Collagen compositions and methods of use thereof

Abstract

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

Description

[0001] References to sequence listings submitted electronically

[0002] The contents of the electronically submitted Sequence Listing XML (name: 4431_090PC01_SequenceListing_ST26.xml; size: 1,309,312 bytes; and creation date: May 30, 2023) submitted with this application are incorporated herein by reference in their entirety. Background Art

[0003] Collagen is one of the most important proteins in the human body and is present in connective tissues such as cartilage, bone, tendon, ligaments 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 produced using recombinant technology.

[0004] Collagen has many known uses. In the cosmetics and skin care industries, for example, skin care compositions comprising collagen can be used to combat the effects of aging and environmental stress on the appearance, elasticity, and thickness of the skin. For example, aging and environmental factors can lead to skin conditions including, but not limited to, fine lines, wrinkles, dry skin, enlarged pores, skin discoloration, reduced elasticity, excess hair, thinning of the skin, purpura, actinic keratosis, pruritus, eczema, acne, rosacea, erythema, telangiectasia, actinic telangiectasia, skin cancer, and hypertrophic rosacea.

[0005] Separately, dermal hyaluronic acid (HA) fillers are commonly used for soft tissue augmentation. Other age-related conditions that may benefit from treatment with collagen and hyaluronic acid include, but are not limited to, arthritis, osteoarthritis, eye problems, cell adhesions, and similar conditions. Although there are many skin care products and dermal fillers on the market that improve the appearance of the skin, many consumers are hesitant to use chemically synthesized products, which they believe are environmentally unfriendly or unsafe. It is very desirable to find a miscible product comprising HA and collagen that retains the benefits of both components, but natural collagen is incompatible with HA—each tends to separate during storage and use. Similar to HA, there is a need in the art for miscible collagen blends of polymers for delivering therapeutic agents. Summary of the invention

[0006] In some embodiments, 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 recombinantly produced collagen. In some embodiments, the collagen is a recombinantly produced collagen fragment.

[0007] In some embodiments, the composition has an isoelectric point (pi) of about 4.0 to about 12.0. In some embodiments, the composition has a pI of about 4.0 to about 5.0. In some embodiments, the composition has a pI of about 6.0 to about 7. In some embodiments, the composition has a pI of about 9.0 to about 10.0. In some embodiments, the composition has a pI of about 9.0 to about 9.7.

[0008] In some embodiments, the collagen has a molecular weight of about 5 kDa to about 150 kDa.

[0009] In some embodiments, the hyaluronic acid has a molecular weight of about 3kDa to about 10,000kDa. In some embodiments, the hyaluronic acid has a molecular weight of about 20kDa to about 2,000kDa. In some embodiments, the hyaluronic acid has a molecular weight of about 1,000kDa to at least about 5,000kDa. In some embodiments, the hyaluronic acid has a molecular weight of about 2,000kDa to at least about 3,000kDa. In some embodiments, the hyaluronic acid has a molecular weight of about 1,000kDa. In some embodiments, the hyaluronic acid has a molecular weight of about 50kDa.

[0010] In some embodiments, the collagen does not contain hydroxyproline (Hyp) residues. In some embodiments, the collagen contains about 1% to about 20% Hyp residues based on the total number of prolines in the collagen. In some embodiments, the collagen does not contain hydroxyl groups. In some embodiments, the collagen contains about 1 to about 120 hydroxyl groups. In some embodiments, the collagen does not contain amine groups. In some embodiments, the collagen contains about 1 to about 100 amine groups. In some embodiments, the collagen does not contain carboxylic acid groups. In some embodiments, the collagen contains about 1 to about 80 carboxylic acid groups.

[0011] In some embodiments, the hyaluronic acid is cross-linked. In some embodiments, the hyaluronic acid is not cross-linked. In some embodiments, the hyaluronic acid is a biphasic cross-linked hyaluronic acid. In some embodiments, the hyaluronic acid is a monophasic cross-linked hyaluronic acid.

[0012] In some embodiments, the hyaluronic acid solution has a pH in the range of about 5.0 to about 12. In some embodiments, the hyaluronic acid has a pH in the range of about 9.0 to about 12. In some embodiments, the hyaluronic acid has a pH in the range of about 5.2 to about 7.2.

[0013] In some embodiments, the ratio of the hyaluronic acid to the collagen (w / w) is in the range of about 1:9 to 9:1.

[0014] In some embodiments, the collagen is cross-linked. In some embodiments, the collagen is not cross-linked.

[0015] In some embodiments, the hyaluronic acid is crosslinked by a chemical crosslinker, ultraviolet (UV) radiation, microwave radiation, or heating. In some embodiments, the collagen is crosslinked by a chemical crosslinker, ultraviolet (UV) radiation, microwave radiation, or heating.

[0016] In some embodiments, the composition has a viscosity of about 1 centipoise to about 100,000 centipoise.

[0017] In some embodiments, the composition has a viscosity of about 10,000 centipoise to about 80,000 centipoise.

[0018] In some embodiments, the composition has a viscosity of about 50,000 centipoise to about 70,000 centipoise.

[0019] In some embodiments, the composition is a solution. In some embodiments, the composition is a gel. In some embodiments, the composition is a foam.

[0020] In some embodiments, the present disclosure provides films comprising the compositions disclosed herein.

[0021] In some embodiments, the present disclosure provides one or more fibers comprising a composition disclosed herein.

[0022] In some embodiments, the present disclosure provides coatings prepared from the compositions disclosed herein.

[0023] In some embodiments, the present disclosure provides powders prepared from the compositions disclosed herein.

[0024] In some embodiments, the present disclosure provides a slurry prepared from a composition disclosed herein.

[0025] In some embodiments, the present disclosure provides one or more particles prepared from a composition disclosed herein.

[0026] In some embodiments, the collagen is non-hydroxylated. In some embodiments, the recombinant collagen fragment is non-hydroxylated.

[0027] 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.

[0028] In some embodiments, the composition further comprises a pharmaceutically active agent. In some embodiments, the composition further comprises a cosmetically active agent.

[0029] In some embodiments, the collagen or the recombinant collagen fragment has at least about 85% sequence identity with the amino acid sequence shown in SEQ ID NO: 1. In some embodiments, the collagen or the recombinant collagen fragment has at least about 85% sequence identity with the amino acid sequence shown in any one of SEQ ID NO: 2 to SEQ ID NO: 1002.

[0030] In some embodiments, the collagen or the recombinant collagen is fermented or chemically synthesized.

[0031] In some embodiments, the present disclosure provides methods of increasing collagen production in a cell, comprising administering or applying an effective amount of a composition disclosed herein.

[0032] In some embodiments, the present disclosure provides a method of medical or cosmetic treatment comprising administering or applying an effective amount of a composition disclosed herein.

[0033] In some embodiments, the treatment comprises filling wrinkles, increasing viscoelasticity, or increasing volume of the face or body. In some embodiments, the treatment comprises forming a cosmetic implant in the patient. In some embodiments, the treatment comprises tissue augmentation.

[0034] In some embodiments, the composition is applied to the face, eye, nose, nasolabial fold, neck, collarbone, arm, abdomen, blood vessel, bladder, urethra, nerve, cartilage, bone, breast, vocal cord, vocal folds, tympanic membrane, joint, intervertebral disc, buttocks, hip, calf, thigh, flank, or hand of a subject.

[0035] In some embodiments, treatment comprises treatment of an inflammatory skin disease or a mucosal disease. In some embodiments, treatment comprises treatment of lipoatrophy.

[0036] 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 wound of the subject. In some embodiments, the wound is a chronic wound, a surgical wound, a superficial wound, an abrasion, or wherein the wound is the result of thermal injury.

[0037] In some embodiments, the present disclosure provides a skin care product comprising a composition disclosed herein, the skin care product being used to reduce the appearance of wrinkles, even out skin tone, provide moisture, reduce the appearance of dark circles under the eyes, increase the collagen content of the skin, increase skin density, improve skin firmness and elasticity, improve the appearance of fine lines and wrinkles, smooth skin texture, increase skin radiance and brightness, improve the appearance of sagging skin, whiten the skin, sun care, makeup, hair care, fragrance, oral care, or any combination thereof.

[0038] In some embodiments, the present disclosure provides a dermal filler comprising a composition disclosed herein.

[0039] In some embodiments, the dermal filler is a heat stable, sterile, 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 over a temperature range of up to at least about 135° C. In some embodiments, the dermal filler is terminally sterilized. In some embodiments, the dermal filler is injectable.

[0040] 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 polylactic acid, aliphatic polyesters, polyorthoesters, polyoxyesters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyesteramides, hydrolyzable amines, polyalkylene oxides, poloxamers, functionalized isoflavonoids, amino acids, solid homopolymers of poly(ε-caprolactone), solid homopolymers of poly(p-dioxanone), solid homopolymers of poly(trimethylene carbonate), solid copolymers of a plurality of ε-caprolactone repeating units and a third lactone repeating unit, solid copolymers 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 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, or any combination thereof.

[0041] In some embodiments, the dermal filler further comprises a pharmaceutically active agent. In some embodiments, the pharmaceutically active agent comprises an anesthetic.

[0042] In some embodiments, the present disclosure provides for use of a composition, skin care product, or dermal filler disclosed herein for medical or cosmetic treatment of a subject in need thereof.

[0043] In some embodiments, the present disclosure provides a method of preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid, the method comprising: dissolving the hyaluronic acid in a NaOH solution; adding the collagen; and adjusting the pH by adding HCl.

[0044] 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, the method comprising: dissolving the hyaluronic acid in a NaOH solution; adjusting the pH by adding HCl; and adding collagen.

[0045] 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, the method 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.

[0046] In some embodiments, the method further comprises: cross-linking the hyaluronic acid and collagen in the solution, optionally by adding a coupling agent.

[0047] In some embodiments, the method further comprises cross-linking the hyaluronic acid and collagen in the solution, optionally by adding a coupling agent prior to adjusting the pH with HCl and adding the collagen.

[0048] In some embodiments, the method further comprises: adding sodium chloride salt to cause gel formation; and allowing the gel to solidify. In some embodiments, the method further comprises sterilizing the gel.

[0049] In some embodiments, the solution comprises NaCl.

[0050] In some embodiments, the method further comprises adding calcium hydroxyapatite, carboxymethyl cellulose, polylactic acid, polylactic acid, aliphatic polyesters, polyorthoesters, polyoxyesters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyesteramides, hydrolyzable amines, polyalkylene oxides, poloxamers, functionalized isoflavones, amino acids, solid homopolymers of poly(ε-caprolactone), solid homopolymers of poly(p-dioxanone), solid homopolymers of poly(trimethylene carbonate), solid copolymers of multiple ε-caprolactone repeating units and a third lactone repeating unit, solid copolymers of multiple trimethylene carbonate repeating units and a second lactone repeating unit; wherein 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 any combination thereof, or any combination thereof.

[0051] In some embodiments, the method further comprises adding an anesthetic. In some embodiments, the method further comprises dispensing the gel into a syringe under sterile conditions.

[0052] In some embodiments, the present disclosure provides a kind of compositions comprising collagen and polyvinylpyrrolidone, wherein this collagen is completely miscible with this polyvinylpyrrolidone.In some embodiments, this polyvinylpyrrolidone has the molecular weight of about 10kDa to about 400kDa.In some embodiments, this polyvinylpyrrolidone has the molecular weight of about 50kDa.

[0053] In some embodiments, the collagen is completely miscible with the polyacrylamide. In some embodiments, the polyacrylamide has a molecular weight of about 5,000 kDa to about 6,000 kDa. In some embodiments, the polyacrylamide has a molecular weight of about 30 kDa to about 50 kDa.

[0054] In some embodiments, the present disclosure provides a composition comprising collagen and polyethylene oxide, wherein the collagen is completely miscible with the polyethylene oxide. In some embodiments, the collagen is completely miscible with poly(2-oxazoline). In some embodiments, the collagen is completely miscible with polyethyleneimine.

[0055] 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 250kDa and a degree of substitution of about 1.00 to about 1.25. In some embodiments, the carboxymethylcellulose has a molecular weight of about 250kDa and a degree of substitution of about 0.70 to about 0.90. In some embodiments, the carboxymethylcellulose has a molecular weight of about 250kDa and a degree of substitution of about 0.79. In some embodiments, the carboxymethylcellulose has a degree of substitution of about 0.87.

[0056] In some embodiments, the carboxymethyl cellulose has a high viscosity.

[0057] 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.

[0058] In some embodiments, the present disclosure provides a composition comprising collagen and acetylated hyaluronate. In some embodiments, the collagen and the acetylated hyaluronate are completely miscible.

[0059] In some embodiments, the present disclosure provides a composition comprising collagen and zinc hyaluronate. In some embodiments, the collagen has a sequence identity of 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% identity to the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the collagen has a sequence identity of at least about 98% to the amino acid sequence set forth in SEQ ID NO: 1.

[0060] 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 shown in any one of SEQ ID NO: 975 to SEQ ID NO: 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 uncross-linked HA.

[0061] In some embodiments, the present disclosure provides a composition comprising 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% amino acid sequence identity with the amino acid sequence shown in any one of SEQ ID NO:975 to SEQ ID NO:1002, and polyvinyl pyrrolidone, and polyvinyl pyrrolidone, wherein the recombinant collagen fragment is completely miscible with the polyvinyl pyrrolidone.

[0062] 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 with SEQ ID NO:1, and / or one or more collagen fragment sequence variants having at least about 85% amino acid sequence identity with the amino acid sequence shown in any one of SEQ ID NO:975 to SEQ ID NO:1002, and sodium carboxymethyl cellulose, wherein the recombinant collagen fragments are completely miscible with the sodium carboxymethyl cellulose.

[0063] In some embodiments, the present disclosure provides a film comprising recombinantly produced collagen fragments and hyaluronic acid. In some embodiments, the film is visually transparent. In some embodiments, the film is turbid. In some embodiments, the film shows phase separation.

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

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

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

[0067] In some embodiments, the present disclosure provides a slurry comprising recombinantly produced collagen fragments and hyaluronic acid.

[0068] In some embodiments, the present disclosure provides one or more particles comprising recombinantly produced collagen fragments and hyaluronic acid.

[0069] In some embodiments, the present disclosure provides a foam comprising recombinantly produced collagen fragments and hyaluronic acid.

[0070] In some embodiments, the present disclosure provides a composition comprising collagen and sodium alginate, wherein the collagen is completely miscible with the sodium alginate. In some embodiments, the sodium alginate has a viscosity of about 4-12 centipoise for a 1% aqueous solution at 25°C.

[0071] In some embodiments, the present disclosure provides a composition comprising collagen and sodium alginate, wherein the collagen is completely miscible with the sodium alginate. In some embodiments, the sodium alginate has a viscosity equal to or greater than 2,000 centipoise for a 2% aqueous solution at 25°C. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Figures 1A to 1C is a series of photographs showing the combination of uncross-linked HA and collagen. Figure 1A It was shown that collagen from natural animal sources is immiscible with 50 kDa uncross-linked HA. Figure 1B The recombinant collagen fragments were shown to be miscible with 50 kDa non-cross-linked HA. Figure 1C The recombinant collagen fragments were shown to be miscible with 1,000 kDa uncross-linked HA.

[0073] FIG. 2A to FIG. 2C is a series of photographs showing that fluorescently labeled recombinant collagen fragments form homogeneous miscible blends with monophasic and biphasic cross-linked HA. Figure 2A 1,000 kDa uncross-linked HA was shown to be miscible with labeled recombinant collagen. Figure 2B It shows that the single-phase uncross-linked HA is miscible with the recombinant collagen. Figure 2C The biphasic uncross-linked HA was shown to be miscible with the recombinant collagen.

[0074] FIG. 3A to FIG. 3C is a series of photographs showing that certain percentages of hydroxylation on recombinant collagen fragments reduce their miscibility with uncross-linked HA. Figure 3A 50 kDa non-cross-linked HA is shown to be miscible with 0% hydroxylated recombinant collagen fragments. Figure 3B Small-scale phase separation is shown in a mixture of 50 kDa non-cross-linked HA and 22% hydroxylated recombinant collagen fragments, which do not separate into two distinct phases. Figure 3C It was shown that 50 kDa uncross-linked HA was immiscible with the 44% hydroxylated recombinant collagen fragment.

[0075] Figure 4A A standard curve is used to convert UV absorbance to protein concentration by showing the relationship between absorbance (Y-axis) and protein concentration (X-axis).

[0076] Figure 4B Shown is the concentration of protein released from a solution of protein in PBS (Y-axis) over time (days) (X-axis) compared to a formulation of HA and protein in PBS. FIG. 4A to FIG. 4B The measurements were taken from the outside of a dialysis membrane with a molecular weight cutoff (MWCO) of 300 kDa.

[0077] Figure 5A A standard curve is used to convert absorbance to protein concentration by showing the relationship between absorbance (Y-axis) and protein concentration (X-axis).

[0078] Figure 5B Shown is the change in protein concentration released in PBS (Y-axis) over time (days) (X-axis) compared to protein released in HA formulations. Figure 4A and Figure 4B Same, FIG. 5A to FIG. 5B The measurements were taken from the outside of the dialysis membrane with a 300 kDa MWCO.

[0079] Fig. 6A A standard curve is used to convert absorbance to protein concentration by showing the relationship between absorbance (Y-axis) and protein concentration (X-axis).

[0080] Figure 6B The protein concentration plotted versus time point is shown to give a trend of the amount of protein remaining inside the PBS solution and HA formulation after dialysis. In this experiment, the data was taken from inside the starting solution, while Figures 4-5 were taken from outside the 300 kDa MWCO dialysis membrane.

[0081] Fig. 7A A standard curve is used to convert absorbance to protein concentration by showing the relationship between absorbance (Y-axis) and protein concentration (X-axis).

[0082] Figure 7B Shown is the concentration of protein released in PBS solution (Y-axis) compared to protein released in HA formulations over time (days) (X-axis). DETAILED DESCRIPTION

[0083] definition

[0084] The indefinite articles "a" and "an" describing an element or component mean that there is one or at least one of these elements or components. Although these articles are usually used to indicate that the modified noun is a singular noun, unless otherwise specified in a specific context, as used herein, the articles "a" and "an" also include plural forms. Similarly, unless otherwise specified in a specific context, as used herein, the definite article "the" also indicates that the modified noun can be singular or plural.

[0085] As used herein, the term "about" used with a numerical value means "within 10% of the stated value," unless expressly stated otherwise. For example, "about 5 weight %" means 4.5 weight % to 5.5 weight %.

[0086] The term "modified" as applied to collagen and collagen fragments refers to collagen fragments comprising an amino acid sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity or similarity to the amino acid sequence of a biologically active molecule. In some embodiments, the modified collagen fragment comprises an amino acid sequence having 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% identity to the amino acid sequence of a native sequence or a previously engineered sequence. The modified sequence can comprise the addition, disappearance, replacement or their combination of the amino acid sequence of the molecule of natural molecule or previous engineering approaches.For example, compared with the native collagen sequence, the modified collagen fragment can be combined with or lack 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid residues.This type of selection can be carried out to change the looseness or compactness of recombinant collagen.The hydroxylation degree of collagen is relevant to the looseness or compactness of collagen triple helix.The modified collagen fragment can also comprise the chemical modification to polypeptide, such as crosslinking between cysteine ​​residues, or hydroxylated or glycosylated residues. As used herein, the terms "variant" and "sequence variant" refer to a polypeptide sequence having 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% identity with the amino acid sequence shown in SEQ ID NO:1.

[0087] 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 nontoxic to recipients at the dosages and concentrations employed.

[0088] As used herein, the term "tissue repair" refers to the restoration of tissue architecture and function after injury in the context of damaged tissue healing. Tissue regeneration refers to a type of healing in which new growth restores parts of damaged tissue to a normal state.

[0089] As used herein, "hyaluronic acid" and "HA" refer to acidic polysaccharides with different molecular weights and their corresponding salts composed of residues of D-glucuronic acid and N-acetyl-D-glucosamine, including but not limited to sodium hyaluronate. Unless otherwise specified, the composition comprising HA described herein should be understood as a combination comprising hyaluronic acid and sodium hyaluronate. 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 intraoral fluid of the eye, and in human umbilical cord tissue. "Hyaluronic acid" may refer to a whole series of polysaccharides or their degradation fractions with alternating residues of D-glucuronic acid and N-acetyl-D-glucosamine with different 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 β-1,4 glycosidic bonds and β-1,3 glycosidic bonds. Hyaluronic acid is also referred to as hyaluronan, hyaluronate or HA.

[0090] As used herein, the terms "sodium carboxymethylcellulose", "carboxymethylcellulose" and "CMC" are used interchangeably and refer to polymers of different molecular weight ranges that are derivatives of cellulose in which carboxymethyl groups are bound to some of the monomers that make up the cellulose backbone. Unless otherwise specified, compositions described herein that contain carboxymethylcellulose should be understood to contain a combination of carboxymethylcellulose and sodium carboxymethylcellulose.

[0091] As used herein, the term "polyvinylpyrrolidone" refers to polymers with different molecular weight ranges made from the monomer N-vinylpyrrolidone. Polyvinylpyrrolidone (PVP) is also known as povidone and povidone.

[0092] As used herein, the term "polyacrylamide" refers to polymers of varying molecular weight ranges made from the monomer acrylamide. Polyacrylamide (PAM) is also known as poly(2-acrylamide).

[0093] As used herein, the term "polyethylene oxide" refers to polymers with different molecular weight ranges made from the monomer ethylene oxide. Polyethylene oxide (PEO) is also known as polyethylene glycol (PEG) or polyoxyethylene.

[0094] As used herein, the term "poly(2-oxazoline)" refers to polymers having different molecular weight ranges made from the monomer 2-oxazoline and its derivatives such as 2-methyl-2-oxazoline and 2-ethyl-2-oxazoline.

[0095] As used herein, the term "polyethyleneimine" refers to polymers with different molecular weight ranges made from the monomer aziridine. Polyethyleneimine (PEI) is also known as polyaziridine.

[0096] As used herein, the term "miscible" refers to a substance that is soluble in another substance so that when the two components are mixed in water or in a buffer system, a molecularly uniform mixture is formed. An exemplary buffer system is phosphate buffered saline (PBS). For example, a collagen described as being miscible with HA refers to when a collagen solution dissolved in water or a buffer system is mixed with a HA solution (e.g., a solution of 1% to 10% HA) dissolved in water, the aqueous mixture of the collagen solution and the HA solution will form a uniform mixture at room temperature. The miscibility of collagen and hyaluronic acid in the composition may depend on the ratio of hyaluronic acid to collagen, the molecular weight of the collagen and hyaluronic acid used in the composition, and the presence of other additional components (such as salts) that affect the compatibility of collagen and hyaluronic acid in the composition. Temperature may also affect the separation of hyaluronic acid and collagen in the composition. A collagen described as being immiscible with hyaluronic acid refers to a mixture of a collagen solution and a HA solution that will not form a stable, transparent, uniform mixture at room temperature. On the contrary, the solution may show turbidity or phase separation. Solid films prepared with completely miscible HA and collagen solutions may remain completely miscible as a solid film that appears transparent and uniform, or may exhibit small-scale phase separation indicated by turbidity. Transparency can be readily determined by one of ordinary skill in the art and can be measured, for example, using a haze meter.

[0097] "Completely miscible" collagen can apply to the property of collagen to form a homogeneous aqueous mixture with any substance or polymer (e.g., polyvinyl pyrrolidone (PVP), polyacrylamide (PAM), polyethylene oxide (PEO), poly(2-oxazoline), polyethyleneimine (PEI), carboxymethyl cellulose, chondroitin sulfate, acetylated hyaluronate and / or zinc hyaluronate), and is not limited to HA.

[0098] "Viscosity" refers to the measurement of a liquid's resistance to flow at a given temperature. Viscosity can be measured using a variety of methods and instruments known in the art, including but not limited to rheometers and viscometers. Viscosity can also represent the hardness of a liquid at a given temperature.

[0099] At 25° C. and a shear rate of 0.1 / sec, the compositions disclosed herein may have a viscosity of at least about 1 centipoise (cps), at least about 10 centipoise, at least about 100 centipoise, at least about 1000 centipoise, at least about 10,000 centipoise, at least about 20,000 centipoise, at least about 50,000 centipoise, at least about 100,000 centipoise, or more. In some embodiments, the viscous carrier in the compositions of the present disclosure may have a viscosity of about 1 centipoise, about 10 centipoise, about 100 centipoise, about 1000 centipoise, about 10,000 centipoise, about 20,000 centipoise, about 50,000 centipoise, about 100,000 centipoise, or more at 25° C. and a shear rate of 0.1 / sec.

[0100] As used herein, the term "isoelectric point" or "pi" refers to the pH value at which the total net charge of a macromolecule (such as a protein) is zero. There can be many charged groups in a protein, 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 total net charge of the protein will be negative, while at a pH below the isoelectric point, the total net charge of the protein will be positive. The isoelectric point of a molecule can be determined using isoelectric focusing, which is a well-known technique for separating different molecules by the difference in their isoelectric points. The isoelectric point of a protein can also be determined by electrophoresis. The isoelectric point can also be calculated (see, e.g., Levene and Simms, "Calculation ofisoelectric point" J.Biol.Chem., 1923, pp. 801-813).

[0101] The compositions described herein may have an isoelectric point in the range of about 4.0 to about 12.0, about 4.5 to about 11.5, about 5.0 to about 11.0, about 5.5 to about 10.5, about 6.0 to about 10.0, about 6.5 to about 9.5, about 7.0 to 9.0, about 7.5 to 8.5, or about 8.0. In certain embodiments, the compositions described herein 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.

[0102] As used herein, the term "cross-linked" describes a polymer in which at least one covalent bond is not present in the repeating unit of the polymer, or is present between the repeating units of the polymer. Cross-linking occurs between individual chains or molecules of a polymer; however, intramolecular reactions may occur within a polymer chain to form a macrocyclic structure. The latter is not considered to be cross-linking in the present disclosure. Cross-linking is formed between any two functional groups of a polymer (e.g., at the end, on a side chain, etc.). In certain embodiments, cross-linking is formed between the terminal units of a polymer. In certain embodiments, cross-linking is formed between the side chain units of a polymer. Any type of covalent bond may form a cross-link (e.g., carbon-carbon, carbon-oxygen, carbon-nitrogen, oxygen-nitrogen, sulfur-sulfur, oxygen-phosphorus, etc.). The resulting cross-linked material may be branched, linear, dendritic, etc. In certain embodiments, cross-linking forms a 3D cross-linked network. In a chemical reaction, cross-linking may be formed by any chemical cross-linking agent, thereby producing covalent bonds. Cross-linking may be formed by applying ultraviolet (UV) radiation or microwave radiation to the polymer. Crosslinks may be formed by free radical initiated reactions, for example, using photoinitiators or thermal initiators, or by application of heat.

[0103] Typical chemical crosslinkers are usually various types of difunctional or polyfunctional crosslinkers and can be selected, for example, from DVS (divinyl sulfone) (see US 4,582,865), difunctional or polyfunctional epoxides (see US 4,716,154), carbodiimides or formaldehyde (see GB 2 151 244) in alkaline medium. Other suitable crosslinkers include epoxides or derivatives thereof, in particular 1,4-butanediol diglycidyl ether (BDDE), diepoxyoctane or 1,2-bis(2,3-epoxypropyl)-2,3-ethylene. Known crosslinkers also include divinyl sulfone, glycerol diglycidyl ether and 1,4-butanediyl diglycidyl ether.

[0104] In certain embodiments, compositions according to the present disclosure can react to form a cross-linked macromolecular matrix. Since the cross-linking reaction occurs in an aqueous solution, when the cross-linking reaction forms a hydrogel, the cross-linked macromolecular matrix can be dispersed in an aqueous liquid in the form of a hydrogel. As used herein, the term "hydrogel" refers to a three-dimensional network comprising a hydrophilic polymer that contains a large amount of water. In w / w terms, a hydrogel can, for example, contain 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or even more water. A "hydrogel precursor" is a polymer that is at least partially soluble in an aqueous medium and can be cross-linked to form a hydrogel.

[0105] In some embodiments, the compositions of the present disclosure may exhibit physiologically acceptable molar osmotic pressure concentrations. As used herein, the term "molar osmotic pressure concentration" refers to the concentration of osmotically active solutes in a solution. As used herein, the term "physiologically acceptable molar osmotic pressure concentration" refers to the molar osmotic pressure concentration that is consistent with or characteristic of the normal function of a living organism. Therefore, when a hydrogel disclosed herein is administered to a mammal, the molar osmotic pressure concentration exhibited by its administration does not substantially have long-term or permanent harmful effects. Molar osmotic pressure concentration is expressed as the number of osmoles of osmotically active solutes per liter of solvent (Osmol / L or Osm / L).

[0106] As used herein, "solubility" refers to the amount of a substance dissolved in a given volume of solvent at a specified temperature and pH, such as when a saturated solution is formed. Solubility can be measured, for example, using a shake bottle solubility method (ASTM: E 1148-02, Standard Test Method for Measurements of Aqueous Solubility, Book of Standards Volume 11.05). Solubility can be measured at any pH. Solubility can be tested at any temperature between 20°C and 40°C. For example, solubility can be measured at about pH 7.0-7.4 and about 37°C.

[0107] A. collagen

[0108] The term "collagen" refers to any known type of collagen, including collagen type I to collagen type XXVIII, and any other collagen, whether natural, synthetic, semi-synthetic or recombinant. The term collagen includes collagen, collagen fragments, collagen-like proteins, triple helical collagens, α chains, monomers, gelatin, trimers, and combinations thereof. Collagen includes all collagens, modified collagens, and collagen-like proteins described herein. In a specific embodiment, the term collagen refers to any known type of collagen, which 1) is soluble in HA; and 2) induces type I collagen production, type III collagen production, ECM production, or a combination thereof. The term also encompasses procollagen and collagen-like proteins or collagenous proteins comprising the motif (Gly-XY) n (where n is an integer). The term encompasses molecules of collagen and collagen-like proteins, trimers of collagen molecules, collagen fibrils, and fibers of collagen fibrils. The term also refers to collagen or collagen-like molecules that can be fibrillated through chemical, enzymatic or recombinant modification, and the collagen, collagen-like molecules and collagenous molecules that can be assembled into nanofibers. Natural or engineered recombinant collagen molecules will usually comprise repeated-(Gly-XY)n-sequences. As used herein, collagen is the generic name for the family of at least 28 different collagen types. Various collagen types have been identified in a range of species, including cattle, sheep, pigs, Galliformes, oceans, plants and human collagen.

[0109] Collagen has a unique amino acid composition; almost one-third of its amino acid residues are glycine; the other 15% to 30% are proline or 4-hydroxyproline (Hyp) residues. Hydroxylysine is also a characteristic of collagen. These hydroxylated amino acid residues, formed as a result of post-translational modification, impart stability to collagen through intramolecular hydrogen bonds involving bridging water molecules. The collagen content in a tissue can be determined by measuring its hydroxyproline content. The hydroxyproline content can be measured by methods known in the art, such as by liquid chromatography-mass spectrometry (LC-MS). Collagen contains many repeated iterations of the sequence Gly-XY, where X is typically proline and Y is typically 4-hydroxyproline.

[0110] Collagen variants containing such amino acid residue substitutions can be produced by a variety of 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, wherein hydroxylase genes (recombinant or other) are expressed in host cells for expressing recombinant collagen or its derivatives. Hydroxyproline content can be measured by methods known in the art, such as by liquid chromatography-mass spectrometry (LC-MS) measurement.

[0111] Collagen is the most abundant protein in vertebrates. Vertebrate collagens are a family of proteins produced by several cell types. Within this protein family, collagen types can be distinguished by their chemical composition, different morphological and pathological characteristics, distribution within tissues, and their functions. Although many types of collagen have been described, five main types have been identified. Mammalian skin is usually type I collagen, but other types of collagen (including type III collagen) can also be used to form leather. The term "collagen" encompasses unprocessed (e.g., procollagen), as well as collagen with a triple helical structure that has been post-translationally modified and proteolytically hydrolyzed.

[0112] Any type of collagen, including types I through XXVIII, is suitable for use in the compositions described herein.

[0113] B. Recombinant collagen and recombinant collagen fragments

[0114] As used herein, the term "recombinant collagen" refers to a family of at least 28 different naturally occurring collagen types that are prepared using recombinant technology, including but not limited to collagen types I to XXVIII.

[0115] In some embodiments, the recombinant collagen described herein is a recombinant collagen fragment. The recombinant collagen fragment can be a fragment of the complete amino acid sequence of a native collagen molecule capable of forming procollagen (trimeric collagen), or the fragment can be a modified collagen molecule or a truncated collagen molecule having at least 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% identity or similarity with the native collagen amino acid sequence (or with its fibril-forming region or with a segment substantially comprising [Gly-XY]n). In a specific embodiment, the term recombinant collagen refers to any known type of recombinant collagen, which (1) is soluble in HA; and (2) induces the production of type I collagen and / or type III collagen. The production of type I collagen and / or type III collagen can be determined by assays known in the art, including those described in PCT / US2022 / 027016, the entire contents of which are incorporated herein by reference.

[0116] Exemplary collagen sequences from which fragments can be derived include the amino acid sequences of Col1A1, Col1A2, and Col3A1, such as those described by the following 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 Col1A2) (www.ncbi.nlm.nih.gov / protein / 27806257). NO: 1006; bovine Col3A1) (www.ncbi.nlm.nih.gov / protein / 116003881), which is incorporated herein by reference.

[0117] Genes encoding collagen can be truncated or otherwise modified to add or remove sequences, such as to encode collagen fragments disclosed herein. In addition, genetic modification can be performed to customize the size of polynucleotides or vectors, target the expressed protein to the endoplasmic reticulum or other cells or extracellular compartments, or control the length of the encoded protein. Polynucleotides encoding collagen fragments can be modified. For example, the polynucleotide coding sequence of collagen or collagen fragments can be modified to encode a protein having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100% identity or similarity to a known amino acid sequence. Such modifications can include codon modification or codon optimization of the polynucleotide encoding collagen fragments.

[0118] In some embodiments, the collagen fragments disclosed herein may have a molecular weight of about 1 kDa to about 150 kDa. In some embodiments, the collagen fragments may 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, In some embodiments, the collagen fragment may have a molecular weight of about 100 kDa, about 150 kDa, about 160 kDa, about 170 kDa, about 180 kDa, about 190 kDa, about 200 kDa, about 210 kDa, about 220 kDa, about 230 kDa, about 240 kDa, about 250 kDa, about 260 kDa, about 270 kDa, about 280 kDa, about 290 kDa, about 300 kDa, about 310 kDa, about 320 kDa, about 330 kDa, about 340 kDa, about 350 kDa, about 360 kDa, about 370 kDa, about 38 kDa, about 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 a specific embodiment, the collagen fragment may have a molecular weight of about 10 kDa. In a specific embodiment, the collagen fragment may have a molecular weight of about 10 kDa. In a specific embodiment, the collagen fragment may have a molecular weight of about 20 kDa. In a specific embodiment, the collagen fragment may have a molecular weight of about 30 kDa. In a specific embodiment, the collagen fragment may have a molecular weight of about 40 kDa. In a specific embodiment, the collagen fragment may have a molecular weight of about 50 kDa. In a specific embodiment, the collagen fragment may have a molecular weight of about 60 kDa. In a specific embodiment, the collagen fragment may have a molecular weight of about 100 kDa. In a specific embodiment, the collagen fragment may have a molecular weight of about 140 kDa.

[0119] In some embodiments, the collagen fragments described herein may have an amino acid chain length of about 350 amino acids to about 600 amino acids and may overlap with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the overlapping collagen fragments described herein may 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.

[0120] In some embodiments, the collagen fragments described herein may have an amino acid sequence according to SEQ ID NO: 1. In some embodiments, the collagen fragments may 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.

[0121] The amino acid sequence of SEQ ID NO:1 is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。

[0122] In some embodiments, the collagen fragments described herein may have an amino acid chain length of about 350 amino acids to about 600 amino acids, and may have a sequence identity or similarity to SEQ ID NO:1 of 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%. In some embodiments, the collagen fragments described herein may 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.

[0123] In some embodiments, the recombinant collagen may include a hydrolysate of a collagen fragment, wherein the hydrolysate may have a sequence that is a portion of SEQ ID NO: 1. In some embodiments, the hydrolysate may have at least about 85% sequence identity with a sequence according to one of SEQ ID NO: 2 to SEQ ID NO: 973. In some embodiments, the collagen fragment may be a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002.

[0124] In some embodiments, the lysine, proline or lysine residue and proline residue present in the collagen or collagen fragment are not hydroxylated. In some embodiments, the lysine, proline or lysine residue and proline residue present in the collagen fragment 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 lysine, proline or lysine residue and proline residue in the recombinant collagen fragment can be hydroxylated.

[0125] In certain embodiments, the proline residues in the recombinant collagen fragment may be about 1% to about 20% hydroxylated.

[0126] In some embodiments, the recombinant collagen fragment may be about 12% hydroxylated. In some embodiments, the recombinant collagen fragment may be about 5% hydroxylated.

[0127] In some embodiments, the collagen fragment may be a collagen fragment, a hydrolysate, or a sequence variant having an amino acid sequence shown in Table 1.

[0128] Table 1.

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159] C. Hyaluronic acid

[0160] Hyaluronic acid (HA) is a natural linear carbohydrate polymer that belongs to the class of non-sulfated glycosaminoglycans. Hyaluronic acid is composed of repeating disaccharide units of β-1,3-N-acetylglucosamine and β-1,4-glucuronic acid with a molecular weight (MW) of up to 6 MDa. HA is found in hyaline cartilage, synovial joint fluid, and skin tissue (dermis and epidermis). HA can be extracted from natural tissues (including connective tissue of vertebrates) or prepared by microbial methods.

[0161] A variety of roles of HA in vivo have been identified. Hyaluronic acid plays an important role in biological organisms, providing mechanical support for cells of many tissues (such as skin, tendons, muscles and cartilage). HA is involved in key biological processes, such as tissue wetting and lubrication. It is also suspected to play a role in many physiological functions, such as adhesion, development, cell motility, cancer, angiogenesis and wound healing. Due to the unique physical and biological properties of HA (including viscoelasticity, biocompatibility, biodegradability), HA is widely used in current and developing applications in the fields of cosmetics, beauty, skin care, ophthalmology, rheumatism, drug delivery, wound healing and tissue engineering. The use of HA is limited by the following facts in some of these applications: HA is soluble in water at room temperature, i.e., about 20°C, is rapidly degraded by hyaluronidase in the body, and is difficult to process into biomaterials. Therefore, HA cross-linking has been introduced to improve the physical and mechanical properties of HA and its residence time in the body. The cross-linking of HA can be described as single-phase or biphasic. The single-phase structure is when cross-linking is formed between HA chains, it forms a uniform gel-like structure. Dual-phase structures are structures that have both gel-like HA and free HA particles that are not part of the cross-linked structure. Dual-phase structures are not as smooth as single-phase structures, but historically have been more voluminous.

[0162] US4,582,865 describes a method for preparing a crosslinked gel of HA using divinyl sulfone (DVS) as a crosslinking agent, alone or mixed with other hydrophilic polymers, and the patent is incorporated by reference in its entirety. EP 0 161 887 B1 discloses a method for preparing crosslinked HA or its salts using a multifunctional epoxy compound, and the patent is also incorporated by reference in its entirety. Other bifunctional or multifunctional agents for crosslinking HA by covalent bonds include formaldehyde (see US4,713,448, which is incorporated by reference in its entirety), polyaziridine (see WO 03 / 089476, which is incorporated by reference in its entirety), and L-amino acids or L-amino esters (see WO 2004 / 067575, which is incorporated by reference in its entirety). Carbodiimides have also been reported for crosslinking HA (see US5,017,229 and US6,013,679, each of which is incorporated by reference in its entirety). Completely or partially cross-linked esters of HA with aliphatic alcohols, and salts of such partial esters with inorganic or organic bases are disclosed in U.S. Pat. No. 4,957,744, which is also incorporated by reference in its entirety. Cross-linking HA chains with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride ("EDAC") and adipic acid dihydrazide in a water / acetone mixture is disclosed in US2006 / 0040892 (which is incorporated by reference in its entirety). WO 2006 / 56204 also discloses a method for preparing a cross-linked gel of HA using divinyl sulfone (DVS) as a cross-linking agent, which is also incorporated by reference in its entirety.

[0163] HA is commercially available from various suppliers, such as Pure Health Botanicals (Saint Charles, IL) and DSM (Kaiseraugst, Switzerland).

[0164] D. Compositions containing collagen fragments and sequence variants thereof

[0165] 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 (e.g., acetylated hyaluronate). In some embodiments, the collagen is completely miscible with an acetylated hyaluronate. Other hyaluronates include zinc hyaluronate.

[0166] In some embodiments, the collagen is completely miscible with other polymers for therapeutic applications. In some embodiments, the collagen is completely miscible with polymers in aqueous solution and as a solid film or article. Contemplated articles include, but are not limited to, stents and wound dressings. Such polymers include, but are not limited to, polyvinyl pyrrolidone (PVP), polyacrylamide (PAM), polyethylene oxide (PEO), poly(2-oxazoline), polyethyleneimine (PEI), sodium carboxymethylcellulose, and chondroitin sulfate.

[0167] In some embodiments, the collagen is completely miscible with polyvinylpyrrolidone in aqueous solution and as a solid film or article.

[0168] In some embodiments, the collagen is completely miscible with polyacrylamide in aqueous solution and as a solid film or article.

[0169] In some embodiments, the collagen is completely miscible with polyethylene glycol. In some embodiments, the collagen is completely miscible with poly(2-oxazoline). In some embodiments, the collagen is completely miscible with polyethyleneimine.

[0170] In some embodiments, the collagen is completely miscible with the carboxymethylcellulose.

[0171] In some embodiments, the collagen is completely miscible with chondroitin sulfate.

[0172] In some embodiments, the collagen is completely miscible with the acetylated hyaluronate.

[0173] In some embodiments, the collagen can be mixed with zinc hyaluronate. In some embodiments, the collagen is completely miscible with zinc hyaluronate.

[0174] In some embodiments, the collagen is completely miscible with the sodium alginate salt. In some embodiments, the sodium alginate salt has a viscosity of about 4 centipoise to 12 centipoise for a 1% aqueous solution at 25°C. In some embodiments, the sodium alginate salt has a viscosity equal to or greater than about 2,000 centipoise for a 2% aqueous solution at 25°C.

[0175] In some embodiments, the collagen is a recombinantly produced collagen fragment. In some embodiments, the collagen in the composition can be physically and / or chemically modified. Chemical and physical modifications may include, but are not limited to, hydroxylation, cross-linking, charge modification, linking agents, and modifications known in the art.

[0176] In some embodiments of the compositions disclosed herein, the compositions may further comprise one or more polymers or copolymers, including but not limited to aliphatic polyesters, polyorthoesters, polyoxyesters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyesteramides, hydrolyzable amines, polyalkylene oxides, poloxamers, functionalized isoflavones, amino acids, and combinations thereof.

[0177] In some embodiments of the compositions disclosed herein, the compositions may further comprise one or more polymers or copolymers selected from the group consisting of 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-dioxepane-2-one and poly(1,5-dioxepane-2-one-co-p-dioxanone), poly(lactide-co-1,4-dioxepane-2-one), poly(1,4-dioxepane-2-one) ketone-co-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), poly(dioxanone-co-D,L-lactide-co-caprolactone), poly(dioxanone-co-glycolide-co-caprolactone), poly(dioxanone-co-L-lactide-co-D,L-lactide 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-L-lactide-co-glycolide-co-caprolactone), poly(dioxanone-co-D,L-lactide-co-caprolactone) and combinations thereof.

[0178] In some embodiments, these polymers 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.

[0179] In some embodiments of the compositions disclosed herein, these compositions may also include biodegradable glasses or ceramics, such as calcium phosphate and other biocompatible metal oxides (ie, CaO) or combinations of biodegradable ceramics, glasses, polymers, and copolymers.

[0180] In some embodiments of the compositions disclosed herein, the isoelectric point (pI) of the collagen described herein is 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, 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, and in some embodiments, about 9.2, and in some embodiments, about 9.66. In some embodiments of the compositions disclosed herein, the isoelectric point (pi) of the composition is 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.

[0181] In some embodiments, HA can have a molecular weight that is compatible with biological tissues. In some embodiments, 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.

[0182] In some embodiments, the molecular weight of HA in the compositions described herein may be in the range of about 10 kDa to about 10,000 kDa, about 25 kDa to about 5,000 kDa, or about 50 kDa to about 3,000 kDa. In some embodiments, the compositions disclosed herein comprise 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.

[0183] 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, about 2.5 kDa to about 2,500 kDa, about 10 kDa to about 2,000 kDa, about 40 kDa to about 1,500 kDa, about 100 kDa to about 1,000 kDa.

[0184] In some embodiments, the molecular weight of the 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.

[0185] In some embodiments, the molecular weight of PEO in the compositions described herein can range from about 5 kDa to about 300 kDa, about 50 kDa to about 200 kDa.

[0186] In some embodiments, the molecular weight of the poly(2-ethyl-2-oxazoline) in the compositions described herein can range from about 10 kDa to about 1,000 kDa, about 50 kDa to about 500 kDa.

[0187] In some embodiments, the molecular weight of the PEI in the compositions described herein can range from about 2.5 kDa to about 2,500 kDa.

[0188] In some embodiments, the molecular weight of the carboxymethyl cellulose in the composition described herein may be in the range of about 10kDa to about 300kDa. In some embodiments, the degree of substitution of the carboxymethyl cellulose in the composition may be in the range of about 1.00 to about 1.25. In some embodiments, the carboxymethyl cellulose may have a molecular weight of about 250kDa and a degree of substitution of about 0.70 to about 0.90. In some embodiments, the carboxymethyl cellulose may have a molecular weight of about 250kDa and a degree of substitution of about 0.79. In some embodiments, the carboxymethyl cellulose may have a degree of substitution of about 0.87.

[0189] In some embodiments, the carboxymethyl cellulose in the compositions described herein may have a high viscosity. In some embodiments, the carboxymethyl cellulose in the compositions described herein may have a viscosity of about 100 centipoise to about 5,000 centipoise for a 1% aqueous solution. In some embodiments, the carboxymethyl cellulose in the compositions described herein may have a viscosity of about 1,000 centipoise to about 3,000 centipoise for a 1% aqueous solution. In some embodiments, the carboxymethyl cellulose in the compositions described herein may have a viscosity of about 2,700 centipoise for a 1% aqueous solution.

[0190] In some embodiments, the molecular weight of the chondroitin sulfate in the composition can be in the range of about 20 kDa to about 80 kDa. In some embodiments, the molecular weight of the chondroitin sulfate in the composition can be about 50 kDa.

[0191] In specific embodiments of the compositions disclosed herein, the hyaluronic acid may have a molecular weight of about 3 kDa to about 10,000 kDa. In some embodiments, the hyaluronic acid may have a molecular weight of about 1,000 kDa to about 5,000 kDa. In some embodiments, the hyaluronic acid may have a molecular weight of about 2,000 kDa to about 3,000 kDa.

[0192] In some embodiments of the compositions disclosed herein, the hyaluronic acid may be cross-linked. In some embodiments, the hyaluronic acid is not cross-linked. In some embodiments of the compositions disclosed herein, the collagen may be cross-linked. In some embodiments, the collagen is not cross-linked.

[0193] In some embodiments of the compositions disclosed herein, the hyaluronic acid can be a biphasic cross-linked hyaluronic acid. In some embodiments, the hyaluronic acid can be a monophasic cross-linked hyaluronic acid.

[0194] In some embodiments, the hyaluronic acid solution can have a pH in the range of about 5.0 to about 12.

[0195] 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, the 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 measured value therebetween.

[0196] In some embodiments of the compositions disclosed herein, hyaluronic acid and collagen may be present in a ratio (w / w) ranging from about 1:9 to 9:1. In some embodiments, the ratio (w / w) of hyaluronic acid to collagen 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 (w / w) of hyaluronic acid to collagen may be 1:9. In some embodiments, the ratio (w / w) of hyaluronic acid to collagen may be 1:3. In some embodiments, the ratio (w / w) of hyaluronic acid to collagen may be 2:3. In some embodiments, the ratio (w / w) of hyaluronic acid to collagen may be 1:1. In some embodiments, the ratio (w / w) of hyaluronic acid to collagen 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.

[0197] In some embodiments, the collagen in the composition comprising collagen and HA can be one or more recombinant collagen fragments disclosed herein. In some embodiments, the one or more collagen fragments can be cross-linked with HA. In some embodiments, the one or more collagen fragments can be covalently or non-covalently cross-linked with HA. In some embodiments, cross-linking can be formed by a chemical reaction, which leads to the formation of covalent cross-linking and is initiated by chemical reagents, heating, pressure, pH or radiation. In some embodiments, non-covalent cross-linking can be formed using multivalent metal ions, such as iron, copper, zinc, calcium, magnesium, barium and other chelated metal ions. One or more suitable cross-linking agents such as glutaraldehyde, carbodiimide, difunctional amino acids (such as lysine), protamine or albumin can be used to achieve covalent bonding. Cross-linking can also be produced by amide, ester or ether bonds. The linking agent used for cross-linking HA by covalent bonds is disclosed and discussed elsewhere herein.

[0198] In some embodiments, crosslinking can be formed directly between hyaluronic acid and one or more collagen or recombinant collagen fragments, at the amino group of the collagen molecule and the carboxylic acid group of the hyaluronic acid. In some embodiments, the crosslinking between HA and the collagen molecule can be formed using a linking agent, suitable examples of which are disclosed and discussed elsewhere herein. In some embodiments, the hyaluronic acid molecule can be crosslinked with other hyaluronic acid molecules (in addition to the crosslinking between HA and the collagen molecule, or in place of the crosslinking between HA and the collagen molecule). In further embodiments, one or more collagen molecules can be crosslinked with another collagen molecule (in addition to the crosslinking between HA and the collagen molecule, or in place of the crosslinking between HA and the collagen molecule).

[0199] In some embodiments, the compositions disclosed herein may include a cross-linked macromolecular matrix. In some embodiments, the cross-linked macromolecular matrix may be a uniform hydrogel composition prepared by methods known in the art under conditions where collagen and HA are soluble in aqueous solution.

[0200] In some embodiments of the compositions disclosed herein, collagen may not include hydroxyproline (Hyp) residues. In some embodiments, based on the total number of proline in collagen, collagen may include about 1% to about 20% hydroxyproline (Hyp) residues. In some embodiments, collagen described herein may include 0% to about 20%, about 8% to about 20%, about 10% to about 20%, or about 12% to about 20% hydroxyproline. In some embodiments, compositions disclosed herein may include 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. Hydroxyproline content can be measured by methods known in the art, such as by liquid chromatography-mass spectrometry (LC-MS) measurement.

[0201] 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 may contain from about 1 to about 120 hydroxyl groups. The number of hydroxyl groups may be measured by methods known in the art, such as by mass spectrometry.

[0202] In some embodiments, the collagen does not contain any amine groups. In some embodiments, the collagen may contain about 1 to about 100 amine groups. The number of amine groups can be measured by methods known in the art, such as by mass spectrometry and colorimetric TNBSA reagent assay.

[0203] In some embodiments, the collagen does not contain any carboxylic acid groups. In some embodiments, the collagen may 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.

[0204] The compositions disclosed herein can be prepared by a method comprising cross-linking hyaluronic acid and collagen. In some embodiments, hyaluronic acid and collagen can be cross-linked under suitable conditions.

[0205] In some embodiments, the composition has a viscosity of about 1 centipoise to about 100,000 centipoise. In some embodiments, the composition has a viscosity of about 10,000 centipoise to about 80,000 centipoise. In some embodiments, the composition has a viscosity of about 50,000 centipoise to about 70,000 centipoise.

[0206] In some embodiments, the composition may include collagen that is completely miscible with hyaluronic acid and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient. In some embodiments, the composition may include a recombinant collagen fragment according to SEQ ID NO: 1 and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient. In other embodiments, the composition may 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 at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient, wherein the recombinant collagen fragment is completely miscible with hyaluronic acid. In further embodiments, the composition may 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 and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient, wherein the recombinant collagen fragment is completely miscible with hyaluronic acid. In a further embodiment, the composition may 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, wherein the recombinant collagen fragment has an amino acid sequence as shown in any one of SEQ ID NO: 975 to SEQ ID NO: 1002 and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient, wherein the recombinant collagen fragment is completely miscible with hyaluronic acid.

[0207] In some embodiments, the composition may include collagen that is completely miscible with hyaluronic acid, and at least one excipient suitable for dietary supplements (e.g., nutritional supplements). In some embodiments, the composition may 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% sequence identity or similarity to SEQ ID NO: 1 and at least one excipient suitable for dietary supplements (e.g., nutritional supplements), wherein the recombinant collagen fragment is completely miscible with hyaluronic acid. In another embodiment, the composition may 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 and at least one excipient suitable for a dietary supplement (e.g., a nutritional supplement), wherein the recombinant collagen fragment has an amino acid sequence shown in any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and wherein the recombinant collagen fragment is completely miscible with hyaluronic acid.

[0208] In some embodiments, the composition may include a hydrolysate of collagen fragments and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient, wherein the hydrolysate may have a sequence as part of SEQ ID NO: 1. In some embodiments, the composition includes a hydrolysate having a sequence according to one or more of SEQ ID NO: 2 to SEQ ID NO: 973, and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient. In some embodiments, the composition may include any of the hydrolysates shown in SEQ ID NO: 2 to SEQ ID NO: 973, and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient. In some embodiments, the composition may 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 NO:2 to SEQ ID NO:973, and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient.

[0209] In yet another embodiment, the composition may comprise a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysate having a sequence according to one of SEQ ID NO: 2 to SEQ ID NO: 973, and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient. In yet another embodiment, the composition may comprise a recombinant collagen fragment according to SEQ ID NO: 1, a plurality of hydrolysates having a sequence according to any one of SEQ ID NO: 2 to SEQ ID NO: 973, which may be the same or different, and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient. In certain embodiments, the number of hydrolysates present in the plurality of hydrolysates in the composition may increase with time, temperature, pH, or due to other conditions that would normally cause a recombinant collagen fragment, such as a recombinant collagen fragment according to SEQ ID NO: 1, to hydrolyze or otherwise decompose. In other embodiments, the composition can be stabilized with one or more stabilizers so that the concentration of the recombinant collagen fragment according to SEQ ID NO: 1 and the concentration of each of the multiple fragments remains substantially constant (i.e., does not vary by more than ± 5% as measured by HPLC over a given period of time) or remains constant. In certain embodiments, the recombinant collagen fragment (such as the recombinant collagen fragment according to SEQ ID NO: 1) can be hydrolyzed so that less than about 10%, about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of the unhydrolyzed recombinant fragments remain in the composition as measured by HPLC. In other embodiments, the composition may comprise a mixture of recombinant collagen fragments (e.g., recombinant collagen fragments according to SEQ ID NO:1) and multiple hydrolysates of the recombinant collagen fragments (e.g., multiple collagen fragments according to any one of SEQ ID NO:2 to SEQ ID NO:973), such that the weight of the hydrolysate in the composition is less than about 10%, about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of the weight of the collagen-related protein in the composition.

[0210] In some embodiments, the present disclosure provides a composition comprising one or more recombinant collagen fragments disclosed herein, and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient. In a specific embodiment, the composition may comprise a recombinant collagen fragment sequence variant according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and at least one pharmaceutically acceptable excipient or cosmetically acceptable excipient. In additional embodiments, the composition may 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 excipient or cosmetically acceptable excipient. In further embodiments, the composition may 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 excipient or cosmetically acceptable excipient. In further embodiments, the composition may 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 a dietary supplement (e.g., a nutritional supplement).

[0211] In some embodiments, the composition may include recombinant collagen fragments 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 in SEQ ID NO: 1. In some embodiments, the composition includes recombinant collagen fragments 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 a specific embodiment, the composition includes a recombinant collagen fragment having an amino acid chain length of 528 amino acids. In a specific embodiment, the composition comprises a recombinant collagen fragment having an amino acid chain length of 546 amino acids.

[0212] In some embodiments, the composition may comprise a recombinant collagen fragment sequence variant according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002, the recombinant collagen fragment sequence variant 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 a specific embodiment, the composition comprises a recombinant collagen fragment having an amino acid chain length of 528 amino acids. In a specific embodiment, the composition comprises a recombinant collagen fragment having an amino acid chain length of 546 amino acids.

[0213] In some embodiments, the composition may comprise a recombinant collagen fragment or a sequence variant thereof having an amino acid chain length of about 150 amino acids to about 600 amino acids and may 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, the collagen fragment described herein may 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.

[0214] In some embodiments, the composition may include a hydrolyzate of collagen fragments, wherein the hydrolyzate may have an amino acid chain length of about 10 amino acids to about 75 amino acids. In some embodiments, the composition may include a hydrolyzate of collagen fragments with an amino acid chain length of about 20 amino acids to about 50 amino acids. In some embodiments, the hydrolyzate described herein may 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.

[0215] In some embodiments, said composition may include about 5ppm to about 500ppm of recombinant collagen fragments or their sequence variants. In some embodiments, said composition may include about 5ppm, about 10ppm, about 25ppm, about 50ppm, about 100ppm, about 150ppm, about 200ppm, about 250ppm, about 300ppm, about 350ppm, about 350ppm, about 400ppm, about 450ppm or about 500ppm of recombinant collagen fragments. In some embodiments, said composition may include about 5ppm, about 10ppm, about 25ppm, about 50ppm, about 100ppm, about 150ppm, about 200ppm, about 250ppm, about 300ppm, about 350ppm, about 350ppm, about 400ppm, about 450ppm or about 500ppm of recombinant collagen fragment sequence variants. In some embodiments, the composition may 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 a sequence according to SEQ ID NO: 1. In some embodiments, the composition may 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 hydrolysates of a recombinant collagen fragment. In some embodiments, the composition may 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 hydrolysates of a recombinant collagen fragment having a sequence according to SEQ ID NO: 1. In some embodiments, the composition may 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 according to any one of SEQ ID NO: 987 to SEQ ID NO: 1015.In some embodiments, the composition may include 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 hydrolysates of the recombinant collagen fragment at a concentration of about 5 ppm to about 500 ppm. In some embodiments, the composition may include a mixture of a recombinant collagen fragment or a variant thereof having a sequence according to SEQ ID NO: 1 at a concentration of about 5 ppm to about 500 ppm and one or more hydrolysates of the recombinant collagen fragment at a concentration of about 5 ppm to about 500 ppm.

[0216] In some embodiments, the composition can be prepared from a recombinant collagen fragment solution of about 0.5 wt % to about 25 wt % with about 0.0001 vol % to about 20 vol %. In some embodiments of these embodiments, the composition can include a recombinant collagen fragment solution of about 0.0001 vol %, about 0.2 vol %, about 0.3 vol %, about 0.4 vol %, about 0.5 vol %, about 0.6 vol %, about 0.7 vol %, about 0.8 vol %, about 0.9 vol %, about 1 vol %, about 2 vol %, about 3 vol %, about 4 vol %, about 5 vol %, about 6 vol %, about 7 vol %, about 8 vol %, about 9 vol %, about 10 vol %, about 11 vol %, about 12 vol %, about 13 vol %, about 14 vol %, about 15 vol %, about 16 vol %, about 17 vol %, about 18 vol %, about 19 vol % or about 20 vol %. In some of these embodiments, the recombinant collagen fragment solution comprises 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 the recombinant collagen fragment solution. The recombinant collagen fragment, or 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 the recombinant collagen fragment sequence variant.

[0217] 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 a sequence according to SEQ ID NO:1. 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 one or more hydrolysates of recombinant collagen fragments. 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 the recombinant collagen fragment according to SEQ ID NO: 2 to SEQ ID NO: One or more hydrolysis products of recombinant collagen fragments having one or more sequences in NO:973.In some of these embodiments, the recombinant collagen fragment sequence variant 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%, or about 25 wt%, or about 40 wt% of a recombinant collagen fragment sequence variant solution according to SEQ ID NO: 975 to SEQ ID NO: One or more hydrolysis products of recombinant collagen fragment sequence variants of one or more sequences in IDNO:1002.

[0218] In some of these embodiments, the composition can be prepared using about 0.0001% to about 35% by volume of a mixture of about 0.0005% to about 40% by weight of a recombinant collagen fragment and one or more hydrolysates of the recombinant collagen fragment. In some of these embodiments, the composition can be prepared using about 0.0001% to about 35% by volume of a mixture of about 0.0005% to about 40% by weight of a recombinant collagen fragment having a sequence according to SEQ ID NO: 1 and one or more hydrolysates of the recombinant collagen fragment.

[0219] In some embodiments, the composition can be prepared using about 0.0001% by volume to about 35% by volume of about 0.0005% by weight to about 40% by weight of a solution comprising a mixture of recombinant collagen fragments having a sequence according to SEQ ID NO: 1 and one or more hydrolysates of the recombinant collagen fragment. In some embodiments, the composition can be prepared using about 0.0001% by volume to about 35% by volume of about 0.0005% by weight to about 40% by weight of a solution comprising a mixture of recombinant collagen fragment sequence variants having a sequence according to any one of SEQ ID NO: 2 to SEQ ID NO: 973. In some embodiments, the composition can be prepared using about 0.0001 volume % to about 35 volume % of about 0.0005 weight % to about 40 weight % of a solution comprising a mixture of a recombinant collagen fragment having a sequence according to SEQ ID NO:1 and one or more hydrolysates of the recombinant collagen fragment, and a recombinant collagen fragment sequence variant solution comprising a sequence variant having a sequence according to any one of SEQ ID NO:975 to SEQ ID NO:1002.

[0220] In some of these embodiments, the recombinant collagen fragment and recombinant collagen fragment sequence variant 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%, or about 25 wt%, or about 40 wt% of the recombinant collagen fragment according to SEQ ID NO: 2 to SEQ ID NO: One or more hydrolysis products of recombinant collagen fragments having one or more sequences in NO:973.

[0221] In some embodiments, the composition may include about 0.0005% to about 25% by weight of recombinant collagen fragments, about 0.001% to about 25% by weight of recombinant collagen fragments, about 0.01% to about 25% by weight of recombinant collagen fragments, about 0.1% to about 25% by weight of recombinant collagen fragments, about 0.5% to about 20% by weight of recombinant collagen fragments, about 0.7% to about 17% by weight of recombinant collagen fragments, about 1% to about 15% by weight of recombinant collagen fragments, about 2% to about 12% by weight of recombinant collagen fragments, about 2% to about 10% by weight of recombinant collagen fragments, about 3% to about 9% by weight of recombinant collagen fragments, about 4% to about 8% by weight of recombinant collagen fragments, or about 5% to about 7% by weight of recombinant collagen fragments.

[0222] In some embodiments, the composition may contain about 0.0005%, about 0.001%, about 0.01%, about 0.1%, 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%, about 20%, about 21%, about 22%, about 23%, about 24%, or about 25% by weight of the recombinant collagen fragment. In some embodiments, the composition can include about 0.0005%, about 0.001%, about 0.01%, about 0.1%, 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%, about 20%, about 21%, about 22%, about 23%, about 24%, or about 25% by weight of a protein having a structure according to SEQ ID Collagen fragment with sequence NO:1.

[0223] In some embodiments, the composition may include 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 one or more hydrolysates of recombinant collagen fragments. In some embodiments, the composition can include about 0.0005%, about 0.001%, about 0.01%, about 0.1%, 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%, about 20%, about 21%, about 22%, about 23%, about 24%, or about 25% by weight of a protein having a structure according to SEQ ID One or more hydrolysis products of recombinant collagen fragments having one or more sequences of SEQ ID NO:2 to SEQ ID NO:973.

[0224] In some embodiments, the composition may include 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 mixture of recombinant collagen fragments and one or more hydrolysates of the recombinant collagen fragments. In some embodiments, the cosmetic composition may include about 0.0005%, about 0.001%, about 0.01%, about 0.1%, 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%, about 20%, about 21%, about 22%, about 23%, about 24%, or about 25% by weight of the compound having a residue according to SEQ ID A mixture of a recombinant collagen fragment having the sequence of NO: 1 and one or more hydrolysates of the recombinant collagen fragment.

[0225] In some embodiments, the composition may contain about 0.0005% by weight to about 25% by weight of recombinant collagen fragment sequence variants, about 0.001% by weight to about 25% by weight of recombinant collagen fragment sequence variants, about 0.01% by weight to about 25% by weight of recombinant collagen fragment sequence variants, about 0.1% by weight to about 25% by weight of recombinant collagen fragment sequence variants, about 0.5% by weight to about 20% by weight of recombinant collagen fragment sequence variants, about 0.7% by weight to about 17% by weight of recombinant collagen fragment sequence variants, about 1% by weight to about 15% by weight of recombinant collagen fragment sequence variants, about 2% by weight to about 12% by weight of recombinant collagen fragment sequence variants, about 2% by weight to about 10% by weight of recombinant collagen fragment sequence variants, about 3% by weight to about 9% by weight of recombinant collagen fragment sequence variants, about 4% by weight to about 8% by weight of recombinant collagen fragment sequence variants, or about 5% by weight to about 7% by weight of recombinant collagen fragment sequence variants.

[0226] In some embodiments, the composition may contain about 0.0005%, about 0.001%, about 0.01%, about 0.1%, 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%, about 20%, about 21%, about 22%, about 23%, about 24%, or about 25% by weight of the recombinant collagen fragment sequence variant. In some embodiments, the composition can include about 0.0005%, about 0.001%, about 0.01%, about 0.1%, 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%, about 20%, about 21%, about 22%, about 23%, about 24%, or about 25% by weight of a protein having a structure according to SEQ ID A collagen fragment sequence variant of any one of SEQ ID NO:975 to SEQ ID NO:1002.

[0227] In some embodiments, the composition may include 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 A mixture of 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 % or about 25 weight % recombinant collagen fragments and one or more recombinant collagen fragment sequence variants. In some embodiments, the cosmetic composition may include about 0.0005%, about 0.001%, about 0.01%, about 0.1%, 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%, about 20%, about 21%, about 22%, about 23%, about 24%, or about 25% by weight of the compound having a residue according to SEQ ID A mixture of a recombinant collagen fragment having a sequence according to SEQ ID NO: 1 and one or more collagen fragment sequence variants having a sequence according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002.

[0228] In some embodiments, the composition may include a recombinant collagen fragment having an amino acid sequence as shown in SEQ ID NO: 1, one or more recombinant collagen fragments having a sequence according to one or more of SEQ ID NO: 2 to SEQ ID NO: 973, and / or one or more collagen fragment sequence variants having a sequence according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and hyaluronic acid, wherein the recombinant collagen fragment or variant is completely miscible with the hyaluronic acid. In some embodiments, the composition may include a recombinant collagen fragment having at least about 40% amino acid sequence identity with SEQ ID NO: 1, and / or one or more collagen fragment sequence variants having at least about 40% amino acid sequence identity with the amino acid sequence shown in any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and hyaluronic acid, wherein the recombinant collagen fragment or variant is completely miscible with the hyaluronic acid. In some embodiments, the hyaluronic acid may be a biphasic cross-linked hyaluronic acid comprising cross-linked HA and uncross-linked HA.

[0229] In some embodiments, the composition may include a recombinant collagen fragment having an amino acid sequence as shown in SEQ ID NO: 1, one or more recombinant collagen fragments having a sequence according to one or more of SEQ ID NO: 2 to SEQ ID NO: 973, and / or one or more collagen fragment sequence variants having a sequence according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and polyvinyl pyrrolidone, wherein the recombinant collagen fragment or variant is completely miscible with the polyvinyl pyrrolidone. In some embodiments, the composition may include one or more recombinant collagen fragments having at least about 40% amino acid sequence identity with SEQ ID NO: 1, and / or one or more collagen fragment sequence variants having at least about 40% amino acid sequence identity with the amino acid sequence shown in any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and polyvinyl pyrrolidone, wherein the recombinant collagen fragment or variant is completely miscible with the polyvinyl pyrrolidone.

[0230] In some embodiments, the composition may include a recombinant collagen fragment having an amino acid sequence as shown in SEQ ID NO: 1, one or more recombinant collagen fragments having a sequence according to one or more of SEQ ID NO: 2 to SEQ ID NO: 973, and / or one or more collagen fragment sequence variants having a sequence according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and sodium carboxymethylcellulose, wherein the recombinant collagen fragment or variant is completely miscible with the sodium carboxymethylcellulose. In some embodiments, the composition may include one or more recombinant collagen fragments having at least about 40% amino acid sequence identity with SEQ ID NO: 1, and / or one or more collagen fragment sequence variants having at least about 40% amino acid sequence identity with the amino acid sequence shown in any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and sodium carboxymethylcellulose, wherein the recombinant collagen fragment or variant is completely miscible with the sodium carboxymethylcellulose.

[0231] 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 range therebetween. In some embodiments, the compositions described herein include 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 range therebetween. In some embodiments, the compositions of the present disclosure include 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 a specific embodiment, the compositions described herein comprise hyaluronic acid at a concentration of about 24 mg / mL. In some embodiments, the disclosure provides a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid.

[0232] In certain embodiments, compositions disclosed herein include one or more collagens disclosed herein or recombinant collagen fragments cross-linked with hyaluronic acid. Collagen can be directly cross-linked with HA, i.e., cross-linked by amide or ester functional groups, or can be connected to HA by a joint. In some embodiments, compositions disclosed herein include a hyaluronic acid component, which includes hyaluronic acid that has been reacted to become a part of a cross-linked macromolecular matrix. In some embodiments, compositions according to the present disclosure include a collagen component, which includes one or more recombinant collagen fragments that have been reacted to become a part of a cross-linked macromolecular matrix. In addition to the crosslinking between hyaluronic acid and collagen, hyaluronic acid and / or collagen can be self-crosslinked.

[0233] In some embodiments, the composition may be a solution. In some embodiments, the composition may be a gel. In some embodiments, the composition may be a film. In some embodiments, the composition may be one or more fibers. In some embodiments, the composition may be a coating. In some embodiments, the composition may be a powder. In some embodiments, the composition may be a slurry. In some embodiments, the composition may be a particle. In some embodiments, the composition may be a foam.

[0234] In some embodiments, the composition described herein also includes a pharmaceutically active agent. Exemplary active agents include, but are not limited to, sunscreens, desquamation agents, moisturizers, decolorants, coloring agents, alpha-hydroxy acids, antimicrobial agents, anti-free radical agents, antifouling agents, anti-inflammatory agents, anesthetics, retinoids, extracts of algae, mushrooms, vegetables, yeasts, bacteria, hydrolyzed proteins, partially hydrolyzed proteins or unhydrolyzed proteins, enzymes, hormones, vitamins and their derivatives, flavonoids and isoflavones, and mixtures thereof. Other exemplary agents include, but are not limited to, anesthetics, such as lidocaine, bupivacaine, carbocaine, and related compounds. Other exemplary agents include, but are not limited to, nonsteroidal anti-inflammatory agents (NSAIDs), such as flurbiprofen, ibuprofen, naproxen, indomethacin, and related compounds. In some embodiments, the composition may also include one or more anti-mitotic drugs, including colchicine, taxol, and related compounds. In some embodiments, the composition may also include one or more topical disinfectants, such as benzoyl peroxide. In some embodiments, the composition may also include one or more polysaccharides produced by microalgae, such as microalginic acid. In some embodiments, the composition may also include one or more bioactive peptides, such as palmitoyl tripeptide-1 and palmitoyl tetrapeptide-7. In some embodiments, the composition may also include one or more polysaccharides produced by microalgae, such as microalginic acid.

[0235] E. How to use

[0236] Dermal fillers

[0237] In some embodiments, the present disclosure provides a dermal filler comprising a composition disclosed herein, the composition comprising collagen and hyaluronic acid. In some embodiments, the dermal fillers disclosed herein may comprise any known type of collagen that is miscible with HA. In some embodiments, the dermal fillers provided herein may comprise any known type of collagen that is completely miscible with HA. In some embodiments, administration of the dermal fillers described herein may increase the production of type I collagen. In some embodiments, administration of the dermal fillers described herein may increase the production of type III collagen. In some embodiments, administration of the dermal fillers described herein may increase the production of any combination of type I collagen, type III collagen, or ECM.

[0238] In some embodiments, the dermal fillers disclosed herein may comprise one or more recombinant collagen fragments disclosed herein. In specific embodiments, the dermal filler may comprise a recombinant collagen fragment according to SEQ ID NO: 1, and hyaluronic acid. In additional embodiments, the dermal filler may 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 SEQ ID NO: 1, and hyaluronic acid. In further embodiments, the composition may comprise 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 a dermal filler comprising a recombinant collagen fragment according to SEQ ID NO: 1 and hyaluronic acid as described herein may increase the production of type I collagen. In some embodiments, administration of a dermal filler comprising a recombinant collagen fragment according to SEQ ID NO: 1 and hyaluronic acid as described herein may increase the production of type III collagen. In some embodiments, administration of a 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 a 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.

[0239] In some embodiments, the dermal filler may comprise a hydrolysate of collagen fragments and hyaluronic acid, wherein the hydrolysate may have a sequence as part of SEQ ID NO: 1. In some embodiments, the composition comprises a hydrolysate having a sequence according to one or more of SEQ ID NO: 2 to SEQ ID NO: 973, and hyaluronic acid. In some embodiments, the composition may comprise any of the hydrolysates shown in SEQ ID NO: 2 to SEQ ID NO: 973, and hyaluronic acid. In some embodiments, the composition may 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 NO:2 to SEQ ID NO:973, and hyaluronic acid.

[0240] In some embodiments, the dermal filler may comprise a hydrolysate of collagen fragments as described herein and hyaluronic acid, wherein the hydrolysate may have a sequence as part of SEQ ID NO: 1, and administration of the dermal filler may increase the production of type I collagen. In some embodiments, administration of a dermal filler comprising a hydrolysate of collagen fragments and hyaluronic acid as described herein may increase the production of type III collagen, wherein the hydrolysate may have a sequence as part of SEQ ID NO: 1. In some embodiments, administration of a dermal filler comprising a hydrolysate of collagen fragments and hyaluronic acid as described herein may increase the production of both type I collagen and type III collagen, wherein the hydrolysate may have a sequence as part of SEQ ID NO: 1.

[0241] In yet another embodiment, the dermal filler may comprise a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysate having a sequence according to one of SEQ ID NO: 2 to SEQ ID NO: 973, and hyaluronic acid. In yet another embodiment, the dermal filler may comprise a recombinant collagen fragment according to SEQ ID NO: 1, a plurality of hydrolysates having a sequence according to any one of SEQ ID NO: 2 to SEQ ID NO: 973, which may be the same or different, and hyaluronic acid. In certain embodiments, the number of hydrolysates present in the plurality of hydrolysates in the dermal filler may increase with time, temperature, pH, or due to other conditions that would normally cause a recombinant collagen fragment (such as a recombinant collagen fragment according to SEQ ID NO: 1) to hydrolyze or otherwise decompose. In other embodiments, the dermal filler may 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 of the plurality of fragments remain substantially constant (i.e., vary by no more than ±5% as measured by HPLC over a given period of time) or remain constant. In certain embodiments, the recombinant collagen fragment (such as a recombinant collagen fragment according to SEQ ID NO: 1) can be hydrolyzed such that less than about 10%, about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of the unhydrolyzed recombinant fragment remains in the composition as measured by HPLC. In other embodiments, the dermal filler may comprise a mixture of recombinant collagen fragments (e.g., recombinant collagen fragments according to SEQ ID NO: 1) and various hydrolysates of the recombinant collagen fragments (e.g., various collagen fragments according to any one of SEQ ID NO: 2 to SEQ ID NO: 973) and hyaluronic acid, such that the weight of the hydrolysate in the composition is less than about 10%, about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of the weight of the collagen-related proteins in the dermal filler.

[0242] In yet another embodiment, the dermal filler may comprise a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysate having a sequence according to one of SEQ ID NO: 2 to SEQ ID NO: 973, and hyaluronic acid. In some embodiments, administration of a dermal filler comprising a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysate having a sequence according to one of SEQ ID NO: 2 to SEQ ID NO: 973, and hyaluronic acid as described herein may increase the production of type I collagen. In some embodiments, administration of a dermal filler comprising a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysate having a sequence according to one of SEQ ID NO: 2 to SEQ ID NO: 973, and hyaluronic acid as described herein may increase the production of type III collagen. In some embodiments, administration of a dermal filler comprising a recombinant collagen fragment according to SEQ ID NO: 1 and hyaluronic acid as described herein may increase the production of both type I and type III collagen.

[0243] In some embodiments, the dermal filler may comprise a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and hyaluronic acid.

[0244] In some embodiments, the dermal filler may further comprise 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, 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, and combinations thereof.

[0245] In some embodiments, the dermal filler may further comprise one or more polymers including, but not limited to, aliphatic polyesters, polyorthoesters, polyoxyesters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyesteramides, hydrolyzable amines, polyalkylene oxides, polyethylene oxide (PEO), poloxamers, functionalized isoflavonoids, amino acids, and combinations thereof.

[0246] In some embodiments, the dermal filler may further comprise one or more polymers selected from the group consisting of 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). -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-dioxepane-2-one and poly(1,5-dioxepane-2-one-co-p-dioxanone), poly(lactide-co-1,4-dioxepane-2-one), poly(1,4-dioxepane-2-one-co-p-dioxanone) Poly(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-L-lactide-co-glycolide-co-caprolactone), poly(dioxanone-co-D,L-lactide-co-glycolide-co-caprolactone) and combinations thereof.

[0247] In some embodiments, the dermal filler may further comprise 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.

[0248] In some embodiments, the dermal filler may also include biodegradable glasses or ceramics, such as calcium phosphate and other biocompatible metal oxides (ie, CaO) or combinations of biodegradable ceramics, glasses, and polymers.

[0249] 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 NO: 975 to SEQ ID NO: 1002 and hyaluronic acid as described herein may 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 NO: 975 to SEQ ID NO: 1002 and hyaluronic acid as described herein may 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 NO: 975 to SEQ ID NO: 1002 and hyaluronic acid may increase the production of both type I collagen and type III collagen.

[0250] In some embodiments, the dermal filler may comprise a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysate having a sequence according to one of SEQ ID NO: 2 to SEQ ID NO: 973, a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and hyaluronic acid. In some embodiments, administration of a dermal filler comprising a recombinant collagen fragment according to SEQ ID NO: 1, a hydrolysate having a sequence according to one of SEQ ID NO: 2 to SEQ ID NO: 973, a collagen fragment sequence variant having an amino acid sequence according to any one of SEQ ID NO: 975 to SEQ ID NO: 1002, and hyaluronic acid as described herein may increase the production of both type I collagen and type III collagen.

[0251] In some embodiments, the dermal filler is a heat-stable, sterile, 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 over a temperature range of up to at least about 135°C. As used herein, the dermal filler is "heat-stable," meaning that the dermal filler is substantially stable when subjected to a heat treatment, such as steam sterilization, at normal pressure or under pressure (autoclaving). 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 present prior to heat treatment. The heat stability of the dermal fillers disclosed herein can prevent degradation of the collagen, collagen fragments and / or collagen variants contained in the dermal filler. In some embodiments, the dermal filler can be terminally sterilized using different sterilization techniques. In some embodiments, the dermal filler can be sterilized by autoclaving, gamma irradiation, electron beam irradiation, or any method known in the art.

[0252] In some embodiments, the dermal filler is injectable. Typically, injectable dermal fillers are administered by subcutaneous or intradermal injection. The term "effective amount" refers to an amount of an injectable dermal filler sufficient to achieve 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 in between.

[0253] In some embodiments, the dermal filler may further comprise calcium phosphate particles. Suitable calcium phosphate particles include, but are not limited to, calcium hydroxyapatite, calcium fluorapatite, calcium chlorapatite, calcium carbonate apatite, tetracalcium phosphate, calcium pyrophosphate, tricalcium phosphate, and octacalcium phosphate particles and combinations thereof. In some embodiments, the dermal filler may further comprise calcium hydroxyapatite particles. In some embodiments, the dermal filler may further comprise carboxymethyl cellulose. In some embodiments, the dermal filler may further comprise polylactic acid.

[0254] In some embodiments, the particles may have a particle size ranging from about 10 μm to about 60 μm, and in specific embodiments, the particles may have a particle size ranging from about 25 μm to about 45 μm. In some embodiments, the particles may have a particle size of less than 60 μm, less than 45 μm, less than 35 μm, or less than 25 μm.

[0255] In some embodiments, the dermal fillers disclosed herein are suitable for injection through a fine gauge needle. In some embodiments, the injectable dermal fillers are suitable for administration with a needle having a size of about 16 gauge (Ga) to about 33 Ga. In some embodiments, a suitable needle has a size of about 22 Ga to about 30 Ga.

[0256] In some embodiments, the dermal fillers disclosed herein can be provided in a ready-to-use pre-filled syringe. Without wishing to be bound by a particular theory, it is believed that in the presently described dermal fillers, the synergistic effect of the components results in a stable suspension of calcium phosphate particles (e.g., calcium hydroxyapatite particles) in the formulation over time without phase separation.

[0257] In some embodiments, the dermal fillers described herein can be injected into any area of ​​the body or face where increased tissue volume is desired. For example, suitable areas of the body and face include glabellar lines, nasolabial folds, mandibular folds, marionette lines, cheek commissures, perioral wrinkles, crow's feet, skin depressions, scars, temples, subcutaneous support of the eyebrows, zygomatic and buccal fat pads, tear troughs, nose, lips, cheeks, areas around the mouth, infraorbital areas, facial asymmetry, jawline, and chin. For example, suitable areas of the body include buttocks, breasts, calves, flanks, or any other area of ​​the body where tissue volume is desired to achieve a cosmetic effect or to address body asymmetry. In addition, an increase in tissue volume can have therapeutic benefits. For example, if a dermal filler described herein is injected into the vocal cords, bladder wall or neck, or any other anatomical tubular structure, sphincter or valve, when enhanced by a dermal filler described herein, enhanced function will be provided, and a therapeutic benefit can be achieved.

[0258] In some embodiments, the dermal fillers disclosed herein further comprise a pharmaceutically active agent. In some embodiments, the pharmaceutically active agent comprises an anesthetic agent. In some embodiments, the anesthetic agent is lidocaine, bupivacaine, carbocaine, or any combination thereof.

[0259] Medical or cosmetic treatments

[0260] In some embodiments, the present disclosure provides a method of medical or cosmetic treatment, the method comprising administering or applying an effective amount of a composition described herein to a subject in need thereof. In some embodiments, the treatment comprises tissue augmentation. In some embodiments, the treatment comprises filling wrinkles or increasing the volume of the face. In some embodiments, the treatment comprises forming a cosmetic implant in a patient.

[0261] The compositions disclosed herein are contemplated for use in any of the following conditions or diseases for which the administration or application of a mixture of collagen and hyaluronic acid may be beneficial. Such conditions include, but are not limited to, orthopedic conditions (e.g., between bone-bone contacts) where internal buffering is desired, osteoarthritis, arthritis, age-related eye conditions, and the like. In some embodiments, the compositions disclosed herein may be administered to a subject's eye, blood vessel, nerve, cartilage, bone, breast, vocal cord, foot or tympanic membrane, joint, intervertebral disc, buttocks, hip, calf, or hand.

[0262] In some embodiments, the treatment comprises treating an inflammatory skin disorder or a mucosal disorder.

[0263] In some embodiments, treatment comprises treating lipoatrophy or other natural or surgically induced loss of tissue volume.

[0264] Increases collagen production in cells

[0265] The inventors have surprisingly found that the collagen fragments described in the present disclosure can induce the production of type I collagen and type III collagen in cells such as fibroblasts. Therefore, when contacting 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.

[0266] Therefore, in some embodiments, the disclosure provides methods for increasing collagen production in cells, which methods include contacting cells with compositions described herein. This contact can consist of directly exposing cells to a composition sufficient to induce these cells to produce a larger amount of collagen. In some embodiments, the method includes applying or applying a composition comprising collagen or collagen fragments, hyaluronic acid and at least one pharmaceutically acceptable excipient. In some embodiments, the composition can be applied to cells in their natural background, such as cells in an organism or tissue. In some embodiments, the composition can be topically applied to the skin. In some embodiments, the composition can be topically applied to human skin. In some embodiments, as described above, the composition can be injected.

[0267] In some embodiments, these cells can be fibroblasts. In some embodiments, these cells can be fibroblasts in their natural background (for example, in human skin). In some embodiments, these cells can be primary human fibroblasts. In some embodiments, these cells can be myocytes, transformed human cells, cardiomyocytes, epithelial cells, endothelial cells, stem cells, induced pluripotent stem cells, intervertebral disc pulposus cells, synoviocytes, osteoblasts, chondrocytes, keratinocytes or corneal cells. In some embodiments, these cells can be cells that produce insulin. In some embodiments, these cells can be pancreatic islet β cells.

[0268] In some embodiments, compositions disclosed herein can be applied to wounds, such as incisions, lacerations, cracks, tearing, scraping, wearing, burning or scratching, so as to increase the collagen production at the wound site and / or its surrounding tissues. As a part of the wound healing process, fibroblasts migrate to the wound site and produce the necessary collagen for wound repair. These cells eventually fill the wound cavity with the network of interwoven collagen lines, which timely arrange themselves into firm bands and form permanent new tissues. Therefore, compositions comprising collagen as described herein, collagen fragments and collagen variants can be applied to wounds, such as incisions, lacerations, cracks, tearing, scraping, wearing, burning or scratching, so as to increase the collagen production at the wound site and / or its surrounding tissues.

[0269] 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.

[0270] In certain embodiments, the present disclosure provides a composition comprising collagen or recombinant collagen fragments, hydrolyzates of recombinant collagen fragments, or a combination thereof and hyaluronic acid applied to a wound, such as an incision, laceration, crack, tear, scrape, abrasion, burn or scratch. In certain embodiments, the wound can be a wound in a human subject. In some embodiments, the method includes providing a composition for promoting collagen production and wound healing in the skin of a subject in need, wherein the composition includes a recombinant collagen fragment as described herein (e.g., a fragment comprising SEQ ID NO: 1). In some embodiments, the method includes providing a composition for promoting collagen production and wound healing in the skin of a subject in need, wherein the composition includes one or more hydrolyzates of recombinant collagen fragments as described herein (e.g., a hydrolyzate comprising one or more of SEQ ID NO: 2 to SEQ ID NO: 973). In some embodiments, the method includes providing a composition as described herein for promoting collagen production and wound healing in the skin of a subject in need thereof, wherein the composition comprises a mixture of recombinant collagen fragments and one or more hydrolysates of the recombinant collagen fragments (e.g., a mixture of fragments comprising SEQ ID NO: 1 and hydrolysates comprising SEQ ID NO: 2 to SEQ ID NO: 973).

[0271] In some embodiments, the method may include administering to the cell a collagen fragment having an amino acid sequence of SEQ ID NO: 1 and hyaluronic acid. In some embodiments, the method may include administering to the cell a hydrolyzate produced by hydrolysis of a collagen fragment having an amino acid sequence of SEQ ID NO: 1 (e.g., administering one or more hydrolyzates having a sequence according to SEQ ID NO: 2 to SEQ ID NO: 973). In some embodiments, the method includes administering to the cell a recombinant collagen fragment as described herein, and one or more hydrolyzates of the recombinant collagen fragment. In some embodiments, the method may include administering to the cell a collagen fragment having an amino acid sequence of SEQ ID NO: 1, and one or more hydrolyzates of the fragment having a sequence according to SEQ ID NO: 2 to SEQ ID NO: 973. In some embodiments, the method may include administering to the cell a collagen fragment having an amino acid sequence of SEQ ID NO: 975 to SEQ ID NO: 1002 and hyaluronic acid.

[0272] 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.

[0273] skin care

[0274] 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 use as a treatment for the eye area, nails and hair. The term "surface appearance" means visual and / or tactile irregularities in the skin and / or scalp, including wrinkles and fine lines, expression lines on the forehead and in the space between the eyebrows, wrinkles and / or fine lines around the mouth and / or sagging in the area around the lips and upper lip area (the area between the upper lip and the nose), uneven skin tone (freckles, actinic lentigo), the appearance and / or visibility of pores, the papery appearance of the skin, imperfections in the micro-relief of the skin such as chickenpox or acne scars, imperfections of oily skin (glowing appearance, etc.). The term "skin texture" can refer to loose, saggy, not too tight, not too elastic skin and / or skin that has sagged.

[0275] 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 produced by the action of stress exerted on the skin by underlying muscles. Age and environmental factors (such as exposure to sunlight) can deepen and perpetuate expression lines. Expression lines are characterized by the presence of grooves between the eyebrows (brow furrows or lion lines) and on the forehead in the areas where the skin muscles are located around the holes formed by the nose (nasal furrows), mouth (perioral lines and so-called bitter lines), and eyes (crow's feet).

[0276] 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 attributed to excess sebum, aging, loss of firmness, relaxation, stress, fatigue, improper hygiene, climatic factors, or any combination thereof. The compositions described herein can tighten pores, making them less visible.

[0277] In some embodiments, the compositions described herein can be used to improve the papery appearance of skin and skin touch behavior. Specifically, older skin can visually take on the appearance of cigarette paper, giving it an appearance similar to papyrus. The papery appearance of skin can be seen on the face and back of the hands of elderly people.

[0278] In some embodiments, the composition described herein can be a composition for protecting, treating or caring for the face, hands, feet or body, for example, a day cream, a night cream, a makeup remover cream, a sunscreen composition, a body milk for skin protection or care, an after-sun milk, a skin care lotion, a gel, a foam, an artificial tanning composition and an after-shave composition. In some embodiments, the compositions described herein can be formulated as, for example, solutions, suspensions, lotions, creams, serums, gels, balms, gels, oils, cream-in-oil, micellar waters, facial sprays, facial serums, concealer or skin tone correcting formulas, toners (water and / or alcohol based), paints, polishes, sticks, pencils, sprays, aerosols, ointments, cleansing liquid detergents, solid sticks, shampoos, conditioners, hair styling products, pastes, foams, powders, mousses, balms, shaving creams, wipes, strips, patches, wound dressings, adhesive bandages, hydrogels, film-forming products, face and skin masks, cosmetics (e.g., foundations, eyeliners, eye shadows), exfoliants, deodorants, and antiperspirants, etc. Exemplary formulations are provided herein.

[0279] In some embodiments, the composition described herein can be a cosmetic composition, and the at least one excipient can be a cosmetically acceptable excipient. A cosmetically acceptable excipient is an excipient suitable for use in a cosmetic product. Exemplary cosmetically acceptable excipients are described below.

[0280] In some embodiments, the cosmetic compositions described herein may include hydrolysates of recombinant collagen fragments, and at least one excipient, such as a cosmetically acceptable excipient. In some embodiments, the cosmetic compositions described herein may include recombinant collagen fragments, one or more hydrolysates of the recombinant collagen fragments, and at least one excipient, such as a cosmetically acceptable excipient.

[0281] In some embodiments, the cosmetic composition may include ingredients commonly used in cosmetic products such as skin care, eye care, nail care, and hair care products. These ingredients may include, but are not limited to, soaps, antimicrobial agents, anti-inflammatory agents, moisturizers, wax alcohols, hydrating agents, moisturizers, penetration enhancers, emulsifiers, natural or synthetic oils, solvents, fats, surfactants, detergents, gelling agents, emollients, antioxidants, fragrances, paints, polishes, fillers, thickeners, waxes, odor absorbers, dyes, colorants, powders, viscosity control agents, analgesics, anesthetics, antipruritics, plant extracts, conditioners, darkening agents or whitening agents, moisturizers, mica, minerals, polyphenols, silicones or silicone derivatives (such as dimethicone), sunscreens, vitamins, botanicals, alcohols (such as denatured alcohols and ethanol), polyols, polyol ethers, and other ingredients listed in the International Cosmetic Ingredient Dictionary and Handbook, 13th edition (2009), the entire contents of which are incorporated herein by reference. In certain embodiments, a given ingredient may perform more than one function and may belong to more than one category.

[0282] In some embodiments, the composition as described herein can be a therapeutic composition, and the at least one excipient can be a therapeutically acceptable excipient. The therapeutic composition can be used to treat one or more symptom(s), 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. An acceptable excipient for treatment is an excipient that can be used as a vehicle or medium for an active substance, and includes excipients that are commonly used in therapeutic compositions (that is, compositions that can be used to treat one or more symptom(s). An exemplary therapeutically acceptable excipient is described below.

[0283] In some embodiments, the composition described herein can be a dietary composition or a dietary supplement, and the at least one excipient can be, for example, a food or beverage additive. "Dietary supplements" are preparations intended to supplement diets, and can be used to provide nutrients or additives that may be missing or may not be consumed in sufficient amounts in a person's diet. In some embodiments, the dietary supplement can be provided in the form of any conventional 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 as soft candies. Suitable excipients include but are not limited to lactose or milk sugar and high molecular weight polyethylene glycols, etc. Exemplary dietary compositions are described below.

[0284] The compositions described herein may also include one or more of the following additional components. Exemplary contemplated additional ingredients are described below; however, the present disclosure is not limited to these exemplary additional ingredients.

[0285] In some embodiments, the compositions described herein may further comprise one or more anti-wrinkle agents. Anti-wrinkle agents are compounds that cause the synthesis and / or increased activity of certain enzymes of the skin when the composition is in contact with a wrinkled skin area (e.g., body or face, including the eye area), which can reduce the appearance of wrinkles and / or fine lines. Exemplary anti-wrinkle agents include, but are not limited to, desquamating agents, anti-glycation agents, nitric oxide synthase inhibitors, muscle relaxants and / or anti-skin wrinkle agents, anti-free radical agents, and mixtures thereof.

[0286] Additional exemplary anti-wrinkle agents that can be included in the compositions described herein include, but are not limited to, 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., tocopherol acetate), niacin and its precursors (e.g., niacinamide), 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 sold under the trade name PRO-XYLANE), plant extracts (e.g., For example, sea fennel extract and olive leaf extract), plant proteins and hydrolysates thereof (e.g., rice or soy protein hydrolysates), algae extracts (e.g., kelp algae extracts), bacterial extracts, sapogenins (e.g., diosgenin), yam extracts (e.g., wild yam extracts), alpha-hydroxy acids, beta-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-methylbutanoylamino}acetic acid), lipopeptides (e.g., MATRIXYL 3000 available from Croda)), lycopene, manganese salts, magnesium salts (e.g., gluconate), and combinations of any of the foregoing.

[0287] Exemplary adenosine derivatives include, but are not limited to, 2'-deoxyadenosine; 2',3'-isopropylidene adenosine; toyocamycin, 1-methyladenosine; N-6-methyladenosine, adenosine N-oxide, 6-methyl-mercaptopurine nucleoside, and 6-chloropurine nucleoside. Other adenosine derivatives include adenosine receptor agonists, including phenylisopropyladenosine ("PIA") / 1-methylisoguanosine, Ns-cyclohexyladenosine (CHA), N6-cyclopentyladenosine (CPA), 2-chloro-Ns-cyclopentyl-adenosine, 2-chloroadenosine, N6-phenyladenosine, 2-phenylaminoadenosine, MECA, Ne-phenylethyladenosine, 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethylformamido-adenosine (CGS-21680), (N-ethylformamido)adenosine-S-(NECA), 5'-(N-cyclopropylformamido)adenosine, DPMA (PD 129,944) and metrifudil.

[0288] In some embodiments, the compositions described herein may include one or more adenosine derivatives that increase the intracellular concentration of adenosine, such as erythro-9-(2-hydroxy-3-nonyl)adenine ("EHNA"), iodotuberculocidin, or a combination thereof. Additional adenosine derivatives contemplated herein include adenosine salts and alkyl esters of adenosine.

[0289] In some embodiments, the composition may also include one or more pearlescent agents. Pearlescent agents are iridescent particles of any shape, especially produced by certain shellfish in their shells. Alternatively, the pearlescent agent may be synthetic, i.e. artificially made. The pearlescent agent may be selected from white pearlescent agents, such as but not limited to mica covered with titanium oxide or with bismuth oxychloride; colored pearlescent agents, such as but not limited to bismuth oxychloride-based pearlescent agents; titanium oxide-coated mica covered with iron oxide; especially titanium oxide-coated mica covered with iron blue or chromium oxide; or titanium oxide-coated mica covered with organic pigments.

[0290] In some embodiments, the composition may further include one or more hydroxy acids. Examples of hydroxy acids include beta hydroxy acids, such as salicylic acid, acetylsalicylic acid, and the like. Other exemplary hydroxy acids suitable for use in the composition include citric acid, glycolic acid, hydroxycaproic acid, hydroxyoctanoic acid, lactic acid, malic acid, tartaric acid, polyhydroxy acids (including gluconolactone), and any combination thereof.

[0291] In some embodiments, the composition may also include one or more emulsifiers. Emulsifiers prevent different ingredients (such as oil and water) from separating in an emulsion. Suitable emulsifiers include, but are not limited to, polysorbate, laureth-4, potassium cetyl sulfate, caprylic glyceride, and any combination thereof.

[0292] In some embodiments, the composition may further comprise one or more chelating agents. Chelating agents bind to metal ions and prevent them from chemically reacting with other substances in the formulation. Suitable chelating agents include, but are not limited to, sodium phytate, disodium ethylenediaminetetraacetate, tetrasodium ethylenediaminetetraacetate, tetrasodium glutamate diacetate, and trisodium ethylenediamine disuccinate.

[0293] In addition to chelating agents, tetrasodium glutamate diacetate may also be used as a stabilizer in any of the compositions described herein.

[0294] In some embodiments, the composition may further comprise one or more antimicrobial agents. Suitable antimicrobial agents include, but are not limited to, caprylyl glyceryl ether, benzalkonium chloride, benzethonium chloride and chloroxylenol (PCMX), tea tree oil, witch hazel oil, rosemary oil, lemon oil, and any combination thereof.

[0295] In some embodiments, the composition may also include one or more humectants (water retaining agents) to improve moisture levels in the skin. Non-limiting examples of suitable humectants 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, tricholoma pubescens extract, creatinine, diglycerol, biosaccharide gum-1, glucosamine salts, glucuronates, glutamates, polyethylene glycol ethers of glycerol (e.g., glycereth 20), glycerol, glycerol monopropoxylate, glycogen, hexylene glycol, honey, hydrogenated starch hydrolysate, hydrolyzed mucopolysaccharides (such as xanthan gum and biosaccharide gum-1), inositol, keratin amino acids, glycosaminoglycans, methoxy PEG 10, methyl gluceth-10 and methyl gluceth-20, methyl glucose, 3-methyl-1,3-butylene glycol, N-acetylglucosamine salts, polyethylene glycol and its derivatives (such as PEG 15 butylene glycol, PEG 4, PEG 5 pentaerythritol, PEG 6, PEG 8, PEG 9), propylene glycol, pentaerythritol, 1,2 pentanediol, PPG-1 glyceryl ether, 2-pyrrolidone-5-carboxylic acid (including its salts and esters), carbohydrate isomers, sericin, silk amino acids, sodium ethyl hyaluronate, sodium hyaluronate, sodium polyaspartate, sodium polyglutamate, caprylyl glycol, sorbitol polyether 20, sorbitol polyether 6, sugars and sugar alcohols and their derivatives, such as glucose, mannose and polyglycerol sorbitol, trehalose, triglycerol, trimethylolpropane, tris (hydroxymethyl) aminomethane salt and yeast extract and mixtures thereof.

[0296] Other humectants suitable for use herein include polyols selected from the group consisting of glycerol, diglycerol, glycerol, erythritol, arabitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, maltitol, mannose, inositol, triethylene glycol, sodium pyrrolidonecarboxylate (PCA), zinc PCA, and derivatives and mixtures thereof.

[0297] In some embodiments, the composition may further include a gelling agent comprising a polyacrylamide-based polymer. In addition to polyacrylamide itself, the polyacrylamide-based polymer may also be a derivative thereof, and may be a mixture of multiple types of polymers, and may also be a copolymer with acrylamide and its derivatives as monomers. The gelling agent may be used to provide a good appearance to the skin, provide a cooling sensation to the skin, and provide a refreshing and non-sticky sensation to the skin. In some embodiments, the gelling agent may include one or more pigments, or one or more fillers, including: inorganic pigments (including extender pigments, coloring pigments and whitening pigments), organic pigments, pearlescent pigments, macromolecular powders, functional pigments, talc, mica, kaolin, calcium carbonate, magnesium carbonate, silicic anhydride, 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 scales, bismuth oxychloride, boron nitride, nylon powder, silk powder, carbomer, tar pigments, natural pigments and titanium oxide, such as amorphous or rutile and / or anatase crystals.

[0298] In some embodiments, the composition may include water-resistant and oil-resistant pigments and / or fillers, and may also include any conventionally used water-repellent and / or oil-proof agent to impart water resistance and oil resistance to the pigment, such as fluorochemicals. Representative fluorochemicals that are conventionally used and can act as water-repellents and oil-proof agents include compounds with perfluoroalkyl groups, such as perfluoroalkyl phosphates, perfluoroalkyl silanes, perfluoroalkyl silazanes, polyhexafluoropropylene oxides, organosiloxanes containing perfluoroalkyl groups, perfluoropolyethers, perfluoroalcohols, perfluoroalkyl acrylate polymers, and their derivatives. Perfluoroalkyl phosphates can provide uniform and stable pigment dispersions in the formulation of the gel composition, and perfluoroalkyl silanes can have excellent compatibility with other cosmetic ingredients. In addition, perfluoroalkyl phosphate-diethanolamine salts produced and marketed by Asahi Glass as AsahiGuard AG530, and perfluoroalkyl silane coupling agents, such as LP-IT and LP-4T of Shin-Etsu Silicone, can be used.

[0299] Representative gelling agents include, but are not limited to, those manufactured and marketed by Sepigel 305, Sepigel 501, and Sepigel 600. Sepigel 305 is a gelling agent containing approximately 40% polyacrylamide, approximately 24% C- 13 -C 14 Isoparaffin and about 6% of laureth-7 (herein, laureth-7 is a polyol having the formula C 12 H 25 --(OCH2CH2) n --OH nonionic surfactants, wherein the average value of n is 7). Sepigel 600 is a mixture of acrylamide / acrylamide-2-propanesulfonate copolymer, isohexadecane and polysorbate 80 (polyoxyethylene sorbitan monooleate (20EO)). Suitable gelling agents comprising polyacrylamide-based polymers are disclosed in EP 0503 853 (Scott Bader Company Ltd.), the disclosure of which is incorporated herein by reference.

[0300] In some embodiments, the composition may also include hyaluronic acid (HA). In some embodiments, HA may be in a non-crosslinked state. In some embodiments, HA may 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 a non-sulfated glycosaminoglycan that is widely distributed in the connective tissue, epithelial tissue, and neural tissue of the human body. Hyaluronan is abundant in different layers of the skin, where it has multiple functions, for example, ensuring good hydration, assisting the organization of the ECM, acting as a filler material; and participating in tissue repair mechanisms. However, with age, the amount of hyaluronan present in the skin decreases.

[0301] In some embodiments, the compositions described herein can be dermal filler compositions, such as injectable dermal filler compositions. The dermal filler compositions can be suitable for use on the face and body, including, for example, around the eyes, on or around the cheeks, on or around the neckline, on or around the hands, on or around the nails, on or around the ears (including on the earlobes), on or around the legs, and on or around the feet.

[0302] In some embodiments, the compositions described herein can be used with a microneedle array, such as an array included in a sheet or patch. The microneedle array can include a plurality of microneedles of sufficient length to penetrate the skin through the stratum corneum and into the living epidermis. In some embodiments, it may be desirable to deliver collagen to the epidermis / dermis junction area for cosmetic or therapeutic purposes.

[0303] In some embodiments, the composition 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, eyes, cheeks, lips, collars and hands. In some embodiments, the microneedle for delivering the composition to the epidermis and dermis in a targeted manner is an injectable microneedle, a drug-coated metal microneedle, or a microneedle with a soluble tip. Exemplary methods and disclosures about microneedles can be found in, for example, Aditya et al., Kinetics of collagen microneedledrug deliverysystem, 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 some embodiments, the compositions described herein may be applied to the skin before, during, or after any skin resurfacing process. Skin resurfacing process refers to a variety of skin care designed to regenerate skin. Skin resurfacing processes include laser skin resurfacing, chemical peeling, microdermabrasion, microdermabrasion, microneedling, and photorejuvenation with intense pulsed light (IPL) technology. In some embodiments, the composition may also include waxy lipids, such as ceramides. Ceramides help create a barrier to prevent permeability, which helps prevent dryness and irritation, and can also protect the epidermis from environmental damage.

[0304] In some embodiments, the composition may further comprise vitamin A or a vitamin A derivative. Examples of vitamin derivatives include, but are not limited to, retinoids such as retinal, retinoic acid, retinoic acid esters, retinol esters, retinol, tretinoin, isotretinoin, adapalene, tazarotene, etc. 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).

[0305] In some embodiments, the composition may further include vitamin C or a derivative thereof, such as ascorbic acid, ascorbic acid esters (eg, tetrahexyldecyl ascorbate), and the like.

[0306] In some embodiments, the composition may further include B vitamins, such as biotin (ie, vitamin B7), niacinamide, and the like.

[0307] In some embodiments, the composition may further comprise vitamin E, such as α-tocopherol, β-tocopherol, γ-tocopherol, and σ-tocopherol and their related corresponding tocotrienols and the like.

[0308] In some embodiments, the composition may further comprise vitamin K and its derivatives.

[0309] Any vitamin, vitamin analog, or derivative thereof that can be suitably formulated as a topical composition is contemplated for the present disclosure.

[0310] In some embodiments, the compositions disclosed herein may also include one or more thickening agents. Thickening agents, i.e., structural synergists, can suspend pigments and / or increase viscosity in the composition. Thickeners and / or structural synergists suitable for the compositions of the present invention include, but are not limited to, organically modified clays, fumed silica, trihydroxystearin, silicone gels or silicone elastomers, ammonium acryloyldimethyltaurate / VP copolymers, acrylates / C10-30 alkyl acrylate crosspolymers, and mixtures thereof.

[0311] Suitable organomodified clays include, but are not limited to, hectorites, bentonites, smectites and organomodified versions of montmorillonite clays (such as those sold under the trade names Those sold from Elementis Specialties, under the trade name Those sold from Sud-Chemie and under the trade name Hydrophilically modified fumed silicas include, but are not limited to, those sold from Southern Clay Products. N20 and T30 grades (Wacker-Chemie AG), and the trade name 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, DOW 9040, 9041, 9045 and 9546 silicone elastomer blends, SFE839 from Momentive Performance Materials TM and Velvesil TM Silicone gel, and from Wacker-Chemie AG RG-100.

[0312] In some embodiments, the compositions disclosed herein may also include one or more fat-soluble / fat-dispersible film-forming agents. Suitable fat-soluble / fat-dispersible film-forming agents for use herein include, but are not limited to, 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-based polymers or copolymers, including silicone and / or fluorinated types (e.g., silicone acrylates from the "KP" series of Shin-Etsu Silicones, and 3M TM Silicone "Plus" polymers VS70 and SA70); polyurethanes (e.g., hydroxy ester triglyceride-derived Poly series); polyesters (e.g., from Inolex Chemical Company series of polymerized polyesters); and mixtures thereof.

[0313] In some embodiments, the compositions disclosed herein may also include one or more colorants. Suitable colorants for use herein include all inorganic and organic pigments / pigments, including mineral or pearlescent pigments suitable for use in cosmetic compositions. Such colorants include those with or without surface coatings or treatments. Colorants can enhance the coloring and / or light scattering and / or light reflection effects of the composition.

[0314] In some embodiments, the compositions disclosed herein may also include one or more sunscreens, such as mineral and / or physical sunscreens.Sunscreens can block UVA and / or UVB radiation.Exemplary UVA sunscreens include but are not limited to avobenzone, terephthalylidene dicamphor sulfonic acid, disodium bisdisulfonazole, disodium phenyl dibenzimidazole tetrasulfonate, diethylamino hydroxybenzoyl hexyl benzoate, bis diethylamino hydroxybenzoyl benzoate, bisbenzoxazolylphenyl ethylhexylamino triazine and combinations thereof.

[0315] Exemplary UVB sunscreens include, but are not limited to, octocrylene, octinoxate, octisalate, homosalate, ensulizole, ethylhexyl triazone, enzacamene, amiloxate, diethylhexyl butyramido triazine, benzalmalonate polysiloxane, padrimate-O, triethanolamine salicylate, cinoxate, p-aminobenzoic acid and derivatives thereof, and combinations thereof.

[0316] Exemplary sunscreens that absorb UVA and UVB radiation include, for example, oxybenzone, meradiate, titanium dioxide, zinc oxide, bis-oxotriazole, bemtrizinol, drometrizoletrisiloxane, sulisobenzone, dioxybenzone, or combinations thereof.

[0317] Specific suitable sunscreens include, but are not limited to, para-aminobenzoic acid, its salts and derivatives thereof (ethyl, isobutyl, glyceryl esters, p-dimethylaminobenzoic acid, anthranilates (i.e., o-aminobenzoate, methyl, menthyl, phenyl, benzyl, phenethyl, linalyl, terpinyl and cyclohexenyl esters), salicylates (amyl, phenyl, benzyl, menthyl, glyceryl and dipropylene glycol esters), cinnamic acid derivatives (methyl and benzyl esters, α-phenylcinnamonitrile, butylcinnamoyl pyruvate), dihydroxycinnamic acid derivatives (umbelliferone, methyl umbelliferone, methyl acetyl-umbelliferone), trihydroxycinnamic acid derivatives (esculetin, methyl esculetin, daphnetin and glucoside, esculetin and daphnetin), hydrocarbons (diphenylbutadiene, stilbene), diisocyanate (dextrin), Benzyl acetone and benzylidene acetophenone, naphthol sulfonates (sodium salts of 2-naphthol-3,3-disulfonic acid and 2-naphthol-6,8-disulfonic acid), β-hydroxynaphthoic acid and its salts, o-hydroxybiphenyl disulfonates and p-hydroxybiphenyl disulfonates, coumarin derivatives (7-hydroxycoumarin, 7-methylcoumarin, 3-phenylcoumarin), diazoles (2-acetyl-3-bromoindazole, phenylbenzoxazole, methylnaphthoxazole, various arylbenzothiazoles), quinine salts (bisulfate, sulfate, chloride, oleate and tannate), quinoline derivatives (8-hydroxyquinolinate, 2-phenylquinoline), hydroxy- or methoxy-substituted benzophenones, uric acid and vilouric acid, tannic acid and its derivatives (e.g. ethyl ether), (butyl carbitol) (6-propyl piperonyl) ether, hydroquinone, benzophenone (hydroxybenzoate, sulfisobenzoate, dihydroxybenzoate, benzoylresorcinol, 2,2,4,4'-tetrahydroxybenzophenone, 2,2'-dihydroxy-4,4'-dimethoxybenzophenone, octabenzoate, 4-isopropyldibenzoylmethane, butyl methoxydibenzoylmethane, etocrylene and 4-isopropyl-dibenzoylmethane, titanium dioxide, iron oxide, zinc oxide and mixtures thereof. Other cosmetically acceptable sunscreen agents and concentrations (percent by weight of the total cosmetic sunscreen composition) include diethanolamine methoxycinnamate (10% or less), ethyl-bis(hydroxypropyl)aminobenzoic acid Esters (5% or less), aminobenzoic acid glyceryl esters (3% or less), 4-isopropyl dibenzoylmethane (5% or less), 4-methylbenzylidene camphor (6% or less), terephthalylidene dicamphor sulfonic acid (10% or less) and sulfisobenzone (also known as benzophenone-4, 10% or less). In some embodiments, the compositions disclosed herein may also include D-aspartic acid and / or D-alanine and any salts thereof. As used herein, the term "derivatives" of D-aspartic acid and D-alanine indicates D-aspartic acid and D-alanine molecules covalently bound to any organic group through their amino, carboxyl or side chains, provided that the effect of D-aspartic acid and D-alanine in promoting collagen production is not impaired.Exemplary organic groups include, but are not limited to, protecting groups such as N-phenylacetyl and 4,4'-dimethoxytrityl (DMT) groups; biopolymers such as proteins, peptides, sugars, lipids, and nucleic acids; synthetic polymers such as polystyrene, polyethylene, polyvinyl, polypropylene, and polyesters; and functional groups such as ester groups. Ester groups may include, for example, aliphatic esters such as methyl and ethyl esters; and aromatic esters.

[0318] In some embodiments, the composition may also include one or more general skin care additives, for example, conditioning agents such as silicones. In some embodiments, the composition may further include one or more shark liver oils, for example, squalane and / or squalene. In some embodiments, the composition may also include one or more polysaccharides produced by microalgae, for example, microalginic acid.

[0319] In some embodiments, the composition may further comprise at least one preservative. In some embodiments, the at least one preservative may be a quaternary ammonium compound, a halogenated phenol, sorbic acid, potassium sorbate, benzoic acid, sodium benzoate, sodium citrate, sodium anisate, capryloyl hydroxamic acid, sodium levulinate, phenoxyethanol, or a combination thereof.

[0320] The pH of the compositions described herein may range from about 4 to about 8, from about 4.7 to about 5.5, from about 5 to about 7, from about 6 to about 7, from about 6.1 to about 6.8, or from about 6.4 to about 6.6.

[0321] soap

[0322] In some embodiments, the cosmetic composition described herein can be a cleaning composition or soap, including traditional soaps and cleansers in the form of solid bars, makeup removers, shower gels, lotions, creams, foaming cream gels, or liquid soaps in the form of gels that can be packaged in tubes, bottles, pump bottles, aerosol shower foams, or foam pump bottles. Soaps can be used in cosmetic methods to clean dirt residues from human keratinous materials in the presence of water, agglomerate to form foam, and the foam and dirt residues formed are removed by rinsing with water, and can be used on any part of the body and face disclosed herein, including, for example, on body skin, face, hands, lips, eyelids, nails, hair, eyelashes, and / or eyebrows.

[0323] Traditionally, solid soaps include alkali metal fatty acid salts and potassium fatty acid soaps, and liquid soaps include four main families of detergent formulations: (1) detergent formulations based on lauryl sulfate; detergent formulations based on α-olefin sulfonates; (3) detergent formulations based on mixtures of synthetic anionic surfactants, amphoteric surfactants and / or nonionic surfactants; (4) mixed formulations based on soap and synthetic surfactants. Liquid soaps typically 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; esters of polyethylene glycol with monobasic acids or stearic acid, such as polyethylene glycol distearate 6000 or mixtures thereof, and the liquid soap is contained in a cosmetically acceptable aqueous medium. However, both solid and liquid soaps may contain any suitable additional ingredients, such as those listed below, in any combination.

[0324] In some embodiments, the cleaning compositions or soaps described herein may also include: one or more cellulose compounds, or polysaccharide compounds having chains of glucose residues connected by β-1,4 bonds in their structure; one or more fatty acids containing straight or branched, 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 and stearic acid, linolenic acid and mixtures thereof; and one or more inorganic 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.

[0325] In some embodiments, the compositions described herein may also include one or more anionic surfactants or salts, including alkali metal salts such as sodium salts, ammonium salts, amine salts, amino alcohol salts, or salts of alkaline earth metals (e.g., magnesium) of the following types: alkyl sulfates, alkyl ether sulfates, alkyl amido 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, all of which have 6 to 24 carbon atoms in the alkyl and acyl groups and phenyl or benzyl in the aryl group; C6-C6 of polyglycoside carboxylic acids. 24Alkyl esters, such as alkyl glucosides citrate, alkyl polyglycoside tartarates and alkyl polyglycoside sulfosuccinates, alkyl sulfosuccinates, acyl isethionates and N-acyl taurates, all of which have 12 to 20 carbon atoms in the alkyl or acyl groups; and / or acyl lactylates in which the acyl groups contain 8 to 20 carbon atoms, and mixtures thereof. In some embodiments, alkyl-D-galactosiduronic acid, polyoxyalkylene (C6-C 24 ) ether carboxylic acid, polyoxyalkylene (C6-C 24 )Aryl(C6-C 24 ) polyoxyalkylene ether carboxylic acid, polyoxyalkylene (C6-C 24 )Alkyl amido ether carboxylic acids, especially those containing 2 to 50 ethylene oxide groups; and their alkali metal salts, ammonium salts, amine salts, amino alcohol salts or alkaline earth metal salts may also be suitable.

[0326] Suitable C6-C 24 Alkyl ether sulfates include alkali metal salts or alkaline earth metal salts, ammonium salts, amine salts or amino alcohol salts, sodium salts and oxyethylated (C 12 -C 14 ) alkyl ether sulfates whose average number of ethylene oxide groups is from 1 to 4 and include sodium laureth sulfate (CTFA name), such as the products sold under the names TEXAPON AOS225UPTEXAPON N702TEXAPON NSW manufactured and marketed by COGNIS or the products sold as EMPICOL ESB3 / FL2, EMPICOL ESB3 / FL3, EMPICOL ESB70 / FL2 by the company Huntsman.

[0327] Suitable amphoteric surfactants include, but are not limited to, derivatives of secondary or tertiary aliphatic amines, wherein the aliphatic group is a straight or branched chain containing 8 to 22 carbon atoms. The amphoteric surfactant may contain at least one water-soluble anionic group, such as a carboxylate, sulfonate, sulfate, phosphate or phosphonate group, (C8-C 20 ) alkyl betaine, sulfo betaine, (C8-C 20 )alkylamido (C6-C8) alkyl betaine or (C8-C 20 )alkylamidoalkyl (C6-C8) sulfobetaines, and mixtures thereof.

[0328] Suitable amine derivatives include those known under the names The products sold are described in U.S. Pat. No. 2,528,378 and U.S. Pat. No. 2,781,354 and are referred to in the CTFA dictionary, 3rd edition, 1982, under the terms amphocarboxyglycinate and amphocarboxypropionate. Other suitable amine derivatives include those concentrated by Rhodia under the trade name Those amine derivatives marketed by C2M and classified in the CTFA dictionary, 5th edition, 1993, under the following names: disodium cocoamphodiacetate, disodium lauroamphodiacetate, disodium caproamphodiacetate, disodium capryloamphodiacetate, disodium cocoamphodipropionate, disodium lauroamphodipropionate, disodium caproamphodipropionate, disodium capryloamphodipropionate, lauroamphodipropionic acid, cocoamphodipropionic acid and cocoamphodiacetate. Suitable alkyl (C8-C 20 ) Betaines include cocoamidopropyl betaine and cocoyl betaine, such as the commercial product MIRATAINE BB / FLA from RHODIA or the commercial product EMPIGEN BB / FL from Huntsman.

[0329] In some embodiments, the cleaning compositions or soaps described herein may be sulfate-free, and may include a sulfate-free surfactant system.

[0330] In some embodiments, the cleaning compositions or soaps described herein may also include one or more thickeners of the type of nonionic cellulose compounds. Suitable cellulose compounds include, but are not limited to, nonionic cellulose ethers, including methylcellulose and ethylcellulose; hydroxyalkyl celluloses, such as hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl cellulose; mixed hydroxyalkyl-alkyl celluloses, such as hydroxypropyl-methyl cellulose, hydroxyethyl-methyl cellulose, hydroxyethyl ethyl cellulose and hydroxybutyl-methyl cellulose and alkyl chain-modified hydroxyalkyl celluloses. Suitable hydroxypropyl methylcelluloses include the commercial products METHOCEL E, F, J and K sold by Dow Coming, and even more particularly METHOCEL E 4MQG or METHOCEL F 4M. Suitable cellulose components may be in crystalline form, microcrystalline form or mixtures thereof.

[0331] In some embodiments, the cleaning compositions or soaps described herein may also include 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 monobasic acid 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 acids such as CARBOPOL 980 and CARBOPOL 1382 produced and marketed by NOVEON, acrylate / acrylonitrile copolymers such as HYPANSS201 produced and marketed by KINGSTON; clays such as smectite, modified or unmodified hectorites such as BENTONE products produced and marketed by Rheox, LAPONITE products produced and marketed by Southern Clay Products, VEEGUM HS products produced and marketed by RT Vanderbilt, and mixtures thereof.

[0332] In some embodiments, the cleaning compositions or soaps described herein may also include one or more nonionic surfactants. These are well-known compounds (see MR PORTER's "Handbook of Surfactants", edited by Blackie & Son (Glasgow and London), 1991, pages 116-178) and may be selected from alcohols, α-diols, (C1-C 20 )alkylphenols or polyethoxylated fatty acids, polypropoxylated fatty acids or polyglycerolated fatty acids, these fatty acids have a fatty chain containing, for example, 8 to 18 atoms, the number of ethylene oxide groups or propylene oxides can be in the range of 2 to 50 and the number of glycerol groups can be in the range of 2 to 30; copolymers of ethylene oxide and propylene; condensates of ethylene oxide and propylene oxide on fatty alcohols; polyethoxylated fatty amides with 2 to 30 mol of ethylene oxide, polyglycerolated fatty amides containing an average of 1 to 5 glycerol groups; polyethoxylated fatty amines with 2 to 30 mol of ethylene oxide, ethoxylated sorbitan fatty acid esters with 2 to 30 mol of ethylene oxide; sucrose fatty acid esters, polyethylene glycol fatty acid esters, (C6-C 24 ) alkyl polyglycosides, N-alkyl (C6-C 24 ) glucosamine derivatives, amine oxides, such as alkyl (C 10 -C 14 ) amine oxide or N-acyl (C 10 -C 14 )-aminopropylmorpholine oxide, and mixtures thereof.

[0333] Other suitable nonionic surfactants include, but are not limited to, alkyl polyglucosides (APGs), maltose esters, polyglycerolated fatty alcohols, glucosamine derivatives (e.g., 2-ethylhexyloxycarbonyl-N-methylglucosamine), and mixtures thereof. Suitable alkyl polyglucosides include those containing an alkyl group containing 6 to 30 carbon atoms and a hydrophilic group (glucoside). Exemplary alkyl polyglucosides include decyl glucoside (alkyl-C9 / C11-polyglucoside (1.4)), including those sold by Kao Chemicals under the name Mydol The product is sold by Cognis under the name Plantaren2000 Products sold and marketed by SEPPIC under the name Oramix NS Products sold by SEPPIC under the name Oramix CG Products marketed by Cognis as Plantaren 1200 and Plantacare Lauryl glucoside and coco-glucoside are marketed, for example, by the company Cognis under the name Plantacare Products for sale.

[0334] Suitable maltose derivatives include those described in document EP-A-566 438, such as O-octanoyl-6'-D-maltose or O-dodecanoyl-6'-D-maltose described in document FR-2 739556.

[0335] In some embodiments, the cleaning composition or soap can be formulated in a cosmetically acceptable aqueous medium. In addition to water, a suitable cosmetically acceptable aqueous medium may also include one or more solvents, such as lower alcohols containing 1 to 6 carbon atoms, such as ethanol; polyols, such as glycerol; 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 composition disclosed herein may be 0.1% to 95% by weight.

[0336] In some embodiments, the cleaning compositions or soaps described herein may also include one or more cationic polymers of the polyquaternium type, which can provide softness and lubricity to the foaming composition. Suitable cationic polymers include polyquaternium 5 such as MERQUAT 5 manufactured and marketed by CALGON, polyquaternium 6 such as SALCARE SC 30 manufactured and marketed by CIBA and MERQUAT 100 manufactured and marketed by CALGON, polyquaternium 7 such as MERQUAT S, MERQUAT 2200 and MERQUAT 550 manufactured and marketed by CALGON and SALCARE SC 10 manufactured and marketed by CIBA, polyquaternium 10 such as Polymer JR 400 manufactured and marketed by Amerchol, polyquaternium 11 such as GAFQUAT 755, GAFQUAT 755N and GAFQUAT 734 manufactured and marketed by ISP, polyquaternium 15 such as ROHM's ROHAGIT KF 720F manufactured and marketed by ROHM, polyquaternium 16 such as BASF's LUVIQUAT FC905, LUVIQUAT FC370, LUVIQUAT HM552 and LUVIQUAT FC550, polyquaternium 22 such as Merquat 280 produced and marketed by Calgon, polyquaternium 28 such as STYLEZE CC10 produced and marketed by ISP, polyquaternium 39 such as MERQUAT PLUS 3330 produced and marketed by Calgon, polyquaternium 44 such as LUVIQUAT CARE sold by BASF, polyquaternium 46 such as LUVIQUATHOLD produced and marketed by BASF, polyquaternium 47 such as MERQUAT 2001 produced and marketed by Calgon, and cationic guar gum such as Jaguar produced and marketed by Rhodia can also be used as cationic polymers.

[0337] In some embodiments, the cleaning compositions or soaps described herein may also include one or more adjuvants or additives used in cosmetic compositions. Suitable adjuvants or additives include, but are not limited to, oils, active agents, fragrances, preservatives, chelating agents, pearlescent agents or sunscreens, pigments, pearlescent agents, inorganic fillers or organic fillers such as talc, kaolin, silica powder or polyethylene, soluble dyes, or any combination thereof.

[0338] Examples of oils include vegetable oils (jojoba oil, avocado oil, sesame oil, sunflower oil, corn oil, soybean oil, safflower oil, grape seed oil), mineral oils (petrolatum, optionally hydrogenated isoparaffins), synthetic oils (isopropyl myristate, cetearyl octanoate, polyisobutene, ethylhexyl palmitate, alkyl benzoates), volatile or non-volatile silicone oils such as polydimethylsiloxane (PDMS) and cyclodimethicone or cyclomethicone, and fluorinated or fluorosilicone oils and mixtures thereof.

[0339] Exemplary active agents include sunscreens, desquamation agents, moisturizing agents, depigmenting agents, coloring agents, alpha-hydroxy acids, antimicrobial agents, anti-free radical agents, anti-fouling agents, anti-inflammatory agents, retinoids, extracts of algae, mushrooms, vegetables, yeast, bacteria, hydrolyzed proteins, partially hydrolyzed proteins or non-hydrolyzed proteins, enzymes, hormones, vitamins and their derivatives, flavonoids and isoflavones, and mixtures thereof.

[0340] Depending on the application chosen, the cleaning compositions or soaps described herein may have a pH in the range of 6 to 10. The pH can conventionally be adjusted to the desired value by adding a base (organic or inorganic), for example ammonia or a primary, secondary or tertiary (poly)amine, such as monoethanolamine, diethanolamine, triethanolamine, isopropanolamine or 1,3-propylenediamine, or by adding an inorganic or organic acid, such as a carboxylic acid, for example citric acid. In the case of shower gels, the pH may vary from 8 to 10.

[0341] In some embodiments, the cleaning compositions described herein may also include optional additives such as colorants, fragrances, antimicrobial agents, preservatives, antioxidants, beads (fragrance, exfoliating or moisturizing beads), mica, glitter, shea butter, shea butter beads, opacifiers, pearlescent agents, and other such ingredients. In some embodiments, the composition has high clarity (about 2NTU to about 25NTU), a target viscosity (about 4,000 centipoise to about 10,000 centipoise) in the range of about 1 / 4 to about 1 / 8 for easy dispensing from an orifice, and a yield value (about 3 Pascals to about 15 Pascals) that allows the composition to suspend multiple additives with uniform distribution and enhanced stability (e.g., about 8 months at 120°F (49°C)), and any mixtures thereof.

[0342] In some embodiments, the cleaning composition or soap may also include one or more moisturizers / emollients. Moisturizers may be included in bar or liquid soap compositions to provide conditioning benefits to the skin. The term "moisturizer" describes a material that imparts a smooth and soft feel to the skin surface.

[0343] There are two ways to reduce water loss from the stratum corneum. One is to deposit an occlusive layer on the skin surface that reduces the evaporation rate. The second method is to add a non-occlusive hygroscopic substance to the stratum corneum that will retain water and make that water available to the stratum corneum to change its physical properties and produce a cosmetically desirable effect. Non-occlusive moisturizers also work by improving the lubricity of the skin.

[0344] Both occlusive and non-occlusive moisturizing agents are contemplated for use in the compositions described herein. Exemplary moisturizing agents include long chain fatty acids, liquid water-soluble polyols, glycerin, propylene glycol, sorbitol, polyethylene glycol, ethoxylated / propoxylated ethers of methyl glucose (e.g., methyl gluceth-20), ethoxylated / propoxylated ethers of lanolin alcohol (e.g., ethoxylated / propoxylated ethers available from Amerchol Co. ), coconut and tallow fatty acids, liquid water-soluble polyols (e.g., glycerol, propylene glycol, butylene glycol, hexylene glycol, polypropylene glycol and polyethylene glycol).

[0345] Non-occlusive moisturizing agents may be naturally present in the stratum corneum of the skin, such as sodium pyrrolidone carboxylate, lactic acid, urea, L-proline, guanidine and pyrrolidone. Examples of other non-occlusive moisturizing agents 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 isostearyl-2-lactylate, sodium octyl lactylate), hydrolyzed proteins and other collagen-derived proteins, aloe gel and acetamide MEA (N-acetylethanolamine). Other examples of occlusive and non-occlusive types of moisturizing agents are disclosed in "Emollients--A Critical Evaluation", Cosmetics & Toiletries, J. Mausner, May 1981, incorporated herein by reference.

[0346] Exemplary occlusive moisturizing agents include petrolatum, mineral oil, beeswax, silicones, 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, apricot kernel oil, walnut oil, palm nut oil, pistachio oil, sesame oil, rapeseed oil, juniper oil, corn oil, peach kernel oil, pine oil, castor oil, soybean oil, avocado oil, sunflower oil, coconut oil, hazelnut oil, olive oil, grapeseed oil, and sunflower seed oil.

[0347] Hair Care

[0348] In some embodiments, the composition is a hair care composition comprising ingredients commonly used in hair care products. These ingredients may include, but are not limited to, cleansers, foaming agents, hydrating agents, surfactants, detergents, gelling agents, fragrances, plant extracts, conditioners, humectants, silicones or silicone derivatives, thickening agents, sunscreens, vitamins, alcohols, polyols, polyol ethers, and other common ingredients in shampoos, conditioners, and fixatives. Hair care products typically include one or more surfactants, one or more viscosity modifiers, one or more preservatives, and one or more fragrances, as well as any of the ingredients listed below or a combination thereof.

[0349] Typically, hair can be damaged and brittle by external atmospheric factors such as light and bad weather, and by mechanical or chemical treatments such as brushing, combing, dyeing, bleaching, perming and / or relaxation, and over time, hair can become dry, rough, dim and / or fragile. In order to overcome these shortcomings, common practice is to use a care composition that properly conditions the hair, thereby giving it satisfactory cosmetic properties, especially in terms of smoothness, gloss, softness, flexibility, brightness, natural feel and good detangling properties. These hair care compositions can be, for example, shampoos, conditioning shampoos, hair conditioners, hair masks, essences, gels, shampoos and emulsifiable pastes, which can be rinsed off or left in the composition. In various embodiments, these compositions typically include a combination of cationic conditioning agents such as cationic surfactants, cationic polymers, silicones and / or fatty substances such as fatty alcohols, so as to give hair satisfactory properties in terms of softness, smoothness and flexibility. Exemplary compositions may include silicone, which is known to improve the cosmetic properties of hair in terms of smoothness and softness (as shown in US Pat. No. 5,374,421, each of which is incorporated herein by reference).

[0350] In some embodiments, the hair care composition may further comprise one or more non-amino polyalkylsiloxanes, one or more oxyethylenated polymers in the presence of a fatty alcohol, one or more non-amino polyalkylsiloxanes comprising 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 may have a molecular weight greater than or equal to 10. 6 In some embodiments, the oxyethylenated polymer may be selected from the group consisting of polymers having the formula H(OCH2CH2) zIn some embodiments, the oxyethylenated polymer may be a PEG-45M (z=45,000), such as the product sold under the name Polyox WSR N 60K by the company Amerchol, and a PEG-90M (z=90 000), and mixtures thereof.

[0351] In some embodiments, the hair care composition may further comprise one or more fatty alcohols. The term "fatty alcohol" means any saturated or unsaturated straight or branched alcohol containing at least 8 carbon atoms and not oxyalkylened. 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, arachidonic acid, erucyl alcohol, cetearyl alcohol (or cetearyl alcohol) and mixtures thereof.

[0352] In some embodiments, the hair care composition may also include one or more conditioning agents, including cationic surfactants, cationic polymers and mixtures thereof. The term "cationic surfactant" means a surfactant that is positively charged when included in the composition described herein. Suitable cationic surfactants may be selected from primary fatty amines, secondary fatty amines or tertiary fatty amines, or salts and quaternary ammonium salts of optionally polyoxyalkylened, and mixtures thereof. An exemplary fatty amine is stearamidopropyl dimethylamine. Exemplary quaternary ammonium salts include tetraalkylammonium salts, including dialkyldimethylammonium salts or alkyltrimethylammonium salts in which the alkyl group contains approximately 16 to 22 carbon atoms, especially behenyltrimethylammonium, distearyldimethylammonium, cetyltrimethylammonium or benzyldimethylstearyl ammonium salts, or on the other hand, including palmitamidopropyltrimethylammonium salts, stearamidopropyltrimethylammonium salts, stearamidopropyldimethylcerotearylammonium salts or stearamidopropyldimethyl (myristyl acetic acid) ammonium salts sold by VanDyk under the name CERAPHYL 70.

[0353] The term "cationic polymer" means any polymer containing cationic groups and / or groups ionizable to cationic groups, which polymer may be non-silicon-containing. Exemplary cationic polymers include any polymer known for styling hair, such as those described in patent application EP-A-0 337 354 and French patent applications FR-A-2 270 846, 2 383 660, 2 598 611, 2 470 596 and 2 519 863, each of which is incorporated by reference in its entirety. Further exemplary cationic polymers include those containing units containing primary, secondary, tertiary and / or quaternary amine groups, which units may form part of the polymer backbone or may be carried by side substituents directly attached thereto. Suitable cationic polymers may have a molecular weight of more than 10. 5 and polyquaternary ammonium types, including those described in French Patents 2 505 348 and 2 542 997, each of which is incorporated herein by reference in its entirety.

[0354] In some embodiments, the hair care composition may also include a "non-sulfate" cleanser, foaming agent or surfactant. Suitable "non-sulfate" agents include, but are not limited to, sodium lauroyl methyl isethionate propylene glycol, sodium methyl oleyl taurate and sodium cocoyl isethionate.

[0355] In some embodiments, the hair care composition may also include any of the following ingredients or mixtures thereof: quaternary ammonium compounds synthesized from rapeseed, quaternary ammonium polymers of vinyl pyrrolidone and / or vinyl imidazole, such as the products sold by BASF under the names LUVIQUAT FC 905, FC 550 and FC 370 and LUVIQUAT Excellence; cationic polysaccharides, including cationic cellulose, including cellulose ether derivatives containing quaternary ammonium groups, cationic cellulose copolymers or cellulose derivatives grafted with water-soluble quaternary ammonium monomers, and cationic galactomannan gums. Exemplary cellulose ether derivatives containing quaternary ammonium groups are described in French Patent 1 492 597. These polymers are also defined in the CTFA dictionary as quaternary ammoniums of hydroxyethyl cellulose that have been reacted with an epoxide substituted with a trimethylammonium group. Cationic cellulose copolymers or cellulose derivatives grafted with water-soluble quaternary ammonium monomers are described in U.S. Pat. No. 4,131,576, such as hydroxyalkyl celluloses such as hydroxymethyl cellulose, hydroxyethyl cellulose or hydroxypropyl cellulose grafted with methacryloylethyltrimethylammonium, methacrylamidopropyltrimethylammonium or dimethyldiallyl ammonium salts. Suitable related celluloses such as C8-C 30Alkyl hydroxyethylcelluloses with fatty chains quaternized, such as the product QUATRISOFT LM 200 (INCI name Polyquaternium-24) sold by the company Amerchol / Dow Chemical and the products CRODACEL QM (INCI name PG-Hydroxyethylcellulose Cocodimonium Chloride), CRODACEL QL (C 12 Alkyl) (INCI name PG-hydroxyethylcellulose lauryl dimonium chloride) and CRODACEL QS (C 18 Alkyl) (INCI name PG-hydroxyethylcellulose stearyl dimonium chloride). Other suitable fatty chain hydroxyethylcellulose derivatives include the commercial products SOFTCAT Polymer SL from Amerchol / Dowchemical, such as SL-100, SL-60, SL-30 and SL-5 with the INCI name Polyquaternium-67. Suitable cationic galactomannan gums are described in U.S. Pat. Nos. 3,589,578 and 4 031 307. Suitable cellulose components may be in crystalline form, microcrystalline form or mixtures thereof.

[0356] In some embodiments, the hair care composition may also contain one or more cationic proteins or cationic protein hydrolysates, polyalkyleneimines (including polyethyleneimines), polymers containing vinylpyridine or vinylpyridinium units, polyamine and epichlorohydrin condensates, quaternary polyureas and chitin derivatives, animal protein hydrolysates carrying trimethylbenzylammonium groups (such as the product sold under the name Crotein BTA by the company Croda and known as benzyltrimethylammonium hydrolyzed animal protein in the CTFA dictionary), protein hydrolysates carrying quaternary ammonium groups on the polypeptide chain, the ammonium group containing at least one alkyl group having 1 to 18 carbon atoms.

[0357] In some embodiments, the hair care composition may also contain one or more quaternized vegetable proteins, such as quaternized vegetable proteins from wheat protein, corn protein or soy protein, for example quaternized wheat proteins, including those sold by the company Croda under the name Hydrotriticum WQ or QM, known in the CTFA dictionary as cocodimonium hydrolyzed wheat protein, sold under the name Hydrotriticum QL, known in the CTFA dictionary as lauryldimonium hydrolyzed wheat protein, or others sold under the name Hydrotriticum QS, known in the CTFA dictionary as stearyldimonium hydrolyzed wheat protein.

[0358] In some embodiments, the hair care composition may also contain one or more polyamines, such as POLYQUART RH sold by Cognis, referred to in the CTFA dictionary under the name polyethylene glycol tallow polyamine. Other suitable polymers include those sold under the name Lupamin by the company BASF, and the products sold under the names Lupamin 9095, Lupamin 5095, Lupamin 1095, Lupamin 9030 and Lupamin 9010.

[0359] In some embodiments, the hair care composition may further comprise one or more 5 The term "fatty substance" means a fatty substance that is liquid at normal temperature (25°C) and atmospheric pressure (1.013*10 5 Pa) insoluble in water (solubility less than 5%, less than 1% or less than 0.1%). Fatty substances are usually soluble in organic solvents under the same temperature and pressure conditions, for example, chloroform, dichloromethane, carbon tetrachloride, ethanol, benzene, toluene, tetrahydrofuran (THF), liquid petrolatum or decamethylcyclopentasiloxane. The liquid fatty substances of the present disclosure may be non-polyoxyethylated and non-polyglycerolated. The term "oil" means an organic compound that is soluble in water at room temperature (25°C) and atmospheric pressure (1.013*10 5 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. The liquid fatty substance may be selected from non-silicone oils, such as, in particular, C6-C 16 Liquid hydrocarbons, liquid hydrocarbons containing more than 16 carbon atoms, non-silicone oils of animal origin, triglycerides of plant origin or synthetic origin, fluorinated oils, liquid fatty acids and / or fatty alcohol esters other than triglycerides, and mixtures thereof. Liquid hydrocarbons may be linear, branched or optionally cyclic, including hexane, cyclohexane, undecane, dodecane, tridecane or isoparaffins, such as isohexadecane, isodecane or isododecane, and mixtures thereof. Suitable linear or branched 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 polyisobutylene (such as PARLEAM) and mixtures thereof. Hydrocarbon-based oils of animal origin may be used, such as fully hydrogenated squalene.

[0360] Exemplary triglycerides of vegetable or synthetic origin may be chosen from liquid fatty acid triglycerides containing from 6 to 30 carbon atoms, for example heptanoic or caprylic triglycerides, or alternatively, more particularly from those present in vegetable oils, for example coconut oil, sunflower oil, corn oil, soybean oil, pith oil, grapeseed oil, sesame oil, hazelnut oil, almond oil, macadamia oil, arara oil, castor oil, avocado oil, jojoba oil, shea butter, or synthetic caprylic / capric triglycerides, such as those sold by the company Stearineries Dubois or those sold under the name MIGLYOL 810, 812 and 818 by the company Dynamit Nobel, and mixtures thereof. Suitable fluoro oils include perfluoromethylcyclopentane and perfluoro-1,3-dimethylcyclohexane, such as FLUTESC PC1 and FLUTESC PC3 from BNFL Fluorochemicals; perfluoro-1,2-dimethylcyclobutane; perfluoroalkanes, such as dodecafluoropentane and tetradecafluorohexane sold under the names PF 5050 and PF5060 by 3M, or bromoperfluorooctane sold under the name FORALKYL by Atochem; nonafluoromethoxybutane and nonafluoroethoxyisobutane; perfluoromorpholine derivatives, such as 4-trifluoromethylperfluoromorpholine sold under the name PF 5052 by 3M.

[0361] Suitable monoesters include dihydroabietyl behenate; octyldodecanol behenate; isocetyl behenate; cetyl lactate; C 12 -C 15 Alkyl 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 nonanoate; Octyl stearate; octyldodecyl erucate; oleyl erucate; ethyl palmitate and isopropyl palmitate; 2-ethylhexyl palmitate, 2-octyldecyl palmitate, alkyl myristates (such as isopropyl myristate, butyl myristate, cetyl myristate, 2-octyldodecyl myristate, myristyl myristate, myristyl stearate), hexyl stearate, butyl stearate, isobutyl stearate; dioctyl malate, hexyl laurate, 2-hexyldecyl laurate and mixtures thereof.

[0362] In some embodiments, the hair care composition may further include diethyl sebacate, diisopropyl sebacate, diisopropyl adipate, di(n-propyl adipate), dioctyl adipate, diisostearyl adipate, dioctyl maleate, glyceryl undecylenate, octyldodecyl stearoyl stearate, pentaerythritol monoricinoleate, pentaerythritol tetraisononanoate, pentaerythritol tetranonanoate, pentaerythritol tetraisostearate, pentaerythritol tetraoctanoate, propylene glycol dicaprylate, propylene glycol dicaprate, tridecyl erucate, triisopropyl citrate, triisostearyl citrate, glyceryl trilactate, glyceryl tricaprylate, trioctyldodecyl citrate, trioleyl citrate, propylene glycol dicaprylate, neopentyl glycol diheptanoate, diethylene glycol diisononanoate, and polyethylene glycol distearate, and mixtures thereof.

[0363] In some embodiments, the hair care composition may further comprise one or more fatty acid esters, one or more sugar esters and / or one or more C6-C 30 (such as C 12 -C 22) fatty acid diester.Term " sugar " means to contain several alcohol functional groups, have or do not have aldehyde or ketone functional group and comprise the compound based on oxygen-containing hydrocarbon of at least 4 carbon atoms.Suitable sugar can comprise monosaccharide, oligosaccharide or polysaccharide, for example sucrose (or sucrose), glucose, galactose, ribose, fucose, maltose, fructose, mannose, arabinose, xylose and lactose, and their derivative, such as alkyl derivative, such as methyl derivative, for example methyl glucose.Suitable ester can comprise oleate, laurate, palmitate, myristate, behenate, cocoate, stearate, linoleate, linolenate, caprate, arachidonic acid ester or their mixture, such as oleate / palmitate, oleate / stearate or palmitate / stearate mixed ester. Suitable mono- and diesters also include mono- or di-oleate, mono- or di-stearate, mono- or di-behenate, mono- or di-olealmitate, mono- or di-linoleate, mono- or di-linolenate or mono- or di-oleostearate of sucrose, glucose or methylglucose, including the product sold under the name GLUCATE DO by the company Amerchol, which is methylglucose dioleate. Other exemplary esters or ester mixtures of sugars and fatty acids that may also be mentioned include: the products sold under the names F160, F140, F110, F90, F70 and SL40 by the company Crodesta, representing sucrose palmitate / stearate formed respectively from 73% of monoesters and 27% of diesters and triesters, from 61% of monoesters and 39% of diesters, triesters and tetraesters, from 52% of monoesters and 48% of diesters, triesters and tetraesters, from 45% of monoesters and 55% of diesters, triesters and tetraesters, from 39% of monoesters and 61% of diesters, triesters and tetraesters and sucrose monolaurate; the products sold under the name Ryoto Sugar Esters, for example with the reference B370 and corresponding to a sucrose behenate formed from 20% of monoesters and 80% of diester-triester-polyesters; sucrose mono-dipalmitate / stearate sold under the name TEGOSOFT PSE by the company Goldschmidt.

[0364] In some embodiments, the hair care composition may further comprise a pH adjuster such as citric acid and / or sodium hydroxide.Any conventional pH adjuster for hair care compositions is contemplated for use herein.

[0365] In various embodiments, the hair care composition may further comprise any of the following ingredients and / or mixtures thereof: sodium lauroyl methyl isethionate (cleansing agent and foaming agent), liquid fatty materials (including silicone oils other than the non-amino polyalkylsiloxanes discussed herein above), and organomodified polysiloxanes comprising at least one functional group selected from amino, aryl and alkoxy. Organopolysiloxanes are defined in more detail in Chemistry and Technology of Silicones (1968), Academic Press, by Walter Noll, the entire contents of which are hereby incorporated by reference. They may be volatile or nonvolatile. Suitable cyclic polydialkylsiloxanes include octamethylcyclotetrasiloxane sold under the name VOLATILE SILICONE 7207 by Union Carbide or under the name SILBIONE 70045V2 by Rhodia, decamethylcyclopentasiloxane sold under the name VOLATILE SILICONE 7158 by Union Carbide and under the name SILBIONE 70045V5 by Rhodia, and mixtures thereof. Cyclic copolymers of the dimethylsiloxane / methylalkylsiloxane type, such as VOLATILE SILICONE FZ 3109 sold by the company Union Carbide, are also suitable. Exemplary cyclic polydialkylsiloxanes with organosilicon compounds include octamethylcyclotetrasiloxane and tetrakis(trimethylsilyl)pentaerythritol (50 / 50) and mixtures of octamethylcyclotetrasiloxane with oxy-1,1'-bis(2,2,2',2',3,3'-hexa-trimethylsiloxy)neopentane, linear volatile polydialkylsiloxanes such as those sold by Toray Silicone under the name SH 200. Silicones belonging to this class are also described in Cosmetics and Toiletries, Vol. 91, No. 76, January, pp. 27-32, Todd & Byers, "Volatile Silicone Fluids for Cosmetics", which is incorporated herein by reference.

[0366] Exemplary suitable non-volatile polydialkylsiloxanes include polydimethylsiloxanes having trimethylsilyl end groups, such as SILBIONE oils of the 47 and 70 047 series or MIRASIL oils sold by Rhodia, for example 70 047V500 000 oil; oils of the MIRASIL series sold by Rhodia; oils of the 200 series from Dow Corning, such as 60 000mm viscosity SILBIONE oils; 2 / s DC200; VISCASIL oils from General Electric and certain oils from the SF series from General Electric (SF 96, SF 18), polydimethylsiloxanes with dimethylsilanol end groups known by the name dimethiconol (CTFA), such as the oils of series 48 from the company Rhodia.

[0367] Exemplary organomodified silicones include polyalkylaryl siloxanes, and products sold under the following names: SILBIONE oils from the 70 641 series from Rhodia; oils from the RHODORSIL 70 633 and 763 series from Rhodia; oils from Dow Corning 556 Cosmetic Grade Fluid from Dow Corning; silicones from the PK series from Bayer, such as the product PK20; silicones from the PN series and PH series from Bayer, such as the products PN1000 and PH1000; certain oils from the SF series from 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 by Genesee or products sold under the names Q2 8220 and Dow Corning 929 or 939 by the company Dow Corning.

[0368] In various embodiments, the hair care composition may further comprise one or more additional surfactants other than 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, monoglycerol 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, acyl glutamates, alkyl sulfosuccinamates, acyl isethionates 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-salt-forming forms of all these compounds; the alkyl and acyl groups of all these compounds (unless otherwise mentioned) typically contain 6 to 24 carbon atoms and the aryl group typically represents a phenyl group. Anionic surfactants in salt form may 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.

[0369] Suitable anionic surfactants also include mild anionic surfactants, i.e. anionic surfactants without sulfate functionality, including polyoxyalkylene alkyl ether carboxylic acids; polyoxyalkylene alkyl aryl ether carboxylic acids; polyoxyalkylene alkyl amide ether carboxylic acids, especially those containing 2 to 50 ethylene oxide groups; alkyl-D-galactosiduronic acids; acyl sarcosinates, acyl glutamates; and alkyl polyglycoside carboxylates, such as those sold under the name AKYPO RLM45CA from Kao.

[0370] Exemplary suitable amphoteric or zwitterionic surfactants can be optionally quaternized aliphatic secondary or tertiary amine derivatives, wherein the aliphatic radical is a straight or branched chain containing 8 to 22 carbon atoms, wherein the amine derivative contains at least one anionic group, for example a carboxylate, sulfonate, sulfate, phosphate or phosphonate group, such as (C8-C 20 ) alkyl betaine, sulfo betaine, (C8-C 20 )alkylamide (C3-C8)alkyl betaine or (C8-C 20) alkylamide (C6-C8) alkyl sulfobetaine. Any suitable aliphatic secondary or tertiary amine derivative may be present in coconut oil or hydrolyzed linseed oil, etc. Representative compounds are classified in the CTFA dictionary, 1993, 5th edition, under the following names: disodium cocoamphodiacetate, disodium lauroamphodiacetate, disodium caproamphodiacetate, disodium capryloamphodiacetate, disodium cocoamphodipropionate, disodium lauroamphodipropionate, disodium caproamphodipropionate, disodium capryloamphodipropionate, lauroamphodipropionic acid, and cocoamphodipropionic acid.

[0371] By way of example, cocoamphodiacetate sold under the name SODIUM C2M Concentrate by the company Rhodia and diethylaminopropyl cocoyl asparagine sold under the name Chimexane HB by the company Chimex are suitable for use in the disclosed compositions.

[0372] Suitable nonionic surfactants are described in MR Porter's Handbook of Surfactants (Glasgow and London), 1991, pp. 116-178, and include fatty alcohols, fatty α-diols, fatty (C1-C 20 ) alkylphenols and fatty acids, these may be ethoxylated, propoxylated or glycerolated and contain at least one fatty chain containing 8 to 18 carbon atoms, the number of ethylene oxide or propylene oxide groups ranging from 1 to 200 and the number of glycerol groups ranging from 1 to 30. Condensates of ethylene oxide and propylene oxide with fatty alcohols, ethoxylated fatty amides having 1 to 30 ethylene oxide units, polyglycerolated fatty amides containing on average 1 to 5 glycerol groups and especially 1.5 to 4 glycerol groups, ethoxylated fatty acid esters of sorbitan containing 1 to 30 ethylene oxide units, fatty acid esters of sucrose, fatty acid esters of polyethylene glycols, (C6-C 24 ) alkyl polyglycosides, ethylene oxide vegetable oil, N-(C6-C 24 ) alkyl glucosamine derivatives, amine oxides such as (C 10 -C 14 ) alkylamine oxide or N-(C 10 -C 14 )Acylaminopropylmorpholine oxide is also suitable for use in the compositions disclosed herein.

[0373] Other suitable nonionic surfactants include alkyl polyglucosides (APGs), maltose esters, polyglycerolated fatty alcohols, glucosamine derivatives (e.g., 2-ethylhexyloxycarbonyl-N-methylglucosamine), and mixtures thereof. Alkyl polyglucosides are those containing an alkyl group containing 6 to 30 carbon atoms and containing a hydrophilic group (glucoside). Exemplary alkyl polyglucosides include decyl glucoside (alkyl-C9 / C11-polyglucoside (1.4)), including those sold by Kao Chemicals under the name Mydol The product is marketed by Cognis under the name Plantaren 2000 Products sold and marketed by SEPPIC under the name Oramix NS Products sold by SEPPIC under the name Oramix CG Products marketed by Cognis as Plantaren 1200 and Plantacare Lauryl glucoside and coco-glucoside are marketed, for example, by the company Cognis under the name Plantacare Products for sale.

[0374] Suitable maltose derivatives include those described in document EP-A-566 438, such as O-octanoyl-6'-D-maltose or O-dodecanoyl-6'-D-maltose described in document FR-2 739 556. Each of these documents is incorporated by reference in its entirety.

[0375] In some embodiments, the collagen in the composition can be chemically modified with a hair colorant to provide a permanent, semi-permanent or temporary hair dye solution. The hair colorant as defined herein can be any dye, pigment, nanoparticle, etc. that can be used to change the color of hair. Suitable hair dyes 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-p-phenylenediamine, N,N-hydroxyethyl-2-nitro-phenylenediamine, 4-nitro-indole, 2-nitro-5-glyceromethylaniline, 3-methylamino-4-nitrophenoxyethanol, p-phenylenediamine, hydrogen peroxide, ammonia, DMDM ​​hydantoin, methylparaben, propylparaben, lead acetate, and resorcinol.

[0376] 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 by water or by a mixture of water and one or more cosmetically acceptable solvents selected from lower alcohols such as ethanol and isopropanol; polyols and polyol ethers, including 2-butoxyethanol, propylene glycol, propylene glycol monomethyl ether, diethylene glycol monoethyl ether and monomethyl ether, and mixtures thereof.

[0377] In some embodiments, the hair care composition may also contain any of the following additives or mixtures thereof: solid fatty substances other than fatty alcohols such as waxes, anionic polymers, nonionic polymers or amphoteric polymers or mixtures thereof, antidandruff agents, antiseborrheic agents, agents for preventing hair loss and / or for promoting hair regrowth, vitamins and provitamins (including panthenol), sunscreens, inorganic or organic pigments, chelating agents, plasticizers, solubilizers, acidulants, inorganic or organic thickeners (particularly polymer thickeners other than oxyethylated polymers), opacifiers or pearlescent agents, antioxidants, hydroxy acids, fragrances and / or preservatives.

[0378] Nail Care

[0379] In some embodiments, the cosmetic composition may include ingredients commonly used in nail care products. Nail care products include, but are not limited to, nail treatments (including nail strengtheners, topcoats, and basecoats), nail polishes, nail polish removers, hand skin care products, foot skin care products, drying agents, and correction pens (including nail polish removers).

[0380] 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 topical treatment of nail infections (including fungal infections), a nail strengthener, a topcoat, a basecoat, a polish remover, or any combination thereof. In some embodiments, the nail treatment composition can be formulated as a topical nail lacquer or nail polish, cream, solution, suspension, lotion, essence, gel, balm, gel, oil, cream-in-oil, and / or liniment for treating hands and / or feet.

[0381] In some embodiments, the nail treatment composition can be a nail strengthener. The nail strengthener can treat fingernails and toenails to harden, strengthen and promote the growth of nails to prevent or minimize breakage, cracking, splitting and peeling, and can include any known composition for preventing and healing sidewall hoof cracks (quarter cracks) while increasing the growth of horse hooves, and the composition has a protein consistency similar to human fingernails and toenails, including: lanolin, tallow, beeswax, rosin, copper acetate and turpentine. In some embodiments, the nail treatment composition may also include titanium dioxide, such as TI-PURE R900 from EIDuPont.

[0382] In some embodiments, the nail treatment composition can be a primer. In some embodiments, the primer can be a liquid composition comprising at least one polymer that provides adhesion, such as a polymer copolymerized by 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, suitable polymers for use as primers include hydroxyethyl methacrylate (HEMA), hydroxypropyl methacrylate (HPMA), ethyl methacrylate (EMA), tetrahydrofurfuryl methacrylate (THFMA), pyromellitic anhydride di(meth)acrylate, pyromellitic anhydride glycerol dimethacrylate, pyromellitic acid dimethacrylate, methacryloyloxyethyl maleate, 2-hydroxyethyl methacrylate / succinate, 1,3-glycerol dimethacrylate / succinate adduct, phthalic acid monoethyl methacrylate, and mixtures thereof. In some aspects, the basecoat layer may also include 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 ingredients are not limiting.

[0383] In some embodiments, the nail treatment composition can be a topcoat, such as a quick-drying topcoat. The topcoat can include a base resin that is cellulose acetate butyrate, a film former that is a methacrylate polymer, a heat-curable or photocurable monomer that is a monofunctional methacrylate and a crosslinker (i.e., a difunctional methacrylate and a trifunctional methacrylate), a thermal initiator or a photoinitiator together in a solution of an aliphatic ester and an alcohol, and a photoreactive coating. Suitable solvents include acetates and alcohols, particularly ethyl acetate, butyl acetate, and isopropanol. The photoreactive coating may include photoreactive monomers, including methacrylate 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, crosslinkers 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 is an oligomeric mixture of phenylacetones, such as a mixture of 2,4,6-trimethylbenzophenone and 4-methylbenzophenone and a mixture of oligomeric-[2-hydroxy-2-methyl-1[4-(1-methylvinyl)phenyl]propanone] and 2-hydroxy-2-methyl-phenylpropanone, which is sold by Sartomer under the names "Esacure KIP 100F" and "Esacure TZT photoinitiator".

[0384] In some embodiments, the nail care composition can be a nail polish or a nail polish composition. In some embodiments, the nail polish or nail polish composition may also include a nitrocellulose film former, a latex film former, a polycarbodiimide film former, a low volatile organic compound (VOC) and a polycarbodiimide film former. Polycarbodiimide may 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 α-methylstyrene-isocyanate. Suitable polycarbodiimide compounds include, but are not limited to, those commercially available from suppliers Nisshinbo (including those known by the name CARBODILITE series V-02, V02-L2, SV-02, E-02, V-10, SW-12G, E-03A), Picassian and 3M.

[0385] In some embodiments, the nail polish or nail polish composition may also 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 may be film-forming latex polymers or non-film-forming latex polymers. In some embodiments, the latex polymer may be a carboxyl functional acrylate latex polymer, such as those produced by homopolymerization or copolymerization of ethylenically unsaturated monomers selected from the following: vinyl monomers, (meth) acrylic acid monomers, (meth) acrylamide monomers, unsaturated monocarboxylic acids and dicarboxylic acids, esters of (meth) acrylic acid monomers and amides of (meth) acrylic acid monomers. As used herein, the term "(meth) acryloyl" and variations thereof mean acryloyl or methacryloyl. (Meth) acrylic acid monomers may be selected from, for example, acrylic acid, methacrylic acid, citraconic acid, itaconic acid, maleic acid, fumaric acid, crotonic acid and maleic anhydride. As a non-limiting example, the ester of the (meth)acrylic acid monomer can be a C1-C8 alkyl (meth)acrylate, 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. Amides of (meth)acrylic monomers can be made, for example, from (meth)acrylamides, especially N-alkyl(meth)acrylamides, especially N—(C1-C12)alkyl(meth)acrylates, such as N-ethyl(meth)acrylamide, N-tert-butyl(meth)acrylamide, N-tert-octyl(meth)acrylamide, N-methylol(meth)acrylamide and N-diacetone(meth)acrylamide, and combinations thereof.

[0386] Vinyl monomers may include, but are not limited to, 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-tert-butyl benzoate, triallyl cyanurate; vinyl halides such as vinyl chloride and vinylidene chloride; aromatic monovinyl or divinyl compounds such as styrene, α-methylstyrene, chlorostyrene, alkylstyrene, divinylbenzene and diallyl phthalate, as well as p-styrenesulfonic acid, vinylsulfonic acid, 2-(meth)acryloyloxyethylsulfonic acid, 2-(meth)acrylamide-2-methylpropylsulfonic acid and mixtures thereof.

[0387] The list of monomers herein is not limiting, and it is understood that any monomer known to those skilled in the art may be used, including acrylic monomers and / or vinyl monomers (including monomers modified with silicone chains).

[0388] In some non-limiting exemplary embodiments, the carboxyl functional acrylate latex polymer can be selected from the following aqueous dispersions: methacrylic acid / ethyl acrylate copolymer (INCI: Acrylates Copolymer, such as LUVIFLEX SOFT from BASF), PEG / PPG-23 / 6 dimethicone citraconate / C10-30 alkyl PEG-25 methacrylate / acrylic acid / methacrylic acid / ethyl acrylate / trimethylolpropane PEG-15 triacrylate copolymer (INCI: Polyacrylate-2 Crosspolymer, such as Fixate Superhold.TM. from Lubrizol), styrene / acrylic acid copolymer (such as Acudyne Shine from Dow Chemical), ethylhexyl acrylate / methyl methacrylate / butyl acrylate / acrylic acid / methacrylic acid copolymer (INCI: Acrylates / Ethylhexyl Acrylate Copolymer, for example Daitosol 5000SJ, DaitoKasei Kogyo), acrylic acid / acrylate copolymer (INCI name: Acrylates Copolymer, such as Daitosol 5000AD, Daito Kasei Kogyo), acrylate copolymers (such as those known under the trade name Dermacryl AQF (AkzoNobel), under the trade name LUVIMER MAE (BASF) or under the trade name BALANCE CR (AKZO NOBEL)), acrylate / hydroxy ester acrylate copolymers (known under the trade name ACUDYNE 180 POLYMER (Dow Chemical)), styrene / acrylate copolymers (from Dow Chemical, known under the trade name Acudyne Bold), styrene / acrylate / ammonium methacrylate copolymers (from Interpolymer, known under the trade name SYNTRAN PC5620 CG) and mixtures thereof.

[0389] In some embodiments, the nail care composition may include a nail polish remover. The nail polish remover may include a polyol compound, including glycerol, glycol, polyglycerol, esters of polyols and mixtures thereof. The glycol may contain 2 to 12 carbon atoms, such as glycerol, propylene glycol, butylene glycol, propylene glycol, hexylene glycol, polyglycerol, dipropylene glycol and diethylene glycol. The ester of suitable polyols includes liquid esters of saturated or unsaturated straight or branched C1-C26 polyols. The example of the ester of suitable polyols includes, but is not limited to, esters of dihydroxy alcohols, trihydroxy alcohols, tetrahydroxy alcohols or pentahydroxy alcohols. The ester of polyols may be a glyceride, such as triglycol, tricitrin, trilactyl, trilactyl, tributyrin, triheptanoin, tricaprylin etc.

[0390] The nail polish remover may also contain a lower alcohol, containing 1 to 8 carbon atoms. The lower alcohol may contain 2 to 6 carbon atoms, such as 2 to 5 carbon atoms. Examples of lower alcohols include, but are not limited to, ethanol, propanol, butanol, amyl alcohol, isopropanol, isobutanol, and isopentanol. The nail polish remover may also contain a high boiling ester compound, including, but not limited to, carbonates, adipates, sebacates, and succinates. Exemplary high boiling point ester compounds 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.

[0391] The nail polish remover may also include a thickening agent, including but not limited to nonionic polymers, anionic polymers, cationic polymers, amphiphilic polymers and amphoteric polymers, and 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 hydroxyl-modified cellulose polymers, such as hydroxyethyl cellulose, for example, those with a molecular weight of more than 500,000 Daltons, such as NATROSOL 250HHR and hydroxypropyl cellulose, such as KLUCEL MF - both available from Ashland, Covington, Ky. The thickening agent may be a polysaccharide such as fructan, dextran, galactan and mannan, or a heteropolysaccharide such as hemicellulose, pullulan or branched polysaccharides such as gum arabic and amylopectin, or a mixed polysaccharide such as starch. The thickening agent may be an acrylic thickening agent (acrylic thickener) or an acrylamide thickening agent (acrylic thickener). Thickening agent can include at least one monomer that performs weak acid function, such as acrylic acid, methacrylic acid, itaconic acid, crotonic acid, maleic acid and / or fumaric acid.Thickening agent can include monomer that performs strong acid function, such as monomer with sulfonic acid type or phosphonic acid type function, such as 2-acrylamide-2-methylpropanesulfonic acid (AMPS).Thickening agent can include crosslinking agent such as methylene bis acrylamide (MBA), ethylene glycol diacrylate, polyethylene glycol dimethacrylate, diacrylamide, cyano methacrylate, vinyloxyethyl acrylate or methacrylate, formaldehyde, glyoxal and glycidyl ether type composition such as ethylene glycol diglycidyl ether or epoxide.Suitable acrylic thickener is disclosed in U.S. Patent Application Publication No. 2004 / 0028637 and No. 2008 / 0196174, both of which are incorporated herein by reference.In some embodiments, thickening agent can include organoclay (hydrophobic treated clay) or hydrophilic clay.

[0392] In some embodiments, thickening agent may include abrasive compound (abrasive system). "Abrasive compound" is a compound that can provide grinding or mechanical stripping. Abrasive particles may include perlite, pumice, zeolite, hydrated silica, calcium carbonate, dicalcium phosphate dihydrate, calcium pyrophosphate, aluminum oxide, sodium bicarbonate, polylactic acid and synthetic polymer materials such as polyethylene, polypropylene, polyethylene terephthalate, polymethyl methacrylate or nylon. In certain embodiments, the abrasive of medium hardness includes perlite, such as the beauty grade perlite that can be obtained from Imerys under the name IMERCARE 270P-Scrub. In certain embodiments, soft abrasive is sugar, ground fruit kernel or husk powder, such as almond, coconut shell or spherical wax (e.g., carnauba jojoba oil (carnauba jojoba)); Argan nut shell powder etc.

[0393] In some embodiments, the nail care composition may also contain additives commonly used in cosmetic compositions and known to technicians, including solvents, preservatives, fragrances, oils, waxes, surfactants, antioxidants, anti-free radical agents, wetting agents, dispersants, defoaming agents, neutralizers, stabilizers, active ingredients selected from essential oils, UV screening agents, sunscreens, moisturizers, vitamins, proteins, ceramides, plant extracts, fibers, etc., and mixtures thereof.

[0394] Therapeutic Compositions

[0395] In some embodiments, the composition described herein can be a therapeutic composition for treating one or more conditions. In some embodiments, the composition described herein can be formulated as a dry film, powder, gel, or water or buffer solution. For example, in some embodiments, the composition described herein can promote wound healing, reduce or prevent the formation of scar tissue, 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 present a large uniform surface area and can be used as a delivery system for collagen and optionally one or more additional therapeutic agents. Injectable collagen materials can also form a scaffold or network that can replace tissue function and support tissue regeneration. In certain embodiments, the composition can be applied topically. In certain embodiments, the composition can be injected percutaneously, intradermally or subcutaneously.

[0396] In some embodiments, the therapeutic compositions described herein may include one or more additional therapeutic and / or prophylactic agents in addition to the collagen fragments described elsewhere herein. The one or more additional therapeutic and / or prophylactic agents may be small molecule active agents or biomolecules, such as enzymes or proteins, polypeptides or nucleic acids.

[0397] Non-limiting examples of additional therapeutic and / or preventive agents include anticancer agents, antimicrobial agents (including antiviral agents, antibacterial agents, antifungal agents and antiparasitic agents), antioxidants, local analgesics, local anesthetics, anti-inflammatory agents, cytokines, immunosuppressants, antiallergic agents, essential nutrients, growth factors (such as fibroblast growth factor, hepatocyte growth factor, platelet-derived growth factor, vascular endothelial cell growth factor and insulin-like growth factor) and combinations thereof. The specific dosage of the additional therapeutic and / or preventive agent can be easily determined by those skilled in the art. See Ansel, Howard C. et al., Pharmaceutical Dosage Forms and Drug Delivery Systems (6th Edition) Williams and Wilkins, Malvern, PA (1995).

[0398] In other embodiments, the compositions described herein can be used in combination with cell delivery, for example, delivery of stem cells, pluripotent cells, somatic cells, and combinations thereof.

[0399] Although the present disclosure contemplates that therapeutic and / or prophylactic agents will be an integral part of the compositions described herein, in some embodiments, the therapeutic and / or prophylactic agents may be administered prior to, concurrently with, or after the administration of the therapeutic compositions described herein. In other words, in some embodiments, the one or more additional therapeutic and / or prophylactic agents may not be integrally incorporated into the compositions of the present invention, but rather provided separately from the compositions of the present invention.

[0400] Non-limiting examples of suitable local anesthetics that can be included in the compositions of the present invention include, but are not limited to, ambucaine, amolanone, amylocaine, benoxinate, benzocaine, betoxycaine, biphenamine, bupivacaine, butacaine, butamben, butanilicaine, butethamine, butoxycaine, carticaine, chloroprocaine, ocaine), cocaethylene, cocaine, cyclomethycaine, dibucaine, dimethysoquin, dimethocaine, diperodon, dycyclonine, ecgonidine, ecgonine, ethyl chloride, etidocaine, eucaine, euprocin, fenalcomine, formcaine, hexylcaine, hydroxybutyric acid,tetracaine), isobutyl para-aminobenzoate, leucaine mesylate, levoxadrol, lidocaine, mepivacaine, meprylcaine, metabutoxycaine, methyl chloride, myrtecaine, naepaine, octacaine, orthocaine, oxethazaine, parethoxycaine, phenacaine, phenol, piperocaine, piridocaine The invention also includes the following: ocaine, polidocanol, pramoxine, prilocaine, procaine, propanocaine, proparacaine, propipocaine, propoxycaine), psuedococaine, pyrrocaine, ropivacaine, salicyl alcohol, tetracaine, tolycaine, trimecaine, zolamine, and combinations thereof.

[0401] 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; monoamines; nitrofurans; penicillins; polypeptides such as bacitracin, colistin, and polymyxin B; quinolones; sulfonamides; and tetracyclines. Additional exemplary antimicrobial agents include iodine, silver compounds, moxifloxacin, ciprofloxacin, levofloxacin, cefazolin, tigecycline, gentamicin, ceftazidime, ofloxacin, gatifloxacin, amphotericin, voriconazole, natamycin.

[0402] Non-limiting examples of suitable anti-inflammatory agents include steroidal active agents, including glucocorticoids, progestins, mineralocorticoids and corticosteroids. Exemplary nonsteroidal anti-inflammatory drugs include ketorolac, ibuprofen, nepafenac, diclofenac, aspirin and naproxen. Other exemplary anti-inflammatory agents include triamcinolone acetonide, fluocinolone acetonide, prednisolone, dexamethasone, loteprendol, fluorometholone and dipotassium glycyrrhizate.

[0403] In some embodiments, the composition may also include one or more additional pharmaceutically active agents. Exemplary medicaments may include nonsteroidal anti-inflammatory agents (NSAIDs), such as flurbiprofen, ibuprofen, naproxen, indomethacin and related compounds. In some embodiments, the composition may also include one or more anti-mitotic drugs, including colchicine, taxol and related compounds. In some embodiments, the composition may also include one or more local disinfectants, such as benzoyl peroxide. In some embodiments, the composition may also include one or more polysaccharides produced by microalgae, such as microalginic acid.

[0404] In some embodiments, the composition may further comprise one or more immunomodulatory drugs. Exemplary immunomodulatory drugs include imiquimod, cyclosporine, tacrolimus, and rapamycin.

[0405] In some embodiments, the composition may also 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, cytokines can induce Treg activation (e.g., IL-25) and inhibit Th17 activation (e.g., IL-10) to minimize rejection.

[0406] In some embodiments, the compositions described herein also include 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 cell can be adipose-derived mesenchymal stem cells. Functional characteristics of mesenchymal stem cells that may be beneficial to wound healing include their 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 through growth factor secretion and matrix remodeling, and exert unique immunomodulatory and anti-inflammatory effects (see, e.g., 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.

[0407] In some embodiments, eukaryotic cells may be responsible for increasing the structural integrity of connective tissue and / or promoting healing. In some embodiments, eukaryotic cells (such as fibroblasts) may be responsible for enhancing or promoting the growth or connection of cells or tissues.

[0408] In some embodiments, the recombinant collagen composition may be in contact with 1) an implanted hair graft and 2) tissue in which the hair graft is implanted. Recipient sites for hair transplantation include the scalp, facial area, axilla or chest area, or pubic area. Specific areas of the facial area include eyebrows, eyelids, beard, sideburns, chin, and cheeks. The recipient site can be any skin area where the subject needs hair to appear. In some embodiments, the contact between the composition described herein and the implanted hair and the surrounding tissue of the recipient site promotes nutrient perfusion from the surrounding tissue into the graft and increases the survival rate of the graft compared to a graft implanted without the composition described herein. In certain embodiments, the contact between the composition described herein and the implanted hair and the contact between the composition described herein and the surrounding tissue of the recipient site promotes vascularization around the implanted graft, so that the survival rate of the graft is increased compared to a graft implanted without the composition described herein.

[0409] Dietary composition

[0410] In some embodiments, the compositions described herein can be dietary compositions that can be used to provide collagen to a subject in need thereof. For example, in some embodiments, consuming the dietary compositions described herein can provide health and / or skin benefits, such as increasing collagen uptake, relieving joint pain, and improving skin health. In certain embodiments, the composition can be in powder, capsule, liquid form, or any other suitable form.

[0411] In some embodiments, the dietary composition may include one or more nutrients such as ascorbic acid, biotin, chromium nicotinate, copper citrate, calcium D-pantothenate, cyanocobalamin, flaxseed, flax, folic acid, oligofructose (fiber), magnesium oxide, manganese citrate, maltodextrin, medium chain triglycerides, flavoring, niacinamide, potassium citrate, potassium iodide, riboflavin, saccharum officinarum), sodium molybdate dihydrate, sodium selenate (selenium), soy protein isolate, stevia leaf extract / stevia, thiamine HCl, tricalcium phosphate, vitamin A palmitate, vitamin D3, xanthan gum, zinc citrate, cellulose gum, guar gum, pyridoxine hydrochloride, salt, tocopherols, antioxidants (e.g., resveratrol, CoQ10, acai berry, lycopene, and pomegranate), natural or artificial sweeteners (e.g., glucose, sucrose, fructose, sugar, cyclamate, aspartamine, sucralose, aspartame, acesulfame K, or sorbitol), flavorings (such as flavoring extracts, volatile oils, chocolate flavorings (e.g., non-caffeinated cocoa or chocolate, chocolate substitutes such as carob), peanut butter flavoring, cookie crumbs, vanilla), or any commercially available flavorings, and any combination thereof.

[0412] Exemplary Formulations

[0413] In some embodiments, as described above, the compositions described herein can be in the form of alcohol-based or water-based toners. Exemplary toner formulations are described below. The collagen, collagen fragments, or stained collagen fragments shown in the following examples can be cross-linked or non-cross-linked. In some embodiments, the HA shown in the following examples can be cross-linked or non-cross-linked.

[0414] Alcohol based toners

[0415] Element: % weight / weight Water (including water) Remaining Ingredients Recombinant collagen fragments disclosed herein About 0.0005% to about 25% Alcohol denaturant 10％-20％ Pentylene glycol 5％-10％ glycerin 1％-5％ Gluconolactone 0.1％-1％ Dipotassium glycyrrhizinate 0.1％-1％ Sodium Citrate 0.1％-1％ Sodium Benzoate 0.1％-1％ HA 0.1％-3％

[0416] Water-based toners

[0417]

[0418] In some embodiments as described above, the compositions described herein can be in the form of a cream, gel, or serum. Exemplary cream, gel, or serum formulations are described below.

[0419] Cream

[0420] Element: % weight / weight Water (including water) Remaining Ingredients Recombinant collagen fragments disclosed herein About 0.0005% to about 25% Cetearyl Alcohol 5％-10％ glycerin 1％-5％ Squalane 1％-5％ Shea Butter 1％-5％ Glyceryl Caprylate 1％-5％ Microcrystalline Cellulose 1％-5％ Glyceryl Citrate Stearate 0.1％-1％ Tocopheryl acetate 0.1％-1％ Cetearyl Glucoside 0.1％-1％ Sodium Stearoyl Glutamate 0.1％-1％ Cellulose Gum 0.1％-1％ Xanthan gum 0.1％-1％ Octanoylhydroxamic acid 0.1％-1％ Sodium Phytate 0.01％-0.1％ HA 0.1％-3％

[0421] gel

[0422] Element: % weight / weight Water (including water) Remaining Ingredients Recombinant collagen fragments disclosed herein About 0.0005% to about 25% Sodium Phytate 0.1％-1％ Sodium hydroxide 0.1％-1％ Carbomer 0.1％-1％ Phenoxyethanol 0.1％-1％ HA 0.1％-3％

[0423] Essence

[0424]

[0425]

[0426] In some embodiments as described above, the compositions described herein can be shampoos or conditioners. Exemplary shampoo or conditioner formulations are described below.

[0427] Shampoo

[0428] Element: % weight / weight Water (including water) Remaining Ingredients Recombinant collagen fragments disclosed herein About 0.0005% to about 25% Cocamidopropyl Betaine 5％-10％ Sodium Lauroyl Methyl Isethionate 5％-10％ Propylene glycol 1％-5％ Sodium Methyl Oleyl Taurate 1％-5％ Sodium Cocoyl Isethionate 1％-5％ Trisodium Ethylenediamine Disuccinate 0.1％-1％ Octanoylhydroxamic acid 0.1％-1％ Panthenol 0.1％-1％ Citric Acid 0.1％-1％ Caprylyl glycol 0.1％-1％ Sodium Benzoate 0.1％-1％ HA 0.1％-3％

[0429] Conditioning Agents

[0430]

[0431]

[0432] In some embodiments as described above, the compositions described herein can be nail care compositions. Exemplary nail care formulations are shown below.

[0433] Nail polish

[0434] Element: % weight / weight Water (including water) Remaining Ingredients Stained recombinant collagen fragments disclosed herein About 0.0005% to about 25% Ethyl acetate 0.1％-1％ Butyl acetate 0.1％-1％ Isopropyl alcohol and diacetone alcohol. 0.1％-1％ Nitrocellulose 0.1％-1％ Acetyl tributyl citrate 0.1％-1％ Adipic acid / neopentyl glycol 0.1％-1％ Neopentyl glycol 0.1％-1％ Plasticizers 0.1％-1％ HA 0.1％-3％

[0435] F. Preparation method

[0436] In some embodiments, the present disclosure provides a method of preparing a composition comprising collagen and hyaluronic acid, wherein the collagen is completely miscible with the hyaluronic acid, the method comprising: a) dissolving the hyaluronic acid in a NaOH solution; b) adjusting the pH by adding HCl; and c) adding collagen.

[0437] 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, the method comprising: (a) dissolving the hyaluronic acid in water, (b) dissolving the collagen in water, and (c) mixing the two solutions. In some embodiments, the method further comprises: d) cross-linking the hyaluronic acid and collagen in the solution, optionally by adding a coupling agent.

[0438] 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, the method comprising: (a) dissolving the hyaluronic acid in water or a NaOH solution, b) optionally cross-linking the hyaluronic acid by adding a coupling agent before lowering the pH with HCl, (c) dissolving the collagen in water, and (d) mixing the two solutions.

[0439] In some embodiments, the method further comprises: e) adding sodium salt to cause gel formation; and f) allowing the gel to solidify. In some embodiments, the method further comprises: g) sterilizing the gel. In some embodiments, the method further comprises: h) dispensing the gel into a syringe under sterile conditions.

[0440] In some embodiments, the present disclosure provides a method of preparing a composition comprising collagen and hyaluronic acid, the method comprising dissolving the hyaluronic acid in a saline buffer, such as PBS, and adding collagen.

[0441] In some embodiments, the present disclosure provides methods of preparing a composition comprising collagen and hyaluronic acid, wherein the solution comprises NaCl.

[0442] In some embodiments, the method further comprises adding calcium hydroxyapatite, carboxymethylcellulose, polylactic acid, or any combination thereof to the composition or gel.

[0443] 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, polyoxyesters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyesteramides, hydrolyzable amines, polyalkylene oxides, polyethylene oxide (PEO), poloxamers, functionalized isoflavones, amino acids, and combinations thereof.

[0444] Other 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). 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-dioxepanone), poly(lactide-co-1,4-dioxepan-2-one) and poly(1,4-dioxepan-2-one-co-p-dioxepanone), 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- cyclohexanone-co-L-lactide-co-glycolide), poly(dioxanone-co-L-lactide-co-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-caprolactone), and combinations thereof.

[0445] In some embodiments, the method further comprises adding 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, 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, and combinations thereof.

[0446] In some embodiments, the method further comprises adding an anesthetic to the composition or gel.

[0447] 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 into a gel as required by the application.

[0448] The methods described herein may include exposing the collagen and collagen fragments described herein to a crosslinking agent (chemical crosslinking agent, such as glutaraldehyde, etc.) or to crosslinking conditions (such as heating, radiation, etc.) such that the collagen does contain non-endogenously applied crosslinks.

[0449] In some embodiments, the method of cross-linking hyaluronic acid and one or more recombinant collagen fragments includes: dissolving hyaluronic acid and collagen in an aqueous solution to form a pre-reaction aqueous solution, wherein the pre-reaction aqueous solution also includes salt or has a low pH; modifying the pre-reaction aqueous solution to form a cross-linked reaction mixture, the cross-linked reaction mixture includes: hyaluronic acid; recombinant collagen fragments; water-soluble coupling agents; and salts; and wherein the cross-linking reaction has a higher pH than the pre-reaction aqueous solution; and reacting the cross-linked reaction mixture to at least cross-link hyaluronic acid and one or more recombinant collagen fragments. In some embodiments, the concentration and ratio of hyaluronic acid and recombinant collagen fragments can vary over a wide range. The respective concentrations are at least about 0.01wt% to 10wt% hyaluronic acid and at least about 0.01wt% to 10wt% recombinant collagen fragments, at least about 0.05wt% to 10wt% hyaluronic acid and at least about 0.1wt% to 10wt% recombinant collagen fragments, at least about 0.05wt% to 5wt% hyaluronic acid and at least about 0.5wt% to 7wt% recombinant collagen fragments, or at least about 0.1wt% to 2wt% hyaluronic acid and at least about 1wt% to 5wt% recombinant collagen fragments. The respective concentrations are about 0.01wt% to 10wt% hyaluronic acid and about 0.01wt% to 10wt% recombinant collagen fragments, about 0.05wt% to 10wt% hyaluronic acid and about 0.1wt% to 10wt% recombinant collagen fragments, about 0.05wt% to 5wt% hyaluronic acid and about 0.5wt% to 7wt% recombinant collagen fragments, or about 0.1wt% to 2wt% hyaluronic acid and about 1wt% to 5wt% recombinant collagen fragments.

[0450] In some embodiments, a base (e.g., NaOH) is first added to hyaluronic acid to adjust the pH to 9-12. A cross-linking agent is added to hyaluronic acid and the reaction is carried out. One or more recombinant collagen fragments (pH 2-3) are then added. Collagen reacts with the cross-linking agent to covalently bind to HA. In some embodiments, when collagen is cross-linked to HA, the cross-linking agent may be slightly consumed (i.e., not completely reacted). A hyaluronic acid solution is added as a lubricant and a cross-linking agent scavenger to react with any excess / unreacted cross-linking agent. The final formulation may have a cross-linked hyaluronic acid core, wherein the collagen is lightly cross-linked and surrounds the hyaluronic acid core. Additional soluble hyaluronic acid consumes residual cross-linking agent and increases lubrication benefits.

[0451] In some embodiments, alkali (e.g., NaOH) can be first added to hyaluronic acid to adjust the pH to 9-12. A cross-linking agent can be added to hyaluronic acid and the reaction can be carried out. The pH can be adjusted to less than 7.5 with acid, and then one or more recombinant collagen fragments (pH 2-3) can be added. Collagen can react with a cross-linking agent to covalently bind to HA. In some embodiments, when collagen is cross-linked with HA, the cross-linking agent can be slightly consumed (i.e., incompletely reacted). A hyaluronic acid solution can be added as a lubricant and a cross-linking agent scavenger to react with any excess / unreacted cross-linking agent. The final formulation can have a cross-linked hyaluronic acid core, in which collagen is lightly cross-linked and injected into the hyaluronic acid core. Additional soluble hyaluronic acid can consume residual cross-linking agents and increase lubrication benefits.

[0452] 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- Any one of bis(2,3-epoxypropyl)-2,3-ethylene, 1-(2,3-epoxypropyl)-2,3-epoxycyclohexane, glycerol triglycidyl ether, resorcinol diglycidyl ether, allyl glycidyl ether, ethylene glycol diglycidyl ether, 1,6-hexanediol diglycidyl ether, neopentyl glycol diglycidyl ether, photocrosslinking agent (such as ethyl eosin), hydrazides (such as dihydrazides, trihydrazides, polyvalent hydrazide compounds). In some embodiments, the linking agent may be an auxiliary crosslinking agent, such as N-hydroxysuccinamide (NHS), NHS (sulfo-NHS), succinyl oxide, and sodium periodate. In some embodiments, the linker can be a carbodiimide linker, such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), 1-ethyl-3-(3-trimethylaminopropyl)carbodiimide (ETC), 1,1'-dithio-p-phenylenebis(carbodiimide), tert-butylcarbodiimide and N,N'-diisopropylcarbodiimide (DIC).

[0453] In some embodiments, the cross-linking reaction is 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 cross-linking step can be carried out using any means known to those of ordinary skill in the art.

[0454] In some embodiments, the cross-linking reaction may 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 cross-linking step may be carried out using any means known to those of ordinary skill in the art. In some embodiments, after cross-linking, the pH may be reduced to less than 7.5 before adding collagen. In water or an optional buffer such as PBS, the amount of collagen added may be about 0.1% by weight to about 25% by weight. Collagen may be added with or without mixing. Without wishing to be bound by a particular theory, it is believed that when collagen is added without mixing, collagen permeates the HA gel by diffusion. In an embodiment in which mixing is employed, any suitable mixing device known to those skilled in the art may be used.

[0455] Those skilled in the art can optimize the cross-linking conditions according to the properties of the hyaluronic acid and collagen fragments, and cross-link to an optimal degree. The degree of cross-linking can be controlled by changing several factors, including the molecular weight of the hyaluronic acid, the molecular weight of the recombinant collagen fragments, the concentration in the reaction mixture, the base concentration, and the ratio of the linking agent.

[0456] Compositions according to the present disclosure may include collagen or fragments thereof produced by yeast or modified yeast for producing collagen fragments. Suitable yeast strains and methods for producing recombinant collagen fragments are described in International Application No. PCT / US2022 / 027016, which is incorporated herein in its entirety.

[0457] Skin Care

[0458] In some embodiments, the compositions described herein can be a skin care composition that can be used to treat an area of ​​skin by topically applying the skin care composition to the area of ​​skin.

[0459] In some embodiments, the compositions described herein may be skin care compositions useful for treating the skin, hair, and nails by consuming the compositions as a dietary supplement.

[0460] In some embodiments, applying 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 tactile perceptible manifestations and any other macroscopic or microscopic effects due to skin aging. Such signs can be induced or caused by intrinsic factors or extrinsic factors (such as chronological age and / or environmental damage). These signs may result from processes including, but not limited to, the development of tissue discontinuities such as wrinkles and coarse and deep wrinkles, fine lines, skin striae, fissures, ridges, large pores (e.g., associated with adnexal structures such as sweat ducts, sebaceous glands, or hair follicles) or unevenness or roughness, loss of skin elasticity (loss and / or inactivation of functional skin elastin), sagging (including puffiness in the eye area and jowls), loss of skin firmness, loss of tautness, loss of skin resilience, discoloration (including eye circles), the appearance of spots, sallowness, hyperpigmented skin areas such as age spots and freckles, keratosis, abnormal differentiation, hyperkeratosis, elastosis, collagen breakdown, and other histological changes in the stratum corneum, dermis, epidermis, skin vasculature (e.g., telangiectasias or spider vessels), and underlying tissues (e.g., fat and / or muscle), particularly those close to the skin.

[0461] In some embodiments, the compositions as described herein may be suitable for use as dermal fillers. Dermal filler compositions may replace lost endogenous matrix polymers, or enhance / boost the function of existing matrix polymers, in order to treat skin conditions caused by aging or injury. Dermal filler compositions may fill wrinkles, lines, creases, scars, and enhance skin tissue, such as plumping up thin lips, or filling sunken eyes or shallow cheeks. Early dermal filler products were typically made of collagen. A common matrix polymer used in modern dermal filler compositions is hyaluronan. Because hyaluronan is natural to the human body, it is generally well tolerated and has a relatively low risk of treating a variety of skin conditions.

[0462] In some embodiments, the compositions described herein may be suitable for use with a microneedle plate or patch. The microneedles may deliver the compositions described herein to any area of ​​the face and body to achieve cosmetic benefits or wound healing benefits.

[0463] In some embodiments, the compositions described herein may be therapeutic compositions useful for reducing or preventing scar tissue formation, promoting healing, promoting tissue regeneration, minimizing local inflammation, minimizing tissue rejection, and / or enhancing integration of skin and / or hair grafts.

[0464] In some embodiments, the present disclosure provides a method of treating a skin condition, the method comprising administering an effective amount of a composition comprising a recombinant collagen fragment to a subject in need thereof. In some embodiments, the skin condition can be fine lines, wrinkles, dry skin, enlarged pores, skin discoloration, decreased elasticity, excess hair, thinning skin, purpura, actinic keratosis, pruritus, eczema, acne, rosacea, erythema, telangiectasia, actinic telangiectasia, skin cancer, or hypertrophic rosacea.

[0465] In some embodiments, the composition can be topically applied to a skin area. In some embodiments, the skin area can be selected from the group consisting of: facial surface, scalp, neck, ears, shoulders, chest (including breasts and / or neckline), arms, hands, legs, stomach, buttocks, groin, back, feet, and combinations thereof.

[0466] In some embodiments, the composition can be topically applied to a facial surface. In some embodiments, the facial surface can be selected from the group consisting of forehead, eye, perioral surface, chin surface, periorbital surface, nasal surface, cheek skin surface, and combinations thereof.

[0467] In some embodiments, the present disclosure provides a method for improving collagen production in skin, the method comprising administering an effective amount of a composition comprising recombinant collagen fragments.

[0468] In some embodiments, the present disclosure provides a skin care product comprising a composition as described herein, the skin care product being used to reduce the appearance of wrinkles, even out skin tone, provide moisture, reduce the appearance of dark circles under the eyes, increase the collagen content of the skin, increase skin density, improve skin firmness and elasticity, improve the appearance of fine lines and wrinkles, smooth skin texture, increase skin radiance and brightness, improve the appearance of sagging skin, whiten skin, sun care, makeup, hair care, perfume, oral care, or any combination thereof.

[0469] Example

[0470] The following examples 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 encompass any and all variations that become apparent as a result of the teachings provided herein.

[0471] Example 1

[0472] 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 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 on a stirring plate for 16 hours. A 5% (w / v) solution of a recombinant collagen fragment containing an amino acid sequence of SEQ ID NO: 1 was prepared by dissolving 5 grams of 50 kDa protein in 100 mL of water. The two solutions were mixed in a 50 mL conical tube at a ratio of 1: 1 (w / w) and mixed at 10 RPM for 16 hours. The conical tubes were placed vertically on a rack overnight before evaluating the stability of the solution. The final solution was turbid and slowly phase separated with white or yellow aggregates settling at the bottom of the tube.

[0473] Example 2

[0474] A mixture comprising polyethylene oxide (PEO) and a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1 was prepared and 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 that shown in Example 1. The final solution achieved phase separation, with a turbid solution at the top and a turbid and light yellow solution at the bottom.

[0475] Example 3

[0476] A mixture of the first polyvinylpyrrolidone (PVP) and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and dried and visually inspected. Example 3 was prepared using the same method as described in Example 1, except that polyvinylpyrrolidone (Sigma; molecular weight 10 kDa) was used instead of polymer 1. The final solution was transparent. Approximately 13 grams (g) of the solution was cast into a 100 mL polytetrafluoroethylene (PTFE) evaporating dish and dried overnight at 45°C. The dried film was visually transparent.

[0477] This example demonstrates that solutions comprising 10 kDa PVP and a recombinant collagen fragment having a molecular weight of 50 kDa and having SEQ ID NO: 1 are completely miscible and can be prepared into films or articles.

[0478] Example 4

[0479] A mixture of a second polyvinylpyrrolidone (PVP) and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and dried and visually inspected. Example 4 was prepared using the method described in Example 1, except that polyvinylpyrrolidone (Sigma; molecular weight of 40 kDa) was used instead of polymer 1. The final solution was clear. The membrane was prepared using the method described in Example 3. The dried membrane was visually clear.

[0480] This example demonstrates that solutions comprising 40 kDa PVP and a recombinant collagen fragment having a molecular weight of 50 kDa having SEQ ID NO: 1 are completely miscible and can be prepared as films or articles.

[0481] Example 5

[0482] A mixture of a third polyvinylpyrrolidone (PVP) and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and dried and visually inspected. Example 5 was prepared using the method described in Example 1, except that polyvinylpyrrolidone (Sigma; molecular weight of 360 kDa) was used instead of polymer 1. The final solution was transparent. The membrane was prepared using the same method as described in Example 3. The dried membrane was visually transparent.

[0483] This example demonstrates that solutions comprising 360 kDa PVP and a recombinant collagen fragment having a molecular weight of 50 kDa having SEQ ID NO: 1 are completely miscible and can be prepared as a membrane.

[0484] Example 6

[0485] A mixture of the first chitosan and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and visually inspected. Example 6 was prepared using the method described in Example 1, except that chitosan (molecular weight range of 50 kDa to 190 kDa; Sigma) was used instead of polymer 1. The pH of the chitosan solution was adjusted to pH 4 using 0.287 mL of acetic acid from Sigma to completely dissolve the solution during mixing. The final solution was phase separated into two layers. The viscosity was visually determined by examining the difference between the two layers. The top layer had a lower viscosity and was more transparent, while the bottom layer had a higher viscosity and was more turbid.

[0486] Example 7

[0487] A mixture of polyethylene glycol and collagen (recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and dried and visually inspected. Example 7 was prepared using the method described in Example 1, except that polyethylene glycol (molecular weight of 35 kDa Sigma) was used instead of polymer 1. The final solution achieved phase separation with a clear, clear solution on the top and a clear, light yellow solution on the bottom.

[0488] Example 8

[0489] A mixture of the first alginic acid and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and 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 in water was equal to or greater than 2,000 centipoise at 25°C. A 2.5wt% alginate solution was prepared by dissolving 2.5g sodium alginate in 97.5mL deionized ("DI") water and mixing at 400RPM for 24 hours on a stirring plate due to the high viscosity of the solution. Viscosity was visually determined by examining the difference between the two layers. The two solutions were mixed in a 50mL conical tube at a ratio of 1:1 (v / v) and mixed at 10RPM for 16 hours. Before evaluating the stability of the solution, the conical tube was placed vertically on a shelf overnight. The final solution was transparent. Films were prepared using the method shown in Example 3. The dried film was visually turbid with a uniform distribution of turbidity across the film.

[0490] Example 9

[0491] Prepare a mixture of a second alginic acid and collagen (a recombinant collagen fragment with a molecular weight of 50kDa with SEQ ID NO: 1) and dry it and visually inspect it. 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 in water was about 4 centipoises to 12 centipoises at 25°C. The stock solutions of sodium alginate and collagen were both 5wt%. The two solutions were mixed in a 50mL conical tube at a ratio of 1:1 (v / v) and mixed for 16 hours at 10RPM. The final solution was transparent. Films were prepared using the method shown in Example 3. The dried film was visually turbid, and the turbidity of the entire film was evenly distributed.

[0492] Example 10

[0493] A mixture of polyacrylamide and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and dried and visually inspected. Example 10 was prepared using the method described in Example 1, except that polyacrylamide (molecular weight range of 5,000 kDa to 6,000 kDa; Sigma) was used instead of polymer 1. The rest of the method was the same as shown in Example 1. The final solution was clear. The membrane was prepared using the same method as described in Example 3. The membrane after drying was visually clear.

[0494] This example demonstrates that solutions comprising 5,000 kDa to 6,000 kDa polyacrylamide and a recombinant collagen fragment having a molecular weight of 50 kDa having SEQ ID NO: 1 are miscible and can be prepared into films or articles.

[0495] Embodiment 11

[0496] A mixture of polyacrylamide and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and dried and visually inspected. Example 11 was prepared using the method described in Example 1, except that polyacrylamide (molecular weight of 40 kDa; Sigma) was used instead of polymer 1. The rest of the method was the same as shown in Example 1. The final solution was transparent. The membrane was prepared using the method described in Example 3, except that an aluminum weighing pan was used. The dried membrane was visually transparent.

[0497] This example demonstrates that solutions comprising 40 kDa polyacrylamide and a recombinant collagen fragment having a molecular weight of 50 kDa and having SEQ ID NO: 1 are miscible and can be prepared into films or articles.

[0498] Example 12

[0499] A mixture of a first polyvinyl alcohol (PVA) and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and visually inspected. Example 12 was prepared using the method described in Example 1, except that PVA (molecular weight range of 89 kDa to 98 kDa; Sigma) was used instead of polymer 1. A 4 wt% PVA solution was prepared by dissolving 2 g of PVA in 48 mL of DI water and mixing at 400 RPM on a stirring plate for 16 hours. The two solutions were mixed in a 50 mL conical tube at a ratio of 1: 1 (w / w) and mixed at 10 RPM for 16 hours. The final solution was phase separated into two layers. The top layer was turbid with a large amount of white powdery aggregates precipitated at the bottom.

[0500] Embodiment 13

[0501] A mixture of a second PVA and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and visually inspected. Example 13 was prepared using the method described in Example 12, except that a 4% PVA (w / v) solution (unknown molecular weight; Fisher Scientific) was used instead of Polymer 1. The rest of the method was the same as shown in Example 12. The final solution was phase separated into two layers. The viscosity was determined visually by inspecting the difference between the two layers. The top layer had a lower viscosity and was more turbid, with small suspended particles. The bottom layer had a higher viscosity and was transparent.

[0502] Embodiment 14

[0503] A mixture of a first hyaluronic acid (HA) and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and 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) (molecular weight of 1,000 kDa, purchased from Pure Health Botanicals (SaintCharles, IL)). A 2.5 wt% HA solution was prepared by dissolving 5 g HA in 195 mL DI water and 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 a recombinant collagen fragment of SEQ ID NO: 1 was prepared as described in Example 1. The HA solution and the collagen solution were mixed in a 50 mL conical tube at a ratio of 2: 1 (w / w) and mixed at 10 RPM for 16 hours to maintain a dry mass ratio of 1: 1. The final solution was transparent. The membrane was prepared using the method described in Example 3. The dried film was visually turbid with a uniform distribution of turbidity across the film.

[0504] Embodiment 15

[0505] A mixture of a second hyaluronic acid (HA) and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) was prepared and dried and visually inspected. Example 15 was prepared using the method in Example 1, except that non-animal-based HA (molecular weight of 50 kDa; Pure Health Botanicals (Saint Charles, IL) and DSM (Kaiseraugst, Switzerland)) was used instead of polymer 1. A 5 wt% HA solution was prepared by dissolving 5 g HA in 95 mL DI water and mixing at 400 RPM on a stirring plate for 16 hours. A 5% (w / v) collagen solution containing a recombinant collagen fragment of SEQ ID NO: 1 was prepared as described in Example 1. The two solutions were mixed in a 50 mL conical tube at a ratio of 1: 1 (w / w) and mixed at 10 RPM for 16 hours. The final solution was transparent. The membrane was prepared using the method described in Example 3. The dried membrane was visually turbid, with a uniform distribution of turbidity throughout the membrane. In addition, an aliquot of the HA and collagen solution mixture was placed in a 4°C refrigerator overnight and the solution remained transparent.

[0506] Example 16

[0507] Prepare the first gelatin and HA mixture and inspect it visually. Prepare Example 16 using the method in Example 15, except that polymer 2 is replaced with a low molecular weight bovine gelatin dietary supplement (Great Lakes Wellness, Grayslake, IL). Prepare a 2.2wt% gelatin solution by dissolving 1g of gelatin in 45mL0.01N HCl and mixing at 400RPM for 16 hours using a high shear impeller due to the high viscosity of the solution. Prepare a 5wt% 50kDa HA solution using the method described in Example 15. Mix the HA solution and the gelatin solution in a 50mL conical tube at a ratio of 1:2.4 (w / w) and mix at 10RPM for 16 hours to maintain a dry mass ratio of 1:1. The final solution is phase separated into two layers. The viscosity is visually determined by examining the difference between the two layers. The top layer has a lower viscosity and is more transparent. The bottom layer has a higher viscosity and is more turbid.

[0508] Embodiment 17

[0509] A second gelatin and HA mixture was prepared and visually inspected. Example 17 was prepared using the method described in Example 16, except that polymer 2 was replaced with gelatin from pig skin (molecular weight 300 kDa; Sigma). A 1.1 wt% gelatin solution was prepared by dissolving 0.5 g of gelatin in 45 mL of 0.01 N HCl and 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 achieved phase separation, with a large amount of white aggregates precipitated at the bottom and dispersed white aggregates suspended or floating at the top.

[0510] Embodiment 18

[0511] HA and type I collagen mixtures were prepared and visually inspected. Example 18 was prepared using the method described in Example 15, except that type I collagen (molecular weight 414kDa; Sigma) was used instead of polymer 2. A 0.7wt% type I collagen solution was prepared by dissolving 0.3g type I collagen in 45mL 0.01N HCl and mixing at 400RPM for 16 hours using a high shear impeller due to the high viscosity of the solution. A 5wt% 50kDa HA solution was prepared using the method described in Example 15. The HA solution and type I collagen solution were mixed together in a 50mL Falcon conical tube at a ratio of 1:7 (w / w) and mixed at 10RPM for 16 hours with a Hula stirrer to maintain a dry mass ratio of 1:1. The final solution achieved phase separation, with a large amount of white fibrils precipitated at the bottom and dispersed white fibrils suspended or floating at the top.

[0512] Embodiment 19

[0513] Prepare a cellulase and HA mixture and inspect it visually. Prepare Example 19 using the method described in Example 15, except that cellulase (Sunson Industry Group Co., Ltd.) is used instead of polymer 2. A 5% (w / v) cellulase solution is prepared by dissolving 2.5g of cellulase in 47.5mL DI water and mixing on a stirring plate at 400RPM for 16 hours. A 5wt% 50kDa HA solution is prepared using the same method described in Example 15. The two solutions are mixed together in a 50mL conical tube at a ratio of 1:1 (w / w) and mixed at 10RPM for 16 hours. The final solution is turbid and a yellow powder is precipitated at the bottom.

[0514] Embodiment 20

[0515] A mixture of HA, chitosan and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having 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 5wt% 50kDa HA solution was prepared by dissolving 5g HA in 95mL DI water and mixing at 400RPM on a stirring plate for 16 hours. A 5% (w / v) collagen solution containing a recombinant collagen fragment of SEQ ID NO: 1 was prepared by dissolving 5 grams of 50kDa protein in 100mL water as described in Example 1. A 5wt% chitosan solution was prepared by dissolving 2.5g chitosan in 47.5mL DI water and mixing at 400RPM on a stirring plate for 16 hours. The pH of the chitosan solution was adjusted to pH 4 using 0.287mL acetic acid from Sigma to completely dissolve the solution during mixing. The three solutions were mixed in a 50 mL conical tube at a ratio of 1:1:1 (w / v) and mixed at 10 RPM for 16 hours. The final solution achieved phase separation, with large yellow aggregates suspended or floating in the low viscosity medium.

[0516] Embodiment 21

[0517] A mixture of hyaluronic acid and a recombinant collagen fragment (hydroxylated to 22%) with a molecular weight of 50kDa having SEQ ID NO: 1 was prepared and visually inspected. The recombinant collagen fragment was hydroxylated by the method described in International Application No. WO2021 / 163485, and the hydroxylation percentage was measured by the method described in International Application No. WO2021 / 163485, the disclosure of which is incorporated herein as a whole. Example 21 was prepared using the method in Example 15, except that polymer 2 was replaced with 22% hydroxylated collagen with a molecular weight of 50kDa. The solid content of the 22% hydroxylated collagen solution was 1.3% and the pH was about 6. A 5wt% 50kDa HA solution (polymer 1) was prepared by dissolving 5g HA in 95mL DI water and mixing at 400RPM on a stirring plate for 16 hours. The two solutions were mixed in a 15mL Falcon tube at a ratio of 1: 1 (w / w) and mixed at 10RPM for 16 hours. The final solution was turbid, indicating that at this level of hydroxylation, the two polymers were immiscible in solution.

[0518] Embodiment 22

[0519] A mixture of hyaluronic acid and a recombinant collagen fragment (hydroxylated to 44%) with a molecular weight of 50kDa having SEQ ID NO: 1 was prepared and visually inspected. The recombinant collagen fragment was hydroxylated by the method described in International Application No. WO2021 / 163485, and the hydroxylation percentage 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 with a molecular weight of 50kDa was used as polymer 2. The solid content of the 44% hydroxylated collagen solution was 1.3% and the pH was about 6. A 5wt% 50kDa HA solution (polymer 1) was prepared by dissolving 5g HA in 95mL DI water and mixing at 400RPM on a stirring plate for 16 hours. The two solutions were mixed in a 15mL conical tube at a ratio of 1: 1 (w / w) and mixed at 10RPM for 16 hours. The final solution was turbid and slowly phase separated with white aggregates settling at the bottom of the tube.

[0520] Embodiment 23

[0521] A mixture of biphasic cross-linked hyaluronic acid (comprising cross-linked HA and uncross-linked HA) and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having 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 of 1.25 mm-2 mm; Beijing Mengbring Bio-Sci-Tec. Co., Ltd.). A 2.4% (w / v) HA solution was used. A 20wt% solution of the recombinant collagen fragment of SEQ ID NO: 1 was prepared (polymer 2) by dissolving 2g 50kDa recombinant collagen in 18mL DI water and mixing at 10RPM for 4 hours. The two solutions were mixed together in a 50mL conical tube at a ratio of 1: 1 (w / w) and mixed at 10RPM for 16 hours. The mixed solution was visually transparent.

[0522] This example demonstrates that solutions comprising biphasic cross-linked HA and a recombinant collagen fragment having a molecular weight of 50 kDa and having SEQ ID NO: 1 are completely miscible.

[0523] Embodiment 24

[0524] A mixture of single-phase cross-linked hyaluronic acid and collagen (a recombinant collagen fragment with a molecular weight of 50kDa with SEQ ID NO: 1) was prepared and visually inspected. Example 24 was prepared using the same method as Example 15, except that polymer 1 was replaced with single-phase cross-linked HA (Bloomage Biotechnology Corp. (Jinan, Shandong, China)). A 2% (w / v) single-phase cross-linked HA solution was used. A 20wt% collagen solution containing a recombinant collagen fragment of SEQ ID NO: 1 was prepared by dissolving 2g of 50kDa recombinant collagen in 18mL DI water and mixing at 10RPM for 4 hours. The two solutions were mixed in a 50mL conical tube at a ratio of 1: 1 (w / w) and mixed at 10RPM for 16 hours. The solution was visually transparent.

[0525] This example demonstrates that a solution comprising single-phase cross-linked HA and a recombinant collagen fragment having a molecular weight of 50 kDa and having SEQ ID NO: 1 is completely miscible.

[0526] Embodiment 25

[0527] A mixture of biphasic cross-linked hyaluronic acid (comprising cross-linked HA and uncross-linked HA) and collagen (a recombinant collagen fragment with 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. A 40wt% solution of the recombinant collagen fragment of SEQ ID NO: 1 was prepared by dissolving 4g of 50kDa recombinant collagen in 16mL DI water and mixing at 10RPM for 4 hours. The two solutions were mixed in a 1mL Eppendorf tube with a pipette at a ratio of HA: collagen of 1:3, 1:1, 10:3 and 100:3 (w / w), and mixed at 10RPM for 16 hours. The solution was visually transparent.

[0528] This example demonstrates that solutions comprising biphasic cross-linked hyaluronic acid and a recombinant collagen fragment having a molecular weight of 50 kDa and having SEQ ID NO: 1 are completely miscible.

[0529] Embodiment 26

[0530] A mixture of 5% hyaluronic acid and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having 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 5wt% 50 kDa HA solution was adjusted to pH 5.2-7.2 with sodium hydroxide before adding 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 ratio of 1: 1 (w / w) and mixed at 10 RPM for 16 hours. The solution was visually transparent. The film was prepared using the method of Example 3. The dried film was visually turbid, and the turbidity was evenly distributed throughout the film.

[0531] This example demonstrates that solutions comprising 5% hyaluronic acid and a recombinant collagen fragment having a molecular weight of 50 kDa and having SEQ ID NO: 1 at pH 5.2-7.2 are completely miscible.

[0532] Embodiment 27

[0533] Mixtures of 5% hyaluronic acid and collagen (a recombinant collagen fragment with a molecular weight of 50 kDa having SEQ ID NO: 1) were prepared at different pH values ​​and visually inspected. Example 27 was prepared using the method described in Example 15, except that the pH of the 5wt% 50 kDa HA solution was adjusted to pH 9, 10, 11, and 12 with sodium hydroxide before adding the recombinant collagen fragment of SEQ ID NO: 1. The two solutions were mixed in a 20 mL glass bottle at a ratio of 1: 1 (w / w) and mixed at 400 RPM for 30 minutes with a stirring rod due to the lower viscosity of the alkaline solution. The solution was then readjusted to pH 7 ± 1 with concentrated hydrochloric acid while mixing at 400 RPM with a stirring rod. Before and after the pH was adjusted to 7 ± 1, the solution was visually transparent.

[0534] This example demonstrates that solutions containing 5% hyaluronic acid and a recombinant collagen fragment having a molecular weight of 50 kDa and having SEQ ID NO: 1 at pH 9, 10, 11, 12 are completely miscible.

[0535] Embodiment 28

[0536] Hyaluronic acid and collagen mixtures were prepared and visually inspected. Example 28 was prepared using the method of Example 15, except that 2% Solution (Geltor) was used instead of polymer 1. A 2 wt% 50 kDa HA solution 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 bottle and mixed on a stir plate at 400 RPM for 1 hour. The final solution was visually clear.

[0537] This example demonstrates that the inclusion of 2% The solution and the solution of the recombinant collagen fragment having a molecular weight of 50 kDa having SEQ ID NO: 1 are completely miscible.

[0538] Embodiment 29

[0539] Hyaluronic acid and full-length recombinant collagen and 50kDa recombinant collagen fragment are blended, mixed with HA, and visually inspected. Full-length recombinant collagen and 50kDa recombinant collagen fragment are blended and mixed with HA. The solution is visually inspected. The blend of 50kDa recombinant collagen fragment and full-length recombinant collagen and / or hydrolyzed recombinant collagen (or any combination of these three kinds) is mixed with HA, and visually inspected. Then, the blend of full-length recombinant collagen and 50kDa recombinant collagen fragment and the blend of full-length recombinant collagen and hydrolyzed full-length recombinant collagen are mixed with HA. The solution is visually inspected.

[0540] Embodiment 30

[0541] A solution of poly(2-ethyl-2-oxazoline) (PEOx) 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 PEOx (molecular weight 50 kDa; Sigma) was used instead of polymer 1. The remainder of the method was the same as that shown in Example 1. The final solution was visually transparent, indicating that the polymer was miscible in the solution. The membrane was prepared using the same method as described in Example 3, except that an aluminum weighing pan was used. The dried membrane visually appeared to phase separate during the drying process.

[0542] Embodiment 31

[0543] 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 PEI (molecular weight of 100 kDa; Polysciences (Warrington, Pennsylvania)) was used instead of polymer 1. A 5 wt % polyethyleneimine solution was prepared by dissolving 0.5 g PEI in 9.5 g Milli-Q water and stirring at 450 RPM with a stirring bar at 80° C. for 3 hours, or until the solution was completely transparent. The solution was then cooled to room temperature and the pH was adjusted to 7 using 12N HCl. The remainder of the method was the same as that shown in Example 1. The solution was transparent, indicating that the polymers were miscible in the solution. The membrane was prepared using the method shown in Example 3. The dried membrane was visually opaque.

[0544] Embodiment 32

[0545] A mixture of the first sodium carboxymethylcellulose (NaCMC) and collagen (a recombinant collagen fragment with a molecular weight of 50kDa having SEQ ID NO: 1) was prepared and dried and visually inspected. Example 32 was prepared using the method described in Example 1, except that sodium carboxymethylcellulose (molecular weight of about 250kDa, degree of substitution of 1.22; Sigma) was used instead of polymer 1. A 5wt% sodium carboxymethylcellulose solution was prepared by dissolving 0.5g sodium carboxymethylcellulose in 9.5g DI water and mixing at 5RPM in a conical tube for 16 hours. The two solutions were mixed in a 50mL conical tube at a ratio of 1:1 (v / v) and mixed at 10RPM for 16 hours. The final solution was a transparent light yellow color, indicating that the solution was miscible. The membrane was prepared using the method shown in Example 3, except that an aluminum weighing pan was used. The dried membrane was visually turbid, and the turbidity of the entire membrane was evenly distributed.

[0546] Embodiment 33

[0547] A mixture of a second sodium carboxymethylcellulose and collagen (a recombinant collagen fragment with a molecular weight of 50kDa having SEQ ID NO: 1) was prepared and dried and visually inspected. Example 33 was prepared using the same method as described in Example 1, except that sodium carboxymethylcellulose (molecular weight of about 2...

Claims

1. A composition comprising collagen and hyaluronic acid, wherein the collagen and the hyaluronic acid are completely miscible.

2. The composition of claim 1, wherein the collagen is recombinantly produced collagen.

3. The composition of claim 1 or 2, wherein the collagen is a recombinantly produced collagen fragment.

4. The composition of any one of claims 1-3, wherein the composition has an isoelectric point (pI) of about 4.0 to about 12.

0.

5. The composition of claim 4, wherein the composition has a pI of about 4.0 to about 5.

0.

6. The composition of claim 4, wherein the composition has a pI of about 6.0 to about 7.

7. The composition of claim 4, wherein the composition has a pI of about 9.0 to about 10.

0.

8. The composition of claim 7, wherein the composition has a pI of about 9.0 to about 9.

7.

9. The composition of claims 1-8, wherein the collagen has a molecular weight of about 5 kDa to about 150 kDa.

10. The composition of any one of claims 1-9, wherein the hyaluronic acid has a molecular weight of about 3 kDa to about 10,000 kDa.

11. The composition of claim 10, wherein the hyaluronic acid has a molecular weight of about 20 kDa to about 2,000 kDa.

12. The composition of claim 11, wherein the hyaluronic acid has a molecular weight of about 1,000 kDa to at least about 5,000 kDa.

13. The composition of claim 11 or 12, wherein the hyaluronic acid has a molecular weight of about 2,000 kDa to at least about 3,000 kDa.

14. The composition of any one of claims 11-13, wherein the hyaluronic acid has a molecular weight of about 1,000 kDa.

15. The composition of any one of claims 11-13, wherein the hyaluronic acid has a molecular weight of about 50 kDa.

16. The composition of any one of claims 1-15, wherein the collagen does not contain hydroxyproline (Hyp) residues.

17. The composition of any one of claims 1-15, wherein the collagen comprises from about 1% to about 20% Hyp residues based on the total number of prolines in the collagen.

18. The composition of any one of claims 1-15, wherein the collagen does not contain hydroxyl groups.

19. The composition of any one of claims 1-15, wherein the collagen comprises from about 1 to about 120 hydroxyl groups.

20. The composition of any one of claims 1-19, wherein the collagen does not contain amine groups.

21. The composition of any one of claims 1-19, wherein the collagen comprises from about 1 to about 100 amine groups.

22. The composition of any one of claims 1-21, wherein the collagen does not contain carboxylic acid groups.

23. The composition of any one of claims 1-21, wherein the collagen comprises from about 1 to about 80 carboxylic acid groups.

24. The composition of any one of claims 1-23, wherein the hyaluronic acid is cross-linked.

25. The composition of any one of claims 1-23, wherein the hyaluronic acid is not cross-linked.

26. The composition according to any one of claims 1-25, wherein the hyaluronic acid is biphasic cross-linked hyaluronic acid.

27. The composition of any one of claims 1-25, wherein the hyaluronic acid is a single-phase cross-linked hyaluronic acid.

28. The composition of any one of claims 1-27, wherein the hyaluronic acid solution has a pH in the range of about 5.0 to about 12.

29. The composition of claim 28, wherein the hyaluronic acid has a pH in the range of about 9.0 to about 12.

30. The composition of claim 28 or 29, wherein the hyaluronic acid has a pH in the range of about 5.2 to about 7.

2.

31. The composition of any one of claims 1-30, wherein the ratio (w / w) of the hyaluronic acid to the collagen is in the range of about 1:9 to 9:

1.

32. The composition of any one of claims 1-31, wherein the collagen is cross-linked.

33. The composition of any one of claims 1-31, wherein the collagen is not cross-linked.

34. The composition of claim 32, wherein the hyaluronic acid is cross-linked by a chemical cross-linking agent, ultraviolet (UV) radiation, microwave radiation, or heating.

35. The composition of claim 32, wherein the collagen is cross-linked by a chemical cross-linking agent, ultraviolet (UV) radiation, microwave radiation, or heating.

36. The composition of any one of claims 1-35, wherein the composition has a viscosity of about 1 centipoise to about 100,000 centipoise.

37. The composition of claim 36, wherein the composition has a viscosity of about 10,000 centipoise to about 80,000 centipoise.

38. The composition of claim 36 or 37, wherein the composition has a viscosity of about 50,000 centipoise to about 70,000 centipoise.

39. The composition of any one of claims 1-38, wherein the composition is a solution.

40. The composition of any one of claims 1-38, wherein the composition is a gel.

41. The composition of any one of claims 1-38, wherein the composition is a foam.

42. A film comprising the composition of any one of claims 1-38.

43. One or more fibers comprising the composition of any one of claims 1-38.

44. A coating prepared from the composition of any one of claims 1-38.

45. A powder prepared from the composition of any one of claims 1-38.

46. ​​A slurry prepared from the composition of any one of claims 1-38.

47. One or more particles prepared from the composition of any one of claims 1-38.

48. The composition of claim 1, wherein the collagen is non-hydroxylated.

49. The composition of any one of claims 2-47, wherein the recombinant collagen fragment is non-hydroxylated.

50. The composition of claim 1, wherein the collagen is hydroxylated.

51. The composition of any one of claims 2-47, wherein the recombinant collagen fragment is hydroxylated.

52. The composition of claim 50 or 51, wherein the recombinant collagen fragment is about 1% to about 20% hydroxylated.

53. The composition of claim 52, wherein the recombinant collagen fragment is about 12% hydroxylated.

54. The composition of claim 52, wherein the recombinant collagen fragment is 5% hydroxylated.

55. The composition of any one of claims 1-54, further comprising a pharmaceutically active agent.

56. A composition according to any one of claims 1-55, further comprising a cosmetically active agent.

57. The composition of any one of claims 1-56, wherein the collagen or the recombinant collagen fragment has at least about 85% sequence identity with the amino acid sequence shown in SEQ ID NO:

1.

58. The composition of any one of claims 1-56, wherein the collagen or the recombinant collagen fragment has at least about 85% sequence identity with the amino acid sequence shown in any one of SEQ ID NO:2 to SEQ ID NO:1002.

59. The composition of any one of claims 1-58, wherein the collagen or the recombinant collagen is fermented or chemically synthesized.

60. A method of increasing collagen production in a cell, the method comprising administering or applying an effective amount of a composition according to any one of claims 1-59 to a subject in need thereof.

61. A method of medical or cosmetic treatment, the method comprising administering or applying an effective amount of a composition according to any one of claims 1 to 59 to a subject in need thereof.

62. The method of claim 61, wherein the treatment comprises filling wrinkles, increasing viscoelasticity, or increasing volume of the face or body.

63. The method of claim 61, wherein the treatment comprises forming a cosmetic implant in the patient.

64. The method of claim 61, wherein the treatment comprises tissue augmentation.

65. The method of any one of claims 60-64, wherein the composition is applied to the face, eye, nose, nasolabial fold, neck, collarbone, arm, abdomen, blood vessel, bladder, urethra, nerve, cartilage, bone, breast, vocal cord, vocal folds, tympanic membrane, joint, intervertebral disc, buttocks, hip, calf, thigh, flank, or hand of the subject.

66. The method of claim 61, wherein the treatment comprises treating an inflammatory skin disease or a mucosal disease.

67. The method of claim 61, wherein the treating comprises treating lipoatrophy.

68. 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 1-59 to the wound of the subject.

69. The method of claim 68, wherein the wound is a chronic wound, a surgical wound, a superficial wound, an abrasion, or wherein the wound is the result of thermal injury.

70. A skin care product comprising a composition according to any one of claims 1-59, the skin care product being used to reduce the appearance of wrinkles, even out skin tone, provide moisture, reduce the appearance of dark circles under the eyes, increase the collagen content of the skin, increase skin density, improve skin firmness and elasticity, improve the appearance of fine lines and wrinkles, smooth skin texture, increase skin radiance and brightness, improve the appearance of sagging skin, whiten the skin, sun care, makeup, hair care, perfume, oral care, or any combination thereof.

71. A dermal filler comprising the composition of any one of claims 1-59.

72. The dermal filler of claim 71 , wherein the dermal filler is a heat stable, sterile, soft tissue filler.

73. The dermal filler of claim 72, wherein the dermal filler is thermally stable up to at least about 125°C.

74. The dermal filler of claim 72 or 73, wherein the dermal filler is thermally stable over a temperature range up to at least about 135°C.

75. The dermal filler according to any one of claims 71-74, wherein the dermal filler is terminally sterilized.

76. The dermal filler of any one of claims 71-75, wherein the dermal filler is injectable.

77. The dermal filler of any one of claims 71-76, further comprising carboxymethylcellulose.

78. The dermal filler of any one of claims 71-76, further comprising calcium hydroxyapatite.

79. The dermal filler of any one of claims 71-78, further comprising polylactic acid, aliphatic polyesters, polyorthoesters, polyoxyesters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyesteramides, hydrolyzable amines, polyalkylene oxides, poloxamers, functionalized isoflavones, amino acids, solid homopolymers of poly(ε-caprolactone), solid homopolymers of poly(p-dioxanone), solid homopolymers of poly(trimethylene carbonate), solid copolymers of a plurality of ε-caprolactone repeating units and a third lactone repeating unit, solid copolymers 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 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, or any combination thereof.

80. The dermal filler of any one of claims 71-79, further comprising a pharmaceutically active agent.

81. The dermal filler of claim 80, wherein the pharmaceutically active agent comprises an anesthetic.

82. Use of the composition of any one of claims 1-59, the skin care product of claim 70, or the dermal filler of any one of claims 71-81 for the medical or cosmetic treatment of a subject in need thereof.

83. A method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen and the hyaluronic acid are completely miscible, the method comprising: a) dissolving hyaluronic acid in NaOH solution; b) adding collagen; as well as c) The pH was adjusted by adding HCl.

84. A method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen and the hyaluronic acid are completely miscible, the method comprising: a) dissolving hyaluronic acid in NaOH solution; b) adjusting the pH by adding HCl; as well as c) Adding collagen.

85. A method for preparing a composition comprising collagen and hyaluronic acid, wherein the collagen and the hyaluronic acid are completely miscible, the method comprising: a) dissolving hyaluronic acid in water to form a hyaluronic acid solution; b) dissolving collagen in water to form a collagen solution; as well as c) mixing the hyaluronic acid solution and the collagen solution.

86. The method of any one of claims 83-85, further comprising: d) optionally cross-linking the hyaluronic acid and the collagen in the solution by adding a coupling agent.

87. The method of any one of claims 83-86, further comprising cross-linking the hyaluronic acid and the collagen in the solution, optionally by adding a coupling agent prior to adjusting the pH with HCl and adding the collagen.

88. The method of any one of claims 83-87, further comprising: e) adding sodium chloride salt to cause gel formation; as well as f) allowing the gel to solidify.

89. The method of claim 88, further comprising: g) Sterilizing the gel.

90. The method of any one of claims 83-89, wherein the solution comprises NaCl.

91. The method of any one of claims 83-90, further comprising adding calcium hydroxyapatite, carboxymethyl cellulose, polylactic acid, polylactic acid, aliphatic polyesters, polyorthoesters, polyoxyesters, polyanhydrides, polycarbonates, polyurethanes, polyamides, polyesteramides, hydrolyzable amines, polyalkylene oxides, poloxamers, functionalized isoflavones, amino acids, solid homopolymers of poly(ε-caprolactone), solid homopolymers of poly(p-dioxanone), solid homopolymers of poly(trimethylene carbonate), multiple ε-caprolactones, and / or poly(propylene glycol). - a solid copolymer of a caprolactone repeating unit and a third lactone repeating unit, 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, and / or any combination thereof, or any combination thereof.

92. The method of any one of claims 83-91 further comprising adding an anesthetic.

93. The method of any one of claims 89-92, further comprising dispensing the gel into a syringe under sterile conditions.

94. A composition comprising collagen and polyvinylpyrrolidone, wherein the collagen is completely miscible with the polyvinylpyrrolidone.

95. The composition of claim 94, wherein the polyvinylpyrrolidone has a molecular weight of about 10 kDa to about 400 kDa.

96. The composition of claim 94 or 95, wherein the polyvinylpyrrolidone has a molecular weight of about 50 kDa.

97. A composition comprising collagen and polyacrylamide, wherein the collagen is completely miscible with the polyacrylamide.

98. The composition of claim 97, wherein the polyacrylamide has a molecular weight of about 5,000 kDa to about 6,000 kDa.

99. The composition of claim 97, wherein the polyacrylamide has a molecular weight of about 30 kDa to about 50 kDa.

100. A composition comprising collagen and polyethylene oxide, wherein the collagen is completely miscible with the polyethylene oxide.

101. A composition comprising collagen and poly(2-oxazoline), wherein the collagen is completely miscible with the poly(2-oxazoline).

102. A composition comprising collagen and polyethyleneimine, wherein the collagen is completely miscible with the polyethyleneimine.

103. A composition comprising collagen and sodium carboxymethylcellulose, wherein the collagen is completely miscible with the carboxymethylcellulose.

104. The composition of claim 103, wherein the carboxymethyl cellulose has a molecular weight of about 250 kDa and a degree of substitution of about 1.00 to about 1.

25.

105. The composition of claim 103 or 104, wherein the carboxymethyl cellulose has a molecular weight of about 250 kDa and a degree of substitution of about 0.70 to about 0.

90.

106. The composition of any one of claims 103-105, wherein the carboxymethyl cellulose has a molecular weight of about 250 kDa and a degree of substitution of about 0.

79.

107. The composition of any one of claims 103-105, wherein the carboxymethyl cellulose has a degree of substitution of about 0.

87.

108. The composition of any one of claims 103-107, wherein the carboxymethyl cellulose has a high viscosity.

109. A composition comprising collagen and chondroitin sulfate, wherein the collagen is completely miscible with the chondroitin sulfate.

110. The composition of claim 109, wherein the chondroitin sulfate has a molecular weight of about 50 kDa.

111. A composition comprising collagen and acetylated hyaluronate.

112. The composition of claim 111, wherein the collagen is completely miscible with the acetylated hyaluronate.

113. A composition comprising collagen and zinc hyaluronate.

114. according to the composition described in any one of claims 94-113, wherein said collagen is with SEQ The amino acid sequence shown in ID NO: 1 has a sequence identity of 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%.

115. according to the composition described in any one of claims 94-114, wherein said collagen is with SEQ The amino acid sequence shown in ID NO: 1 has at least about 98% sequence identity.

116. A composition comprising a recombinant collagen fragment having at least about 85% amino acid sequence identity to SEQ ID NO: 1, and / or a recombinant collagen fragment having at least about 85% amino acid sequence identity to SEQ ID NO: 975 to SEQ ID One or more collagen fragment sequence variants having at least about 85% amino acid sequence identity with the amino acid sequence shown in any one of NO:1002, and hyaluronic acid, wherein the recombinant collagen fragment is completely miscible with the hyaluronic acid.

117. The composition of claim 116, wherein the hyaluronic acid is a cross-linked HA and biphasic cross-linked hyaluronic acid with uncross-linked HA.

118. A composition comprising 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% amino acid sequence identity with the amino acid sequence shown in any one of SEQ ID NO:975 to SEQ ID NO:1002, and polyvinyl pyrrolidone, and polyvinyl pyrrolidone, wherein the recombinant collagen fragment is completely miscible with the polyvinyl pyrrolidone.

119. A composition comprising one or more recombinant collagen fragments 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% amino acid sequence identity with the amino acid sequence shown in any one of SEQ ID NO:975 to SEQ ID NO:1002, and sodium carboxymethyl cellulose, wherein the recombinant collagen fragments are completely miscible with the sodium carboxymethyl cellulose.

120. A membrane comprising recombinantly produced collagen fragments and hyaluronic acid.

121. The film of claim 120, wherein the film is visually transparent.

122. The film of claim 120, wherein the film is turbid.

123. The film of claim 120, wherein the film exhibits phase separation.

124. A gel comprising recombinantly produced collagen fragments and hyaluronic acid.

125. One or more fibers comprising recombinantly produced collagen fragments and hyaluronic acid.

126. A coating comprising recombinantly produced collagen fragments and hyaluronic acid.

127. A slurry comprising recombinantly produced collagen fragments and hyaluronic acid.

128. One or more particles comprising recombinantly produced collagen fragments and hyaluronic acid.

129. A foam comprising recombinantly produced collagen fragments and hyaluronic acid.

130. A composition comprising collagen and sodium alginate, wherein the collagen is completely miscible with the sodium alginate.

131. The composition of claim 130, wherein the sodium alginate salt has a viscosity of about 4 centipoise to 12 centipoise for a 1% aqueous solution at 25°C.

132. A composition comprising collagen and sodium alginate, wherein the collagen is completely miscible with the sodium alginate.

133. The composition of claim 130, wherein the sodium alginate salt has a viscosity equal to or greater than 2,000 centipoise for a 2% aqueous solution at 25°C.

Images