen fragment can have a molecular weight of about 40 kDa. In certain embodiments, the collagen fragment can have a molecular weight of about 50 kDa. In certain embodiments, the collagen fragment can have a molecular weight of about 60 kDa. In certain embodiments, the collagen fragment can have a molecular weight of about 100 kDa. In certain embodiments, the collagen fragment can have a molecular weight of about 140 kDa.

[0103] In some embodiments, the collagen fragments described herein can have an amino acid chain length of about 350 amino acids to about 600 amino acids and can overlap with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the overlapping collagen fragments described herein can have a length of about 150 amino acids, about 200 amino acids, about 250 amino acids, about 300 amino acids, 350 amino acids, about 370 amino acids, about 390 amino acids, about 400 amino acids, about 420 amino acids, about 440 amino acids, about 460 amino acids, about 480 amino acids, about 500 amino acids, about 510 amino acids, about 520 amino acids, about 530 amino acids, about 540 amino acids, about 550 amino acids, about 560 amino acids, about 570 amino acids, about 580 amino acids, about 590 amino acids or about 600 amino acids.

[0104] In some embodiments, the collagen fragments described herein can have an amino acid sequence according to SEQ ID NO: 1. In some embodiments, the collagen fragment can have at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 9%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% sequence identity or similarity to SEQ ID NO: 1.

[0105] The amino acid sequence of SEQ ID NO: 1 is as follows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。

[0106] In some embodiments, the collagen fragments described herein can have an amino acid chain length of from about 350 amino acids to about 600 amino acids and can have at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or 100% sequence identity or similarity to SEQ ID NO: 1. In some embodiments, the collagen fragments described herein can have a length of about 150 amino acids, about 200 amino acids, about 250 amino acids, about 300 amino acids, about 350 amino acids, about 370 amino acids, about 390 amino acids, about 400 amino acids, about 420 amino acids, about 440 amino acids, about 460 amino acids, about 480 amino acids, about 500 amino acids, about 510 amino acids, about 520 amino acids, about 530 amino acids, about 540 amino acids, about 550 amino acids, about 560 amino acids, about 570 amino acids, about 580 amino acids, about 590 amino acids or about 600 amino acids.

[0107] In some embodiments, the recombinant collagen can include hydrolysis products of the collagen fragments, and the hydrolysis products can have a sequence that is part of SEQ ID NO: 1. In some embodiments, the hydrolysis products can have at least about 85% sequence identity to the sequences set forth in any one of SEQ ID NOs: 2-973. In some embodiments, the collagen fragment can be a collagen fragment sequence variant having the amino acid sequence set forth in any one of SEQ ID NOs: 975-1002.

[0108] In some embodiments, the lysine, proline, or lysine and proline residues present in collagen or collagen fragments are hydroxylated. In some embodiments, the lysine, proline, or lysine and proline residues present in collagen fragments are not hydroxylated. In some embodiments, about 1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% (or any intermediate value or sub-range) of the lysine, proline, or lysine and proline residues in the recombinant collagen fragment may be hydroxylated.

[0109] In certain embodiments, the proline residues in the recombinant collagen fragment can be hydroxylated from about 1% to about 20%.

[0110] In some embodiments, the recombinant collagen fragment can be hydroxylated at about 12%. In some embodiments, the recombinant collagen fragment can be hydroxylated at about 5%.

[0111] In some embodiments, the collagen fragment can be a collagen fragment, hydrolysis product, or sequence variant having the amino acid sequence set forth in Table 1.

[0112]

Table 1-1

[0113]

Table 1-2

[0114]

Table 1-3

[0115]

Table 1-4

[0116]

Table 1-5

[0117]

Table 1-6

[0118]

Table 1-7

[0119]

Table 1-8

[0120]

Table 1-9

[0121]

Table 1-10

[0122]

Table 1-11

[0123]

Table 1-12

[0124]

Table 1-13

[0125]

Table 1-14

[0126]

Table 1-15

[0127]

Table 1-16

[0128]

Table 1-17

[0129]

Table 1-18

[0130]

Table 1-19

[0131]

Table 1-20

[0132]

Table 1-21

[0133]

Table 1-22

[0134]

Table 1-23

[0135]

Table 1-24

[0136]

Table 1-25

[0137]

Table 1-26

[0138]

Table 1-27

[0139]

Table 1-28

[0140] C. Hyaluronic acid Hyaluronic acid (HA) is a natural linear carbohydrate polymer belonging to the class of non-sulfated glycosaminoglycans. It is composed of beta-1,3-N-acetylglucosamine and beta-1,4-glucuronic acid repeating disaccharide units with a molecular weight (MW) up to 6 MDa. HA is present in vitreous cartilage, synovial joint fluid, and skin tissue, both the dermis and epidermis. HA can be extracted from natural tissues including vertebrate connective tissues or prepared by microbiological methods.

[0141] Many roles of HA in the body have been identified. It plays an important role in organisms as a mechanical support for cells in many tissues such as skin, tendon, muscle, and cartilage. HA is involved in important biological processes such as tissue wetting and lubrication. It is also presumed to have roles in many physiological functions, such as adhesion, development, cell motility, cancer, neovascularization, and wound healing. Due to the unique physical and biological properties of HA, including viscoelasticity, biocompatibility, and biodegradability, HA is used in current and developing a wide range of applications within the scope of cosmetics, aesthetics, skin care, ophthalmology, rheumatology, drug delivery, wound healing, and tissue engineering. The use of HA in some of these applications is limited by the fact that HA is soluble in water at room temperature, i.e., about 20 °C, rapidly degraded by hyaluronidase in the body, and difficult to process into biomaterials. Therefore, cross-linking of HA has been introduced to improve the physical and mechanical properties of HA and its in vivo residence time. Cross-linking of HA can be described as single-phase or two-phase. The single-phase structure is the case where cross-links are formed between HA chains, which creates a uniform gel-like structure. The two-phase structure is the case where HA is made to be gel-like and has free HA particles in a mixture that is not part of the cross-linked structure. This type of structure is not as smooth as the single-phase, but historically gives more volume.

[0142] U.S. Patent No. 4,582,865, which is incorporated herein by reference in its entirety, describes a method for preparing crosslinked gels of HA alone or crosslinked gels of HA mixed with other hydrophilic polymers using divinyl sulfone (DVS) as a crosslinking agent. The preparation of crosslinked HA or its salts using polyfunctional epoxy compounds is disclosed in European Patent No. 0 161 887 B1, which is also incorporated herein by reference in its entirety. Other bifunctional or polyfunctional reagents used to crosslink HA via covalent linkages include formaldehyde (see U.S. Patent No. 4,713,448, which is incorporated herein by reference in its entirety), polyaziridine (see International Publication No. 03 / 089476, which is incorporated herein by reference in its entirety), and L-amino acids or L-amino esters (see International Publication No. 2004 / 067575, which is incorporated herein by reference in its entirety). Carbodiimides have also been reported for the crosslinking of HA (see U.S. Patent Nos. 5,017,229 and 6,013,679, which are incorporated herein by reference in their entireties). Completely or partially crosslinked esters of HA with aliphatic alcohols, and salts of such partial esters with inorganic or organic bases, are disclosed in U.S. Patent No. 4,957,744, which is also incorporated herein by reference in its entirety. Crosslinking of HA chains with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride ("EDAC") and adipic acid dihydrazide in a water / acetone mixture is disclosed in U.S. Patent Application Publication No. 2006 / 0040892 (which is incorporated herein by reference in its entirety). International Publication No. 2006 / 56204 (which is also incorporated herein by reference in its entirety) also discloses a method for preparing crosslinked gels of HA using divinyl sulfone (DVS) as a crosslinking agent.

[0143] HA is commercially available from various suppliers such as Pure Health Botanicals (Saint Charles, IL) and DSM (Kaiseraugst, Switzerland).

[0144] D. Compositions Containing Collagen Fragments and Sequence Variants Thereof The present disclosure provides a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid. In some embodiments, the collagen is completely miscible with a hyaluronate, such as acetylated hyaluronate. In some embodiments, the collagen is completely miscible with acetylated hyaluronate. Other hyaluronates include zinc hyaluronate.

[0145] In some embodiments, the collagen is completely miscible with other polymers used for therapeutic applications. In some embodiments, the collagen can be completely miscible with the polymer both in an aqueous solution and as a solid film or article. Articles contemplated include, but are not limited to, scaffolds and wound dressings. Such polymers include, but are not limited to, polyvinylpyrrolidone (PVP), polyacrylamide (PAM), poly(ethylene oxide) (PEO), poly(2-oxazoline), polyethyleneimine (PEI), sodium carboxymethyl cellulose, and chondroitin sulfate.

[0146] In some embodiments, the collagen is completely miscible with polyvinylpyrrolidone both in an aqueous solution and as a solid film or article.

[0147] In some embodiments, the collagen is completely miscible with polyacrylamide both in an aqueous solution and as a solid film or article.

[0148] In some embodiments, the collagen is completely miscible with poly(ethylene oxide glycol). In some embodiments, the collagen is completely miscible with poly(2-oxazoline). In some embodiments, the collagen is completely miscible with polyethyleneimine.

[0149] In some embodiments, the collagen is completely miscible with carboxymethyl cellulose.

[0150] In some embodiments, collagen is completely miscible with chondroitin sulfate.

[0151] In some embodiments, collagen is completely miscible with acetylated hyaluronate.

[0152] In some embodiments, collagen can be mixed with zinc hyaluronate. In some embodiments, collagen is completely miscible with zinc hyaluronate.

[0153] In some embodiments, collagen is completely miscible with sodium alginate. In some embodiments, sodium alginate has a viscosity of about 4 - 12 cps for a 1% aqueous solution at 25°C. In some embodiments, sodium alginate has a viscosity of about 2,000 cps or more for a 2% aqueous solution at 25°C.

[0154] In some embodiments, collagen is a recombinantly produced collagen fragment. In some embodiments, the collagen in the composition can be physically and / or chemically modified. Chemical modifications and physical modifications can include, but are not limited to, hydroxylation, crosslinking, charge modification, linking agents, and modifications known in the art.

[0155] In some embodiments of the compositions disclosed herein, the composition can further include one or more polymers or copolymers including, but not limited to, aliphatic polyesters, polyorthoesters, polyoxy esters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyesteramides, hydrolyzable amines, polyalkylene oxides, poloxamers, functionalized isoflavonoids, amino acids, and combinations thereof.

[0156] In some embodiments of the compositions disclosed herein, the composition is poly(hydroxyethyl methacrylate), poly(sulfobetaine), poly(dimethylsiloxane), poly(caprolactone), poly(p-dioxanone), poly(trimethylene carbonate), poly(glycolide), poly(L-lactide), poly(D,L-lactide), poly(L-lactide-co-glycolide), poly(D,L-lactide-co-glycolide), poly(L-lactide-co-caprolactone), poly(L-lactide-co-D,L-lactide), poly(caprolactone-co-trimethylene carbonate), poly(lactide-co-trimethylene carbonate), poly(caprolactone-co-p-dioxanone), poly(trimethylene carbonate-p-dioxanone), poly(caprolactone-co-lactide), poly(lactide-co-1,5-dioxepan-2-one) and poly(1,5-dioxepan-2-one-co-p-dioxanone), poly(lactide-co-1,4-dioxepan-2-one), poly(1,4-dioxepan-2-one-co-p-dioxanone), poly(glycolide-co-caprolactone-co-dioxanone), poly(lactide-co-caprolactone-co-dioxanone), poly(dioxanone-co-D,L-lactide), poly(dioxanone-co-glycolide), poly(dioxanone-co-caprolactone), poly(dioxanone-co-L-lactide), poly(dioxanone-co-D,L-lactide-co-caprolactone), poly(dioxanone-co-glycolide-co-caprolactone), poly(dioxanone-co-L-lactide-co-D,L-lactide), poly(dioxanone-co-L-lactide-co-glycolide), poly(dioxanone-co-L-lactide-do-caprolactone), poly(dioxanone-co-D,L-lactide-co-glycolide), poly(dioxanone-co-D,L-lactide-co-caprolactone), poly(dioxanone-co-glycolide-co-caprolactone, poly(dioxanone-co-L-lactide-co-glycolide-co-caprolactone), poly(dioxanone-co-D,It may further comprise one or more polymers or copolymers selected from the group consisting of L-lactide-co-glycolide-co-caprolactone, and combinations thereof.,

[0157] In some embodiments, the polymer can include PEG copolymers such as PEG / poly(L-lactide), PEG / poly(D,L-lactide), PEG / poly(caprolactone), PEG / poly(L-lactide-co-glycolide), PEG / poly(sulfobetaine methacrylate), PEG diacrylate, and PEG dimethacrylate.,

[0158] In some embodiments of the compositions disclosed herein, the composition can further comprise a biodegradable glass or ceramic such as calcium phosphate, and other biocompatible metal oxides (i.e., CaO), or a combination of biodegradable ceramics, glasses, polymers and copolymers.,

[0159] In some embodiments of the compositions disclosed herein, the collagen described herein has an isoelectric point (pI) of about 4.5 to about 12, such as about 5.0 to about 11.5, about 5.5 to about 11.0, about 6.0 to about 10.5, about 6.5 to about 10.0, about 7.0 to about 9.5, about 7.5 to about 9.0, about 8.0 to about 8.5, about 8.5 to about 11.5, about 9 to about 11, about 9 to about 10.5, about 9 to about 10, about 9 to about 9.5, in some embodiments about 9.2, and in some embodiments about 9.66. In some embodiments of the compositions disclosed herein, the composition has an isoelectric point (pI) of about 1 to about 12.0, about 4.0 to about 5.0, about 5.0 to about 6.0, about 6.0 to about 7.0, about 7.0 to about 8.0, about 8.0 to about 9.0, about 9.0 to about 10.0, or about 9.0 to about 9.5, or about 9.5 to about 10.0.,

[0160] In some embodiments, HA can have a molecular weight compatible with living tissue. In some embodiments, the collagen can have a molecular weight of 5 kDa to 150 kDa, 10 kDa to 120 kDa or about 30 kDa to about 60 kDa.,

[0161] In some embodiments, the molecular weight of HA in the compositions described herein can range from about 10 kDa to about 10,000 kDa, from about 25 kDa to about 5,000 kDa, or from about 50 kDa to about 3,000 kDa. In some embodiments, the compositions disclosed herein contain hyaluronic acid or a salt thereof having an average molecular weight in the range of about 300 kDa to about 3,000 kDa, about 400 kDa to about 2,500 kDa, about 500 kDa to about 2,000 kDa, and about 500 kDa to about 1,000 kDa.

[0162] In some embodiments, the molecular weight of PVP in the compositions described herein can range from about 2.0 kDa to about 6,000 kDa, from about 2.5 kDa to about 2,500 kDa, from about 10 kDa to about 2,000 kDa, from about 40 kDa to about 1,500 kDa, from about 100 kDa to about 1,000 kDa.

[0163] In some embodiments, the molecular weight of PAM in the compositions described herein can range from about 10 kDa to about 7,000 kDa and can be purchased from various sources such as Sigma - Aldrich.

[0164] In some embodiments, the molecular weight of PEO in the compositions described herein can range from about 5 kDa to about 300 kDa, from about 50 kDa to about 200 kDa.

[0165] In some embodiments, the molecular weight of poly(2 - ethyl - 2 - oxazoline) in the compositions described herein can range from about 10 kDa to about 1,000 kDa, from about 50 kDa to about 500 kDa.

[0166] In some embodiments, the molecular weight of PEI in the compositions described herein can range from about 2.5 kDa to about 2,500 kDa.

[0167] In some embodiments, the molecular weight of carboxymethyl cellulose in the compositions described herein can range from about 10 kDa to about 300 kDa. In some embodiments, the degree of substitution of carboxymethyl cellulose in the compositions can range from about 1.00 to about 1.25. In some embodiments, carboxymethyl cellulose can have a molecular weight of about 250 kDa and a degree of substitution of about 0.70 to about 0.90. In some embodiments, carboxymethyl cellulose can have a molecular weight of about 250 kDa and a degree of substitution of about 0.79. In some embodiments, carboxymethyl cellulose can have a degree of substitution of about 0.87.

[0168] In some embodiments, the carboxymethyl cellulose in the compositions described herein can have a high viscosity. In some embodiments, the carboxymethyl cellulose in the compositions described herein can have a viscosity of about 100 to about 5,000 cps for a 1% aqueous solution. In some embodiments, the carboxymethyl cellulose in the compositions described herein can have a viscosity of about 1,000 to about 3000 cps for a 1% aqueous solution. In some embodiments, the carboxymethyl cellulose in the compositions described herein can have a viscosity of about 2,700 cps for a 1% aqueous solution.

[0169] In some embodiments, the molecular weight of chondroitin sulfate in the compositions can range from about 20 kDa to about 80 kDa. In some embodiments, the molecular weight of chondroitin sulfate in the compositions can be about 50 kDa.

[0170] In certain embodiments of the compositions disclosed herein, hyaluronic acid can have a molecular weight of about 3 kDa to about 10,000 kDa. In some embodiments, hyaluronic acid can have a molecular weight of about 1,000 kDa to about 5,000 kDa. In some embodiments, the hyaluronic acid can have a molecular weight of about 2,000 kDa to about 3,000 kDa.

[0171] In some embodiments of the compositions disclosed herein, hyaluronic acid can be cross-linked. In some embodiments, hyaluronic acid is not cross-linked. In some embodiments of the compositions disclosed herein, collagen can be cross-linked. In some embodiments, collagen is not cross-linked.

[0172] In some embodiments of the compositions disclosed herein, hyaluronic acid can be biphasic cross-linked hyaluronic acid. In some embodiments, hyaluronic acid can be monophasic cross-linked hyaluronic acid.

[0173] In some embodiments, hyaluronic acid in solution can have a pH in the range of about 5.0 to about 12.

[0174] In some embodiments, the hyaluronic acid solution can have a pH in the range of about 5.2 to about 7.2. In some embodiments, hyaluronic acid can have a pH of about 5.0, about 5.2, about 5.5, about 6.0, about 6.5, about 7.0, about 7.2, about 7.5, about 8.0, about 8.5, about 9.0, about 9.5, about 10.0, about 10.5, about 11, about 11.5 or about 12, or any scale of pH therebetween.

[0175] In some embodiments of the compositions disclosed herein, hyaluronic acid and collagen may be present in a ratio (w / w) in the range of about 1:9 to 9:1. In some embodiments, the ratio of hyaluronic acid to collagen (w / w) may be about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, or about 9:1. In some embodiments, the ratio of hyaluronic acid to collagen (w / w) may be 1:9. In some embodiments, the ratio of hyaluronic acid to collagen (w / w) may be 1:3. In some embodiments, the ratio of hyaluronic acid to collagen (w / w) may be 2:3. In some embodiments, the ratio of hyaluronic acid to collagen (w / w) may be 1:1. In some embodiments, the ratio of hyaluronic acid to collagen (w / w) may be 3:2. In some embodiments, the ratio of hyaluronic acid to collagen (w / w) may be 3:1. In some embodiments, the ratio of hyaluronic acid to collagen (w / w) may be 9:1.

[0176] In some embodiments, the collagen in a composition comprising collagen and HA can be one or more recombinant collagen fragments disclosed herein. In some embodiments, one or more collagen fragments can be cross-linked to HA. In some embodiments, one or more collagen fragments can be cross-linked to HA either covalently or non-covalently. In some embodiments, cross-linking results in the formation of covalent cross-links and can be formed by a chemical reaction initiated by a chemical agent, heat, pressure, pH, or irradiation. In some embodiments, non-covalent cross-links can be formed using polyvalent metal ions such as, for example, iron, copper, zinc, calcium, magnesium, barium, and other chelating metal ions. Covalent linkage can be achieved using one or more suitable cross-linking agents such as, for example, glutaraldehyde, carbodiimide, bifunctional amino acids (e.g., lysine), protamine, or albumin. It is also possible to effect cross-linking by amide, ester, or ether linkages. The linking agents used to cross-link HA via covalent linkage are disclosed and discussed elsewhere in this specification.

[0177] In some embodiments, cross-linking can be formed directly between hyaluronic acid and one or more collagens or recombinant collagen fragments at the amino groups of the collagen molecules and the carboxylic acid groups of the hyaluronic acid. In some embodiments, cross-links between HA and collagen molecules can be formed using a linking agent, suitable examples of which are disclosed and discussed elsewhere in this specification. In some embodiments, hyaluronic acid molecules can be cross-linked to other hyaluronic acid molecules (in addition to, or instead of, cross-links between HA and collagen molecules). In still further embodiments, one or more of the collagen molecules can be cross-linked to another collagen molecule (in addition to, or instead of, cross-links between HA and collagen molecules).

[0178] In some embodiments, the compositions disclosed herein can include a crosslinked polymer matrix. In some embodiments, the crosslinked polymer matrix can be a homogeneous hydrogel composition prepared by methods known in the art under conditions where collagen and HA are soluble in an aqueous solution.

[0179] In some embodiments of the compositions disclosed herein, the collagen may not contain hydroxyproline (Hyp) residues. In some embodiments, the collagen can contain from about 1% to about 20% hydroxyproline (Hyp) residues, based on the total number of prolines in the collagen. In some embodiments, the collagen described herein may not contain about 20%, about 8% to about 20%, about 10% to about 20%, or about 12% to about 20% hydroxyproline. In some embodiments, the compositions disclosed herein can contain about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% hydroxyproline. The hydroxyproline content can be measured by methods known in the art, for example, by liquid chromatography - mass spectrometry (LC - MS).

[0180] In some embodiments of the compositions disclosed herein, the collagen does not contain any hydroxyl groups. In some embodiments of the compositions disclosed herein, the collagen can contain from about 1 to about 120 hydroxyl groups. The number of hydroxyl groups can be measured by methods known in the art, for example, by mass spectrometry.

[0181] In some embodiments, the collagen does not contain any amine groups. In some embodiments, the collagen can contain from about 1 to about 100 amine groups. The number of amine groups can be measured by methods known in the art, for example, by mass spectrometry and colorimetric TNBSA reagent assay.

[0182] In some embodiments, collagen does not contain any carboxylic acid groups. In some embodiments, collagen can contain from about 1 to about 80 carboxylic acid groups. The number of carboxylic acid groups can be measured by methods known in the art.

[0183] The compositions disclosed herein can be prepared by a method that includes crosslinking hyaluronic acid and collagen. In some embodiments, hyaluronic acid and collagen can be crosslinked under suitable conditions.

[0184] In some embodiments, the composition has a viscosity of from about 1 cps to about 100,000 cps. In some embodiments, the composition has a viscosity of from about 10,000 cps to about 80,000 cps. In some embodiments, the composition has a viscosity of from about 50,000 cps to about 70,000 cps.

[0185] In some embodiments, the composition can include collagen that is fully miscible with hyaluronic acid and at least one pharmaceutically or cosmetically acceptable excipient. In some embodiments, the composition can include a recombinant collagen fragment according to SEQ ID NO: 1, and at least one pharmaceutically or cosmetically acceptable excipient. In a further embodiment, the composition can include a recombinant collagen fragment having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 87.5%, at least about 90%, at least about 92.5%, at least about 95%, at least about 97.5%, at least about 98%, at least about 99%, or 100% sequence identity or similarity to SEQ ID NO: 1, wherein the recombinant collagen fragment is fully miscible with hyaluronic acid, and at least one pharmaceutically or cosmetically acceptable excipient. In yet a further embodiment, the composition can include a recombinant collagen fragment having about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% identity or similarity to SEQ ID NO: 1, wherein the recombinant collagen fragment is fully miscible with hyaluronic acid, and at least one pharmaceutically or cosmetically acceptable excipient.In a further embodiment, the composition can comprise a recombinant collagen fragment having about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% identity or similarity to SEQ ID NO: 1, having the amino acid sequence set forth in any one of SEQ ID NOS: 975 - 1002, being completely miscible with hyaluronic acid, and at least one pharmaceutically or cosmetically acceptable excipient.

[0186] In some embodiments, the composition can comprise collagen that is completely miscible with hyaluronic acid and at least one excipient suitable for use in dietary supplements, such as nutritional supplements. In some embodiments, the composition can comprise a recombinant collagen fragment having about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% sequence identity or similarity to SEQ ID NO: 1, being completely miscible with hyaluronic acid, and at least one excipient suitable for use in dietary supplements, such as nutritional supplements. In a further embodiment, the composition can comprise a recombinant collagen fragment having about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% sequence identity or similarity to SEQ ID NO: 1, having the amino acid sequence set forth in any one of SEQ ID NOS: 975 - 1002, being completely miscible with hyaluronic acid, and at least one excipient suitable for use in dietary supplements, such as nutritional supplements.

[0187] In some embodiments, the composition can include a hydrolyzate of a collagen fragment, the hydrolyzate having a sequence that is part of SEQ ID NO: 1, and at least one pharmaceutically acceptable or cosmetically acceptable excipient. In some embodiments, the composition can include a hydrolyzate having a sequence set forth in one or more of SEQ ID NOs: 2-973, and at least one pharmaceutically acceptable or cosmetically acceptable excipient. In some embodiments, the composition can include any of the hydrolyzates set forth in SEQ ID NOs: 2-973, and at least one pharmaceutically acceptable or cosmetically acceptable excipient. In some embodiments, the composition can include a recombinant collagen fragment having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 87.5%, at least about 90%, at least about 92.5%, at least about 95%, at least about 97.5%, at least about 98%, at least about 99%, or 100% sequence identity or similarity to one of SEQ ID NOs: 2-973, and at least one pharmaceutically acceptable or cosmetically acceptable excipient.

[0188] In yet another embodiment, the composition can comprise a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolyzate having a sequence according to one of SEQ ID NOs: 2-973, and at least one pharmaceutically acceptable or cosmetically acceptable excipient. In yet further embodiments, the composition can comprise a recombinant collagen fragment according to SEQ ID NO: 1, a plurality of hydrolyzates having the same or different sequences according to any of SEQ ID NOs: 2-973, and at least one pharmaceutically acceptable or cosmetically acceptable excipient. In certain embodiments, the number of hydrolyzates present among the plurality of hydrolyzates in the composition can increase over time as a result of temperature, pH, or other conditions that cause hydrolysis or other degradation of a recombinant collagen fragment such as the recombinant collagen fragment according to SEQ ID NO: 1. In other embodiments, the composition can be stabilized with one or more stabilizers such that the concentration of the recombinant collagen fragment according to SEQ ID NO: 1 and the concentration of each fragment among the plurality of fragments remains substantially constant (i.e., varies by no more than ±5% by HPLC over a given period) or remains constant. In certain embodiments, a recombinant collagen fragment such as the recombinant collagen fragment according to SEQ ID NO: 1 can be hydrolyzed such that less than about 10%, about 10% - about 20%, about 20% - about 30%, about 30% - about 40%, about 40% - about 50%, about 50% - about 60%, about 60% - about 70%, about 70% - about 80%, about 80% - about 90%, or about 90% - about 100% non-hydrolyzed recombinant fragment remains in the composition as measured by HPLC. In other embodiments, the composition can comprise a mixture of a recombinant collagen fragment (e.g., a recombinant collagen fragment according to SEQ ID NO: 1) and a plurality of hydrolyzates of the recombinant collagen fragment (e.g., a plurality of collagen fragments according to any of SEQ ID NOs: 2-973) such that the weight of the hydrolyzates in the composition is less than about 10%, about 10% - about 20%, about 20% - about 30%, about 30% - about 40%, about 40% - about 50%, about 50% - about 60%, about 60% - about 70%, about 70% - about 80%, about 80% - about 90%, or about 90% - about 100% of the weight of the collagen-related proteins in the composition.

[0189] In some embodiments, the present disclosure provides a composition comprising one or more recombinant collagen fragment sequence variants disclosed herein and at least one pharmaceutically acceptable or cosmetically acceptable excipient. In certain embodiments, the composition can comprise a recombinant collagen fragment sequence variant as set forth in any one of SEQ ID NOs: 975-1002 and at least one pharmaceutically acceptable or cosmetically acceptable excipient. In further embodiments, the composition can comprise a recombinant collagen fragment sequence variant having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 87.5%, at least about 90%, at least about 92.5%, at least about 95%, at least about 97.5%, at least about 98%, or at least about 99% sequence identity or similarity to SEQ ID NO: 1 and at least one pharmaceutically acceptable or cosmetically acceptable excipient. In still further embodiments, the composition can comprise a recombinant collagen fragment sequence variant having about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity or similarity to SEQ ID NO: 1 and at least one pharmaceutically acceptable or cosmetically acceptable excipient. In still further embodiments, the composition can comprise a recombinant collagen fragment sequence variant having about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity or similarity to SEQ ID NO: 1 and at least one excipient suitable for use in dietary supplements, such as nutritional supplements.

[0190] In some embodiments, the composition can include a recombinant collagen fragment having an amino acid chain length of about 150 amino acids, about 200 amino acids, about 250 amino acids, about 300 amino acids, about 350 amino acids to about 600 amino acids that overlap with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the composition includes a recombinant collagen fragment having a length of about 350 amino acids, about 370 amino acids, about 390 amino acids, about 400 amino acids, about 420 amino acids, about 440 amino acids, about 460 amino acids, about 480 amino acids, about 500 amino acids, about 510 amino acids, about 520 amino acids, about 530 amino acids, about 540 amino acids, about 550 amino acids, about 560 amino acids, about 570 amino acids, about 580 amino acids, about 590 amino acids, or about 600 amino acids. In certain embodiments, the composition includes a recombinant collagen fragment having an amino acid chain length of 528 amino acids. In certain embodiments, the composition includes a recombinant collagen fragment having an amino acid chain length of 546 amino acids.

[0191] In some embodiments, the composition can include a recombinant collagen fragment sequence variant as set forth in any one of SEQ ID NOs: 975 to 1002, having a length of about 150 amino acids, about 200 amino acids, about 250 amino acids, about 300 amino acids, about 350 amino acids, about 370 amino acids, about 390 amino acids, about 400 amino acids, about 420 amino acids, about 440 amino acids, about 460 amino acids, about 480 amino acids, about 500 amino acids, about 510 amino acids, about 520 amino acids, about 530 amino acids, about 540 amino acids, about 550 amino acids, about 560 amino acids, about 570 amino acids, about 580 amino acids, about 590 amino acids, or about 600 amino acids. In certain embodiments, the composition includes a recombinant collagen fragment having an amino acid chain length of 528 amino acids. In certain embodiments, the composition includes a recombinant collagen fragment having an amino acid chain length of 546 amino acids.

[0192] In some embodiments, the composition can comprise a recombinant collagen fragment having an amino acid chain length of from about 150 amino acids to about 600 amino acids or a sequence variant thereof, and can have at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 87.5%, at least about 90%, at least about 92.5%, at least about 95%, at least about 97.5%, at least about 98%, at least about 99% or 100% sequence identity or similarity to SEQ ID NO: 1. In some embodiments, such collagen fragments described herein can have a length of about 150 amino acids, about 200 amino acids, about 250 amino acids, about 300 amino acids, about 350 amino acids, about 370 amino acids, about 390 amino acids, about 400 amino acids, about 420 amino acids, about 440 amino acids, about 460 amino acids, about 480 amino acids, about 500 amino acids, about 510 amino acids, about 520 amino acids, about 530 amino acids, about 540 amino acids, about 550 amino acids, about 560 amino acids, about 570 amino acids, about 580 amino acids, about 590 amino acids, or about 600 amino acids.

[0193] In some embodiments, the composition can comprise a hydrolysis product of a collagen fragment, and the hydrolysis product can have an amino acid chain length of from about 10 amino acids to about 75 amino acids. In some embodiments, the composition can comprise a hydrolysis product of a collagen fragment having an amino acid chain length of from about 20 amino acids to about 50 amino acids. In some embodiments, the hydrolysis products described herein can have a length of about 10 amino acids, about 15 amino acids, about 20 amino acids, about 25 amino acids, about 30 amino acids, about 35 amino acids, about 40 amino acids, about 45 amino acids, about 50 amino acids, about 55 amino acids, about 60 amino acids, about 65 amino acids, about 70 amino acids, or about 75 amino acids.

[0194] In some embodiments, the composition can comprise from about 5 ppm to about 500 ppm of a recombinant collagen fragment or a sequence variant thereof. In some embodiments, the composition can comprise about 5 ppm, about 10 ppm, about 25 ppm, about 50 ppm, about 100 ppm, about 150 ppm, about 200 ppm, about 250 ppm, about 300 ppm, about 350 ppm, about 350 ppm, about 400 ppm, about 450 ppm, or about 500 ppm of a recombinant collagen fragment. In some embodiments, the composition can comprise about 5 ppm, about 10 ppm, about 25 ppm, about 50 ppm, about 100 ppm, about 150 ppm, about 200 ppm, about 250 ppm, about 300 ppm, about 350 ppm, about 350 ppm, about 400 ppm, about 450 ppm, or about 500 ppm of a recombinant collagen fragment sequence variant. In some embodiments, the composition can comprise about 5 ppm, about 10 ppm, about 25 ppm, about 50 ppm, about 100 ppm, about 150 ppm, about 200 ppm, about 250 ppm, about 300 ppm, about 350 ppm, about 350 ppm, about 400 ppm, about 450 ppm, or about 500 ppm of a recombinant collagen fragment having the sequence according to SEQ ID NO: 1. In some embodiments, the composition can comprise about 5 ppm, about 10 ppm, about 25 ppm, about 50 ppm, about 100 ppm, about 150 ppm, about 200 ppm, about 250 ppm, about 300 ppm, about 350 ppm, about 350 ppm, about 400 ppm, about 450 ppm, or about 500 ppm of one or more hydrolysis products of a recombinant collagen fragment. In some embodiments, the composition can comprise about 5 ppm, about 10 ppm, about 25 ppm, about 50 ppm, about 100 ppm, about 150 ppm, about 200 ppm, about 250 ppm, about 300 ppm, about 350 ppm, about 350 ppm, about 400 ppm, about 450 ppm, or about 500 ppm of one or more hydrolysis products of a recombinant collagen fragment having the sequence according to SEQ ID NO: 1.In some embodiments, the composition can comprise a recombinant collagen sequence variant as set forth in any one of SEQ ID NOs: 987 to 1015 at about 5 ppm, about 10 ppm, about 25 ppm, about 50 ppm, about 100 ppm, about 150 ppm, about 200 ppm, about 250 ppm, about 300 ppm, about 350 ppm, about 350 ppm, about 400 ppm, about 450 ppm, or about 500 ppm. In some embodiments, the composition can comprise a mixture of a recombinant collagen fragment or a variant thereof at a concentration of about 5 ppm to about 500 ppm and one or more hydrolysis products of the recombinant collagen fragment at a concentration of about 5 ppm to about 500 ppm. In some embodiments, the composition can comprise a mixture of a recombinant collagen fragment having a sequence according to SEQ ID NO: 1 or a variant thereof at a concentration of about 5 ppm to about 500 ppm and one or more hydrolysis products of the recombinant collagen fragment at a concentration of about 5 ppm to about 500 ppm.

[0195] In some embodiments, the composition can be prepared using a recombinant collagen fragment solution that is from about 0.0001 volume % to about 20 volume % and from about 0.5 weight % to about 25 weight %. In some of these embodiments, the composition can comprise from about 0.0001%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20 volume % of the recombinant collagen fragment solution. In some of these embodiments, the recombinant collagen fragment solution comprises from about 0.5 weight %, about 0.6 weight %, about 0.7 weight %, about 0.8 weight %, about 0.9 weight %, about 1 weight %, about 2 weight %, about 3 weight %, about 4 weight %, about 5 weight %, about 6 weight %, about 7 weight %, about 8 weight %, about 9 weight %, about 10 weight %, about 11 weight %, about 12 weight %, about 13 weight %, about 14 weight %, about 15 weight %, about 16 weight %, about 17 weight %, about 18 weight %, about 19 weight %, about 20 weight %, about 21 weight %, about 22 weight %, about 23 weight %, about 24% or about 25 weight % of recombinant collagen fragments, or from about 0.5 weight %, about 0.6 weight %, about 0.7 weight %, about 0.8 weight %, about 0.9 weight %, about 1 weight %, about 2 weight %, about 3 weight %, about 4 weight %, about 5 weight %, about 6 weight %, about 7 weight %, about 8 weight %, about 9 weight %, about 10 weight %, about 11 weight %, about 12 weight %, about 13 weight %, about 14 weight %, about 15 weight %, about 16 weight %, about 17 weight %, about 18 weight %, about 19 weight %, about 20 weight %, about 21 weight %, about 22 weight %, about 23 weight %, about 24 weight %, about 25% weight % or about 40 weight % of recombinant collagen fragment sequence variants.

[0196] In some of these embodiments, the recombinant collagen fragment solution comprises about 0.0001 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, about 25 wt%, or about 40 wt% of collagen fragments having the sequence according to SEQ ID NO: 1. In some of these embodiments, the recombinant collagen fragment solution comprises one or more hydrolysis products of recombinant collagen fragments of about 0.0001 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, about 25 wt%, or about 40 wt%. In some of these embodiments, the recombinant collagen fragment solution comprises one or more hydrolysis products of recombinant collagen fragments of about 0.0001 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt%, or about 40 wt% having the sequence described in one or more of SEQ ID NOs: 2-973.In some of these embodiments, the recombinant collagen fragment sequence variant solution comprises one or more recombinant collagen fragment sequence variants having the sequences set forth in any one of SEQ ID NOs: 975 to 1002, in an amount of about 0.0001 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt% or about 25 wt% or about 40 wt%.

[0197] In some of these embodiments, the composition can be prepared using a mixture of about 0.0001 vol% to about 35 vol% of about 0.0005 wt% to about 40 wt% of a recombinant collagen fragment and one or more hydrolysis products of the recombinant collagen fragment. In some of these embodiments, the composition can be prepared using a mixture of about 0.0001 vol% to about 35 vol% of about 0.0005 wt% to about 40 wt% of a recombinant collagen fragment having the sequence according to SEQ ID NO: 1 and one or more hydrolysis products of the recombinant collagen fragment.

[0198] In some embodiments, the composition can be prepared using a mixture of a recombinant collagen fragment having the sequence according to SEQ ID NO: 1 and one or more hydrolysis products of the recombinant collagen fragment, in an amount of about 0.0001% to about 35% by volume and about 0.0005% to about 40% by weight. In some embodiments, the composition can be prepared using a solution containing a mixture of recombinant collagen fragment sequence variants having the sequences described in any one of SEQ ID NOs: 2 to 973, in an amount of about 0.0001% to about 35% by volume and about 0.0005% to about 40% by weight. In some embodiments, the composition can be prepared using a solution containing a mixture of a recombinant collagen fragment having the sequence according to SEQ ID NO: 1 and one or more hydrolysis products of the recombinant collagen fragment, in an amount of about 0.0001% to about 35% by volume and about 0.0005% to about 40% by weight, and a recombinant collagen fragment sequence variant solution containing a sequence variant containing the sequence according to any one of SEQ ID NOs: 975 to 1002.

[0199] In some of these embodiments, the recombinant collagen fragment and the recombinant collagen fragment sequence variant solution contain one or more hydrolysis products of the recombinant collagen fragment having the sequences described in one or more of SEQ ID NOs: 2 to 973, in an amount of about 0.0001% by weight, about 0.2% by weight, about 0.3% by weight, about 0.4% by weight, about 0.5% by weight, about 0.6% by weight, about 0.7% by weight, about 0.8% by weight, about 0.9% by weight, about 1% by weight, about 2% by weight, about 3% by weight, about 4% by weight, about 5% by weight, about 6% by weight, about 7% by weight, about 8% by weight, about 9% by weight, about 10% by weight, about 11% by weight, about 12% by weight, about 13% by weight, about 14% by weight, about 15% by weight, about 16% by weight, about 17% by weight, about 18% by weight, about 19% by weight, about 20% by weight, about 21% by weight, about 22% by weight, about 23% by weight, about 24% by weight or about 25% by weight or about 40% by weight.

[0200] In some embodiments, the composition can comprise from about 0.0005 wt% to about 25 wt% of recombinant collagen fragments, from about 0.001 wt% to about 25 wt% of recombinant collagen fragments, from about 0.01 wt% to about 25 wt% of recombinant collagen fragments, from about 0.1 wt% to about 25 wt% of recombinant collagen fragments, from about 0.5 wt% to about 20 wt% of recombinant collagen fragments, from about 0.7 wt% to about 17 wt% of recombinant collagen fragments, from about 1 wt% to about 15 wt% of recombinant collagen fragments, from about 2 wt% to about 12 wt% of recombinant collagen fragments, from about 2 wt% to about 10 wt% of recombinant collagen fragments, from about 3 wt% to about 9 wt% of recombinant collagen fragments, from about 4 wt% to about 8 wt% of recombinant collagen fragments, or from about 5 wt% to about 7 wt% of recombinant collagen fragments.

[0201] In some embodiments, the composition can comprise about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt% of recombinant collagen fragments. In some embodiments, the composition can comprise about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt% of collagen fragments having the sequence according to SEQ ID NO: 1.

[0202] In some embodiments, the composition can include one or more hydrolysis products of recombinant collagen fragments at about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt%. In some embodiments, the composition can include one or more hydrolysis products of recombinant collagen fragments having a sequence according to one or more of SEQ ID NOs: 2-973 at about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt%.

[0203] In some embodiments, the composition can comprise a mixture of recombinant collagen fragments and one or more hydrolysis products of the recombinant collagen fragments at about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt%. In some embodiments, the cosmetic composition can comprise a mixture of a recombinant collagen fragment having the sequence according to SEQ ID NO: 1 and one or more hydrolysis products of the recombinant collagen fragment at about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt%.

[0204] In some embodiments, the composition can comprise from about 0.0005 wt% to about 25 wt% of a recombinant collagen fragment sequence variant, from about 0.001 wt% to about 25 wt% of a recombinant collagen fragment sequence variant, from about 0.01 wt% to about 25 wt% of a recombinant collagen fragment sequence variant, from about 0.1 wt% to about 25 wt% of a recombinant collagen fragment sequence variant, from about 0.5 wt% to about 20 wt% of a recombinant collagen fragment sequence variant, from about 0.7 wt% to about 17 wt% of a recombinant collagen fragment sequence variant, from about 1 wt% to about 15 wt% of a recombinant collagen fragment sequence variant, from about 2 wt% to about 12 wt% of a recombinant collagen fragment sequence variant, from about 2 wt% to about 10 wt% of a recombinant collagen fragment sequence variant, from about 3 wt% to about 9 wt% of a recombinant collagen fragment sequence variant, from about 4 wt% to about 8 wt% of a recombinant collagen fragment sequence variant, or from about 5 wt% to about 7 wt% of a recombinant collagen fragment sequence variant.

[0205] In some embodiments, the composition can comprise from about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt% of a recombinant collagen fragment sequence variant. In some embodiments, the composition can comprise from about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt% of a collagen fragment sequence variant having a sequence according to any one of SEQ ID NOs: 975 to 1002.

[0206] In some embodiments, the composition can comprise from about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt% of recombinant collagen fragments and one or more recombinant collagen fragment sequence variants. In some embodiments, the composition can comprise a mixture comprising from about 0.0005 wt%, about 0.001 wt%, about 0.01 wt%, about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, or about 25 wt% of a recombinant collagen fragment having the sequence according to SEQ ID NO: 1 and one or more collagen fragment sequence variants having the sequence according to any one of SEQ ID NOs: 975 - 1002.

[0207] In some embodiments, the composition can include a recombinant collagen fragment having the amino acid sequence set forth in SEQ ID NO: 1, one or more recombinant collagen fragments having a sequence according to one or more of SEQ ID NOs: 2 to 973, and / or one or more collagen fragment sequence variants having a sequence according to any one of SEQ ID NOs: 975 to 1002, and hyaluronic acid, wherein the recombinant collagen fragment or variant is completely miscible with hyaluronic acid. In some embodiments, the composition can include a recombinant collagen fragment having at least about 40% amino acid sequence identity to SEQ ID NO: 1 and / or one or more collagen fragment sequence variants having at least about 40% amino acid sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 975 to 1002, and hyaluronic acid, wherein the recombinant collagen fragment or variant is completely miscible with hyaluronic acid. In some embodiments, the hyaluronic acid can be a biphasic cross-linked hyaluronic acid comprising cross-linked HA and non-cross-linked HA.

[0208] In some embodiments, the composition can include a recombinant collagen fragment having the amino acid sequence set forth in SEQ ID NO: 1, one or more recombinant collagen fragments having a sequence according to one or more of SEQ ID NOs: 2 to 973, and / or one or more collagen fragment sequence variants having a sequence according to any one of SEQ ID NOs: 975 to 1002, and polyvinylpyrrolidone, wherein the recombinant collagen fragment or variant is completely miscible with polyvinylpyrrolidone. In some embodiments, the composition can include one or more recombinant collagen fragments having at least about 40% amino acid sequence identity to SEQ ID NO: 1 and / or one or more collagen fragment sequence variants having at least about 40% amino acid sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 975 to 1002, and polyvinylpyrrolidone, wherein the recombinant collagen fragment or variant is completely miscible with polyvinylpyrrolidone.

[0209] In some embodiments, the composition can include a recombinant collagen fragment having the amino acid sequence set forth in SEQ ID NO: 1, one or more recombinant collagen fragments having a sequence according to one of SEQ ID NOs: 2-973, and / or one or more collagen fragment sequence variants having a sequence according to any one of SEQ ID NOs: 975-1002, and sodium carboxymethyl cellulose, wherein the recombinant collagen fragment or variant is completely miscible with sodium carboxymethyl cellulose. In some embodiments, the composition can include one or more recombinant collagen fragments having at least about 40% amino acid sequence identity to SEQ ID NO: 1, and / or one or more collagen fragment sequence variants having at least about 40% amino acid sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 975-1002, and sodium carboxymethyl cellulose, wherein the recombinant collagen fragment or variant is completely miscible with sodium carboxymethyl cellulose.

[0210] In some embodiments, the compositions described herein comprise hyaluronic acid at a concentration of at least about 1 mg / mL to at least about 100 mg / mL, at least about 1 mg / mL to at least about 50 mg / mL, at least about 5 mg / mL to at least about 25 mg / mL, at least about 6 mg / mL to at least about 24 mg / mL, at least about 7 mg / mL to at least about 23 mg / mL, at least about 8 mg / mL to at least about 22 mg / mL, at least about 9 mg / mL to at least about 21 mg / mL, or at least about 10 mg / mL to at least about 20 mg / mL, and any concentration ranges therebetween. In some embodiments, the compositions described herein comprise hyaluronic acid at a concentration of about 1 mg / mL to about 100 mg / mL, about 1 mg / mL to about 50 mg / mL, about 5 mg / mL to about 25 mg / mL, about 6 mg / mL to about 24 mg / mL, about 7 mg / mL to about 23 mg / mL, about 8 mg / mL to about 22 mg / mL, about 9 mg / mL to about 21 mg / mL, or about 10 mg / mL to about 20 mg / mL, and any concentration ranges therebetween. In some embodiments, the compositions of the present disclosure comprise hyaluronic acid at a concentration of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL or about 100 mg / mL. In some embodiments, the compositions described herein comprise hyaluronic acid at a concentration of about 20 mg / mL to about 25 mg / mL. In certain embodiments, the compositions described herein comprise hyaluronic acid at a concentration of about 24 mg / mL. In some embodiments, the present disclosure provides a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid.

[0211] In certain embodiments, the compositions disclosed herein include one or more collagens or recombinant collagen fragments disclosed herein crosslinked to hyaluronic acid. The collagen can be crosslinked directly to HA, i.e., via an amide or ester functional group, or can be linked to HA via a linker. In some embodiments, the compositions disclosed herein include a hyaluronic acid component that includes hyaluronic acid that has reacted to become part of a crosslinked polymer matrix. In some embodiments, the compositions according to the present disclosure include a collagen component that includes one or more recombinant collagen fragments that have reacted to become part of a crosslinked polymer matrix. In addition to crosslinking between hyaluronic acid and collagen, hyaluronic acid and / or collagen can be self-crosslinked.

[0212] In some embodiments, the composition can be a solution. In some embodiments, the composition can be a gel. In some embodiments, the composition can be a film. In some embodiments, the composition can be one or more fibers. In some embodiments, the composition can be a coating. In some embodiments, the composition can be a powder. In some embodiments, the composition can be a slurry. In some embodiments, the composition can be particles. In some embodiments, the composition can be a foam.

[0213] In some embodiments, the compositions described herein further comprise a pharmaceutically active agent. Exemplary active agents include sunscreen, anti-dandruff agents, moisturizers, depigmenting agents, pro-pigmentants, alpha-hydroxy acids, antibacterial agents, radical scavengers, anti-pollution agents, anti-inflammatory agents, anesthetics, retinoids, algae, mushrooms, vegetables, yeast, bacterial extracts, hydrolyzed, partially hydrolyzed, or non-hydrolyzed proteins, enzymes, hormones, vitamins, and derivatives thereof, flavonoids and isoflavones, and mixtures thereof. Further exemplary pharmaceutical agents include anesthetics such as lidocaine, bupivacaine, mepivacaine, and related compounds, but are not limited thereto. Further exemplary pharmaceutical agents include non-steroidal anti-inflammatory drugs (NSAIDs) such as flurbiprofen, ibuprofen, naproxen, indomethacin, and related compounds, but are not limited thereto. In some embodiments, the composition can further comprise one or more drugs including colchicine, taxol, and related compounds. In some embodiments, the composition can further comprise one or more topical disinfectants such as, for example, benzoyl peroxide. In some embodiments, the composition can further comprise one or more polysaccharides produced by microalgae such as, for example, alguronic acid. In some embodiments, the composition can further comprise one or more bioactive peptides such as, for example, palmitoyl tripeptide-1 and palmitoyl tetrapeptide-7. In some embodiments, the composition can further comprise one or more polysaccharides produced by microalgae such as, for example, alguronic acid.

[0214] E. Method of Use Dermal Filler In some embodiments, the present disclosure provides a dermal filler comprising a composition comprising collagen and hyaluronic acid disclosed herein. In some embodiments, the dermal filler disclosed herein can comprise any known type of collagen that is miscible with HA. In some embodiments, the dermal filler provided herein can comprise any known type of collagen that is completely miscible with HA. In some embodiments, administration of the dermal filler described herein can increase the production of type I collagen. In some embodiments, administration of the dermal filler described herein can increase the production of type III collagen. In some embodiments, administration of the dermal filler described herein can increase the production of any combination of type I collagen, type III collagen, or ECM.

[0215] In some embodiments, the dermal fillers disclosed herein can include one or more recombinant collagen fragments disclosed herein. In certain embodiments, the dermal filler can include a recombinant collagen fragment according to SEQ ID NO:1 and hyaluronic acid. In further embodiments, the dermal filler can include a recombinant collagen fragment having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 87.5%, at least about 90%, at least about 92.5%, at least about 95%, at least about 97.5%, at least about 98%, at least about 99% or 100% sequence identity or similarity to SEQ ID NO:1, and hyaluronic acid. In still further embodiments, the composition can include a recombinant collagen fragment having about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% identity or similarity to SEQ ID NO:1, and hyaluronic acid. In some embodiments, administration of the dermal filler comprising a recombinant collagen fragment according to SEQ ID NO:1 and hyaluronic acid as described herein can increase the production of type I collagen. In some embodiments, administration of the dermal filler comprising a recombinant collagen fragment according to SEQ ID NO:1 and hyaluronic acid as described herein can increase the production of type III collagen. In some embodiments, administration of the dermal filler comprising a recombinant collagen fragment according to SEQ ID NO:1 and hyaluronic acid as described herein can increase the production of ECM. In some embodiments, administration of the dermal filler comprising a recombinant collagen fragment according to SEQ ID NO:1 and hyaluronic acid as described herein can increase the production of any combination of type I collagen, type III collagen, and ECM.

[0216] In some embodiments, the dermal filler can comprise a hydrolyzate of a collagen fragment, and the hydrolyzate can have a sequence that is part of SEQ ID NO: 1 and hyaluronic acid. In some embodiments, the composition comprises a hydrolyzate having a sequence set forth in any one of SEQ ID NOs: 2-973 and hyaluronic acid. In some embodiments, the composition can comprise any of the hydrolyzates set forth in SEQ ID NOs: 2-973 and hyaluronic acid. In some embodiments, the composition can comprise a recombinant collagen fragment having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 87.5%, at least about 90%, at least about 92.5%, at least about 95%, at least about 97.5%, at least about 98%, at least about 99% or 100% sequence identity or similarity to any one of SEQ ID NOs: 2-973, and hyaluronic acid.

[0217] In some embodiments, the dermal filler can comprise a hydrolyzate of a collagen fragment, and the hydrolyzate can have a sequence that is part of SEQ ID NO: 1 as described herein and hyaluronic acid, and can increase the production of type I collagen. In some embodiments, administration of a dermal filler comprising a hydrolyzate of a collagen fragment as described herein, wherein the hydrolyzate can have a sequence that is part of SEQ ID NO: 1, and hyaluronic acid can increase the production of type III collagen. In some embodiments, administration of a dermal filler comprising a hydrolyzate of a collagen fragment as described herein, which can have a sequence that is part of SEQ ID NO: 1, and hyaluronic acid can increase the production of both type I collagen and type III collagen.

[0218] In yet another embodiment, the dermal filler can comprise a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysis product having a sequence according to one of SEQ ID NOs: 2-973, and hyaluronic acid. In still further embodiments, the dermal filler can comprise a recombinant collagen fragment according to SEQ ID NO: 1, a plurality of hydrolysis products having the same or different sequences according to any of SEQ ID NOs: 2-973, and hyaluronic acid. In certain embodiments, the number of hydrolysis products present among the plurality of hydrolysis products in the dermal filler can increase over time, with temperature, pH, or as a result of other conditions that typically cause hydrolysis or other degradation of recombinant collagen fragments such as the recombinant collagen fragment according to SEQ ID NO: 1. In other embodiments, the dermal filler can be stabilized with one or more stabilizers such that the concentration of the recombinant collagen fragment according to SEQ ID NO: 1 and the concentration of each fragment among the plurality of fragments remains substantially constant (i.e., varies by no more than ±5% by HPLC over a given period) or remains constant. In certain embodiments, a recombinant collagen fragment such as the recombinant collagen fragment according to SEQ ID NO: 1 can be hydrolyzed such that when measured by HPLC, less than about 10%, about 10% - about 20%, about 20% - about 30%, about 30% - about 40%, about 40% - about 50%, about 50% - about 60%, about 60% - about 70%, about 70% - about 80%, about 80% - about 90%, or about 90% - about 100% of the non-hydrolyzed recombinant fragment remains in the composition. In other embodiments, the dermal filler can comprise a mixture of a recombinant collagen fragment (e.g., a recombinant collagen fragment according to SEQ ID NO: 1) and a plurality of hydrolysis products of the recombinant collagen fragment (e.g., a plurality of collagen fragments according to any of SEQ ID NOs: 2-973) such that the weight of the hydrolysis products in the composition is less than about 10%, about 10% - about 20%, about 20% - about 30%, about 30% - about 40%, about 40% - about 50%, about 50% - about 60%, about 60% - about 70%, about 70% - about 80%, about 80% - about 90%, or about 90% - about 100% of the weight of the collagen-related proteins in the dermal filler, and hyaluronic acid.

[0219] In yet another embodiment, the dermal filler can include a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysis product having a sequence according to one of SEQ ID NOs: 2 to 973, and hyaluronic acid. In some embodiments, administration of the dermal filler described herein, which includes a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysis product having a sequence according to one of SEQ ID NOs: 2 to 973, and hyaluronic acid, can increase the production of type I collagen. In some embodiments, administration of the dermal filler described herein, which includes a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysis product having a sequence according to one of SEQ ID NOs: 2 to 973, and hyaluronic acid, can increase the production of type III collagen. In some embodiments, administration of the dermal filler described herein, which includes a recombinant collagen fragment according to SEQ ID NO: 1 and hyaluronic acid, can increase the production of both type I collagen and type III collagen.

[0220] In some embodiments, the dermal filler can include a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NOs: 975 to 1002, and hyaluronic acid.

[0221] In some embodiments, the dermal filler can further include a bioabsorbable polymer selected from the group consisting of a solid homopolymer of poly(ε-caprolactone), a solid homopolymer of poly(p-dioxanone), a solid homopolymer of poly(trimethylene carbonate), a solid copolymer of a plurality of ε-caprolactone repeating units and a third lactone repeating unit, and a solid copolymer of a plurality of trimethylene carbonate repeating units and a second lactone repeating unit; the third lactone repeating unit is selected from the group consisting of a glycolide repeating unit, a lactide repeating unit, a trimethylene carbonate repeating unit, a p-dioxanone repeating unit, a 1,4-dioxepan-2-one repeating unit, a 1,5-dioxepan-2-one repeating unit, and combinations thereof.

[0222] In some embodiments, the dermal filler can further include one or more polymers including, but not limited to, aliphatic polyesters, polyorthoesters, polyoxy esters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyesteramides, hydrolyzable amines, polyalkylene oxides, polyethylene oxide (PEO), poloxamers, functionalized isoflavonoids, amino acids, and combinations thereof.

[0223] In some embodiments, the dermal filler is poly(hydroxyethyl methacrylate), poly(sulfobetaine), poly(dimethylsiloxane), poly(caprolactone), poly(p-dioxanone), poly(trimethylene carbonate), poly(glycolide), poly(L-lactide), poly(D,L-lactide), poly(L-lactide-co-glycolide), poly(D,L-lactide-co-glycolide), poly(L-lactide-co-caprolactone), poly(L-lactide-co-D,L-lactide), poly(caprolactone-co-trimethylene carbonate), poly(lactide-co-trimethylene carbonate), poly(caprolactone-co-p-dioxanone), poly(trimethylene carbonate-p-dioxanone), poly(caprolactone-co-lactide), poly(lactide-co-1,5-dioxepan-2-one) and poly(1,5-dioxepan-2-one-co-p-dioxanone), poly(lactide-co-1,4-dioxepan-2-one), poly(1,4-dioxepan-2-one-co-p-dioxanone), poly(glycolide-co-caprolactone-co-dioxanone), poly(lactide-co-caprolactone-co-dioxanone), poly(dioxanone-co-D,L-lactide), poly(dioxanone-co-glycolide), poly(dioxanone-co-caprolactone), poly(dioxanone-co-L-lactide), poly(dioxanone-co-D,L-lactide-co-caprolactone), poly(dioxanone-co-glycolide-co-caprolactone), poly(dioxanone-co-L-lactide-co-D,L-lactide), poly(dioxanone-co-L-lactide-co-glycolide), poly(dioxanone-co-L-lactide-do-caprolactone), poly(dioxanone-co-D,L-lactide-co-glycolide), poly(dioxanone-co-D,L-lactide-co-caprolactone), poly(dioxanone-co-glycolide-co-caprolactone), poly(dioxanone-co-L-lactide-co-glycolide-co-caprolactone), poly(dioxanone-co-D,It can further include one or more polymers selected from the group consisting of L-lactide-co-glycolide-co-caprolactone, and combinations thereof.,

[0224] In some embodiments, the dermal filler can further include PEG in combination with one or more polymers provided herein, examples of which include, but are not limited to, PEG / poly(L-lactide), PEG / poly(D,L-lactide), PEG / poly(caprolactone), PEG / poly(L-lactide-co-glycolide), PEG / poly(sulfobetaine methacrylate), PEG diacrylate, and PEG dimethacrylate.

[0225] In some embodiments, the dermal filler can further include a biodegradable glass or ceramic such as calcium phosphate, and other biocompatible metal oxides (i.e., CaO), or a combination of biodegradable ceramics, glasses, and polymers.

[0226] In some embodiments, administration of a dermal filler comprising a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NOs: 975-1002 and hyaluronic acid as described herein can increase the production of type I collagen. In some embodiments, administration of a dermal filler comprising a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NOs: 975-1002 and hyaluronic acid as described herein can increase the production of type III collagen. In some embodiments, administration of a dermal filler comprising a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NOs: 975-1002 and hyaluronic acid can increase the production of both type I and type III collagen.

[0227] In some embodiments, the dermal filler can include a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysis product having a sequence according to one of SEQ ID NOs: 2-973, a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NOs: 975-1002, and hyaluronic acid. In some embodiments, administration of a dermal filler comprising a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysis product having a sequence according to one of SEQ ID NOs: 2-973, a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NOs: 975-1002, and hyaluronic acid as described herein can increase the production of both type I and type III collagen.

[0228] In some embodiments, the dermal filler is a heat-stable, sterilizable soft tissue filler. In some embodiments, the dermal filler is heat-stable up to at least about 125°C. In some embodiments, the dermal filler is heat-stable at temperatures in the range up to at least about 135°C. As used herein, a dermal filler is "heat-stable" which means that the dermal filler is substantially stable when subjected to heat treatment such as steam sterilization (autoclaving) under normal or elevated pressure. The stability of the dermal fillers disclosed herein can be evaluated by methods known in the art. The heat-stable dermal fillers disclosed herein can retain one or more desired rheological properties that were present prior to heat treatment. The heat stability of the dermal fillers disclosed herein can prevent the degradation of collagen, collagen fragments, and / or collagen variants contained within the dermal filler. In some embodiments, the dermal filler can be ultimately sterilized using different sterilization techniques. In some embodiments, the dermal filler can be sterilized by autoclaving, by gamma irradiation, by e-beam irradiation, or by any method known in the art.

[0229] In some embodiments, the dermal filler is injectable. Typically, an injectable dermal filler is administered by subcutaneous or intradermal injection. The term "effective amount" refers to an amount of the injectable dermal filler sufficient to produce a beneficial or desired cosmetic or therapeutic result. In some embodiments, the dermal filler is injected in a volume of about 0.1 mL, about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, about 0.6 mL, about 0.7 mL, about 0.8 mL, about 0.9 mL, about 1.0 mL, about 1.5 mL, about 2.0 mL, about 2.5 mL, about 3.0 mL, about 3.5 mL, about 4.0 mL, about 4.5 mL, about 6.0 mL, about 6.5 mL, about 7.0 mL, about 7.5 mL, about 8.0 mL, about 8.5 mL, about 9.0 mL, about 9.5 mL, about 10.0 mL, about 20.0 mL, about 30.0 mL, about 40.0 mL, about 50.0 mL, about 60.0 mL, about 70.0 mL, about 80.0 mL, about 90.0 mL, about 100.0 mL, and any amount therebetween.

[0230] In some embodiments, the dermal filler can further comprise calcium phosphate particles. Suitable calcium phosphate particles include, but are not limited to, calcium hydroxyapatite, calcium fluoroapatite, calcium chloroapatite, calcium carbonate apatite, tetracalcium phosphate, calcium pyrophosphate, tricalcium phosphate, and octacalcium phosphate particles, and combinations thereof. In some embodiments, the dermal filler can further comprise calcium hydroxyapatite particles. In some embodiments, the dermal filler can further comprise carboxymethyl cellulose. In some embodiments, the dermal filler can further comprise polylactic acid.

[0231] In some embodiments, the particles can have a particle size in the range of about 10 μm to about 60 μm, and in certain embodiments about 25 μm to about 45 μm. In some embodiments, the particles can have a particle size less than 60 μm, less than 45 μm, less than 35 μm, or less than 25 μm.

[0232] In some embodiments, the dermal fillers disclosed herein are suitable for injection via a fine gauge needle. In some embodiments, the injectable dermal filler is suitable for administration using a needle having a size of from about 16 gauge (Ga) to about 33 Ga. In some embodiments, the composition has a size of from about 22 to about 30.

[0233] In some embodiments, the dermal fillers disclosed herein may be provided in a prefilled syringe. Without wishing to be bound by any particular theory, it is believed that the synergistic effect of the components in the dermal fillers described herein results in a formulation in which calcium phosphate particles, such as calcium hydroxyapatite particles, are stably suspended over time without undergoing phase separation.

[0234] In some embodiments, the dermal fillers described herein can be injected into any area of the body or face where an increase in tissue volume is desired. For example, suitable areas of the body and face include the glabellar line, nasolabial fold, mental groove, marionette line, lateral commissure, perioral wrinkles, crow's feet, skin depressions, scars, forehead, subcutaneous support of the eyebrows, cheekbones and buccal fat pads, tear troughs, nose, lips, cheeks, perioral region, infraorbital region, facial asymmetry, jaw line, and chin. For example, suitable areas of the body include the buttocks, breasts, calves, flanks, or any other area of the body where tissue volume is desired for aesthetic purposes or to address body asymmetry. Further, an increase in tissue volume can have therapeutic benefits. For example, when the dermal fillers described herein are injected into the vocal cords, bladder wall or neck, or any other anatomical tubular structure, sphincter or valve, a therapeutic benefit can be achieved that provides enhanced function when enhanced by the dermal fillers described herein.

[0235] In some embodiments, the dermal fillers disclosed herein further comprise a pharmaceutically active agent. In some embodiments, the pharmaceutically active agent comprises an anesthetic. In some embodiments, the anesthetic is lidocaine, bupivacaine, mepivacaine, or any combination thereof.

[0236] Medical or cosmetic treatment In some embodiments, the disclosure provides a method of medical or cosmetic treatment comprising administering or applying to a subject in need thereof an effective amount of the compositions described herein. In some embodiments, the treatment comprises tissue enhancement. In some embodiments, the treatment comprises filling facial wrinkles or increasing deposition. In some embodiments, the treatment comprises forming a cosmetic implant in a patient.

[0237] The compositions disclosed herein are contemplated for any condition or disease for which administration or application of a mixture of collagen and hyaluronic acid may be beneficial. Such conditions include, but are not limited to: orthopedic conditions where internal cushioning is desired (e.g., between bone-to-bone contact), osteoarthritis, arthritis, age-related eye conditions, etc. In some embodiments, the compositions disclosed herein can be administered to the eyes, blood vessels, nerves, cartilage, bone, breasts, vocal cords, feet, or tympanic membranes, joints, intervertebral discs, buttocks, hip joints, gastrocnemius, or hands of a subject.

[0238] In some embodiments, the treatment comprises treating an inflammatory skin disease or mucosal disease.

[0239] In some embodiments, the treatment comprises treating lipodystrophy or other natural or surgical loss of tissue volume.

[0240] Increase in collagen production in cells The inventors have surprisingly discovered that the collagen fragments described in the disclosure can induce the production of type I and type III collagen in cells such as fibroblasts. Thus, when contacted with cells such as fibroblasts or other suitable cells, the collagen fragments described herein (i.e., collagen fragments having the amino acid sequence of SEQ ID NO: 1, or compositions comprising such fragments and hyaluronic acid) can advantageously induce collagen formation in the cells with which they are contacted.

[0241] Thus, in some embodiments, the present disclosure provides a method of increasing collagen production in a cell, the method comprising contacting the cell with a composition described herein. Such contact can consist of directly exposing the cell to an amount of the composition sufficient to induce the cell to produce increased amounts of collagen. In some embodiments, the method comprises administering or applying a composition comprising collagen or a collagen fragment, hyaluronic acid, and at least one pharmaceutically acceptable excipient. In some embodiments, the composition can be applied to the cells in their natural context, such as in a living organism or tissue. In some embodiments, the composition can be applied topically to a skin region. In some embodiments, the composition can be applied topically to human skin. In some embodiments, as described above, the composition can be injected.

[0242] In some embodiments, the cell can be a fibroblast. In some embodiments, the cell can be a fibroblast in its natural context (e.g., within human skin). In some embodiments, the cell can be a primary human fibroblast. In some embodiments, the cell can be a muscle cell, a transformed human cell, a cardiomyocyte, an epithelial cell, an endothelial cell, a stem cell, an induced pluripotent stem cell, an intervertebral disc nucleus pulposus cell, a synovial cell, an osteoblast, a chondrocyte, a keratinocyte, or a corneal stromal cell. In some embodiments, the cell can be a cell that produces insulin. In some embodiments, the cell can be a pancreatic beta cell.

[0243] In some embodiments, the compositions disclosed herein can be applied to wounds such as incisions, lacerations, gashes, tears, excoriations, abrasions, burns, or scratches to increase collagen production in the wound site and / or the surrounding tissue. As part of the wound healing process, fibroblasts migrate to the wound site and produce the collagen necessary for wound repair. The cells eventually fill the wound cavity with a network of interwoven collagen threads and eventually arrange themselves into a firm band to form permanent new tissue. Thus, the compositions containing the collagen, collagen fragments, and collagen variants described herein can be applied to wounds such as incisions, lacerations, gashes, tears, excoriations, abrasions, burns, or scratches to increase collagen production in the wound site and / or the surrounding tissue.

[0244] In some embodiments, the wound is a chronic wound. Chronic wounds include, but are not limited to, diabetic foot ulcers, venous leg ulcers, and pressure ulcers.

[0245] In certain embodiments, the present disclosure provides a method of applying a composition comprising collagen or recombinant collagen fragments, hydrolysis products of recombinant collagen fragments, or combinations thereof, and hyaluronic acid, to a wound such as an incision, laceration, tear, scratch, abrasion, burn, or scald. In certain embodiments, the wound can be a wound in a human subject. In some embodiments, the method comprises providing a composition for collagen production and wound healing and promoting collagen production and wound healing in the skin of a subject in need thereof, the composition comprising a recombinant collagen fragment described herein (e.g., a fragment comprising SEQ ID NO: 1). In some embodiments, the method comprises providing a composition for collagen production and wound healing and promoting collagen production and wound healing in the skin of a subject in need thereof, the composition comprising one or more hydrolysis products of a recombinant collagen fragment described herein (e.g., a hydrolysis product comprising one or more of SEQ ID NOs: 2-973). In some embodiments, the method comprises providing a composition for collagen production and wound healing and promoting collagen production and wound healing in the skin of a subject in need thereof, the composition comprising a mixture of a recombinant collagen fragment described herein and one or more hydrolysis products of the recombinant collagen fragment (e.g., a mixture of a fragment comprising SEQ ID NO: 1 and a hydrolysis product comprising SEQ ID NOs: 2-973).

[0246] In some embodiments, the method can include administering to the cells a collagen fragment having the amino acid sequence of SEQ ID NO: 1 and hyaluronic acid. In some embodiments, the method can include administering to the cells a hydrolysis product resulting from the hydrolysis of a collagen fragment having the amino acid sequence of SEQ ID NO: 1 (e.g., administering one or more hydrolysis products having the sequences according to SEQ ID NOs: 2 to 973). In some embodiments, the method includes administering to the cells a recombinant collagen fragment described herein and one or more hydrolysis products of the recombinant collagen fragment. In some embodiments, the method can include administering to the cells a collagen fragment having the amino acid sequence of SEQ ID NO: 1 and one or more hydrolysis products of the fragment having the sequences according to SEQ ID NOs: 2 to 973. In some embodiments, the method can include administering to the cells a collagen fragment having any one of the amino acid sequences of SEQ ID NOs: 975 to 1002 and hyaluronic acid.

[0247] In some embodiments, the method can increase the production of type I collagen. In some embodiments, the method can increase the production of type III collagen. In some embodiments, the method can increase the production of ECM. In some embodiments, the method can increase the production of any combination of type I collagen, type III collagen, and ECM.

[0248] Skin care In some embodiments, the compositions described herein can be used to improve the aesthetic appearance of the skin and / or its appendages, such as the surface appearance and / or texture of the skin. In some embodiments, the compositions described herein can be formulated for use on the body and face, hands, and feet, including the treatment of the eye area, nails, and hair. The term "surface appearance" means visual and / or tactile irregularities on the skin and / or scalp, including wrinkles and fine lines, expression lines in the area between the forehead and eyebrows, wrinkles and / or fine lines around the mouth, and / or looseness in the area around the lips and the upper lip area (the area located between the upper lip and the nose), unevenness in skin color (liver spots, senile lentigines), the appearance and / or visibility of pores, a paper-like appearance of the skin, defects in skin microrelief such as varicella scars or acne scars, and defects in seborrheic skin (such as a shiny appearance). The term "skin texture" can mean loose, non-tightened, non-tensed, non-elastic skin, and / or sagging skin.

[0249] In some embodiments, the compositions described herein can be used to improve the aesthetic appearance of the skin, including improving the appearance of expression lines. Expression lines are caused by the effect of stress exerted on the skin by underlying muscles. Environmental factors such as age and exposure to sunlight can deepen expression lines and make them permanent. Expression lines are characterized by the presence of grooves around openings formed by the nose (nasolabial groove), mouth (perioral lines and so-called bitter lines), and eyes (crow's feet) where the surrounding skin muscles are located, as well as in the area between the eyebrows (glabella or lion's wrinkle) and the forehead.

[0250] In some embodiments, the compositions described herein can be used to improve the aesthetic appearance of the skin and / or the visibility of pores. The visibility of pores can be due to excessive sebum, aging, loss of tension, looseness, stress, fatigue, improper hygiene, climatic factors, or any combination thereof. The compositions described herein can tighten pores and make them less visible.

[0251] In some embodiments, the compositions described herein can be used to improve the appearance of the skin like paper and the behavior of the skin upon touch. In particular, older skin can visually exhibit the appearance of cigarette paper and may give an appearance similar to that of a single piece of blotting paper. The appearance like that of skin paper can be seen on the face and backs of the hands of the elderly.

[0252] In some embodiments, the compositions described herein can be compositions for protecting, treating, or caring for the face, hands, feet, or body, such as, for example, day creams, night creams, makeup remover creams, anti-sunlight compositions, body milks for skin protection or care, after-sun milks, skin care lotions, gels, foams, artificial tanning compositions, and compositions for after shaving. In some embodiments, the compositions described herein can be, for example, solutions, suspensions, lotions, creams, serums, gels, balms, gels, oils, oil-in-cream, micellar water, face mists, face essences, blemish balms or color-correcting formulas, toners (water and / or alcohol-based), paints, finishing agents, sticks, pencils, sprays, aerosols, ointments, cleansing liquid detergents, solid bars, shampoos, hair conditioners, hair styling products, pastes, foams, powders, mousses, balms, shaving creams, wipes, strips, patches, wound dressings, band-aids, hydrogels, film-forming products, face and skin masks, cosmetics (e.g., foundations, eyeliners, eyeshadows), exfoliators, deodorants, and antiperspirants, etc., and can be formulated as such. Exemplary formulations are provided herein.

[0253] In some embodiments, the compositions described herein can be cosmetic compositions, and at least one excipient can be a cosmetically acceptable excipient. A cosmetically acceptable excipient is an excipient suitable for use in cosmetics. Exemplary cosmetically acceptable excipients are described below.

[0254] In some embodiments, the cosmetic compositions described herein can include a hydrolyzed product of a recombinant collagen fragment and at least one excipient, such as a cosmetically acceptable excipient. In some embodiments, the cosmetic compositions described herein can include a recombinant collagen fragment, one or more hydrolyzed products of the recombinant collagen fragment, and at least one excipient, such as a cosmetically acceptable excipient.

[0255] In some embodiments, the cosmetic compositions can include ingredients commonly used in cosmetics, such as skin care, eye care, nail care, and hair care products. These ingredients include, but are not limited to, soap, antibacterial agents, anti-inflammatory agents, humectants, waxy alcohols, hydrating agents, moisturizers, penetration enhancers, emulsifiers, natural or synthetic oils, solvents, fats, surfactants, detergents, gelling agents, skin softeners, antioxidants, fragrances, paints, brighteners, fillers, thickeners, waxes, odor absorbers, dyes, colorants, powders, viscosity regulators, analgesics, anesthetics, anti-itch agents, plant extracts, conditioning agents, darkening or whitening agents, water retention agents, mica, minerals, polyphenols, silicones or silicone derivatives, such as dimethicone, sunscreens, vitamins, plant-derived pharmaceuticals, alcohols, such as denatured alcohol and ethanol, polyols, polyol ethers, and other ingredients listed in the International Cosmetic Ingredient Dictionary and Handbook, 13th Ed. (2009) (incorporated herein by reference in its entirety). Other ingredients listed in (2009) (incorporated herein by reference in its entirety) may be included, but are not limited to these. In certain embodiments, a given ingredient can perform two or more functions and can belong to two or more classes.

[0256] In some embodiments, the compositions described herein can be therapeutic compositions, and at least one excipient can be a pharmaceutically acceptable excipient. The therapeutic compositions can be useful for treating one or more conditions such as reducing or preventing the formation of scar tissue, promoting healing, promoting tissue regeneration, minimizing local inflammation, minimizing tissue rejection, and / or enhancing skin and / or hair graft integration. A pharmaceutically acceptable excipient is an excipient that can act as a vehicle or medium for the active substance and includes excipients commonly used in therapeutic compositions, i.e., compositions useful for treating one or more conditions. Exemplary pharmaceutically acceptable excipients are described below.

[0257] In some embodiments, the compositions described herein can be dietary compositions or dietary supplements, and at least one excipient can be, for example, a food or beverage additive. A “dietary supplement” is a preparation intended to supplement the diet and can be useful for providing nutrients or additives that may be lacking or not consumed in sufficient amounts in a person's diet. In some embodiments, the dietary supplement can be in the form of any commonly used solid or liquid dosage form for oral administration, including but not limited to capsules, tablets, pills, powders, granules or powders, soft gelatin capsules and hard gelatin capsules, and / or can be provided as gummies. Suitable excipients include, but are not limited to, lactose or milk sugar and high molecular weight polyethylene glycol. Exemplary dietary compositions are described below.

[0258] The compositions described herein can also include one or more of the following additional components. Exemplary contemplated additional components are described below. However, the present disclosure is not limited to these exemplary additional components.

[0259] In some embodiments, the compositions described herein can further include one or more anti-wrinkle agents. An anti-wrinkle agent is a compound that, when the composition contacts an area of wrinkled skin on the body or face, including for example the area around the eyes, results in an increase in the synthesis and / or activity of certain skin enzymes, thereby reducing the appearance of wrinkles and / or fine lines. Exemplary anti-wrinkle agents include, but are not limited to, exfoliants, anti-glycation agents, nitric oxide synthase inhibitors, muscle relaxants and / or anti-skin contraction agents, agents against free radicals, and mixtures thereof.

[0260] Further exemplary anti-wrinkle agents that can be included in the compositions described herein include adenosine and its derivatives, retinol and its derivatives (e.g., retinyl palmitate), ascorbic acid and its derivatives (e.g., magnesium ascorbyl phosphate and ascorbyl glucoside), tocopherol and its derivatives (e.g., tocopheryl acetate), nicotinic acid and its precursors (e.g., nicotinamide), ubiquinone, glutathione and its precursors (e.g., L-2-oxothiazolidine-4-carboxylic acid), C-glycoside compounds (also known as C-glycosyl compounds) and their derivatives (e.g., β-C-xylosyl derivatives having the trade name PRO-XYLANE), plant extracts (e.g., rock sanfire extract and olive leaf extract), vegetable proteins and their hydrolysates (e.g., rice or soybean protein hydrolysates), algal extracts (e.g., kelp extract), bacterial extracts, saponins (e.g., diosgenin), yam (Dioscorea) extracts (e.g., natural yam extract), α-hydroxy acids, β-hydroxy acids (e.g., salicylic acid and 5-(n-octanoyl)salicylic acid), oligopeptides and pseudodipeptides and their acylated derivatives (e.g., {2-[acetyl(3-(trifluoromethyl)phenyl)amino]-3-methylbutyrylamino}acetic acid), lipopeptides, e.g., MATRIXYL 3000 available from Croda, lycopene, manganese, magnesium salts (e.g., gluconate), and any combination of the foregoing, but not limited thereto.

[0261] Exemplary adenosine derivatives include, but are not limited to, 2'-deoxyadenosine; 2',3'-iso-propylidene adenosine; toyocamycin, 1-methyladenosine; N-6-methyladenosine, adenosine N-oxide, 6-methyl-mercaptopurine riboside, and 6-chloropurine riboside. Other adenosine derivatives include phenylisopropyladenosine (PIA) / 1-methylisoguanosine, Ns-cyclohexyladenosine (CHA), N6-cyclopentyladenosine (CPA), 2-chloro-Ns-cyclopentyl-adenosine, 2-chloroadenosine, N6-phenyladenosine, 2-phenylaminoadenosine, MECA, Ne-phenethyladenosine, 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethylcarboxamide-adenosine (CGS-21680), (N-ethylcarboxamide)adenosine-S-(NECA), 5'-(N-cyclopropylcarboxamide)adenosine, DPMA (PD 129,944), and metrifudil, including adenosine receptor agonists.

[0262] In some embodiments, the compositions described herein can include one or more adenosine derivatives that increase the intracellular concentration of adenosine, such as erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), iodotubercidin, or combinations thereof. Additional adenosine derivatives contemplated herein include adenosine salts and alkyl esters of adenosine.

[0263] In some embodiments, the composition can further comprise one or more nacreous pearlescent agents. The nacreous pearlescent agents are specifically iridescent particles of any shape produced within the shells by certain crustaceans. Alternatively, the nacreous pearlescent agents can be synthesized, i.e., artificial. The nacreous pearlescent agents can be, but are not limited to, white nacreous pearlescent agents such as mica coated with titanium oxide or bismuth oxychloride, nacreous pearlescent agents based on bismuth oxychloride, titanium oxide coated mica coated with iron oxide, specifically titanium oxide coated mica coated with ferric blue or chromium oxide, or colored nacreous pearlescent agents such as titanium oxide coated mica coated with an organic pigment.

[0264] In some embodiments, the composition can further comprise one or more hydroxy acids. Examples of hydroxy acids include beta-hydroxy acids such as salicylic acid and acetylsalicylic acid. Additional exemplary hydroxy acids suitable for use in the composition include citric acid, glycolic acid, hydroxycaproic acid, hydroxycaprylic acid, lactic acid, malic acid, tartaric acid, polyhydroxy acids including gluconolactone, and any combination thereof.

[0265] In some embodiments, the composition can further comprise one or more emulsifiers. The emulsifier prevents different components (such as oil and water) from separating in the emulsion. Suitable emulsifiers include, but are not limited to, polysorbate, laureth-4, potassium lauryl sulfate, glyceryl caprylate, and any combination thereof.

[0266] In some embodiments, the composition can further comprise one or more chelating agents. The chelating agent binds to metal ions and prevents them from chemically reacting with other substances in the formulation. Suitable chelating agents include, but are not limited to, sodium phytate, disodium EDTA, tetrasodium EDTA, tetrasodium glutamate diacetate, and trisodium ethylenediamine disuccinate.

[0267] In addition to the chelating agent, disodium glutamate diacetate can also act as a stabilizer in any of the compositions described herein.

[0268] In some embodiments, the composition can further comprise one or more antibacterial agents. Suitable antibacterial agents include, but are not limited to, caprylyl glyceryl ether, benzalkonium chloride, benzethonium chloride, and chloroxylenol (PCMX), tea tree oil, witch hazel, rosemary oil, lemon oil, and any combination thereof.

[0269] In some embodiments, the composition can further comprise one or more humectants (moisture retainers) to improve the level of moisture in the skin. Non-limiting examples of humectants suitable for use in the compositions described herein are described in WO 98 / 22085, WO 98 / 18444, and WO 97 / 01326, and include amino acids and their derivatives, such as proline and arginine aspartate, 1,3-butylene glycol, propylene glycol, pentylene glycol, water, imosemyl extract, creatinine, diglycerol, biosaccharide gum-1, glucamine salts, glucuronates, glutamate salts, polyethylene glycol ethers of glycerin (e.g., glycereth 20), glycerin, glycerol monopropoxylate, glycogen, hexylene glycol, honey, hydrolyzed hydrogenated starch, hydrolyzed mucopolysaccharides (such as xanthan gum and biosaccharide gum-1), inositol, keratin amino acids, glycosaminoglycans, methoxy PEG 10, methyl gluceth-10 and -20, methyl glucose, 3-methyl-1,3-butanediol, N-acetylglucosamine salts, polyethylene glycol and its derivatives (PEG 15 butanediol, PEG 4, PEG 5 pentaerythitol, PEG 6, PEG 8, PEG 9, etc.), propanediol, pentaerythitol, 1,2 pentanediol, PPG-1 glyceryl ether, 2-pyrrolidone-5-carboxylic acid (including its salts and esters), isomerized sugar, sericin, silk amino acids, sodium acetyl hyaluronate, sodium hyaluronate, sodium poly-aspartate, sodium polyglutamate, caprylyl glycol, sorbeth 20, sorbeth 6, sugars and sugar alcohols and their derivatives, such as glucose, mannose and polyglycerol sorbitol, trehalose, triglycerol, trimethyolpropane, tris(hydroxymethyl)aminomethane salts, and yeast extract, and mixtures thereof.

[0270] Additional humectants suitable for use in the present specification include polyhydric alcohols selected from the group consisting of glycerin, diglycerin, glycerol, erythritol, arabitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, maltitol, mannose, inositol, triethylene glycol, sodium pyrrolidone carboxylate (pyrrolidone carboxylic acid, PCA), zinc PCA, and derivatives and mixtures thereof.

[0271] In some embodiments, the composition can further include a gelling agent comprising a polyacrylamide-based polymer. The polyacrylamide-based polymer can be polyacrylamide itself, its derivatives, a mixture of multiple types of polymers, or a copolymer of acrylamide and its derivatives as monomers. The gelling agent can be used to provide a good appearance to the skin, give a cooling sensation to the skin, and provide a non-sticky and refreshing feeling to the skin. In some embodiments, the gelling agent includes one or more pigments or one or more fillers, such as extender pigments, coloring pigments, and whitening pigments, including inorganic pigments, organic pigments, nacreous pigments, polymer powders, functional pigments, talc, mica, kaolin, calcium carbonate, magnesium carbonate, anhydrous silicic acid, aluminum silicate, magnesium silicate, calcium silicate, aluminum oxide, barium sulfate, red iron oxide, yellow iron oxide, black iron oxide, chromium oxide, ultramarine blue, Prussian blue, carbon black, zinc oxide, mica titanium, fish scale foil, bismuth oxychloride, boron nitride, nylon powder, silk powder, carbomer, tar pigments, natural pigments, and titanium oxide, for example, including amorphous or rutile and / or anatase type crystals.

[0272] In some embodiments, the composition can include pigments and / or fillers that are resistant to water and oil, and can further include any conventionally used water repellent and / or oil repellent agent, such as a fluorine compound, to impart water repellency and oil repellency to the pigments. Representative fluorine compounds that have been conventionally used and can act as water repellent and oil repellent agents include compounds having perfluoroalkyl groups, such as perfluororalkyl phosphate, perfluoroalkylsilane, perfluoroalkylsilazane, polyhexafluoropropylene oxide, perfluoroalkyl group-containing organosiloxane, perfluoropolyether, perfluoroalcohol, perfluoroalkyl acrylate polymer, and derivatives thereof. Perfluororalkyl phosphate can provide uniform and stable dispersion of the pigments within the formulation of the gel composition, and perfluoroalkylsilane can have very good compatibility with other cosmetic components. Also, perfluororalkyl phosphate - diethanolamine salt commercially available as AsahiGuard AG530 by Asahi Glass, and perfluoroalkylsilane coupling agents such as LP-IT and LP-4T of Shin-Etsu Silicone can be used.

[0273] Representative gelling agents include, but are not limited to, those commercially available under the trade names Sepigel 305, Sepigel 501, and Sepigel 600 by Seppic. Sepigel 305 contains approximately 40% polyacrylamide, approximately 24% -C 13 ~C 14 isoparaffin, and approximately 6% laureth-7 (where laureth-7 is the formula C 12 H 25 --(OCH2CH2) n--OH (wherein n has an average value of 7), which is a mixture containing a nonionic surfactant. Sepigel 600 is a mixture of acrylamide / acrylamide-2-propanesulfonic acid copolymer, isododecane, and polysorbate 80 (polyoxyethylene sorbitan monooleate (20 EO)). Suitable gelling agents containing polyacrylamide-based polymers are disclosed in European Patent No. 0503853 (Scott Bader Company Ltd.), the disclosure of which is incorporated herein by reference.

[0274] In some embodiments, the composition can further comprise hyaluronic acid (HA). In some embodiments, the HA can be in an uncrosslinked state. In some embodiments, the HA can be in a crosslinked state. Similar to collagen, HA is an important structural component of human tissue. Hyaluronan, also known as hyaluronic acid (HA), is an unsulfated glycosaminoglycan that is widely distributed throughout the human body in connective tissue, epithelial tissue, and nerve tissue. Hyaluronan is abundant in different layers of the skin and has multiple functions, such as ensuring good hydration, assisting in the organization of the ECM, acting as a filler material, and participating in tissue repair mechanisms. However, with aging, the amount of hyaluronan present in the skin decreases.

[0275] In some embodiments, the composition described herein can be a dermal filler composition, such as an injectable dermal filler composition. The dermal filler composition can be suitable for use on the face and body, including, for example, around the eyes, above or around the cheeks, above or around the décolletage, above or around the hands, above or around the nails, above or around the ears including the earlobes, above or around the legs, and above or around the feet.

[0276] In some embodiments, the compositions described herein can be used with a microneedle array, such as an array contained in a sheet or patch. The microneedle array can include a plurality of microneedles that are long enough to penetrate the skin through the stratum corneum and into the viable epidermis. In some embodiments, it may be desirable for cosmetic or therapeutic purposes to deliver collagen to the region of the epidermal / dermal junction.

[0277] In some embodiments, the compositions described herein can be used with a microneedle sheet or patch. Microneedles and microneedle patches are suitable for delivering collagen to the epidermis and dermis of human skin on the face and body, including, for example, the eyes, cheeks, lips, décolletage, and hands. In some embodiments, the microneedles for delivering the composition to the epidermis and dermis in a targeted manner are injectable microneedles, drug-coated metal microneedles, or microneedles with dissolvable tips. Exemplary methods and disclosures regarding microneedles can be found, for example, in Aditya et al., Kinetics of collagen microneedle drug delivery system, Journal of Drug Delivery Science and Technology, vol. 52, pp. 618 - 623 (August 2019) and Sun et al., Transdermal Delivery of Functional Collagen Via Polyvinylpyrrolidone Microneedles, Ann. Biomed. Eng., 43(12):2978 - 2990 (2015), each of which is incorporated by reference in its entirety. In some embodiments, the compositions described herein can be applied to the skin before, during, or after any skin resurfacing procedure. Skin resurfacing procedures refer to various skin treatments designed to rejuvenate the skin. Skin resurfacing procedures include laser skin resurfacing, chemical peels, microdermabrasion, dermabrasion, microneedling, and Pulsed Light (IPL) technology photorejuvenation. In some embodiments, the composition can further comprise a waxy lipid, such as a ceramide. Ceramide helps to form a barrier to prevent permeability, which helps to prevent dryness and irritation and can also protect the epidermis from environmental damage.

[0278] In some embodiments, the composition can further comprise vitamin A or a vitamin A derivative. Examples of vitamin derivatives include retinoids such as retinal, retinoic acid, retinates, retinyl esters, retinol, tretinoin, isotretinoin, adapalene, tazarotene, etc., but are not limited thereto. The term "retinoid" includes cis and trans derivatives of retinoids (e.g., all-trans-retinoic acid, 13-cis-retinoic acid, 13-trans-retinoic acid, and 9-cis-retinoic acid).

[0279] In some embodiments, the composition can further comprise vitamin C or a derivative thereof, such as ascorbic acid, ascorbates (e.g., ascorbic acid tetrahexyldecyl), etc.

[0280] In some embodiments, the composition can further comprise vitamin B, such as biotin (i.e., vitamin B7), niacinamide, etc.

[0281] In some embodiments, the composition can further comprise vitamin E, such as α-, β-, γ-, and σ-tocopherols and their related corresponding tocotrienols, etc.

[0282] In some embodiments, the composition can further comprise vitamin K and its derivatives.

[0283] Any vitamin, vitamin analog, or derivative thereof that can be suitably formulated as a topical composition is contemplated for the present disclosure.

[0284] In some embodiments, the compositions disclosed herein can further include one or more thickeners. A thickener, i.e., a structure builder, can suspend pigments and / or increase the viscosity in the composition. Suitable thickeners and / or structure builders for the present composition include, but are not limited to, organically modified clays, fumed silica, trihydroxystearin, silicone gels or silicone elastomers, ammonium acryloyldimethyltaurate / VP copolymer, acrylate / C10-30 alkyl acrylate crosspolymer, and mixtures thereof.

[0285] Suitable organically modified clays include, but are not limited to, organically modified forms of hectorite, bentonite, smectite, and montmorillonite clays (such as those sold under the trade names BENTONE® by Elementis Specialties, TIXO-GEL® by Sud-Chemie, and CLAYTONE® by Southern Clay Products). Suitable hydrophobically modified fumed silicas include the WACKER HDK® N20 and T30 grades (Wacker-Chemie AG), and the hydrophilic grades sold under the trade name AEROSIL® (Evonik). Silicone gels or silicone elastomers include, but are not limited to, the "KSG" thickening series (KSG-15, KSG-16, KSG-18, KSG-41, KSG-42, KSG-43, KSG-44) from Shin-Etsu Silicones, the DOW CORNING® 9040, 9041, 9045, and 9546 silicone elastomer blends from Dow Corning, the SFE839™ from Momentive Performance Materials, and the Velvesil™ silicone gels, and the WACKER-BELSIL® RG-100 from Wacker-Chemie AG.

[0286] In some embodiments, the compositions disclosed herein can further comprise one or more fat-soluble / lipid-dispersible film-forming agents. Suitable fat-soluble / lipid-dispersible film-forming agents for use herein include organic silicone resins (e.g., trimethylsiloxysilicates such as SRI 000 from GE Silicones), and copolymers of organic silicone resins (e.g., diisostearyl trimethylolpropane siloxysilicates such as SF1318 from GE Silicones); fluorinated silicone resins; acrylic and / or vinyl polymers or copolymers containing silicone and / or fluorinated types (e.g., the "KP" series of silicone acrylates from Shin-Etsu Silicones, and 3M (trademark) Silicones "Plus" Polymer VS70 and SA70); polyurethanes (e.g., the Poly derm® series derived from hydroxyester triglycerides from Alzo International); polyesters (e.g., the Lexorez® series of polymeric polyesters from Inolex Chemical Company); and mixtures thereof, but are not limited thereto.

[0287] In some embodiments, the compositions disclosed herein can further comprise one or more colorants. Suitable colorants for use herein include all inorganic and organic colorants / pigments, including mineral or pearl pigments suitable for use in cosmetic compositions. Such colorants include those with or without surface coatings or treatments. The colorants can enhance the coloring, and / or light scattering, and / or light reflection effects of the composition.

[0288] In some embodiments, the compositions disclosed herein can further include one or more sunscreens, such as mineral sunscreens and / or physical sunscreens. Sunscreens can block UVA and / or UVB radiation. Exemplary UVA sunscreen agents include, but are not limited to, avobenzone, terephthalylidene dicamphor sulfonic acid, bis-disulzol disodium, phenyl dibenzimidazole tetrasulfonic acid disodium, diethylamino hydroxybenzoyl hexyl benzoate, bis-diethylamino hydroxybenzoyl benzoate, bis-benzooxazolyl phenyl ethylhexyl aminotriazine, and combinations thereof.

[0289] Exemplary UVB sunscreen agents include, but are not limited to, octocrylene, octinoxate, octisalate, homosalate, ensulizole, ethylhexyl triazone, enzacamene, amyloglucoside, diethylhexyl butamido triazine, benzylidene malonate polysiloxane, padimate-O, trolamine salicylate, cinoxate, p-aminobenzoic acid, and derivatives thereof, and combinations thereof.

[0290] Exemplary sunscreen agents that absorb both UVA and UVB radiation include, for example, oxybenzone, meradimate, titanium dioxide, zinc oxide, bis-octrizole, bemotrizinol, droxmetrizole trisiloxane, sulisobenzone, dioxybenzone, or combinations thereof.

[0291] Specific suitable sunscreens include p-aminobenzoic acid, its salts and its derivatives (ethyl, isobutyl, glyceryl esters, p-dimethylaminobenzoic acid, anthranilates (i.e., o-aminobenzoates, methyl, menthyl, phenyl, benzyl, phenylethyl, linalyl, terpinyl, and cyclohexenyl esters), salicylates (amyl, phenyl, benzyl, menthyl, glyceryl, and dipropylene glycol esters), cinnamic acid derivatives (methyl and benzyl esters, alpha-cinnamoyl nitrile, butyl cinnamoyl pyruvate), dihydroxycinnamic acid derivatives (umbelliferone, methyl umbelliferone, methyl acetounbelliferone), trihydroxycinnamic acid derivatives (esculetin, methyl esculetin, daphnetin, and glucosides, esculin and daphnin), hydrocarbons (diphenylbutadiene, stilbene), dibenzalacetone and benzalacetophenone, naphthol sulfonates (sodium salts of 2-naphthol-3,3-disulfonic acid and 2-naphthol-6,8-disulfonic acid), cihydroxynaphthoic acid and its salts, o and p-hydroxybiphenyl disulfonates, coumarin derivatives (7-hydroxy, 7-methyl, 3-phenyl), diazoles (2-acetyl-3-bromindazole, phenylbenzoxazole, methylnaphthoxazole, various arylbenzothiazoles), quinine salts (bisulfate, sulfate, chloride, oleate, and tannate), quinoline derivatives (8-hydroxyquinoline salts, 2-phenylquinoline), hydroxy- or methoxy-substituted benzophenones, uric acid and vilouric acid, tannic acid and its derivatives (e.g., hexaethyl ether), (butylcarbitol)(6-propylpiperonyl) ether, hydroquinone, benzophenones (oxybenzene, sulisobenzone, dioxybenzene, benzoresorcinol, 2,2,4,4'-tetrahydroxybenzophenone, 2,2'-dihydroxy4,4'-dimethoxybenzophenone, octabenzone, 4-isopropyldibenzoylmethane, butyl methoxydibenzoylmethane, etocrylene, and 4-isopropyl-dibenzoylmethane), titanium dioxide, iron oxide, zinc oxide, and mixtures thereof, but are not limited thereto. Other cosmetically acceptable sunscreens and concentrations (weight percent of the total cosmetic sunscreen composition) include diethanolamine methoxycinnamate (10% or less), ethyl-bis(hydroxypropyl)aminobenzoate (5% or less), glyceryl aminobenzoate (3% or less), 4-isopropyldibenzoylmethane (5% or less), 4-methylbenzylidene camphor (6% or less), terephthalylidene dicamphor sulfonic acid (10% or less), and sulisobenzone (also known as benzophenone-4, 10% or less). In some embodiments, the compositions disclosed herein can further comprise D-aspartic acid and / or D-alanine and any salts thereof. As used herein, the term "derivative" of D-aspartic acid and D-alanine refers to D-aspartic acid and D-alanine molecules covalently bonded to any organic group via their amino, carboxyl, or side chains, provided that the effect of D-aspartic acid and D-alanine on promoting collagen production is not impaired. Exemplary organic groups include protecting groups such as N-phenylacetyl group and 4,4'-dimethoxytrityl (DMT) group; biopolymers such as proteins, peptides, saccharides, lipids, and nucleic acids; synthetic polymers such as polystyrene, polyethylene, polyvinyl, polypropylene, and polyester; and functional groups such as ester groups, but are not limited thereto. The ester groups may include, for example, aliphatic esters such as methyl ester and ethyl ester; and aromatic esters.

[0292] In some embodiments, the composition can further comprise one or more common skin care additives such as a conditioning agent, e.g., silicone. In some embodiments, the composition can further comprise one or more shark liver oils, e.g., squalane and / or squalene. In some embodiments, the composition can further comprise one or more polysaccharides produced by microalgae, e.g., alguronic acid.

[0293] In some embodiments, the composition can further comprise at least one preservative. In some embodiments, the at least one preservative can be a quaternary ammonium compound, a halogenated phenol, sorbic acid, potassium sorbate, benzoic acid, sodium benzoate, sodium citrate, sodium anisate, caprylic hydroxamic acid, sodium levulinate, phenoxyethanol, or a combination thereof.

[0294] The compositions described herein can have a pH in the range of about 4 to about 8, about 4.7 to about 5.5, about 5 to about 7, about 6 to about 7, about 6.1 to about 6.8, or about 6.4 to 6.6.

[0295] Soap In some embodiments, the cosmetic compositions described herein can be a cleansing composition or a soap, including conventional soap in the form of a solid bar, and a liquid soap in the form of a cleaner, makeup remover, body wash, milk, cream, foam cream gel, or gel that can be packaged in a tube, bottle, pump bottle, aerosol shower foam, or foam pump bottle. Soap is used in a beauty process for washing away dirt residues of human keratinous substances in the presence of water, can be made into a mass to form foam, and the formed foam and dirt residues are removed by rinsing with water and can be used on any part of the body and face disclosed herein, including, for example, the skin of the body, face, hands, lips, eyelids, nails, hair, eyelashes, and / or eyebrows.

[0296] Traditionally, examples of solid soaps include alkali metal fatty acid salts and potassium fatty acid soaps, and examples of liquid soaps include four major families of detergent formulations: (1) those based on lauryl sulfate, (2) those based on alpha-olefin sulfonate, (3) those based on mixtures of synthetic anionic, amphoteric, and / or nonionic surfactants, and (4) mixed formulations based on soaps and synthetic surfactants. Liquid soaps generally contain a thickening system selected from, for example, electrolytes such as sodium chloride, potassium chloride, or potassium sulfate; alkanolamides such as cocamide DEA or cocamide MEA; polyethylene glycol and esters of monoacids or stearic acid such as polyethylene glycol distearate 6000, or mixtures thereof, and are contained in a cosmetically acceptable aqueous medium. However, both solid soaps and liquid soaps can contain any suitable additional ingredients, such as those listed below herein, in any combination.

[0297] In some embodiments, the cleaning composition or soap described herein can further comprise one or more cellulose-based compounds, or polysaccharide compounds having chains of glucose residues linked by β-1,4 bonds in their structure, one or more fatty acids including straight-chain or branched-chain saturated or unsaturated alkyl chains having 6 to 30 carbon atoms or 12 to 22 carbon atoms, one or more fatty acids including lauric acid, myristic acid, palmitic acid, stearic acid, linolenic acid, and mixtures thereof, and one or more mineral bases including alkali metal hydroxides (sodium hydroxide and potassium hydroxide), metal hydroxides, or ammonia, or organic bases such as triethanolamine, monoethanolamine, monoisopropanolamine, N-methylglucamine, lysine, and arginine.

[0298] In some embodiments, the compositions described herein include sodium salts, ammonium salts, amine salts, amino alcohol salts, or alkaline earth metal salts of the following types, for example, alkali metal salts containing magnesium: alkyl sulfates, alkyl ether sulfates, alkyl amide ether sulfates, alkyl aryl polyether sulfates, monoglyceride sulfates; alkyl sulfonates, alkyl amide sulfonates, alkyl aryl sulfonates, α-olefin-sulfonates, paraffin-sulfonates; alkyl sulfosuccinates, alkyl ether sulfosuccinates, alkyl amide-sulfosuccinates; alkyl sulfoacetates; acyl sarcosinates; and acyl glutamates, alkyl groups and acyl groups of all these compounds having 6 to 24 carbon atoms, and aryl groups representing phenyl or benzyl groups, alkyl glucoside citrates, alkyl polyglycoside tartrates, and polyglycoside carboxylic acids such as alkyl polyglycoside sulfosuccinates C6~C 24 alkyl esters, alkylsulfosuccinamates, acyl isethionates, and N-acyl taurates, alkyl groups or acyl groups of all these compounds having 12 to 20 carbon atoms, and / or acyl lactylates in which the acyl group contains 8 to 20 carbon atoms, and mixtures thereof, may further include one or more anionic surfactants. In some embodiments, alkyl-D-galacturonic acid, polyoxyalkylenated (C6~C 24 ) ether carboxylic acids, polyoxyalkylenated (C6~C 24 ) aryl (C6~C 24 ) polyoxyalkylenated ether carboxylic acids, polyoxyalkylenated (C6~C 24 ) alkyl amide ether carboxylic acids, specifically those containing 2 to 50 ethylene oxide groups; and their alkali metal, ammonium, amine, amino alcohol, or alkaline earth metal salts may also be suitable.

[0299] Suitable C6 - C containing 1 to 30 ethylene oxide groups 24 Examples of alkyl ether sulfates include alkali metal or alkaline earth metal, ammonium, amine or amino alcohol salts, sodium salts, and oxyethylenated (C 12 - C 14 ) alkyl ether sulfates having an average of 1 to 4 ethylene oxide groups and including sodium lauryl sulfate (CTFA name), such as commercially available products sold under the names TEXAPON AOS 225 UP, TEXAPON N702, TEXAPON NSW by COGNIS, or EMPICOL ESB3 / FL2, EMPICOL ESB3 / FL3, EMPICOL ESB70 / FL2 sold by Huntsman.

[0300] Suitable amphoteric surfactants include, but are not limited to, derivatives of secondary or tertiary aliphatic amines in which the aliphatic group is straight - chain or branched - chain and contains 8 to 22 carbon atoms. The amphoteric surfactant can contain at least one water - soluble anionic group, such as a carboxylate, sulfonate, sulfate, phosphate or phosphonate group, (C8 - C 20 ) alkyl betaines, sulfobetaines, (C8 - C 20 ) alkylamide (C6 - C8) alkyl betaines or (C8 - C 20 ) alkylamide alkyl (C6 - C8) sulfobetaines, and mixtures thereof.

[0301] Suitable amine derivatives include those described in U.S. Patent No. 2,528,378 and U.S. Patent No. 2,781,354, and products sold under the name MIRANOL®, which were submitted under the names amphocarboxy-glycinate and amphocarboxypropionate in the CTFA Dictionary, 3rd Edition, 1982. Additional suitable amine derivatives include those classified under the names disodium cocoamphodiacetate, disodium lauroamphodiacetate, disodium capryloamphodiacetate, disodium capryloamphodiacetate, disodium cocoamphodipropionate, disodium lauroamphodipropionate, disodium capryloamphodipropionate, disodium capryloamphodipropionate, lauroamphodipropionate, cocoamphodipropionate, and cocoamphodiacetate in the CTFA Dictionary, 5th Edition, 1993, and sold by Rhodia under the trade name MIRANOL® C2M concentrated. Suitable alkyl (C8-C 20 ) betaines include cocamidopropyl betaine and coco betaine, such as the commercially available products MIRATAINE BB / FLA from RHODIA or EMPIGEN BB / FL from Huntsman.

[0302] In some embodiments, the cleaning compositions or soaps described herein may be sulfate-free and may include a sulfate-free surfactant system.

[0303] In some embodiments, the cleaning compositions or soaps described herein can further include one or more thickeners of the nonionic cellulose-based compound type. Suitable cellulose-based compounds include nonionic cellulose ethers including methylcellulose and ethylcellulose; hydroxyalkylcelluloses such as hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose; mixed hydroxyalkyl-alkylcellulose celluloses such as hydroxypropyl-methylcellulose, hydroxyethyl-methylcellulose, hydroxyethyl ethylcellulose, and hydroxybutyl-methylcellulose, and hydroxyalkylcelluloses modified with alkyl chains, but are not limited thereto. Suitable hydroxypropylmethylcelluloses include the commercially available products METHOCEL E, F, J, and K sold by Dow Coming, and more specifically METHOCEL E 4MQG or METHOCEL F 4M. Suitable cellulose-based components can be in crystalline form, microcrystalline form, or a combination thereof.

[0304] In some embodiments, the cleaning composition or soap described herein may further comprise one or more additional thickeners, including electrolytes such as sodium chloride, potassium chloride, or potassium sulfate; alkanolamides such as cocamide DEA or cocamide MEA; esters of polyethylene glycol and a monoacid or stearic acid such as polyethylene glycol distearate 6000, or mixtures thereof; polysaccharide biopolymers such as xanthan gum, guar gum, alginate; synthetic polymers such as polyacrylic acid like CARBOPOL 980, CARBOPOL 1382 commercially available from NOVEON, acrylate / acrylonitrile copolymers such as HYPAN SS201 commercially available from KINGSTON; clays such as smectite, modified or unmodified hectorite such as BENTONE products commercially available from Rheox, LAPONITE products commercially available from Southern Clay Products, VEEGUM HS products commercially available from RT Vanderbilt, and mixtures thereof.

[0305] In some embodiments, the cleaning composition or soap described herein may further comprise one or more nonionic surfactants. These are well-known compounds (see reference to "Handbook of Surfactants" by MR PORTER, Blackie & Son editions (Glasgow and London), 1991, pp 116 - 178), alcohols, alpha-diols, (C1 - C 20) Alkylphenol, or, for example, a fatty acid having a fatty chain containing 8 to 18 atoms, which may have 2 to 50 ethylene oxide groups or propylene oxide groups, and may have 2 to 30 glycerol groups, polyethoxylated, polypropoxylated, or polyglycerolized fatty acid, a copolymer of ethylene oxide and propylene, a condensate of ethylene oxide and propylene oxide on a fatty alcohol; a polyethoxylated fatty amide having 2 to 30 moles of ethylene oxide, a polyglycerolized fatty amide containing an average of 1 to 5 glycerol groups; a polyethoxylated fatty amine having 2 to 30 moles of ethylene oxide, an ethoxylated sorbitan fatty acid ester having 2 to 30 moles of ethylene oxide; sucrose fatty acid ester, polyethylene glycol fatty acid ester, (C6~C 24 ) alkylpolyglycoside, N-alkyl (C6~C 24 ) glucamine derivative, amine oxide, for example, an oxide of alkyl (C 10 ~C 14 ) amine or an oxide of N-acyl (C 10 ~C 14 )-aminopropylmorpholine, and may be selected from mixtures thereof.

[0306] Suitable additional nonionic surfactants include, but are not limited to, alkyl polyglucosides (APG), maltose esters, polyglycerolized fatty alcohols, glucamine derivatives such as 2-ethylhexyl oxycarbonyl-N-methylglucamine, and mixtures thereof. Suitable alkyl polyglucosides include those containing an alkyl group having 6 to 30 carbon atoms and a hydrophilic group (glucoside). Exemplary alkyl polyglucosides include products sold under the name Mydol 10 (registered trademark) by Kao Chemicals, products sold under the name Plantaren 2000 UP (registered trademark) by Cognis, and products sold under the name Oramix NS 10 (registered trademark) by SEPPIC, including decyl glucoside (alkyl-C9 / C11-polyglucoside (1.4)), and products sold under the name Oramix CG 110 (registered trademark) by SEPPIC, including caprylyl / capryl glucoside; lauryl glucoside sold by Cognis under the names Plantaren 1200 N (registered trademark) and Plantacare 1200 (registered trademark), and cocoglucoside, such as products sold under the name Plantacare 818 / UP (registered trademark) by Cognis.

[0307] Suitable maltose derivatives include those described in document EP-A-566 438, such as O-octanoyl-6'-D-maltose, and O-dodecanoyl-6'-D-maltose described in document FR-2 739 556.

[0308] In some embodiments, the cleaning composition or soap can be formulated in a cosmetically acceptable aqueous medium. Suitable cosmetically acceptable aqueous media can include, in addition to water, one or more solvents such as lower alcohols containing 1 to 6 carbon atoms, such as ethanol; polyols such as glycerin; glycols such as butylene glycol, isoprene glycol, propylene glycol, polyethylene glycol such as PEG-8, sorbitol, sugars such as glucose, fructose, maltose, lactose, sucrose, and mixtures thereof. The amount of solvent in the compositions disclosed herein can range from 0.1 to 95% by weight.

[0309] In some embodiments, the cleaning compositions or soaps described herein can further include one or more cationic polymers of the polyquaternium type that can provide flexibility and lubricity to the foaming composition. Suitable cationic polymers include polyquaternium 5, such as the product MERQUAT 5 commercially available from CALGON, polyquaternium 6, such as the product SALCARE SC 30 commercially available from CIBA, and the product MERQUAT 100 commercially available from CALGON, polyquaternium 7, such as the products MERQUAT S, MERQUAT 2200, and MERQUAT 550 commercially available from CALGON, and the product SALCARE SC 10 commercially available from CIBA, polyquaternium 10, such as the product Polymer JR400 commercially available from Amerchol, polyquaternium 11, such as the products GAFQUAT 755, GAFQUAT 755N, and GAFQUAT 734 commercially available from ISP, polyquaternium 15, such as the product ROHAGIT KF 720 F commercially available from ROHM, polyquaternium 16, such as the products LUVIQUAT FC905, LUVIQUAT FC370, LUVIQUAT HM552, and LUVIQUAT FC550 commercially available from BASF, polyquaternium 22, such as the product Merquat 280 sold by Calgon, polyquaternium 28, such as the product STYLEZE CC10 commercially available from ISP, polyquaternium 39, such as the product MERQUAT PLUS 3330 commercially available from Calgon, polyquaternium 44, such as the product LUVIQUAT CARE sold by BASF, polyquaternium 46, such as the product LUVIQUAT HOLD commercially available from BASF, polyquaternium 47, such as MERQUAT 2001 commercially available from Calgon, and cationic guar, such as the product Jaguar commercially available from Rhodia, can also be used as a cationic polymer.

[0310] In some embodiments, the cleaning compositions or soaps described herein can further include one or more adjuvants or additives used in cosmetic compositions. Suitable adjuvants or additives include, but are not limited to, oils, surfactants, fragrances, preservatives, sequestering agents, pearlescent or opacifying agents, pigments, pearlescent agents, mineral or organic fillers such as talc, kaolin, silica powder, or polyethylene, soluble dyes, or any combination thereof.

[0311] Examples of oils include vegetable oils (jojoba, avocado, sesame, sunflower, corn, soybean, safflower, grape seed), mineral oils (petrolatum, optionally hydrogenated isoparaffin), synthetic oils (isopropyl myristate, cetearyl octanoate, polyisobutylene, ethylhexyl palmitate, alkyl benzoate), volatile or non-volatile silicone oils such as polydimethylsiloxane (PDMS), and cyclodimethylsiloxane or cyclomethicone, and fluorinated oils or fluorosilicone oils, and mixtures thereof.

[0312] Exemplary surfactants include sunscreens, anti-dandruff agents, moisturizers, depigmenting agents, pro-pigmentants, alpha-hydroxy acids, antibacterial agents, anti-radical agents, anti-staining agents, anti-inflammatory agents, retinoids, algae, mushrooms, vegetables, yeast, bacterial extracts, hydrolyzed, partially hydrolyzed, or non-hydrolyzed proteins, enzymes, hormones, vitamins, and their derivatives, flavonoids and isoflavonoids, and mixtures thereof.

[0313] The cleaning compositions or soaps described herein can have a pH in the range of 6 - 10 depending on the selected use. Adjustment of the pH to the desired value can be carried out in the conventional manner by adding a base (organic or inorganic) to the composition, such as ammonia, or a primary, secondary, or tertiary (poly)amine, such as monoethanolamine, diethanolamine, triethanolamine, isopropanolamine, or 1,3 - propanediamine, or by adding a mineral or organic acid, such as a carboxylic acid, such as citric acid. In the context of shower gels, the pH can vary from 8 - 10.

[0314] In some embodiments, the cleaning compositions described herein can further include optional additives such as colorants, fragrances, antibacterial agents, preservatives, antioxidants, beads (for fragrance, exfoliation, or moisturization), mica, glitter, shea butter, shea butter beads, opacifiers, pearlescers, and other such ingredients. In some embodiments, the composition has a high transparency (about 2 - about 25 NTU), a target viscosity (about 4,000 - about 10,000 centipoise) to facilitate dispensing from an orifice in the range of about 1 / 4 - about %, and a yield value (about 3 - about 15 Pascals) that enables the composition to suspend various additives with a uniform distribution and improved stability (e.g., about 8 months at 120°F (49°C)), and any combination thereof.

[0315] In some embodiments, the cleaning composition or soap can further include one or more moisturizers / skin softeners. A moisturizer can be included in a bar or liquid soap composition to provide a conditioning effect on the skin. The term "moisturizer" refers to a substance that gives a smooth and soft feel to the skin surface.

[0316] There are two ways to reduce water loss from the stratum corneum. One is to deposit an occlusive layer on the surface of the skin that reduces the rate of evaporation. The second method is to add a non-occlusive hygroscopic substance that retains water to the stratum corneum, make this water available to the stratum corneum, change its physical properties, and produce an aesthetically desirable effect. Non-occlusive humectants also function by improving the lubricity of the skin.

[0317] Both occlusive and non-occlusive humectants are contemplated for use in the compositions described herein. Exemplary humectants include long-chain fatty acids, liquid water-soluble polyols, glycerin, propylene glycol, sorbitol, polyethylene glycol, ethoxylated / propoaxylated ethers of methylglucose (e.g., methylglucose sesquistearate), ethoxylated / propoaxylated ethers of lanolin alcohol (e.g., Solulan-75® available from Amerchol Co.), coconut and tallow fatty acids, liquid water-soluble polyols (e.g., glycerin, propylene glycol, butylene glycol, hexylene glycol, polypropylene glycol, and polyethylene glycol).

[0318] Non-occlusive humectants can occur naturally in the stratum corneum of the skin, such as sodium pyrrolidone carboxylic acid, lactic acid, urea, L-proline, guanidine, and pyrrolidone. Examples of other non-occlusive humectants include hexadecyl, myristyl, isodecyl, or isopropyl esters of adipic acid, lactic acid, oleic acid, stearic acid, isostearic acid, myristic acid, or linoleic acid, and many of their corresponding alcohol esters (sodium isostearoyl-2-lactate, sodium caprylyl lactate), hydrolyzed proteins and other collagen-derived proteins, aloe vera gel, and acetamide MEA (N-acetyl ethanolamine). Other examples of both occlusive and non-occlusive humectants are disclosed in "Emollients--A Critical Evaluation", J. Mausner, Cosmetics & Toiletries, May 1981, which is incorporated herein by reference.

[0319] Exemplary occlusive emollients include petrolatum, mineral oil, beeswax, silicone, lanolin and oil-soluble lanolin derivatives, saturated and unsaturated fatty alcohols such as behenyl alcohol, squalene, and squalane, and various animal and vegetable oils such as almond oil, peanut oil, wheat germ oil, linseed oil, jojoba oil, anise seed oil, walnut, palm nut, pistachio nut, sesame seed, rapeseed, cade oil, corn oil, peach kernel oil, castor seed oil, pine oil, castor oil, soybean oil, avocado oil, safflower oil, coconut oil, hazelnut oil, olive oil, grape seed oil, and sunflower seed oil.

[0320] Hair care In some embodiments, the composition can be a hair care composition comprising components commonly used in hair care products. These components can include, but are not limited to, detergents, foaming agents, humectants, surfactants, cleaners, gelling agents, fragrances, plant extracts, conditioning agents, water retention agents, silicones or silicone derivatives, thickeners, sunscreens, vitamins, alcohols, polyols, polyol ethers, and other components commonly used in shampoos, conditioners, and styling agents. Hair care products generally include one or more surfactants, one or more viscosity modifiers, one or more preservatives, and one or more fragrances, and any of the components listed below, or combinations thereof.

[0321] Generally, hair can be damaged and embrittled by the action of external atmospheric agents such as light and bad weather, as well as by mechanical or chemical treatments such as brushing, combing, bleaching, permanent waving, and / or hair straightening. However, it can also be damaged and embrittled by mechanical or chemical treatments such as brushing, combing, dyeing, bleaching, permanent waving, and / or relaxing. Over time, hair can become dry, hard, dull, and / or brittle. To overcome these drawbacks, it is common practice to use hair care compositions that properly condition the hair and impart satisfactory cosmetic properties to the hair, particularly with respect to the properties of smoothness, gloss, softness, suppleness, lightness, natural feel, and good detangling ease. These hair care compositions can be, for example, shampoos, conditioning shampoos, hair conditioners, masks, serums, gels, hair lotions, and creams, which can be rinse-out or leave-in compositions. In various embodiments, these compositions generally include a combination of cationic conditioning agents, such as cationic surfactants, cationic polymers, silicones, and / or fatty substances, such as fatty alcohols, to impart satisfactory cosmetic properties to the hair with respect to softness, smoothness, and suppleness. Exemplary compositions can include silicones, which are known to improve the cosmetic properties of hair with respect to smoothness and softness (as shown in U.S. Patent No. 5,374,421, each of which is incorporated herein by reference).

[0322] In some embodiments, the hair care composition can further include one or more non-aminopolyalkylsiloxanes, one or more oxyethylenated polymers in the presence of a fatty alcohol, one or more non-aminopolyalkylsiloxanes containing at least one alkyl chain having at least 12 carbon atoms, one or more oxyethylenated polymers, and / or one or more fatty alcohols. Exemplary oxyethylenated polymers are 10 6It can have the above weight average molecular weight. In some embodiments, the oxyethylenated polymer is of the formula H(OCH2CH2) z OH, where z is an integer of 30,000 or more. In certain embodiments, z can range from 5,000 to 120,000, or from 40,000 to 95,000. In some embodiments, the oxyethylenated polymer can be PEG-45M (z = 45,000), for example, a product sold under the name Polyox WSR N 60 K by Amerchol, and PEG-90M (z = 90,000), and mixtures thereof.

[0323] In some embodiments, the hair care composition can further comprise one or more fatty alcohols. The term "fatty alcohol" means any saturated or unsaturated straight-chain or branched-chain alcohol containing at least 8 carbon atoms and not being oxyalkylenated. Exemplary fatty alcohols are solid at room temperature (25 °C) and atmospheric pressure (1.013 * × 105 Pa). Exemplary fatty alcohols include cetyl alcohol, stearyl alcohol, oleyl alcohol, behenyl alcohol, linoleyl alcohol, palmitoleyl alcohol, arachidonyl alcohol, erucyl alcohol, cetylstearyl (or cetearyl) alcohol, and mixtures thereof.

[0324] In some embodiments, the hair care composition can further comprise one or more conditioning agents including a cationic surfactant, a cationic polymer, and mixtures thereof. The term "cationic surfactant" means a surfactant that is positively charged when contained in the compositions described herein. Suitable cationic surfactants can be optionally polyoxyalkylenated primary, secondary, or tertiary fatty amines, or salts thereof, and quaternary ammonium salts, and mixtures thereof. Exemplary fatty amines are stearamidopropyldimethylamine. Examples of quaternary ammonium salts include tetraalkylammonium salts, such as dialkyldimethylammonium or alkyltrimethylammonium salts having alkyl groups containing from about 16 to 22 carbon atoms, specifically behenyltrimethylammonium, distearyldimethylammonium, cetyltrimethylammonium, or benzyldimethylstearylammonium salts, or on the other hand, palmitylamidopropyltrimethylammonium salts, stearamidopropyltrimethylammonium salts, stearamidopropyldimethylcetearylammonium salts, or stearamidopropyldimethyl(myristyl acetate)ammonium salts sold under the name CERAPHYL 70 by Van Dyk.

[0325] The term "cationic polymer" means any polymer containing a cationic group and / or a group that can be ionized to a cationic group, which can be non-silica-based. Exemplary cationic polymers include any known for styling hair, such as those described in European Patent Application No. 0337354 (A), as well as French Patent Applications Nos. 2270846 (A), 2383660, 2598611, 2470596, and 2519863, each of which is incorporated herein by reference in its entirety. Additional exemplary cationic polymers include those containing units with primary, secondary, tertiary, and / or quaternary amine groups that can form part of the main polymer chain or be carried by side chain substituents directly attached thereto. Suitable cationic polymers can have a weight average molecular weight greater than 10 5 and include polyamines, polyaminoamides, and polyquaternary ammonium-type polymers, including those described in French Patents Nos. 2505348 and 2542997, each of which is incorporated herein by reference in its entirety.

[0326] In some embodiments, the hair care composition can further comprise a "non-sulfate" detergent, foaming agent, or surfactant. Suitable "non-sulfate" agents include, but are not limited to, sodium lauroyl methyl isethionate, sodium methyl oleoyl taurate, and sodium cocoyl isethionate.

[0327] In some embodiments, the hair care composition can further comprise any of the following ingredients or mixtures thereof: quaternary ammonium compounds synthetically derived from rapeseed, quaternary polymers of vinyl pyrrolidone and / or vinyl imidazole, such as products sold under the names LUVIQUAT FC 905, FC 550, and FC 370, and LUVIQUAT Excellence by BASF, cationic celluloses including cellulose ether derivatives containing a quaternary ammonium group, cationic cellulose copolymers or cellulose derivatives grafted with a water-soluble quaternary ammonium monomer, and cationic polysaccharides including cationic galactomannan gum. Exemplary cellulose ether derivatives containing a quaternary ammonium group are described in French Patent No. 1492597. These polymers are also defined in the CTFA dictionary as quaternary ammoniums of hydroxyethyl cellulose reacted with an epoxide substituted with a trimethylammonium group. Cationic cellulose copolymers or cellulose derivatives grafted with a water-soluble quaternary ammonium monomer are described in U.S. Patent No. 4,131,576, such as hydroxyalkyl celluloses, for example, hydroxymethyl, hydroxyethyl, or hydroxypropyl cellulose grafted with methacryloylethyltrimethylammonium, methacrylamidopropyltrimethylammonium, or dimethyldiallylammonium salts. Suitable associative celluloses, for example, C8-C 30 alkyl hydroxyethyl cellulose quaternized with a fatty chain, such as the product QUATRISOFT LM 200 (INCI name, polyquaternium-24) sold by Amerchol / Dow Chemical, and the products CRODACEL QM (INCI name, PG-hydroxyethyl cellulose coco dimonium chloride), CRODACEL QL (C 12 alkyl) (INCI name, PG-hydroxyethyl cellulose lauryl dimonium chloride), and CRODACEL QS (C 18(Alkyl)(INCI name, PG-Hydroxyethylcellulose Stearyldimonium Chloride). Other suitable fatty chain hydroxyethylcellulose derivatives include commercially available products such as SOFTCAT Polymer SL of INCI name, Polyquaternium-67, such as SL-100, SL-60, SL-30, and SL-5 manufactured by Amerchol / Dow chemical. Suitable cationic galactomannan gums are described in U.S. Patent Nos. 3,589,578 and 4,031,307. Suitable cellulose-based components can be in crystalline form, microcrystalline form, or a combination thereof.

[0328] In some embodiments, the hair care composition further comprises one or more cationic proteins or cationic protein hydrolysates, polyalkyleneimines including polyethyleneimine, polymers containing vinyl pyridine or vinyl pyridinium units, condensates of polyamines and epichlorohydrin, quaternary polyurethanes and chitin derivatives, animal protein hydrolysates carrying a trimethylbenzylammonium group, for example, a product sold by Croda under the name Crotein BTA and referred to as benzyltrimonium hydrolyzed animal protein in the CTFA dictionary, and protein hydrolysates carrying a quaternary ammonium group on the polypeptide chain, wherein the ammonium group comprises at least one alkyl radical having 1 to 18 carbon atoms.

[0329] In some embodiments, the hair care composition further comprises one or more quaternized plant proteins, for example, wheat, corn, or soy proteins, such as quaternized wheat proteins (Hydrotriticum WQ or QM, referred to as cocodimonium hydrolyzed wheat protein in the CTFA dictionary, Hydrotriticum QL, referred to as laurdimonium hydrolyzed wheat protein in the CTFA dictionary, or Hydrotriticum QS, referred to as stearydimonium hydrolyzed wheat protein in the CTFA dictionary, sold by Croda).

[0330] In some embodiments, the hair care composition can further comprise one or more polyamines, such as POLYQUART R H sold by Cognis, which is referred to by the name polyethylene glycol tallow polyamine in the CTFA dictionary. Additional suitable polymers include, in particular, those sold by BASF under the name Lupamin, as well as products sold under the names Lupamin 9095, Lupamin 5095, Lupamin 1095, Lupamin 9030, and Lupamin 9010.

[0331] In some embodiments, the hair care composition can further comprise one or more fatty substances that are liquid at room temperature (25 °C) and atmospheric pressure (1.013 * 10 5 Pa). The term "fatty substance" means an organic compound that is insoluble in water (solubility less than 5%, less than 1%, or less than 0.1%) at normal temperature (25 °C) and atmospheric pressure (1.013 * 10 5 Pa). Fatty substances are generally soluble in organic solvents such as chloroform, dichloromethane, carbon tetrachloride, ethanol, benzene, toluene, tetrahydrofuran (THF), liquid petrolatum, or decamethylcyclopentasiloxane under the same temperature and pressure conditions. The liquid fatty substances of the present disclosure can be non-polyoxyethylenated and non-polyglycerolized. The term "oil" means a "fatty substance" that is liquid at room temperature (25 °C) and atmospheric pressure (1.013 * 10 5 Pa). The term "non-silicone oil" means an oil that does not contain any silicon atoms (Si), and the term "silicone oil" means an oil that contains at least one silicon atom. Liquid fatty substances are non-silicone oils, for example, specifically, C6-C 16It may be selected from liquid hydrocarbons, liquid hydrocarbons containing more than 16 carbon atoms, non-silicone oils of animal origin, triglycerides of plant or synthetic origin, fluorinated oils, liquid fatty acids and / or fatty alcohol esters other than triglycerides, and mixtures thereof. The liquid hydrocarbons may include hexane, cyclohexane, undecane, dodecane, tridecane, or isoparaffins such as isohexadecane, isodecane, or isododecane, and mixtures thereof, and may be straight-chain, branched-chain, or optionally cyclic. Suitable straight-chain or branched-chain liquid hydrocarbons of mineral or synthetic origin containing more than 16 carbon atoms may be selected from liquid paraffin, petrolatum, liquid petrolatum, mineral oil, polydecene, and hydrogenated polyisobutene such as PARLEAM, and mixtures thereof. Hydrocarbon-based oils of animal origin such as perhydrosqualene may be used.

[0332] Exemplary triglycerides of plant or synthetic origin can be selected from liquid fatty acid triglycerides containing 6 to 30 carbon atoms, such as heptanoic acid or octanoic acid triglycerides, or more specifically, vegetable oils such as coconut oil, sunflower oil, corn oil, soybean oil, marrow oil, grape seed oil, sesame seed oil, hazelnut oil, apricot oil, macadamia oil, arare oil, castor oil, avocado oil, jojoba oil, shea butter oil, or synthetic caprylic / capric acid triglycerides, such as those sold by Stearineries Dubois, or those sold under the names MIGLYOL 810, 812, and 818 by Dynamit Nobel, and mixtures thereof. Suitable fluorinated oils include perfluoromethylcyclopentane and perfluoro-1,3-dimethylcyclohexane, such as FLUTEC PC1 and FLUTEC PC3 by BNFL Fluorochemicals; perfluoro-1,2-dimethylcyclobutane; perfluoroalkanes, such as dodecafluoropentane and tetradecafluorohexane sold under the names PF 5050 and PF 5060 by 3M, or bromoperfluorooctyl sold under the name FORALKYL by Atochem; nonafluoromethoxybutane and nonafluoroethoxyisobutane; perfluoromorpholine derivatives, such as 4-trifluoromethylperfluoromorpholine sold under the name PF 5052 by 3M.

[0333] Suitable monoesters include dihydroabietyl behenate; octyldodecyl behenate; isocetyl behenate; cetyl lactate; C 12 ~C 15Alkyl lactate; Isostearyl lactate; Lauryl lactate; Linoleyl lactate; Oleyl lactate; (Iso)stearyl octanoate; Isocetyl octanoate; Octyl octanoate; Cetyl octanoate; Decyl oleate; Isocetyl isostearate; Isocetyl laurate; Isocetyl stearate; Isodecyl octanoate; Isodecyl oleate; Isononyl isononanoate; Isostearyl palmitate; Methyl acetyl ricinoleate; Myristyl stearate; Octyl isononanoate; 2-Ethylhexyl isononanoate; Octyl palmitate; Octyl pelargonate; Octyl stearate; Octyldodecyl erucate; Oleyl erucate; Ethyl and isopropyl palmitate; 2-Ethylhexyl palmitate; 2-Octyldecyl palmitate; Alkyl myristate, for example, isopropyl myristate, butyl, cetyl, 2-octyldodecyl, myristyl, or stearyl; Hexyl stearate; Butyl stearate; Isobutyl stearate; Dioctyl malate; Hexyl laurate; 2-Hexyldecyl laurate; and mixtures thereof.

[0334] In some embodiments, the hair care composition can further comprise diethyl sebacate, diisopropyl sebacate, diisopropyl adipate, di(n-propyl) adipate, dioctyl adipate, diisostearyl adipate, dioctyl maleate, glyceryl undecylenate, octyldodecyl stearoyl stearate, pentaerythrityl monolinoleate, pentaerythrityl tetraisononanoate, pentaerythrityl tetra-pelargonate, pentaerythrityl tetraisostearate, pentaerythrityl tetraoctanoate, propylene glycol dicaprylate, propylene glycol dicaprate, tridecyl erucate, triisopropyl citrate, triisostearyl citrate, glyceryl trilactate, glyceryl trioctanoate, trioctyldodecyl citrate, trioleyl citrate, propylene glycol dioctanoate, neopentyl glycol diheptanoate, diethylene glycol diisononanoate, and polyethylene glycol distearate, and mixtures thereof.

[0335] In some embodiments, the hair care composition is C6-C 30 one or more fatty acid esters, one or more sugar esters, and / or one or more diesters, such as C 12 -C 22It can further contain fatty acids. The term "sugar" means an oxygen-containing hydrocarbon compound containing several alcohol functional groups, with or without an aldehyde or ketone functional group, and containing at least 4 carbon atoms. Suitable sugars include monosaccharides, oligosaccharides, or polysaccharides, such as sucrose (or saccharose), glucose, galactose, ribose, fucose, maltose, fructose, mannose, arabinose, xylose, and lactose, and their derivatives, such as alkyl derivatives, such as methyl derivatives, such as methyl glucose. Suitable esters include oleate, laurate, palmitate, myristate, behenate, cocoate, stearate, linoleate, linolenate, caprate, arachidonate, or mixtures thereof, such as oleate / palmitate, oleate / stearate, or palmitate / stearate mixed esters. Also, suitable mono- and di-esters include mono- or di-oleates, stearates, behenates, oleopalmitates, linoleates, linolenates, or oleostearates of sucrose, glucose, or methyl glucose, including products sold under the name GLUCATE DO by Amerchol, which is methyl glucose dioleate.Additional exemplary esters or mixtures of esters of sugars and fatty acids that can be mentioned include products sold by Crodesta under the names F160, F140, F110, F90, F70, and SL40 (respectively, sucrose palmitate / stearate formed from 73% monoesters and 27% diesters and triesters, from 61% monoesters and 39% diesters, triesters, and tetraesters, from 52% monoesters and 48% diesters, triesters, and tetraesters, from 45% monoesters and 55% diesters, triesters, and tetraesters, from 39% monoesters and 61% diesters, triesters, and tetraesters, and sucrose monolaurate); products sold under the name Ryoto Sugar Esters (for example, corresponding to sucrose behenate with reference number B370 and formed from 20% monoesters and 80% diester - triester - polyester); and mono - dipalmitate / stearate sucrose sold by Goldschmidt under the name TEGOSOFT PSE.

[0336] In some embodiments, the hair care composition can further comprise a pH modifier such as citric acid and / or sodium hydroxide. Any pH modifier commonly used in hair care compositions is contemplated for use herein.

[0337] In various embodiments, the hair care composition can further comprise any of the following ingredients and / or mixtures thereof: sodium lauroyl methyl isethionate (a detergent and foaming agent), a liquid fatty substance comprising a silicone oil different from the non-aminopolyalkylsiloxanes described above herein, and an organomodified polysiloxane comprising at least one functional group selected from an amino group, an aryl group, and an alkoxy group. Organopolysiloxanes are more particularly defined in Walter Noll’s Chemistry and Technology of Silicones (1968), Academic Press, the entirety of which is incorporated herein by reference. They may be either volatile or non-volatile. Suitable cyclic polydialkylsiloxanes include octamethylcyclotetrasiloxane sold under the name VOLATILE SILICONE 7207 by Union Carbide or SILBIONE 70045 V2 by Rhodia, decamethylcyclopentasiloxane sold under the name VOLATILE SILICONE 7158 by Union Carbide and SILBIONE 70045 V5 by Rhodia, and mixtures thereof. Cyclocopolymers of the dimethylsiloxane / methylalkylsiloxane type such as VOLATILE SILICONE FZ 3109 sold by Union Carbide are also suitable. Exemplary cyclic polydialkylsiloxanes having an organosilicon compound include octamethylcyclotetrasiloxane and tetra(trimethylsilyl)pentaerythritol (50 / 50), and mixtures of octamethylcyclotetrasiloxane and oxy-1,1'-bis(2,2,2',2',3,3'-hexatrimethylsilyloxy)neopentane, linear volatile polydialkylsiloxanes, for example, those sold under the name SH 200 by Toray Silicone.Silicones falling into this category are also described in the paper published in Cosmetics and Toiletries, Vol. 91, January 76, pp. 27 - 32, Todd & Byers, "Volatile Silicone Fluids for Cosmetics", which is incorporated herein by reference.

[0338] Exemplary suitable non - volatile polydialkylsiloxanes include polydimethylsiloxane having trimethylsilyl end groups, for example, SILBIONE oils or MIRASIL oils of series 47 and 70047 sold by Rhodia, for example, oil 70047 V 500000; oils of the MIRASIL series sold by Rhodia; oils of the 200 series manufactured by Dow Corning, for example, DC200 having a viscosity of 60000 mm 2 / sec; VISCASIL oils manufactured by General Electric, and certain oils of the SF series (SF 96, SF 18) manufactured by General Electric, polydimethylsiloxane having dimethylsilanol end groups known by the name dimethiconol (CTFA), for example, oils of series 48 manufactured by Rhodia.

[0339] Exemplary organomodified silicones include polyalkylaryl siloxanes and products sold under the following names: SILBIONE oils of the 70 641 series manufactured by Rhodia; oils of the RHODORSIL 70 633 and 763 series manufactured by Rhodia; Dow Corning 556 Cosmetic Grade Fluid, an oil manufactured by Dow Corning; silicone of the PK series manufactured by Bayer, such as product PK20; silicone of the PN and PH series manufactured by Bayer, such as products PN1000 and PH1000; certain oils of the SF series manufactured by General Electric, such as SF 1023, SF 1154, SF 1250, and SF 1265; products sold under the names GP 4 Silicone Fluid and GP 7100 from Genesee, or products sold by Dow Corning under the names Q2 8220 and Dow Corning 929 or 939.

[0340] In various embodiments, the hair care composition can further comprise one or more additional surfactants different from the cationic surfactants described above, including anionic surfactants, amphoteric or zwitterionic surfactants, nonionic surfactants, and mixtures thereof. Exemplary anionic surfactants include alkyl sulfates, alkyl ether sulfates, alkyl amide ether sulfates, alkyl aryl polyether sulfates, monoglyceride sulfates, alkyl sulfonates, alkyl amide sulfonates, alkyl aryl sulfonates, α-olefin sulfonates, paraffin sulfonates, alkyl sulfosuccinates, alkyl ether sulfosuccinates, alkyl amide sulfosuccinates, alkyl sulfonoacetates, acyl sarcosinates, acyl glutamates, alkylsulfosuccinamate, acyl isothionates, and N-(C1-C4)alkyl N-acyl taurates, salts of alkyl monoesters and polyglycoside-polycarboxylic acids, acyl lactylates, D-galactoside uronates, alkyl ether carboxylates, alkyl aryl ether carboxylates, alkyl amide ether carboxylates; and the corresponding non-chlorinated forms of all of these compounds. The alkyl groups and acyl groups (unless otherwise specified) of all of these compounds generally contain from 6 to 24 carbon atoms, and the aryl group generally represents a phenyl group. Salt forms of anionic surfactants can include alkali metal salts, such as sodium or potassium salts, sodium salts, ammonium salts, amine salts, and amino alcohol salts or alkaline earth metal salts, such as magnesium salts. Exemplary amino alcohol salts include monoethanolamine, diethanolamine, and triethanolamine salts, monoisopropanolamine, diisopropanolamine, or triisopropanolamine salts, 2-amino-2-methyl-1-propanol salts, 2-amino-2-methyl-1,3-propanediol salts, and tris(hydroxymethyl)aminomethane salts.

[0341] Suitable anionic surfactants include weakly anionic surfactants, namely, polyoxyalkylenated alkyl ether carboxylic acids; polyoxyalkylenated alkyl aryl ether carboxylic acids; polyoxyalkylenated alkyl amide ether carboxylic acids, specifically those containing 2 to 50 ethylene oxide groups; alkyl-D-galacturonate; acyl sarcosinates, acyl glutamates; and alkyl polyglycoside carboxylates, for example, those sold under the name AKYPO RLM 45 CA by Kao, and also include anionic surfactants having no sulfate functional group.

[0342] Exemplary suitable amphoteric or zwitterionic surfactants are those in which the aliphatic group is linear or branched and contains 8 to 22 carbon atoms, and the amine derivative has at least one anionic group, for example, a carboxylate, sulfonate, sulfate, phosphate, or phosphonate group, for example, (C8-C 20 ) alkyl betaines, sulfobetaines, (C8-C 20 ) alkylamide (C3-C8) alkyl betaines, or (C8-C 20 ) alkylamide (C6-C8) alkyl sulfobetaines, and can be optionally quaternized secondary or tertiary aliphatic amine derivatives. Any suitable secondary or tertiary aliphatic amine derivative can be present in coconut oil or hydrolyzed linseed oil, etc. Representative compounds are classified under the names disodium cocoamphodiacetate, disodium lauroamphodiacetate, disodium capryloamphodiacetate, disodium capryloamphodiacetate, disodium cocoamphodipropionate, disodium lauroamphodipropionate, disodium capryloamphodipropionate, disodium capryloamphodipropionate, lauroamphodipropionic acid, and cocoamphodipropionic acid in the CTFA Dictionary, 5th Edition, 1993.

[0343] As an example, cocoamphodiacetate sold under the trade name MIRANOL C2M Concentrate by Rhodia, and sodium diethylaminopropylcocospartamidate sold under the name Chimexane HB by Chimex, are suitable for use in the disclosed compositions.

[0344] Suitable nonionic surfactants are described in Handbook of Surfactants by M.R. Porter, published by Blackie & Son (Glasgow and London), 1991, pp. 116 - 178, and include fatty alcohols, fatty α-diols, fatty (C1 - C 20 ) alkylphenols and fatty acids, which may be ethoxylated, propoxylated, or glycerolated, and contain at least one fatty chain having from 8 to 18 carbon atoms, a number of ethylene oxide or propylene oxide groups in the range of 1 to 200, and a number of glycerol groups in the range of 1 to 30. Condensates of ethylene oxide and propylene oxide with fatty alcohols, ethoxylated fatty amides having from 1 to 30 ethylene oxide units, polyglycerolized fatty amides containing on average from 1 to 5, specifically from 1.5 to 4, glycerol groups, ethoxylated fatty esters of sorbitan containing from 1 to 30 ethylene oxide units, fatty esters of sucrose, fatty esters of polyethylene glycol, (C6 - C 24 ) alkyl polyglycosides, oxyethylenated vegetable oils, N-(C6 - C 24 ) alkyl glucamine derivatives, amine oxides such as (C 10 - C 14 ) alkylamine oxides, or N-(C 10 - C 14 ) acylaminopropylmorpholine oxides are also suitable for use in the compositions disclosed herein.

[0345] Suitable additional nonionic surfactants include alkyl polyglucosides (APG), maltose esters, polyglycerolized fatty alcohols, glucamine derivatives such as 2-ethylhexyl oxycarbonyl-N-methylglucamine, and mixtures thereof. Alkyl polyglucosides contain an alkyl group having 6 to 30 carbon atoms and a hydrophilic group (glucoside). Exemplary alkyl polyglucosides include products sold under the name Mydol 10 (registered trademark) by Kao Chemicals, products sold under the name Plantaren 2000 UP (registered trademark) by Cognis, and products sold under the name Oramix NS 10 (registered trademark) by SEPPIC, including decyl glucoside (alkyl-C9 / C11-polyglucoside (1.4)), and products sold under the name Oramix CG 110 (registered trademark) by SEPPIC, including caprylyl / capryl glucoside; lauryl glucoside sold under the names Plantaren 1200 N (registered trademark) and Plantacare 1200 (registered trademark) by Cognis, and coco glucoside, for example, products sold under the name Plantacare 818 / UP (registered trademark) by Cognis.

[0346] Suitable maltose derivatives include those described in document EP-A-566 438, for example, O-octanoyl-6'-D-maltose, and O-dodecanoyl-6'-D-maltose described in document FR-2 739 556. Each of these documents is hereby incorporated by reference in its entirety.

[0347] In some embodiments, the collagen protein in the composition can be chemically modified with a hair colorant to provide a permanent, semi-permanent, or temporary hair dye solution. A hair colorant as defined herein can be any dye, pigment, nanoparticle, etc. that can be used to change the color of hair. Suitable hair colorants include, but are not limited to, dyes such as 4-hydroxypropylamino-3-nitrophenol, 4-amino-3-nitrophenol, 2-amino-6-chloro-4-nitrophenol, 2-amino-6-chloro-4-nitrophenol, 2-nitro-paraphenylenediamine, N,N-hydroxy-ethyl-2-nitro-phenylenediamine, 4-nitro-indole, 2-nitro-5-glycerylmethylaniline, 3-methylamino-4-nitrophenoxyethanol, para-phenylenediamine, hydrogen peroxide, ammonia, DMDM hydantoin, methylparaben, propylparaben, lead acetate, and resorcinol.

[0348] In some embodiments, the hair care composition can be formulated in a cosmetically acceptable medium. The term "cosmetically acceptable medium" means a medium that is compatible with human keratin fibers such as hair. The cosmetically acceptable medium can be formed from water or a mixture of water and one or more cosmetically acceptable solvents selected from lower alcohols such as ethanol and isopropanol; 2-butoxyethanol, propylene glycol, propylene glycol monomethyl ether, diethylene glycol monoethyl ether, and polyols and polyol ethers including monomethyl ether, and mixtures thereof.

[0349] In some embodiments, the hair care composition can further comprise any of the following additives, or mixtures thereof: solid fatty substances different from fatty alcohols, such as waxes, anionic, nonionic, or amphoteric polymers or mixtures thereof, anti-dandruff agents, anti-seborrheic agents, agents for preventing hair loss and / or promoting hair growth, vitamins and provitamins including panthenol, sunscreens, mineral or organic pigments, sequestering agents, plasticizers, solubilizers, acidifying agents, mineral or organic thickeners, especially polymer thickeners different from oxyethylenated polymers, opacifying or pearlescent agents, antioxidants, hydroxy acids, fragrances, and / or preservatives.

[0350] Nail care In some embodiments, the cosmetic composition can include components commonly used in nail care products. Nail care products include nail treatments such as nail hardeners, top coats and base coats, manicures, manicure removers, hand skin care, foot skin care, drying agents, and correction pens including manicure removers.

[0351] In some embodiments, the nail care composition can be a nail treatment composition. The nail treatment composition can include a composition for treating ingrown nails or nail deformities, a composition for the topical treatment of nail infections including fungal infections, a nail hardener, a top coat, a base coat, a manicure remover, or any combination thereof. In some embodiments, the nail treatment composition can be formulated as a topical nail lacquer or manicure, cream, solution, suspension, lotion, serum, gel, balm, gel, oil, oil-in-cream, and / or scrub for treating the hands and / or feet.

[0352] In some embodiments, the nail treatment composition can be a nail strengthener. The nail strengthener can treat fingernails and toenails to both promote nail hardening, strengthening, and growth and prevent or minimize breakage, cracking, splitting, and peeling, and any known composition used to enhance hoof growth and prevent and heal hoof cracks while having a protein concentration similar to that of human fingernails and toenails: it can include lanolin, butter, beeswax, rosin, copper acetate, and turpentine oil. In some embodiments, the nail treatment composition can further include titanium dioxide, for example, TI-PURE R900 manufactured by E.I.DuPont.

[0353] In some embodiments, the nail treatment composition can be a base coat. In some embodiments, the base coat is a liquid composition comprising at least one polymer that provides adhesion, for example, a polymer copolymerized from methyl methacrylate (MMA) and methacrylic acid (MAA) to form a polymer composed of polymethyl methacrylate (PMMA) and polymethacrylic acid (PMAA), wherein the MAA monomer fraction can vary from 0 to 100%. In some embodiments, polymers suitable for use as a base coat include hydroxyethylmethacrylate (HEMA), hydroxypropylmethacrylate (HPMA), ethyl methacrylate (EMA), tetrahydrofurfuryl methacrylate (THFMA), pyromellitic dianhydride di(meth)acrylate, pyromellitic dianhydride glyceryl dimethacrylate, pyromellitic dimethacrylate, methacryloyloxyethyl maleate, 2-hydroxyethyl methacrylate / succinate, 1,3-glycerol dimethacrylate / succinate adduct, monoethyl methacrylate phthalate, and mixtures thereof. In some aspects, the base coat can further comprise a non-reactive, solvent-soluble, film-forming polymer such as a cellulose ester, for example, cellulose acetate alkylate, cellulose acetate butyrate, or cellulose acetate propionate. The above exemplary components are not limiting.

[0354] In some embodiments, the nail treatment composition can be a top coat, such as a fast-drying top coat. The top coat can include a base resin that is cellulose acetate butyrate, a film former that is a methacrylate polymer, a monofunctional methacrylate thermosetting or photocurable monomer, and a crosslinking agent (i.e., difunctional and trifunctional methacrylates), a thermal initiator or a photoinitiator together in a solution of aliphatic esters and alcohols, and a photoreactive coating. Suitable solvents include acetates and alcohols, particularly ethyl acetate, butyl acetate, and isopropyl alcohol. The photoreactive coating can include photoreactive monomers, such as cyclohexyl methacrylate, n-decyl methacrylate, 2-ethylhexyl methacrylate, ethyl methacrylate, hydroxypropyl methacrylate, isobornyl methacrylate, 2-methoxyethyl methacrylate; difunctional and trifunctional methacrylate monomers, crosslinking agents such as diurethane dimethacrylate, ethylene glycol dimethacrylate, 1,10-decanediol dimethacrylate, 1,6-hexanediol dimethacrylate, and trimethylolpropane trimethacrylate. Commercially available photoinitiators suitable for use include, but are not limited to: benzoin methyl ether, 2-hydroxy-2-methyl-1-phenyl-1-propanone (“Darocur 1173”), diethoxyacetophenone, and benzyl diketal. The photoinitiator can be an oligomeric mixture of phenylpropanone, such as a mixture of 2,4,6-trimethylbenzophenone and 4-methylbenzophenone, and a mixture of oligo-[2-hydroxy-2-methyl-1[4-(1-methylvinyl)phenyl]propanone] and 2-hydroxy-2-methyl-phenylpropanone, which are sold by Sartomer under the names “Esacure KIP 100F” and “Esacure TZT Photoinitiator”.

[0355] In some embodiments, the nail care composition can be a manicure or nail enamel composition. In some embodiments, the manicure or nail enamel composition can further include a nitrocellulose film former, a latex film former, a polycarbodiimide film former, a low volatile organic compound (VOC), and a polycarbodiimide film former. The polycarbodiimide can include a polymer having a plurality of carbodiimide groups attached to the polymer backbone. For example, U.S. Patent No. 5,352,400, the disclosure of which is incorporated herein by reference, discloses polymers and copolymers derived from alpha-methylstyrene-isocyanate. Suitable polycarbodiimide compounds include those commercially available from suppliers such as Nisshinbo (including those known by the names CARBODILITE series, V-02, V02-L2, SV-02, E-02, V-10, SW-12G, E-03A), Picassian, and 3M, but are not limited thereto.

[0356] In some embodiments, the nail polish or nail enamel composition can further include one or more latex polymers, including carboxyl-functional acrylate latex polymers, carboxyl-functional polyurethane latex polymers, carboxyl-functional silicone latex polymers, carboxyl-functional non-acrylate latex polymers, and mixtures thereof. In various embodiments, suitable latex polymers can be film-forming latex polymers or non-film-forming latex polymers. In some embodiments, the latex polymer can be a carboxyl-functional acrylate latex polymer, for example, obtained by homopolymerization or copolymerization of ethylenically unsaturated monomers selected from vinyl monomers, (meth)acrylic monomers, (meth)acrylamide monomers, mono- and dicarboxylic unsaturated acids, esters of (meth)acrylic monomers, and amides of (meth)acrylic monomers. The term "(meth)acrylic" and variations thereof, as used herein, mean acrylic or methacrylic. The (meth)acrylic monomer can be selected, for example, from acrylic acid, methacrylic acid, citraconic acid, itaconic acid, maleic acid, fumaric acid, crotonic acid, and maleic anhydride. Esters of the (meth)acrylic monomer include, as non-limiting examples, C1-C8 alkyl (meth)acrylates, such as methyl (meth)acrylate, ethyl (meth)acrylate, propyl (meth)acrylate, isopropyl (meth)acrylate, butyl (meth)acrylate, tert-butyl (meth)acrylate, pentyl (meth)acrylate, isopentyl (meth)acrylate, neopentyl (meth)acrylate, hexyl (meth)acrylate, isohexyl (meth)acrylate, 2-ethylhexyl (meth)acrylate, cyclohexyl (meth)acrylate, isohexyl (meth)acrylate, heptyl (meth)acrylate, isoheptyl (meth)acrylate, octyl (meth)acrylate, isooctyl (meth)acrylate, allyl (meth)acrylate, and combinations thereof.(Meth)acrylamide monomers can be prepared, for example, from (meth)acrylamide, and in particular N-alkyl(meth)acrylamide, specifically N-(C1-C12)alkyl(meth)acrylate, for example, N-ethyl(meth)acrylamide, N-t-butyl(meth)acrylamide, N-t-octyl(meth)acrylamide, N-methylol(meth)acrylamide, and N-diacetone(meth)acrylamide, and any combination thereof.

[0357] Examples of vinyl monomers include vinyl cyanide compounds such as acrylonitrile and methacrylonitrile; vinyl esters such as vinyl formate, vinyl acetate, vinyl propionate, vinyl neodecanoate, vinyl pivalate, vinyl benzoate, and vinyl t-butylbenzoate, triallyl cyanurate; vinyl halides such as vinyl chloride and vinylidene chloride; aromatic mono- or divinyl compounds such as styrene, alpha-methylstyrene, chlorostyrene, alkylstyrene, divinylbenzene, and diallyl phthalate, and para-styrenesulfonic acid, vinylsulfonic acid, 2-(meth)acryloyloxyethylsulfonic acid, 2-(meth)acrylamide-2-methylpropylsulfonic acid, and mixtures thereof, but are not limited thereto.

[0358] It should be understood that the list of monomers herein is not limiting and that any monomer known to those skilled in the art, including acrylic and / or vinyl monomers (including monomers modified with silicone chains), can be used.

[0359] In some non-limiting, exemplary embodiments, the carboxyl-functional acrylate latex polymer may be selected from an aqueous dispersion of a methacrylic acid / ethyl acrylate copolymer (INCI: acrylate copolymer, e.g., LUVIFLEX SOFT by BASF), a PEG / PPG-23 / 6 dimethicone citraconate / C10-30 alkyl PEG-25 methacrylate / acrylic acid / methacrylic acid / ethyl acrylate / trime thylolpropane PEG-15 triacrylate copolymer (INCI: polyacrylate-2 crosspolymer, e.g., Fixate Superhold™ by Lubrizol), a styrene / acrylic copolymer (e.g., Acudyne Shine by Dow Chemical), an ethylhexyl acrylate / methyl methacrylate / butyl acrylate / acrylic acid / methacrylic acid copolymer (INCI: acrylate / ethylhexyl acrylate copolymer, e.g., Daitosol 5000SJ, Daito Kasei Kogyo), an acrylic / acrylate copolymer (INCI name: acrylate copolymer, e.g., Daitosol 5000AD, Daito Kasei Kogyo), an acrylate copolymer known by the trade name Dermacryl AQF (Akzo Nobel), the trade name LUVIMER MAE (BASF), or the trade name BALANCE CR (AKZO NOBEL), an acrylate / hydroxyester acrylate copolymer, e.g., the one known by the trade name ACUDYNE 180 POLYMER (Dow Chemical), a styrene / acrylate copolymer known from the trade name Acudyne Bold by Dow Chemical, a styrene / acrylate / ammonium methacrylate copolymer known from the trade name SYNTRAN PC5620 CG by Interpolymer, and mixtures thereof.

[0360] In some embodiments, the nail care composition can include a nail polish remover. The nail polish remover can include a polyhydric alcohol compound including glycerin, glycol, polyglycerin, esters of polyhydric alcohols, and mixtures thereof. The glycol can contain 2 to 12 carbon atoms, such as, for example, glycerin, propylene glycol, butylene glycol, propane diol, hexylene glycol, polyglycerin, dipropylene glycol, and diethylene glycol. Suitable esters of polyhydric alcohols include liquid esters of saturated or unsaturated, straight-chain or branched-chain C1-C26 polyhydric alcohols. Examples of suitable esters of polyhydric alcohols include, but are not limited to, esters of dihydroxy, trihydroxy, tetrahydroxy, or pentahydroxy alcohols. Esters of polyhydric alcohols can be glyceryl esters such as glyceryl triglycolate, glyceryl tricitrate, glyceryl trilactate, glyceryl trilactate, glyceryl tributanoate, glyceryl triheptanoate, glyceryl trioctanoate, and the like.

[0361] The nail polish remover can further contain a lower carbon alcohol having 1 to 8 carbon atoms. The lower carbon alcohol can contain 2 to 6 carbon atoms, for example, 2 to 5 carbon atoms. Examples of the lower carbon alcohol include, but are not limited to, ethanol, propanol, butanol, pentanol, isopropanol, isobutanol, and isopentanol. The nail polish remover can further contain a high-boiling ester compound including, but not limited to, carbonic acid esters, adipates, sebacates, and succinates. Examples of the high-boiling ester compound include, but are not limited to, alkylene carbonates such as propylene carbonate, dimethyl succinate, diethyl succinate, dimethyl glutarate, diethyl glutarate, dimethyl sebacate, diethyl sebacate, diisopropyl sebacate, bis(2-ethylhexyl) sebacate, dimethyl adipate, diisopropyl adipate, di-n-propyl adipate, dioctyl adipate, bis(2-ethylhexyl) adipate, diisostearyl adipate, ethyl maleate, bis(2-ethylhexyl) maleate, triisopropyl citrate, triisocetyl citrate, triisostearyl citrate, trioctyldodecyl citrate, and trioleyl citrate.

[0362] The nail polish remover can further contain a thickener including nonionic, anionic, cationic, amphiphilic, and amphoteric polymers, as well as other known rheology modifiers, such as cellulose-based thickeners, such as hydroxyethyl cellulose, hydroxypropyl cellulose, methyl cellulose, and ethyl hydroxyethyl cellulose. Certain notable cellulose derivatives include those having a molecular weight exceeding 500,000 daltons, such as hydroxyl-modified cellulose polymers, such as hydroxyethyl cellulose, such as NATROSOL 250 HHR and hydroxypropyl cellulose, such as KLUCEL MF (both available from Ashland, Covington, Ky.). The thickener can be a polysaccharide such as fructan, glucan, galactan, and mannan, or a heteropolysaccharide such as hemicellulose, pullulan, or a branched polysaccharide such as gum arabic and amylopectin, or a mixed polysaccharide such as starch. The thickener can be an acrylic thickening agent (acrylic thickener) or an acrylamide thickening agent (acrylamide thickener). The thickener can contain at least one monomer that serves a weak acid function, such as acrylic acid, methacrylic acid, itaconic acid, crotonic acid, maleic acid, and / or fumaric acid. The thickener can contain a monomer that serves a strong acid function, such as a monomer having a sulfonic acid type or phosphonic acid type function, such as 2-acrylamido-2-methylpropane sulfonic acid (AMPS). The thickener can contain a crosslinking agent such as a glycidyl ether type composition such as methylene bisacrylamide (MBA), ethylene glycol diacrylate, polyethylene glycol dimethacrylate, diacrylamide, cyanomethacrylate, vinyloxyethacrylate or methacrylate, formaldehyde, glyoxal, and ethylene glycol diglycidyl ether, or an epoxide.Suitable acrylic thickeners are disclosed in U.S. Patent Application Publication Nos. 2004 / 0028637 and 2008 / 0196174, both of which are incorporated herein by reference. In some embodiments, the thickener can include an organoclay (hydrophobically treated clay) or a hydrophilic clay.

[0363] In some embodiments, the thickener can include a polishing compound (polishing system). A "polishing compound" is a compound that can provide polishing or mechanical exfoliation. The polishing particles can include perlite, pumice, zeolite, hydrated silica, calcium carbonate, dicalcium phosphate dihydrate, calcium pyrophosphate, alumina, sodium bicarbonate, polylactic acid, and synthetic polymer materials such as polyethylene, polypropylene, polyethylene terephthalate, polymethyl methacrylate, or nylon. In certain embodiments, a moderately hard abrasive includes perlite such as cosmetic grade perlite available from Imerys under the name IMERCARE 270P-Scrub. In certain embodiments, a soft abrasive is sugar, ground fruit pits or shell powder, such as apricot pits, coconut shells, or spherical waxes (e.g., carnauba jojoba); argon shell powder, etc.

[0364] In some embodiments, the nail care composition can further include additives commonly used in cosmetic compositions and known to those skilled in the art, selected from solvents, preservatives, fragrances, oils, waxes, surfactants, antioxidants, agents for combating free radicals, wetting agents, dispersants, defoamers, neutralizing agents, stabilizers, active ingredients selected from essential oils, ultraviolet light blockers, sunscreens, moisturizers, vitamins, proteins, ceramides, plant extracts, fibers, etc., and mixtures thereof.

[0365] Therapeutic composition In some embodiments, the compositions described herein can be therapeutic compositions useful for treating one or more conditions. In some embodiments, the compositions described herein can be formulated as dry films, powders, gels, or solutions in water or buffer. For example, in some embodiments, application of the compositions described herein can promote wound healing, reduce or prevent scar tissue formation, promote tissue regeneration, minimize local inflammation, minimize tissue rejection, and / or enhance graft integration. In some embodiments, the composition can be formulated as an injectable material, such as a hydrogel. Collagen hydrogels exhibit a large, relatively uniform surface area and can function as a delivery system for collagen and optionally one or more additional therapeutic agents. Injectable collagen materials can also form a scaffold or network capable of both replacing tissue function and supporting tissue regeneration. In certain embodiments, the composition can be applied topically. In certain embodiments, the composition can be injected intradermally, subcutaneously, or into the skin.

[0366] In some embodiments, the therapeutic compositions described herein can include one or more additional therapeutic and / or prophylactic agents other than the collagen fragments described elsewhere herein. The one or more additional therapeutic and / or prophylactic agents can be small molecule active agents or biomolecules such as enzymes or proteins, polypeptides, or nucleic acids.

[0367] Non-limiting examples of additional therapeutic and / or prophylactic agents include anti-cancer agents, antibacterial agents (including antiviral, antibacterial, antifungal, and antiparasitic agents), antioxidants, topical analgesics, topical anesthetics, anti-inflammatory agents, cytokines, immunosuppressants, anti-allergy agents, essential nutrients, growth factors (such as fibroblast growth factor, hepatocyte growth factor, platelet-derived growth factor, vascular endothelial growth factor, and insulin-like growth factor), and combinations thereof. The specific dosage of the additional therapeutic and / or prophylactic agent can be readily determined by one of ordinary skill in the art. See Ansel, Howard C. et al., Pharmaceutical Dosage Forms and Drug Delivery Systems (6th ed.) Williams and Wilkins, Malvern, PA (1995).

[0368] In other embodiments, the compositions described herein can be used in combination with cell delivery, e.g., delivery of stem cells, pluripotent cells, somatic cells, and combinations thereof.

[0369] The present disclosure contemplates that the therapeutic and / or prophylactic agent is an integral part of the compositions described herein, but in some embodiments, the therapeutic and / or prophylactic agent can be administered before, in conjunction with, or after administration of the therapeutic composition described herein. In other words, in some embodiments, one or more additional therapeutic and / or prophylactic agents need not be incorporated integrally into the composition, but may be provided separately from the composition.

[0370] Non-limiting examples of suitable local anesthetics that may be included in the composition of the present invention include ambucaine, amolanone, amylocaine, benoxinate, benzocaine, betoxycaine, bupivacaine, butacaine, butamben, butanilicaine, butetamine, butoxycaine, carticaine, chloroprocaine, cocaethylene, cocaine, cyclomethycaine, dibucaine, dimethisoquin, dimethocaine, diperodon, dicyclomine, ecgonidine, ecgonine, ethyl chloride, etidocaine, beta-eucaine, euprocin, phenacolin, hormocaine, hexylcaine, hydroxytetracaine, isobutyl p-aminobenzoate, leucinocaine mesylate, levoxadrol, lidocaine, mepivacaine, meprilocaine, metabutoxycaine, methyl chloride, myrtocaine, naepaine, octocaine, orthocaine, oxethazaine, parethoxycaine, phenacaine, phenol, piperocaine, pyridocaine, polidocanol, pramoxine, prilocaine, procaine, propanocaine, propalacaine, propicocaine, propoxycaine, pseudococaine, pyrocaine, ropivacaine, salicyl alcohol, tetracaine, tricaine, trimecaine, xylamine, and any combination thereof, but are not limited thereto.

[0371] Non-limiting examples of suitable antiviral agents include ganciclovir and acyclovir. Non-limiting examples of suitable antibiotic agents include aminoglycosides such as streptomycin, amikacin, gentamicin, and tobramycin, ansamycins such as geldanamycin and herbimycin, carbacephems, carbapenems, cephalosporins, glycopeptides such as vancomycin, teicoplanin, and telavancin, lincosamides, lipopeptides such as daptomycin, macrolides such as azithromycin, clarithromycin, dirithromycin, and erythromycin, monobactams, nitrofurans, penicillins, polypeptides such as bacitracin, colistin, and polymyxin B, quinolones, sulfonamides, and tetracyclines. Additional exemplary antibacterial agents include iodine, silver compounds, moxifloxacin, ciprofloxacin, levofloxacin, cefazolin, tigecycline, gentamicin, ceftazidime, ofloxacin, gatifloxacin, amphotericin, voriconazole, and natamycin.

[0372] Non-limiting examples of suitable anti-inflammatory agents include steroid active agents, including glucocorticoids, progestins, mineralocorticoids, and corticosteroids. Exemplary non-steroidal anti-inflammatory drugs include ketorolac, ibuprofen, nepafenac, diclofenac, aspirin, and naproxen. Other exemplary anti-inflammatory agents include triamcinolone acetonide, fluocinolone acetonide, prednisolone, dexamethasone, loteprednol, fluorometholone, and dipotassium glycyrrhizinate.

[0373] In some embodiments, the composition can further include one or more additional pharmaceutically active agents. Exemplary pharmaceuticals can include non-steroidal anti-inflammatory drugs (NSAIDs), such as flurbiprofen, ibuprofen, naproxen, indomethacin, and related compounds. In some embodiments, the composition can further include one or more mitotic inhibitors, including colchicine, taxol, and related compounds. In some embodiments, the composition can further include one or more topical disinfectants, such as benzoyl peroxide. In some embodiments, the composition can further include one or more polysaccharides produced by microalgae, such as alguronic acid.

[0374] In some embodiments, the composition can further include one or more immunomodulatory agents. Exemplary immunomodulatory agents include imiquimod, cyclosporine, tacrolimus, and rapamycin.

[0375] In some embodiments, the composition can further include one or more cytokines. Exemplary suitable cytokines include, but are not limited to, IL-10, TGF-β, IL-25, and IL-35. In certain embodiments, the cytokine can induce Treg activation (e.g., IL-25) and suppress Thl7 activation (e.g., IL-10) to minimize rejection.

[0376] In some embodiments, the compositions described herein further comprise at least one eukaryotic cell type. Some exemplary eukaryotic cell types include stem cells, mesenchymal stem cells, pancreatic islet cells, keratinocytes, fibroblasts, melanocytes, adipocytes, immune cells such as T lymphocytes, B lymphocytes, natural killer cells, and dendritic cells, or combinations thereof. In some embodiments, the stem cells can be adipose-derived mesenchymal stem cells. Functional characteristics of mesenchymal stem cells that can be beneficial for wound healing include the ability to migrate to sites of injury or inflammation, participate in the regeneration of damaged tissue, stimulate the proliferation and differentiation of resident progenitor cells, promote the recovery of damaged cells via growth factor secretion and matrix remodeling, and exert unique immunomodulatory and anti-inflammatory effects (see, for example, Phinney OG et al., Stem Cells, 25:2896-2902 (2007), Chamberlain G et al, Stem Cells, 25:2739-2749 (2007), Dazzi F et al., Curr Opin Oncol. 19:650-655 (2007). Each of these references is incorporated by reference in its entirety).

[0377] In some embodiments, the eukaryotic cells can be involved in increasing the structural integrity of connective tissue and / or promoting healing. In some embodiments, eukaryotic cells such as fibroblasts can be involved in enhancing or promoting the growth or attachment of cells or tissues.

[0378] In some embodiments, the recombinant collagen composition can contact 1) the transplanted hair graft and 2) the tissue into which the hair graft is transplanted. Recipient sites for the hair grafts include the scalp, facial area, axillary or chest area, or genital area. Specific areas of the facial area include the eyebrows, eyelids, mustache, sideburns, jaw, and cheeks. The recipient site can be any area of the skin where the appearance of hair is desired by the subject. In some embodiments, the contact between the composition described herein and the transplanted hair having the surrounding tissue of the recipient site promotes nutrient perfusion from the surrounding tissue to the graft, increasing the survival rate of the graft as compared to grafts transplanted without using the composition described herein. In certain embodiments, the contact between the composition described herein and the transplanted hair and between the composition described herein and the surrounding tissue of the recipient site promotes angiogenesis around the transplanted graft, resulting in an increased survival rate of the graft as compared to grafts transplanted without using the composition described herein.

[0379] Food composition In some embodiments, the composition described herein can be a food composition useful for providing it to a subject in need of collagen. For example, and in some embodiments, ingestion of the food composition described herein can provide health and / or skin benefits such as an increase in collagen intake, reduction of joint pain, and improvement of skin health. In certain embodiments, the composition can be in the form of a powder, capsule, liquid, or any other suitable form.

[0380] In some embodiments, the food composition can include one or more nutritional components such as ascorbic acid, biotin, chromium nicotinate, copper citrate, calcium D-pantothenate, cyanocobalamin, flaxseed, linum usitatissimum, folic acid, fructooligosaccharide (fiber), magnesium oxide, manganese citrate, maltodextrin, medium-chain triglyceride, flavoring, niacinamide, potassium iodide citrate, riboflavin, sugarcane (saccharum officinarum), sodium molybdate dihydrate, sodium selenite (selenium), soy protein isolate, stevia leaf extract / Stevia rebaudiana, thiamine HCl, tricalcium phosphate, vitamin A palmitate, vitamin D3, xanthan gum, zinc citrate, cellulose gum, pyridoxine hydrochloride, salt, tocopherol, antioxidants such as resveratrol, CoQ10, acai berry, lycopene, and pomegranate, natural or artificial sweeteners such as glucose, sucrose, fructose, saccharide, cyclamate, aspartame, sucralose, aspartame, acesulfame K, or sorbitol, flavorings such as flavor extracts, volatile oils, chocolate flavorings (e.g., caffeine-free cocoa or chocolate, chocolate substitutes such as soybeans), peanut butter flavorings, cookie crumbs, vanilla, or any commercially available flavorings, and any combination thereof.

[0381] Exemplary formulations In some embodiments, as described above, the compositions described herein can be in the form of an alcoholic or aqueous toner. Exemplary toner formulations are shown below. The collagen, collagen fragments, or stained collagen fragments described in the following examples may or may not be cross-linked. In some embodiments, the HA described in the following examples may or may not be cross-linked.

[0382] [Table 2]

[0383]

Table 3

[0384] In some of the above embodiments, the compositions described herein can be in the form of a cream, gel, or serum. Exemplary cream, gel, and serum formulations are described below.

[0385]

Table 4

[0386]

Table 5

[0387]

Table 6

[0388] In some of the embodiments as described above, the compositions described herein can be a shampoo or conditioner. Exemplary shampoo or conditioner formulations are described below.

[0389]

Table 7

[0390]

Table 8

[0391] In some of the embodiments as described above, the compositions described herein can be a nail care composition. Exemplary nail care formulations are described below.

[0392]

Table 9

[0393] F. Manufacturing Method In some embodiments, the present disclosure provides a method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with hyaluronic acid, and the method includes: a) dissolving hyaluronic acid in an NaOH solution; b) adjusting the pH by adding HCl; and c) adding collagen.

[0394] In some embodiments, the present disclosure provides a method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with hyaluronic acid, and the method includes: (a) dissolving hyaluronic acid in water; (b) dissolving collagen in water; and (c) mixing the two solutions. In some embodiments, the method further includes d) optionally crosslinking the hyaluronic acid and collagen in the solution by adding a coupling agent.

[0395] In some embodiments, the present disclosure provides a method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with hyaluronic acid, and the method includes: (a) dissolving hyaluronic acid in water or an NaOH solution; (b) optionally crosslinking the hyaluronic acid by adding a coupling agent before reducing the pH with HCl; (c) dissolving collagen in water; and (d) mixing the two solutions.

[0396] In some embodiments, the method further includes: e) adding a sodium salt to form a gel; and f) solidifying the gel. In some embodiments, the method further includes g) sterilizing the gel. In some embodiments, the method further includes h) dispensing the gel into a syringe under aseptic conditions.

[0397] In some embodiments, the present disclosure provides a method for preparing a composition comprising collagen and hyaluronic acid, the method comprising dissolving hyaluronic acid in a salt buffer such as PBS and adding collagen.

[0398] In some embodiments, the present disclosure provides a method for preparing a composition comprising collagen and hyaluronic acid, the method comprising a solution comprising NaCl.

[0399] In some embodiments, the method further comprises adding calcium hydroxyapatite, carboxymethylcellulose, polylactic acid, or any combination thereof to the composition or gel.

[0400] In some embodiments, the method further comprises adding any suitable biocompatible polymer known in the art. Suitable polymers include, but are not limited to, aliphatic polyesters, polyorthoesters, polyoxy esters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyester amides, hydrolyzable amines, polyalkylene oxides, polyethylene oxide (PEO), poloxamers, functionalized isoflavonoids, amino acids, and combinations thereof.

[0401] Examples of additional suitable polymers include, but are not limited to: poly(hydroxyethyl methacrylate), poly(sulfobetaine), poly(dimethylsiloxane), poly(caprolactone), poly(p-dioxanone), poly(trimethylene carbonate), poly(glycolide), poly(L-lactide), poly(D,L-lactide), poly(L-lactide-co-glycolide), poly(D,L-lactide-co-glycolide), poly(L-lactide-co-caprolactone), poly(L-lactide-co-D,L-lactide), poly(caprolactone-co-trimethylene carbonate), poly(lactide-co-trimethylene carbonate), poly(caprolactone-co-p-dioxanone), poly(trimethylene carbonate-co-p-dioxanone), poly(caprolactone-co-lactide), poly(lactide-co-1,5-dioxepan-2-one), and poly(1,5-dioxepan-2-one-co-p-dioxanone), poly(lactide-co-1,4-dioxepan-2-one), and poly(1,4-dioxepan-2-one-co-p-dioxanone), poly(glycolide-co-caprolactone-co-dioxanone), poly(lactide-co-caprolactone-co-dioxanone), poly(dioxanone-co-D,L-lactide), poly(dioxanone-co-glycolide), poly(dioxanone-co-caprolactone), poly(dioxanone-co-L-lactide), poly(dioxanone-co-D,L-lactide-co-caprolactone), poly(dioxanone-co-glycolide-co-caprolactone), poly(dioxanone-co-L-lactide-co-D,L-lactide), poly(dioxanone-co-L-lactide-co-glycolide), poly(dioxanone-co-L-lactide-do-caprolactone), poly(dioxanone-co-D,L-lactide-co-glycolide), poly(dioxanone-co-D,L-lactide-co-caprolactone), poly(dioxanone-co-glycolide-co-caprolactone), poly(dioxanone-co-L-lactide-co-glycolide-co-caprolactone), poly(dioxanone-co-D,L-lactide-co-glycolide-co-caprolactin), and combinations thereof.

[0402] In some embodiments, the method further comprises adding a bioabsorbable polymer selected from the group consisting of a solid homopolymer of poly(e-caprolactone), a solid homopolymer of poly(p-dioxanone), a solid homopolymer of poly(trimethylene carbonate), a solid copolymer of a plurality of e-caprolactone repeating units and a third lactone repeating unit, and a solid copolymer of a plurality of trimethylene carbonate repeating units and a second lactone repeating unit; the third lactone repeating unit is selected from the group consisting of a glycolide repeating unit, a lactide repeating unit, a trimethylene carbonate repeating unit, a p-dioxanone repeating unit, a 1,4-dioxepan-2-one repeating unit, a 1,5-dioxepan-2-one repeating unit, and combinations thereof.

[0403] In some embodiments, the method further comprises adding an anesthetic to the composition or gel.

[0404] In some embodiments, the method further comprises drying the composition by any suitable option such as oven drying, freeze drying, or spray drying. In some embodiments, the dried product can be a coating, film, or powder, which can be rehydrated in the gel as desired for application.

[0405] The methods described herein can include exposing the collagen and collagen fragments described herein to a crosslinking agent (chemical crosslinking agent such as glutaraldehyde, etc.) or crosslinking conditions (such as heat, radiation, etc.) such that the collagen contains non-endogenously imposed crosslinks.

[0406] In some embodiments, a method of crosslinking hyaluronic acid and one or more recombinant collagen fragments comprises dissolving hyaluronic acid and collagen in an aqueous solution to form an aqueous pre-reaction solution, wherein the aqueous pre-reaction solution further comprises a salt or has a low pH; modifying the aqueous pre-reaction solution to form a crosslinking reaction mixture comprising hyaluronic acid; recombinant collagen fragments; a water-soluble coupling agent; and a salt, wherein the crosslinking reaction has a higher pH than the aqueous pre-reaction solution; and reacting the crosslinking reaction mixture to thereby crosslink at least hyaluronic acid and one or more recombinant collagen fragments. In some embodiments, the concentrations and ratios of hyaluronic acid and recombinant collagen fragments can vary within a wide range. Each concentration is at least about 0.01-10 wt% for hyaluronic acid and at least about 0.01-10 wt% for recombinant collagen fragments, at least about 0.05-10 wt% for hyaluronic acid and at least about 0.1-10 wt% for recombinant collagen fragments, at least about 0.05-5 wt% for hyaluronic acid and at least about 0.5-7 wt% for recombinant collagen fragments, or at least about 0.1-2 wt% for hyaluronic acid and at least about 1-5 wt% for recombinant collagen fragments. Each concentration is about 0.01-10 wt% for hyaluronic acid and about 0.01-10 wt% for recombinant collagen fragments, about 0.05-10 wt% for hyaluronic acid and about 0.1-10 wt% for recombinant collagen fragments, about 0.05-5 wt% for hyaluronic acid and about 0.5-7 wt% for recombinant collagen fragments, or about 0.1-2 wt% for hyaluronic acid and about 1-5 wt% for recombinant collagen fragments.

[0407] In some embodiments, to adjust the pH to 9 - 12, a base (e.g., NaOH) is first added to hyaluronic acid. A crosslinking agent is added to the hyaluronic acid and the reaction is allowed to proceed. Then, one or more recombinant collagen fragments (pH 2 - 3) are added. The collagen reacts with the crosslinking agent and covalently binds to the HA. In some embodiments, the crosslinking agent may be slightly consumed (i.e., not fully reacted) when crosslinking the collagen to the HA. A hyaluronic acid solution is added as a lubricant and crosslinking agent scavenger to react with any excess / unreacted crosslinking agent. The final formulation can have a core of crosslinked hyaluronic acid with collagen that slightly crosslinks to the hyaluronic acid core. Additional soluble hyaluronic acid consumes the residual crosslinking agent and adds lubrication benefits.

[0408] In some embodiments, a base (e.g., NaOH) can be first added to hyaluronic acid to adjust the pH to 9 - 12. A crosslinking agent can be added to the hyaluronic acid and the reaction is allowed to proceed. The pH can be adjusted to less than 7.5 using an acid, and then one or more recombinant collagen fragments (pH 2 - 3) can be added. The collagen can react with the crosslinking agent and covalently bind to the HA. In some embodiments, the crosslinking agent may be slightly consumed (i.e., not fully reacted) when crosslinking the collagen to the HA. A hyaluronic acid solution can be added as a lubricant and crosslinking agent scavenger to react with any excess / unreacted crosslinking agent. The final formulation can have a core of crosslinked hyaluronic acid, and the collagen is slightly crosslinked to the hyaluronic acid core. Additional soluble hyaluronic acid can consume the residual crosslinking agent and add lubrication benefits.

[0409] Suitable linking agents for covalent crosslinking include, but are not limited to, ethylene glycol diglycidyl ether or 1,4-butanediol diglycidyl ether, divinyl sulfone, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), glycol ether diglycidyl ether, polyethylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, 1,6-hexanediol diglycidyl ether, 1,2,7,8-diepoxyoctane, divinyl sulfone. 1,2-bis(2,3-epoxypropyl)-2,3-ethylene, 1-(2,3-epoxypropyl)-2,3-epoxycyclohexane, glycerin triglycidyl ether, resorcinol diglycidyl ether, glycidyl allyl ether, ethylene glycol diglycidyl ether, 1,6-hexanediol diglycidyl ether, any one of neopentyl glycol diglycidyl ether, a photocrosslinking reagent such as ethyl eosin, a hydrazide such as bis-hydrazide, a trihydrazide or a polyhydrazide compound. In some embodiments, the linking agent can be an auxiliary crosslinking agent such as N-hydroxy-succinamide (NHS), NHS (sulfonate group-NHS), succinyl oxide, and sodium periodate. In some embodiments, the linking agent can be a carbodiimide class of linking agents such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), 1-ethyl-3-(3-trimethylaminopropyl)carbodiimide (ETC), 1,1'-2 sulfur-p-phenylene 2(carbodiimide), tert-butyl carbodiimide, and N,N'-diisopropylcarbodiimide (DIC).

[0410] In some embodiments, the crosslinking reaction occurs at a temperature of about 0 °C to about 100 °C, about 10 °C to about 90 °C, about 20 °C to about 90 °C, about 20 °C to about 80 °C, about 30 °C to about 70 °C or about 40 °C to about 60 °C. The crosslinking step can be carried out using any means known to those skilled in the art.

[0411] In some embodiments, the crosslinking reaction can be carried out at a temperature of about 0 °C to about 100 °C, about 10 °C to about 90 °C, about 20 °C to about 90 °C, about 20 °C to about 80 °C, about 30 °C to about 70 °C, or about 40 °C to about 60 °C. The crosslinking step can be carried out using any means known to those skilled in the art. In some embodiments, after crosslinking, the pH can be reduced to less than 7.5 before the addition of collagen. The addition of collagen can be about 0.1 wt% to about 25 wt% in water or optionally in a buffer such as PBS. Collagen can be added with or without mixing. Without being bound by a particular theory, when collagen is added without mixing, it is thought that the collagen penetrates the HA gel by diffusion. In embodiments where mixing is used, any suitable mixing device known to those skilled in the art can be used.

[0412] Those skilled in the art can optimize the crosslinking conditions according to the properties of hyaluronic acid and collagen fragments and perform crosslinking to an optimal degree. The degree of crosslinking can be controlled by varying several factors including the molecular weight of hyaluronic acid, the molecular weight of the recombinant collagen fragment, the concentration in the reaction mixture, the alkali concentration, and the ratio of the crosslinking agent.

[0413] The compositions according to the present disclosure can include collagen or fragments thereof produced by yeast or modified yeast used to produce collagen fragments. Suitable yeast strains and methods for producing recombinant collagen fragments therein are described in International Application No. PCT / US2022 / 027016, which is hereby incorporated by reference in its entirety.

[0414] Skin care In some embodiments, the compositions described herein can be skin care compositions that can be used to treat a skin area by topically applying the skin care composition to the skin area.

[0415] In some embodiments, the compositions described herein can be skin care compositions that can be used to treat the skin, hair, and nails by ingesting the composition as a dietary supplement.

[0416] In some embodiments, administration of the compositions described herein to the skin can improve or maintain skin quality and reduce or eliminate signs of aging. Signs of aging include, but are not limited to, all external visual and tactilely recognizable signs, as well as any other macroscopic or microscopic effects due to skin aging. Such signs can be induced or caused by endogenous or exogenous factors (e.g., aging over time and / or environmental damage). These signs include discontinuities in texture such as wrinkles and coarse deep wrinkles, fine lines, skin lines, cracks, bumps, enlarged pores (e.g., associated with accessory structures such as sweat gland ducts, sebaceous glands, or hair follicles), or unevenness or roughness, loss of skin elasticity (loss and / or inactivation of functional skin elastin), sagging (including swelling in the eye area and jowls), loss of skin firmness, loss of tightness, loss of skin bounce due to deformation, discoloration (including dark circles under the eyes), large freckles, poor complexion, hyperpigmented skin areas such as age spots and lentigines, keratosis, abnormal differentiation, hyperkeratosis, elastosis, collagenolysis, and other histological changes in the keratin, dermis, epidermis, cutaneous vasculature (e.g., telangiectasia or spider veins), and underlying tissues (e.g., fat and / or muscle), particularly tissues adjacent to the skin, and can result from processes including, but not limited to, these.

[0417] In some embodiments, the compositions described herein may be suitable for use as a dermal filler. Dermal filler compositions can replace lost endogenous matrix polymers or enhance / facilitate the function of existing matrix polymers in order to treat skin conditions due to aging or damage. Dermal filler compositions can fill wrinkles, fine lines, creases, scars, for example, to plump thin lips, or to fill sunken eyes or hollow cheeks to strengthen the skin tissue. Initial dermal filler products were generally made from collagen. One common matrix polymer used in modern dermal filler compositions is hyaluronic acid. Since hyaluronic acid is natural to the human body, it is generally well tolerated and is a treatment with relatively low risk for a wide variety of skin conditions.

[0418] In some embodiments, the compositions described herein may be suitable for use with a microneedle sheet or patch. The microneedles can deliver the compositions described herein to any area of the face and body to achieve a cosmetic or wound healing effect.

[0419] In some embodiments, the compositions described herein can be therapeutic compositions used to reduce or prevent the formation of scar tissue, promote healing, promote tissue regeneration, minimize local inflammation, minimize tissue rejection, and / or enhance skin and / or hair graft integration.

[0420] In some embodiments, the present disclosure provides a method of treating a skin condition, the method comprising administering to a subject in need thereof an effective amount of a composition comprising a recombinant collagen fragment. In some embodiments, the skin condition can be fine lines, wrinkles, dry skin, enlarged pore size, skin pigmentation disorders, loss of elasticity, unwanted hair, thinning of the skin, purpura, actinic keratosis, itching, eczema, acne, rosacea, erythema, telangiectasia, actinic telangiectasia, skin cancer, or nasal polyps.

[0421] In some embodiments, the composition can be administered topically to a skin area. In some embodiments, the skin area can be selected from the group consisting of the face, scalp, neck, ears, shoulders, chest (including breasts and / or décolletage), arms, hands, legs, abdomen, buttocks, groin, back, feet, and combinations thereof.

[0422] In some embodiments, the composition can be administered topically to the face. In some embodiments, the face can be selected from the group consisting of the forehead, eyes, perioral surface, jaw surface, periorbital surface, nasal surface, cheek skin surface, and combinations thereof.

[0423] In some embodiments, the present disclosure provides a method for improving collagen production in the skin, the method comprising administering an effective amount of a composition comprising a recombinant collagen fragment.

[0424] In some embodiments, the present disclosure provides a skin care product comprising the composition described herein for use in reducing the appearance of wrinkles, evening skin tone, providing moisture, reducing the appearance of dark circles under the eyes, increasing the collagen content of the skin, increasing skin density, improving skin firmness and elasticity, improving the appearance of fine lines and wrinkles, smoothing the texture of the skin, increasing skin glow and brightness, improving the appearance of sagging skin, whitening the skin, or any combination thereof.

Examples

[0425] The examples presented below are provided for illustrative purposes only and the embodiments described herein should in no way be construed as being limited to these examples. Rather, the embodiments should be construed to include any and all variations that become apparent as a result of the teachings provided herein.

[0426] Example 1 Polyethylene oxide (PEO) with a molecular weight of 400 kDa was purchased from Sigma (Sigma-Aldrich, St. Louis, MO) (“Polymer 1”). A recombinant collagen fragment with a molecular weight of 50 kDa having the amino acid sequence of SEQ ID NO: 1 was prepared in yeast as described in PCT / US2022 / 027016, which is incorporated herein by reference in its entirety (“Polymer 2”). A 5 wt% polyethylene oxide (PEO) solution was prepared by dissolving 2.5 g of PEO in 47.5 mL of deionized (DI) water and mixing at 400 RPM for 16 hours on a stirring plate. A 5% (w / v) solution containing the recombinant collagen fragment having the amino acid sequence of SEQ ID NO: 1 was generated by dissolving 5 grams of the 50 kDa protein in 100 mL of water. The two solutions were mixed in a 50 mL conical tube at a 1:1 (w / w) ratio and mixed at 10 RPM for 16 hours. Prior to evaluating solution stability, the conical tube was placed vertically on a rack overnight. The final solution was turbid and slowly phase-separated, with white or yellowish aggregates settling to the bottom of the tube.

[0427] Example 2 A mixture containing polyethylene oxide (PEO) and a recombinant collagen fragment with a molecular weight of 50 kDa having the amino acid sequence of SEQ ID NO: 1 was prepared, dried, and visually inspected. PEO with a molecular weight of 100 kDa was purchased from Sigma (“Polymer 1”). The remainder of the method was the same as in Example 1. The final solution phase-separated, with a turbid solution at the top and a slightly yellowish turbid solution at the bottom.

[0428] Example 3 A mixture of a first polyvinylpyrrolidone (PVP) and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 3 was prepared using the same method as described in Example 1, except that Polymer 1 was replaced with polyvinylpyrrolidone (Sigma; molecular weight 10 kDa). The final solution was clear. Approximately 13 grams (g) of the solution was poured into a 100 mL polytetrafluoroethylene (PTFE) evaporating dish and dried overnight at 45 °C. The dried film was visually clear.

[0429] This example demonstrates that a solution containing 10 kDa PVP and a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1 is completely miscible and can be prepared as a film or an article.

[0430] Example 4 A mixture of a second polyvinylpyrrolidone (PVP) and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 4 was prepared using the method described in Example 1, except that Polymer 1 was replaced with polyvinylpyrrolidone (Sigma; molecular weight 40 kDa). The final solution was clear. A film was prepared using the method described in Example 3. The dried film was visually clear.

[0431] This example demonstrates that a solution containing 40 kDa PVP and a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1 is completely miscible and can be prepared as a film or an article.

[0432] Example 5 A mixture of a third polyvinylpyrrolidone (PVP) and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 5 was prepared using the method described in Example 1 except that Polymer 1 was replaced with polyvinylpyrrolidone (Sigma; molecular weight 360 kDa). The final solution was clear. A film was prepared using the same method as in Example 3. The dried film was visually transparent.

[0433] This example demonstrates that a solution containing 360 kDa PVP and a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1 is completely miscible and can be prepared as a film.

[0434] Example 6 A mixture of a first chitosan and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared and visually inspected. Example 6 was prepared using the method described in Example 1 except that Polymer 1 was replaced with chitosan (molecular weight range of 50 kDa to 190 kD; Sigma). To completely solubilize the solution during mixing, 0.287 mL of acetic acid from Sigma was used to adjust the pH of the chitosan solution to pH 4. The final solution phase-separated into two layers. Viscosity was determined visually by examining the difference between the two layers. The upper layer had a lower viscosity and was more transparent, while the lower layer had a higher viscosity and was more turbid.

[0435] Example 7 A mixture of polyethylene glycol and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 7 was prepared using the method described in Example 1 except that Polymer 1 was replaced with polyethylene glycol (molecular weight 35 kDa Sigma). The final solution phase-separated, with a clear and clear solution at the top and a clear and slightly yellow solution at the bottom.

[0436] Example 8 A mixture of the first alginic acid and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 8 was prepared using the method described in Example 1, except that Polymer 1 was replaced with a medium-viscosity sodium alginate salt derived from brown algae (Sigma). The viscosity of 2% sodium alginate salt in water at 25 °C was 2,000 cps or more. 2.5 g of sodium alginate salt was dissolved in 97.5 mL of deionized ("DI") water, and a 2.5 wt% alginic acid solution was prepared by mixing at 400 RPM for 24 hours on a stirring plate due to the high viscosity of the solution. The viscosity was determined visually by examining the difference between two layers. Two solutions were mixed in a 50 mL conical tube at a ratio of 1:1 (v / v) and mixed at 10 RPM for 16 hours. The conical tube was placed vertically on a rack overnight before evaluating the solution stability. The final solution was clear. A film was prepared using the method described in Example 3. The dried film was visually cloudy with a haze uniformly dispersed throughout the film.

[0437] Example 9 A mixture of the second alginic acid and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 9 was prepared using the method described in Example 1, except that Polymer 1 was replaced with a low-viscosity sodium alginate salt derived from brown algae (Sigma). The viscosity of 1% sodium alginate salt in water at 25 °C was about 4 - 12 cps. Both the sodium alginate salt and collagen stock solutions were 5 wt%. Two solutions were mixed in a 50 mL conical tube at a ratio of 1:1 (v / v) and mixed at 10 RPM for 16 hours. The final solution was clear. A film was prepared using the method described in Example 3. The dried film was visually cloudy with a haze uniformly dispersed throughout the film.

[0438] Example 10 A mixture of polyacrylamide and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 10 was prepared using the method described in Example 1, except that Polymer 1 was replaced with polyacrylamide (molecular weight range of 5,000 kDa to 6,000 kDa; Sigma). The remaining part of the method was the same as in Example 1. The final solution was transparent. A film was prepared using the same method as described in Example 3. The dried film was visually transparent.

[0439] This example demonstrates that a solution containing 5,000 kDa to 6,000 kDa polyacrylamide and a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1 is miscible and can be prepared as a film or an article.

[0440] Example 11 A mixture of polyacrylamide and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 11 was prepared using the method described in Example 1, except that Polymer 1 was replaced with polyacrylamide (molecular weight of 40 kDa; Sigma). The remaining part of the method was the same as in Example 1. The final solution was transparent. A film was prepared using the method described in Example 3, except that an aluminum weighing pan was used. The dried film was visually transparent.

[0441] This example demonstrates that a solution containing 40 kDa polyacrylamide and a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1 is miscible and can be prepared as a film or an article.

[0442] Example 12 A mixture of a first polyvinyl alcohol (PVA) and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared and visually inspected. Example 12 was prepared using the method described in Example 1, except that Polymer 1 was replaced with PVA (molecular weight range of 89 kDa to 98 kDa; Sigma). A 4 wt% PVA solution was prepared by dissolving 2 g of PVA in 48 mL of DI water and mixing at 400 RPM for 16 hours on a stirring plate. Two solutions were mixed in a 50 mL conical tube in a 1:1 (w / w) ratio and mixed at 10 RPM for 16 hours. The final solution was phase-separated into two layers. The upper layer was turbid and a large amount of white powdery aggregates had settled at the bottom.

[0443] Example 13 A mixture of a second PVA and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared and visually inspected. Example 13 was prepared using the method described in Example 12, except that Polymer 1 was replaced with a 4% PVA (w / v) solution (unknown molecular weight; Fisher Scientific). The remaining part of the method was the same as in Example 12. The final solution was phase-separated into two layers. The viscosity was determined visually by examining the difference between the two layers. The upper layer had a lower viscosity and was more turbid with small suspended particles. The lower layer had a higher viscosity and was transparent.

[0444] Example 14 A mixture of the first hyaluronic acid (HA) and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 14 was prepared using the method described in Example 1, except that Polymer 1 was replaced with hyaluronic acid (HA) (having a molecular weight of 1,000 kDa purchased from Pure Health Botanicals (Saint Charles, IL)). 5 g of HA was dissolved in 195 mL of DI water, and a 2.5 wt% HA solution was prepared by mixing at 400 RPM for 40 hours using a high-shear impeller due to the high viscosity of the solution. A 5% (w / v) collagen solution containing the recombinant collagen fragment of SEQ ID NO: 1 was produced as described in Example 1. The HA solution and the collagen solution were mixed in a 50 mL conical tube at a 2:1 (w / w) ratio and mixed at 10 RPM for 16 hours to maintain a dry mass ratio of 1:1. The final solution was clear. A film was prepared using the method described in Example 3. The dried film was visually cloudy with a haze uniformly dispersed throughout the film.

[0445] Example 15 A mixture of a second hyaluronic acid (HA) and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 15 was prepared using the method in Example 1 except that Polymer 1 was replaced with a non-animal-based HA (molecular weight 50 kDa; Pure Health Botanicals (Saint Charles, IL) and DSM (Kaiseraugst, Switzerland)). A 5 wt% HA solution was prepared by dissolving 5 g of HA in 95 mL of DI water and mixing at 400 RPM for 16 hours on a stirring plate. A 5% (w / v) collagen solution containing the recombinant collagen fragment of SEQ ID NO: 1 was produced as described in Example 1. The two solutions were mixed in a 50 mL conical tube in a 1:1 (w / w) ratio and mixed at 10 RPM for 16 hours. The final solution was clear. A film was prepared using the method described in Example 3. The dried film was visually cloudy with a haze uniformly dispersed throughout the film. Further, when an aliquot of the mixture of HA and the collagen solution was placed in a refrigerator at 4 °C overnight, the solution remained clear.

[0446] Example 16 A mixture of a first gelatin and HA was prepared and visually inspected. Example 16 was prepared using the method of Example 15 except that Polymer 2 was replaced with a low molecular weight beef gelatin dietary supplement (Great Lakes Wellness, Grayslake, IL). 1 g of gelatin was dissolved in 45 mL of 0.01 N HCl, and a 2.2 wt% gelatin solution was prepared by mixing at 400 RPM for 16 hours using a high shear impeller due to the high viscosity of the solution. A 5 wt%, 50 kDa HA solution was prepared using the method described in Example 15. The HA solution and the gelatin solution were mixed in a 50 mL conical tube in a 1:2.4 (w / w) ratio and mixed at 10 RPM for 16 hours to maintain a 1:1 dry mass ratio. The final solution phase-separated into two layers. The viscosity was determined visually by examining the difference between the two layers. The upper layer had a lower viscosity and was more transparent. The lower layer had a higher viscosity and was more turbid.

[0447] Example 17 A mixture of the second gelatin and HA was prepared and visually inspected. Example 17 was prepared using the method described in Example 16, except that Polymer 2 was replaced with gelatin derived from porcine skin (molecular weight 300 kDa; Sigma). 0.5 g of gelatin was dissolved in 45 mL of 0.01 N HCl, and a 1.1 wt% gelatin solution was prepared by mixing at 400 RPM for 16 hours using a high-shear impeller due to the high viscosity of the solution. A 5 wt%, 50 kDa HA solution was prepared using the method described in Example 15. The HA solution and the gelatin solution were mixed in a 50 mL conical tube at a ratio of 1:4.5 (w / w) and mixed at 10 RPM for 16 hours to maintain a dry mass ratio of 1:1. The final solution phase-separated, with a large number of white aggregates sedimenting at the bottom and scattered white aggregates suspended or floating at the top.

[0448] Example 18 A mixture of HA and type I collagen was prepared and visually inspected. Example 18 was prepared using the method described in Example 15, except that Polymer 2 was replaced with type I collagen (molecular weight 414 kDa; Sigma). 0.3 g of type I collagen was dissolved in 45 mL of 0.01 N HCl, and a 0.7 wt% type I collagen solution was prepared by mixing at 400 RPM for 16 hours using a high-shear impeller due to the high viscosity of the solution. A 5 wt%, 50 kDa HA solution was prepared using the method described in Example 15. The HA solution and the type I collagen solution were mixed together in a 50 mL Falcon conical tube at a ratio of 1:7 (w / w) and mixed at 10 RPM for 16 hours using a Hula mixer to maintain a dry mass ratio of 1:1. The final solution phase-separated, with a large number of white fibrils sedimenting at the bottom and scattered white fibrils suspended or floating at the top.

[0449] Example 19 A mixture of cellulase and HA was prepared and visually inspected. Example 19 was prepared using the method described in Example 15, except that Polymer 2 was replaced with cellulase (Sunson Industry Group Co., Ltd.). A 5% (w / v) cellulase solution was prepared by dissolving 2.5 g of cellulase in 47.5 mL of DI water and mixing at 400 RPM for 16 hours on a stirring plate. A 5 wt%, 50 kDa HA solution was prepared using the same method as described in Example 15. The two solutions were mixed together in a 50 mL conical tube at a 1:1 (w / w) ratio and mixed at 10 RPM for 16 hours. The final solution was turbid and a yellow powder settled to the bottom.

[0450] Example 20 A mixture of HA, chitosan, and collagen (recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared and visually inspected. Example 20 was prepared using the method described in Example 15, except that the chitosan used in Example 6 was added as Polymer 3. A 5 wt%, 50 kDa HA solution was prepared by dissolving 5 g of HA in 95 mL of DI water and mixing at 400 RPM for 16 hours on a stirring plate. As described in Example 1, a 5% (w / v) collagen solution containing the recombinant collagen fragment of SEQ ID NO: 1 was produced by dissolving 5 grams of the 50 kDa protein in 100 mL of water. A 5 wt% chitosan solution was prepared by dissolving 2.5 g of chitosan in 47.5 mL of DI water and mixing at 400 RPM for 16 hours on a stirring plate. To completely solubilize the solution during mixing, 0.287 mL of acetic acid from Sigma was used to adjust the pH of the chitosan solution to pH 4. The three solutions were mixed in a 50 mL conical tube at a 1:1:1 (w / v) ratio and mixed at 10 RPM for 16 hours. The final solution was phase-separated, and large masses of yellow aggregates were suspended or floated in a low-viscosity medium.

[0451] Example 21 A mixture of hyaluronic acid and a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1 (hydroxylated to 22%) was prepared and visually inspected. The recombinant collagen fragment was hydroxylated by the method described in International Publication No. 2021 / 163485, and the percentage of hydroxylation was measured by the method described in International Publication No. 2021 / 163485 (the disclosure of which is incorporated herein by reference in its entirety). Example 22 was prepared using the method in Example 15, except that Polymer 2 was replaced with 22% hydroxylated collagen having a molecular weight of 50 kDa. The 22% hydroxylated collagen solution had a solids content of 1.3% and a pH of about 6. A 5 wt%, 50 kDa HA solution (Polymer 1) was prepared by dissolving 5 g of HA in 95 mL of DI water and mixing at 400 RPM for 16 hours on a stirring plate. The two solutions were mixed in a 15 mL Falcon tube at a 1:1 (w / w) ratio and mixed at 10 RPM for 16 hours. The final solution was turbid, indicating that the two polymers were immiscible in solution at this level of hydroxylation.

[0452] Example 22 A mixture of hyaluronic acid and a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1 (hydroxylated to 44%) was prepared and visually inspected. The recombinant collagen fragment was hydroxylated by the method described in International Application No. WO2021 / 163485, and the percentage of hydroxylation was measured by the method described in International Application No. WO2021 / 163485 (the disclosure of which is incorporated herein in its entirety). Example 22 was prepared using the same method as described in Example 21, except that 44% hydroxylated collagen having a molecular weight of 50 kDa was used as Polymer 2. The 44% hydroxylated collagen solution had a solids content of 1.3% and a pH of approximately 6. A 5 wt%, 50 kDa HA solution (Polymer 1) was prepared by dissolving 5 g of HA in 95 mL of DI water and mixing at 400 RPM on a stirring plate for 16 hours. The two solutions were mixed in a 15 mL conical tube at a 1:1 (w / w) ratio and mixed at 10 RPM for 16 hours. The final solution was turbid, slowly phase-separated, and white aggregates settled to the bottom of the tube.

[0453] Example 23 A mixture of biphasic cross-linked hyaluronic acid (containing cross-linked HA and non-cross-linked HA) and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared and visually inspected. Example 23 was prepared using the method described in Example 15, except that Polymer 1 was replaced with biphasic cross-linked HA (particle size 1.25 - 2 mm; Beijing Mengbring Bio-Sci-Tec.Co.,Ltd.). A 2.4% (w / v) HA solution was used. A solution of 20 wt% recombinant collagen fragment of SEQ ID NO: 1 (Polymer 2) was prepared by dissolving 2 g of 50 kDa recombinant collagen in 18 mL of DI water and mixing at 10 RPM for 4 hours. The two solutions were mixed together in a 50 mL conical tube at a 1:1 (w / w) ratio and mixed at 10 RPM for 16 hours. The mixed solution was visually transparent.

[0454] This example demonstrates that a solution containing biphasic cross-linked HA and a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1 is completely miscible.

[0455] Example 24 A mixture of monophasic cross-linked hyaluronic acid and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and visually inspected. Example 24 was prepared using the same method as in Example 15, except that Polymer 1 was replaced with monophasic cross-linked HA (Bloomage Biotechnology Corp. (Jinan, Shandong, China)). A 2% (w / v) monophasic cross-linked HA solution was used. 2 g of 50 kDa recombinant collagen was dissolved in 18 mL of DI water and mixed at 10 RPM for 4 hours to prepare a 20 wt% collagen solution containing the recombinant collagen fragment of SEQ ID NO: 1. The two solutions were mixed in a 50 mL conical tube at a 1:1 (w / w) ratio and mixed at 10 RPM for 16 hours. The solution was visually clear.

[0456] This example demonstrates that a solution containing monophasic cross-linked HA and a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1 is completely miscible.

[0457] Example 25 A mixture of biphasic cross-linked hyaluronic acid (containing cross-linked HA and non-cross-linked HA) and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and visually inspected. Example 25 was prepared using the method described in Example 24, except that Polymer 1 was replaced with biphasic cross-linked HA (Restylane). A 2.0% (w / v) biphasic cross-linked HA solution was used. 4 g of 50 kDa recombinant collagen was dissolved in 16 mL of DI water and mixed at 10 RPM for 4 hours to prepare a 40 wt% solution of the recombinant collagen fragment of SEQ ID NO: 1. The two solutions were mixed in a 1 mL Eppendorf tube using a pipette at HA:collagen ratios of 1:3, 1:1, 10:3, and 100:3 (w / w) and mixed at 10 RPM for 16 hours. The solution was visually clear.

[0458] This example demonstrates that a solution containing biphasic cross-linked hyaluronic acid and a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1 is completely miscible.

[0459] Example 26 A mixture of 5% hyaluronic acid and collagen (recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) was prepared and visually inspected. Example 26 was prepared using the method described in Example 15, except that the pH of the 5 wt%, 50 kDa HA solution was adjusted to pH 5.2 - 7.2 with sodium hydroxide prior to the addition of the collagen solution containing the recombinant collagen fragment of SEQ ID NO: 1. The two solutions were mixed together in a 50 mL tube at a 1:1 (w / w) ratio and mixed at 10 RPM for 16 hours. The solution was visually clear. A film was prepared using the method of Example 3. The dried film was visually cloudy with a haze uniformly dispersed throughout the film.

[0460] This example demonstrates that a solution containing 5% hyaluronic acid at pH 5.2 - 7.2 and a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1 is completely miscible.

[0461] Example 27 Mixtures of 5% hyaluronic acid and collagen (recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1) were prepared at different pHs and visually inspected. Example 27 was prepared using the method described in Example 15, except that the pH of the 5 wt%, 50 kDa HA solution was adjusted to pH 9, 10, 11, and 12 with sodium hydroxide prior to the addition of the recombinant collagen fragment of SEQ ID NO: 1. The two solutions were mixed in a 20 mL glass vial at a 1:1 (w / w) ratio and, since the viscosity of the basic solution was lower, mixed for 30 minutes at 400 RPM using a stir bar. Then, while mixing at 400 RPM using a stir bar, the solution was readjusted to pH 7 ± 1 with concentrated hydrochloric acid. The solution was visually clear both before and after adjusting the pH to 7 ± 1.

[0462] This example demonstrates that a solution containing 5% hyaluronic acid at pH 9, 10, 11, 12 and a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1 is completely miscible.

[0463] Example 28 A mixture of hyaluronic acid and collagen was prepared and visually inspected. Example 28 was prepared using the method of Example 15, except that Polymer 1 was replaced with a 2% HumaColl21® solution (Geltor). A 2 wt% HA solution with a molecular weight of 50 kDa was prepared using the method described in Example 15. The two solutions were mixed in a 1:1 (w / w) ratio in a 2 mL glass vial and mixed on a stirring plate at 400 RPM for 1 hour. The final solution was visually clear.

[0464] This example demonstrates that a solution containing a 2% HumaColl21® solution and a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1 is completely miscible.

[0465] Example 29 Blend hyaluronic acid with full-length recombinant collagen and a 50 kDa recombinant collagen fragment, mix with HA, and visually inspect. Blend full-length recombinant collagen and a 50 kDa recombinant collagen fragment, and mix with HA. Visually inspect the solution. Blend a 50 kDa recombinant collagen fragment with full-length recombinant collagen and / or hydrolyzed recombinant collagen (or any combination of these three), mix with HA, and visually inspect. Then, blend full-length recombinant collagen and a 50 kDa recombinant collagen fragment, and blend full-length recombinant collagen and hydrolyzed full-length recombinant collagen, and mix with HA. Visually inspect the solution.

[0466] Example 30 A poly(2-ethyl-2-oxazoline) (PEOx) solution was mixed with a solution containing a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1. Example 30 was prepared using the same method as described in Example 1, except that Polymer 1 was replaced with PEOx (molecular weight 50 kDa; Sigma) 1. The remaining part of the method was the same as in Example 1. The final solution was visually transparent, indicating that the polymer was miscible in the solution. A film was prepared using the same method as described in Example 3, except in an aluminum weighing pan. The dried film appeared to phase-separate during drying.

[0467] Example 31 A polyethyleneimine (PEI) solution was mixed with a solution containing a recombinant collagen fragment having a molecular weight of 50 kDa with SEQ ID NO: 1. Example 31 was prepared using the method described in Example 1, except that Polymer 1 was replaced with PEI (molecular weight 100 kDa; Polysciences (Warrington, Pennsylvania)). A 5 wt% polyethyleneimine solution was prepared by dissolving 0.5 g of PEI in 9.5 g of Milli-Q water at 80 °C using a stir bar at 450 RPM for 3 hours or until the solution became completely clear. The solution was then cooled to room temperature and the pH was adjusted to 7 using 12 N HCl. The remaining part of the method was the same as in Example 1. The solution was transparent, indicating that the polymer was miscible in the solution. A film was prepared using the method described in Example 3. The dried film was visually opaque.

[0468] Example 32 A mixture of the first sodium carboxymethyl cellulose (NaCMC) and collagen (a recombinant collagen fragment having a molecular weight of 50 kDa and having SEQ ID NO: 1) was prepared, dried, and visually inspected. Example 32 was prepared using the method described in Example 1, except that Polymer 1 was replaced with sodium carboxymethyl cellulose (molecular weight approximately 250 kDa and degree of substitution 1.22; Sigma)). A 5 wt% sodium carboxymethyl cellulose solution was prepared by dissolving 0.5 g of sodium carboxymethyl cellulo...

Claims

1. A composition comprising (i) recombinantly produced collagen and (ii) one selected from the list consisting of hyaluronic acid, polyvinylpyrrolidone, polyacrylamide, poly(ethylene oxide), poly(2-oxazoline), polyethyleneimine, sodium carboxymethylcellulose, chondroitin sulfate, acetylated hyaluronate, zinc hyaluronate, or sodium alginate salt, wherein the recombinant collagen is completely miscible with the one selected from the list. A composition wherein the recombinant collagen has at least about 85% sequence identity with respect to the amino acid sequence described in Sequence ID No. 1, or at least about 85% sequence identity with respect to the amino acid sequence described in any one of Sequence IDs No. 2 to 1002.

2. The composition according to claim 1, wherein the molecular weight of the recombinant collagen is about 5 kDa to about 150 kDa.

3. The composition according to claim 1, wherein the hyaluronic acid has a molecular weight of about 3 kDa to about 10,000 kDa.

4. The composition according to claim 1, wherein the polyvinylpyrrolidone has a molecular weight of about 10 kDa to about 400 kDa.

5. The composition according to claim 1, wherein the polyacrylamide has a molecular weight of about 30 kDa to about 50 kDa, or about 5,000 kDa to about 6,000 kDa.

6. The composition according to claim 1, wherein the carboxymethylcellulose has a molecular weight of about 250 kDa and a degree of substitution of about 0.70 to about 0.90 or about 1.00 to about 1.

25.

7. The composition according to claim 1, wherein the chondroitin sulfate has a molecular weight of about 50 kDa.

8. A non-therapeutic method comprising administering or applying an effective amount of the composition according to any one of claims 1 to 7 to a subject in need thereof.

9. A composition according to any one of claims 1 to 7, which is used in a treatment comprising administering or applying an effective amount of the composition to a subject requiring the treatment.

10. The composition according to claim 9, wherein the treatment comprises treating an inflammatory skin disease or a mucosal disease.

11. The composition according to claim 9, wherein the treatment comprises treating lipodystrophy.

12. The composition according to claim 9, wherein the treatment comprises treating a wound, the wound being a chronic wound, a surgical wound, a superficial wound, an abrasion, or the wound being the result of thermal injury.

13. A method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid, a) Dissolve hyaluronic acid in NaOH solution; b) Adding collagen; and c) Adjusting the pH by adding HCl; Includes, A method wherein the collagen is recombinant collagen having at least about 85% amino acid sequence identity with SEQ ID NO: 1, and / or one or more collagen fragment sequence variants having at least about 85% sequence identity with the amino acid sequence described in any one of SEQ ID NOs: 975 to 1002.

14. A method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid, a) Dissolving hyaluronic acid in water to form a hyaluronic acid solution; b) Dissolving collagen in water to form a collagen solution; and c) Mixing the hyaluronic acid and collagen solution. Includes, A method wherein the collagen is a recombinant collagen fragment having at least about 85% amino acid sequence identity with SEQ ID NO: 1, and / or one or more collagen fragment sequence variants having at least about 85% sequence identity with the amino acid sequence described in any one of SEQ ID NOs: 975 to 1002.

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