WO2020010171A1 - Dermal compositions and methods of use - Google Patents

Abstract

Disclosed herein are dermal compositions and methods of use. In some embodiments, the composition comprises a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

Description

DERMAL COMPOSITIONS AND METHODS OF USE

CLAIM OF PRIORITY

[0001] This application claims priority to U.S. Provisional Application No.

62/694,514, filed July 6, 2018, U.S. Provisional Application No. 62/694,522, filed July 6, 2018, U.S. Provisional Application No. 62/694,533, filed on July 6, 2018, U.S. Provisional Application No. 62/694,560, filed July 6, 2018, and U.S. Provisional Application No. 62/697,741, filed July 13, 2018, and each of which is incorporated herein by reference in its entirety.

SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on June 28, 2019, is named l47476_02402_SL.txt and is 17,282 bytes in size.

SUMMARY

[0003] Disclosed herein are dermal compositions and methods of use. In some embodiments, the composition comprises a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the dermal filler is selected from hyaluronic acid, collagen, heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, elastin, or a combination thereof. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0004] In some embodiments, a method of stimulating elastogenesis in a subject in need thereof comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0005] In some embodiments, a method of ameliorating a skin condition in a subject in need thereof comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0006] In some embodiments, a method of improving skin texture or appearance in a subject in need thereof comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0007] In some embodiments, a method of treating scars and/or wrinkles on a subject’s skin comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0008] In some embodiments, a method of improving appearance or reducing fine lines and wrinkles on a subject’s skin comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone- iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

DETAILED DESCRIPTION

[0009] As used in this application and in the claims, the singular forms“a,” “an,” and“the” include the plural forms unless the context clearly dictates otherwise. Additionally, the term“includes” means“comprises.”

[0010] The term“about” when immediately preceding a numerical value means a range of plus or minus 10% of that value, e.g,“about 50” means 45 to 55, “about 25,000” means 22,500 to 27,500, etc, unless the context of the disclosure indicates otherwise, or is inconsistent with such an interpretation. For example, in a list of numerical values such as“about 49, about 50, about 55,“about 50” means a range extending to less than half the interval(s) between the preceding and subsequent values, e.g, more than 49.5 to less than 52.5. Furthermore, the phrases“less than about” a value or“greater than about” a value should be understood in view of the definition of the term“about” provided herein.

[0011] The terms“administer,”“administering” or“administration” as used herein refer to either directly administering a compound (also referred to as an agent of interest) or pharmaceutically acceptable salt of the compound (agent of interest) or a composition to a subject.

[0012] The transitional term“comprising,” which is synonymous with “including,”“containing,” or“characterized by,” is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. By contrast, the transitional phrase“consisting of’ excludes any element, step, or ingredient not specified in the claim. The transitional phrase“consisting essentially of’ limits the scope of a claim to the specified materials or steps“and those that do not materially affect the basic and novel characteristic(s)” of the claimed invention. In embodiments or claims where the term comprising is used as the transition phrase, such embodiments can also be envisioned with replacement of the term“comprising” with the terms“consisting of’ or “consisting essentially of”

[0013] An“effective amount” of a composition is a predetermined amount calculated to achieve the desired effect, i.e., to ameliorate, prevent or improve an unwanted condition, disease or symptom of a patient. The activity contemplated by the present methods may include both therapeutic and/or prophylactic treatment, as appropriate. The specific dose of the agent administered according to this invention is to obtain therapeutic and/or prophylactic effects. These will, of course, be determined by the particular circumstances surrounding the case, including, for example, the

compound administered, the route of administration, and the condition being treated.

The effective amount administered may be determined by a physician in the light of the relevant circumstances, including the condition to be treated, the choice of the effective agent to be administered, and the chosen route of administration.

[0014] The term“patient” and“subject” are interchangeable and may be taken to mean any living organism which may be treated with compounds of the present invention. As such, the terms“patient” and“subject” may include, but is not limited to, any non-human mammal, primate or human. In some embodiments, the“patient” or “subject” is a mammal, such as mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, primates, or humans. In some embodiments, the patient or subject is an adult, child or infant. In some embodiments, the patient or subject is a human.

[0015] The term“treating” is used herein, for instance, in reference to methods of treating a skin disorder or a systemic condition, and generally includes the administration of a compound or composition which reduces the frequency of, or delays the onset of, symptoms of a medical condition or enhance the texture, appearance, color, sensation, or hydration of the intended tissue treatment area of the tissue surface in a subject relative to a subject not receiving the compound or composition. This can include reversing, reducing, or arresting the symptoms, clinical signs, and underlying pathology of a condition in a manner to improve or stabilize a subject’s condition. [0016] The term“wt. % (weight percent)” as used herein may be weight-to- weight or weight-to-volume percentages with respect to the total amount of the composition, as applicable.

[0017] Skin aging is a progressive phenomenon, occurs over time and can be affected by lifestyle factors, such as alcohol consumption, tobacco and sun exposure. Aging of the facial skin can be characterized by atrophy, slackening, and fattening. Atrophy corresponds to a massive reduction of the thickness of skin tissue. Slackening of the subcutaneous tissues leads to an excess of skin and ptosis and leads to the appearance of drooping cheeks and eye lids. Fattening refers to an increase in excess weight by swelling of the bottom of the face and neck. These changes are typically associated with dryness, loss of elasticity, and rough texture.

[0018] Skin is composed of a top layer, the epidermis, which is approximately 20 cell layers or about 0.1 mm in thickness, and a lower layer, the dermis, which is from about 1 to about 4 mm in thickness and contains small blood vessels, collagen, elastin and fibroblasts. The dermis provides structural support and nutrients to the epidermis. Aging has been shown to increase cellular heterogeneity of the epidermal layer, however, it has little effect on the thickness of the epidermal layer. The supporting dermis, on the other hand, is known to thin with age and exposure to the sun and environmental contaminants. As the dermal layer provides the support and blood supply for the epidermis, the dermal layer is important in maintaining the elasticity and appearance of the skin. Disruption of the supporting dermis leads directly to sagging and, consequently, furrowing of the epidermis, i.e., the formation of wrinkles.

[0019] Deep wrinkles are also due to continual stretching and contraction of both the dermis and epidermis. Currently, these deep wrinkles or furrows may only be eliminated by plastic surgery or by collagen injections directly beneath the depressed areas. The fine wrinkles that occur with age and prolonged exposure to the sun and other environmental contaminants are the direct result of deterioration of the supporting dermal layer, including loss of elastin. [0020] Elastic libers present in the skin are responsible for its essential elasticity and tonicity. Decrease in elastin causes the skin to sag, allowing line lines, folds and wrinkles to appear and grow. Elastic fibers are highly organized structures consisting of a scaffold of 10 to 12 nm micro fibrils made up of several distinct glycoproteins, and an amorphous core consisting of the unique polymeric protein, elastin, which can be stretched over 150% of its original length and still return to its original size and shape. Elastin polymer is formed after enzymatic cross-linking of multiple molecules of the 70 to73 kDa precursor protein, tropoelastin. Tropoelastin (often referred as a“soluble elastin”) is synthesized by dermal fibroblasts and secreted in association with the 67 kDa elastin-binding protein, which acts as a molecular chaperone, protecting the highly hydrophobic tropoelastin molecules from intracellular self-aggregation and premature degradation, and facilitating their proper assembly on the microfibrillar scaffold in the extracellular space. Tropoelastin molecules are then polymerized into the insoluble elastin via lysine residues. Mature (insoluble) elastin, synthesized almost exclusively during late gestation and early childhood, is

metabolically inert and remains the most durable element of the ECM, that may last over the entire human lifespan in undisturbed tissues. The biosynthesis of elastin begins with the embryonic period and continues through adulthood. With aging, the

replenishment of elastin protein stops, and elastin progressively degenerates and separates into fragments. As a result, the skin progressively loses its elasticity, resulting in fine lines and wrinkles. Thus, there is a need to develop compositions and methods to stimulate elastogenesis.

[0021] Dermal fillers are useful in treating soft tissue condition and in other skin therapies because the fillers can replace lost endogenous matrix polymers, or enhance/facilitate the function of existing matrix polymers, in order to treat these skin conditions. In the past, such compositions have been used in cosmetic applications to fill wrinkles, lines, folds, scars, and to enhance dermal tissue, such as, e.g., to plump thin lips, or fill-in sunken eyes or shallow cheeks. Dermal fillers made from various substances have been used to treat facial wrinkles. Commonly used dermal fillers include hyaluronic acid, collagen, hydroxyapatite, poly-lactic acid, and

polymethylmethacrylate. Collagen based dermal fillers include Zyderm, Zyplast, Cosmoderm, Cosmoplast and Autologen. Polylactic acid dermal fillers include Sculptra™. Calcium hydroxylapatite dermal fillers include Radiesse™. Hyaluronic acid dermal fillers include Hylaform®, Restylane® and Juvederm™.

[0022] It is known that injection of a botulinum toxin into facial muscles can, by weakening the injected muscles, result in a decrease of hyperkinetic wrinkles in the skin overlying the paralyzed muscles. Botulinum toxin has been injected into facial muscles, such as the orbicularis oculis, corrugator supercilii and frontalis muscles for the cosmetic purpose of reducing certain facial wrinkle. A botulinum toxin type A (Allergan, Inc., BOTOX®) has been approved by the U.S. Food and Drug

Administration for the treatment of blepharospasm, strabismus, cervical dystonia, hyperhidrosis and glabellar lines.

[0023] Without wishing to be bound by theory, it is believed that presence of polyphenolic acid in the compositions prolongs the life of the dermal filler

(glycosaminoglycans, collagen, and elastin) and botulinum toxin. In addition,

Applicants observed that povidone-iodine was elastogenic, and combination of povidone-iodine and an elastogenic peptide was synergistic in stimulating elastogenesis. Without wishing to be bound by theory, it is believed that administering povidone- iodine breaks down the skin barrier, and disrupts cell-cell junctions, and facilitates penetration of elastogenic peptides into deeper layers of skin. This may result in enhanced elastogenesis, and improved appearance of skin. Further, Applicants also observed that combination of metformin and glucose stimulated elastogenesis.

[0024] Disclosed herein are dermal compositions for stimulating

elastogenesis. In some embodiments, the composition comprises a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In other embodiments, the composition comprises a polyphenolic compound and a dermal filler. In additional embodiments, the composition comprises a polyphenolic compound and botulinum toxin. In further embodiments, the composition comprises a polyphenolic compound and povidone-iodine. In other embodiments, the composition comprises a polyphenolic compound, povidone-iodine, and metformin.

[0025] In some embodiments, the composition comprises a polyphenolic compound that is present from about 0.01 wt. % to about 10 wt. % of the composition, about 0.01 wt. % to about 8 wt. % of the composition, about 0.01 wt. % to about 6 wt. % of the composition, about 0.01 wt. % to about 4 wt. % of the composition, about 0.01 wt. % to about 2 wt. % of the composition, or about 0.01 wt. % to about 1 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 1 wt. %, about 2 wt. %, about 5 wt. %, about 9 wt. %, about 10 wt. %, and any individual amount or any ranges between any two of these values. Non-limiting examples of polyphenolic acid are gallic acid, cinnamic acid, caffeic acid, ferulic acid, tannic acid, ellagic acid, coumoric acid, and their derivatives. In some embodiments, the

polyphenolic compound is tannic acid, ellagic acid, or combinations thereof.

[0026] In some embodiments, povidone-iodine (also called Betadine®) is present from about 0.01 wt. % to about 10 wt. % of the composition, about 0.01 wt. % to about 8 wt. % of the composition, about 0.01 wt. % to about 6 wt. % of the

composition, about 0.01 wt. % to about 4 wt. % of the composition, about 0.01 wt. % to about 2 wt. % of the composition, or about 0.01 wt. % to about 1 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 1 wt. %, about 2 wt. %, about 5 wt. %, about 9 wt. %, about 10 wt. %, and any individual amount or any ranges between any two of these values. Povidone-iodine is a

polyvinylpyrrolidinone-iodine complex usually contains approximately 85% povidone, 10% iodine, and 5% iodide. In some embodiments, the average molecular weight of polyvinylpyrrolidinone may be low molecular weight ranges, such as 20 kD, 30 kD, 40 kD, 50 kD, or 100 kD. In some embodiments, the average molecular weight of polyvinylpyrrolidinone may be in the range from about 15,000 kD to about 50,000 kD.

[0027] In some embodiments, potassium iodide is present from about 0.01 wt. % to about 10 wt. % of the composition, about 0.01 wt. % to about 8 wt. % of the composition, about 0.01 wt. % to about 6 wt. % of the composition, about 0.01 wt. % to about 4 wt. % of the composition, about 0.01 wt. % to about 2 wt. % of the composition, or about 0.01 wt. % to about 1 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 1 wt. %, about 2 wt. %, about 5 wt. %, about 9 wt. %, about 10 wt. %, and any individual amount or any ranges between any two of these values.

[0028] In some embodiments, the composition comprises metformin present from about 2 wt. % to about 20 wt. % of the composition, about 2 wt. % to about 15 wt. % of the composition, about 2 wt. % to about 10 wt. % of the composition, about 2 wt. % to about 8 wt. % of the composition, about 2 wt. % to about 6 wt. % of the

composition, or about 2 wt. % to about 4 wt. % of the composition. Specific examples include about 2 wt. %, about 4 wt. %, about 5 wt. %, about 10 wt. %, about 12 wt. %, about 15 wt. %, about 20 wt. %, and any individual amount or any ranges between any two of these values. In some embodiments, metformin is present from about 1

microgram/mL to about 100 micrograms/mL, about 1 microgram/mL to about 75 micrograms/mL, about 1 microgram/mL to about 50 micrograms/mL, about 1

microgram/mL to about 25 micrograms/mL, or about 1 microgram/mL to about 10 micrograms/mL. Specific examples include about 1 microgram/mL, about 5

micrograms/mL, about 10 micrograms/mL, about 20 micrograms/mL, about 50 micrograms/mL, about 75 micrograms/mL, about 100 micrograms/mL, and any individual amount or any ranges between any two of these values. In some

embodiments, metformin may be free metformin or metformin salts, such as metformin hydrochloride, phosphate, sulfate, hydrobromide, salicylate, maleate, benzoate, succinate, ethanesulfonate, fumarate and glycolate salts of metformin, metformin orotate, metformin clofibrate, metformin acetylsalicylate, metformin nicotinate, metformin adamantoate, sulfamido aryloxyalkyl carboxylic acid salts of metformin, and combinations thereof.

[0029] In some embodiments, the composition comprises botulinum toxin selected from the group consisting of botulinum toxin type A, botulinum toxin type B, botulinum toxin type C, botulinum toxin type D, botulinum toxin type E, botulinum toxin type F, abobotulinumtoxinA, BTX-A, daxibotulinumtoxinA, incobotulinumtoxinA, onabotulinumtoxinA, rimabotulinumtoxinB, syntaxin, and derivatives thereof, including, but not limited to, engineered recombinant botulinum toxins and fragments thereof.

[0030] In some embodiments, the composition comprises an effective amount of botulinum toxin and the effective amount may be 0.1 units, 0.2 units, 0.3 units, 0.4 units, 0.5 units, 0.6 units, 0.7 units, 0.8 units, 0.8 units, 0.9 units, 1.0 unit, 2 units, 3, units, 4 units, 5 units, 6 units, 7 units, 8 units, 9 units, 10 units, 1 1 units, 12 units, 13 units, 14 units, 15 units, 16 units, 17 units, 18 units, 19 units, 20 units, 25 units, 30 units, 35 units, 40 units, 45 units, 50 units, 100 units, 125 units, 150 units, 175 units, 200 units, 225 units, 250 units, 275 units, 300 units, 325 units, 350 units, 375 units, 400 units, 425 units, 450 units, 475 units, 500 units and any individual amount or any ranges between any two of these values.

[0031] In some embodiments, the composition comprises an effective amount of botulinum toxin and is present in the composition from about 0.1 wt. % to about 25 wt. %, about 0.1 wt. % to about 20 wt. %, about 0.1 wt. % to about 15 wt. %, about 0.1 wt. % to about 10 wt. %, about 0.1 wt. % to about 8 wt. %, about 0.1 wt. % to about 5 wt. %, about 0.1 wt. % to about 4 wt. %, about 0.1 wt. % to about 3 wt. %, about 0.1 wt. % to about 3 wt. %, or about 0.1 wt. % to about 1 wt. %. Specific examples include about 0.1 wt. %, about 0.5 wt. %, about 1 wt. %, about 2 wt. %, about 5 wt. %, about 10 wt. %, about 25 wt. %, and any individual amount or any ranges between any two of these values.

Dermal Fillers

[0032] In some embodiments, the dermal filler is selected from hyaluronic acid, collagen, heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, elastin, or a combination thereof.

[0033] In some embodiments, the composition comprises hyaluronic acid and is present from about 0.01 wt. % to about 85 wt. % of the composition, about 0.01 wt. % to about 70 wt. % of the composition, about 0.01 wt. % to about 60 wt. % of the composition, about 0.01 wt. % to about 40 wt. % of the composition, about 0.01 wt. % to about 20 wt. % of the composition, or about 0.01 wt. % to about 10 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 10 wt. %, about 20 wt. %, about 50 wt. %, about 75 wt. %, about 85 wt. %, and any individual amount or any ranges between any two of these values. As used herein, the term “hyaluronic acid” is synonymous with“hyaluronic acid polymer”,“HA polymer”, “hyaluronan” and“hyaluronate polymer”, and refers to an anionic, non-sulfated glycosaminoglycan polymer comprising disaccharide units, which themselves include D-glucuronic acid and D-N-acetylglucosamine monomers, linked together via alternating b- 1,4 and b-1,3 glycosidic bonds and pharmaceutically acceptable salts thereof. Hyaluronic acid polymers can be purified from animal and non-animal sources. Polymers of hyaluronic acid can range in size from about 5,000 Da to about 20,000,000 Da. Any hyaluronic acid polymer is useful in the compositions disclosed herein with the proviso that the hyaluronic acid improves a condition of the skin. Non-limiting examples of pharmaceutically acceptable salts of hyaluronic acid include sodium hyaluronate, potassium hyaluronate, magnesium hyaluronate, calcium hyaluronate, and combinations thereof.

[0034] In some embodiments, the hyaluronic acid present in the composition may be uncross-linked or cross-linked, or a mixture of both uncross-linked and cross- linked. For example, about 10 wt.% of hyaluronic acid in the composition is cross- linked, about 15 wt.% of hyaluronic acid in the composition is cross-linked, about 25 wt.% of hyaluronic acid in the composition is cross-linked, about 35 wt.% of hyaluronic acid in the composition is cross-linked, about 45 wt.% of hyaluronic acid in the composition is cross-linked, about 50 wt.% of hyaluronic acid in the composition is cross-linked, about 75 wt.% of hyaluronic acid in the composition is cross-linked, or about 85 wt.% of hyaluronic acid in the composition is cross-linked.

[0035] The cross-linking can be performed by any cross-linking agent known in the art, such as proanthocyanidin, a bifimctional epoxide, a carbodiimide,

glutaraldehyde, or a periodate. If the cross-linking agent is a proanthocyanidin, it is typically selected from the group consisting of proanthocyanidin, procyanidin (2H-1- benzopyran-3,4,5,7-tetrol, 2-(3,4-dihydroxyphenyl)-2-((2-(3,4-dihydroxyphenyl)-3,4- dihydro-5 ,7-dihydroxy-2H- 1 -benzopyran-3 -yl)oxy)-3 ,4-dihydro), procyanidin B, procyanidin B2, rhatannin, procyanidol oligomer, procyanidin C, procyanidin B3, procyanidin Bl, selligueain A (8,l4-methano-2H,l4H-l-benzopyrano(7,8-d)

(l,3)benzodioxocin-3,5,l 1, 13,15 -pentol, 4-(3,4-dihydro-3,5,7-trihydroxy-2-(4- hydroxyphenyl)-2H-l-benzopyran-8-yl)-3,4-dihydro-2,8-bis(4-hydroxyphenyl)-, (2R- (2a,3a,4P(2R*,3S*),8p, 14b, 15R*)), geranin A, geranin D, procyanidin B5, procyanidin B5-3'-O-gallate, vitisinol, amurensisin, terminalin, geranin B, 6,8-dihydroxyafzelin, afzelin-3"-O-gallate, geranin C, afzelin, flavangenol, carallidin, mahuannin A, proanthocyanidin Al, proanthocyanidin A2, procyanidin D, and analogues, derivatives, and bioisosteres of these compounds. If the cross-linking agent is a carbodiimide, the carbodiimide is typically selected from the group consisting of l-ethyl-3-(3- dimethylaminopropyl) carbodiimide hydrochloride (EDC), dicyclohexylcarbodiimide (DCC), l-ethyl-3-(4-azonia-4,4-dimethylpentyl) carbodiimide iodide (EAC); 1- cyclohexyl-3 -(2-morpholinyl-(4)-ethyl-carbodiimide metho-p-toluene sulfonate (CMC), and N-benzyl-N'-3-dimethylaminopropyl-carbodiimide hydrochloride (BDC).

Preferably, the carbodiimide is 1 -ethyl-3 -(3 -dimethylaminopropyl) carbodiimide hydrochloride (EDC).

[0036] In some embodiments, the composition comprises collagen and/or collagen fragments that is present from about 0.01 wt. % to about 85 wt. % of the composition, about 0.01 wt. % to about 70 wt. % of the composition, about 0.01 wt. % to about 60 wt. % of the composition, about 0.01 wt. % to about 40 wt. % of the composition, about 0.01 wt. % to about 20 wt. % of the composition, or about 0.01 wt. % to about 10 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 10 wt. %, about 20 wt. %, about 50 wt. %, about 75 wt. %, about 85 wt. %, and any individual amount or any ranges between any two of these values. Collagen can be isolated in a various forms and from a number of sources. Example collagens include collagen type I, collagen type II, collagen type III, collagen type IV, or collagen type Y. The collagen (and fragments thereof) can also be fibrillary collagen or non-fibrillar collagen. In some embodiments, the collagen (and fragments thereof) may be a uncross-linked or cross-linked collagen, or a mixture of uncross-linked and cross-linked collagen, and the cross-linking can be performed by any cross-linking agent disclosed herein. For example, about 10 wt.% of collagen in the composition is cross-linked, about 15 wt.% of collagen in the composition is cross-linked, about 25 wt.% of collagen in the composition is cross-linked, about 35 wt.% of collagen in the composition is cross-linked, about 45 wt.% of collagen in the composition is cross- linked, about 50 wt.% of collagen in the composition is cross-linked, about 75 wt.% of collagen in the composition is cross-linked, or about 85 wt.% of collagen in the composition is cross-linked.

[0037] In some embodiments, the composition comprises heparin that is present from about 0.01 wt. % to about 85 wt. % of the composition, about 0.01 wt. % to about 70 wt. % of the composition, about 0.01 wt. % to about 60 wt. % of the composition, about 0.01 wt. % to about 40 wt. % of the composition, about 0.01 wt. % to about 20 wt. % of the composition, or about 0.01 wt. % to about 10 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 10 wt. %, about 20 wt. %, about 50 wt. %, about 75 wt. %, about 85 wt. %, and any individual amount or any ranges between any two of these values. Heparin may be obtained from any source, and may have molecular weight ranging from 3 Da to 30,000 Da. In some embodiments, the heparin may be uncross-linked or cross-linked, or a mixture of both uncross-linked and cross-linked, and the cross-linking can be performed by any cross- linking agent disclosed herein. For example, about 10 wt.% of heparin in the

composition is cross-linked, about 15 wt.% of heparin in the composition is cross- linked, about 25 wt.% of heparin in the composition is cross-linked, about 35 wt.% of heparin in the composition is cross-linked, about 45 wt.% of heparin in the composition is cross-linked, about 50 wt.% of heparin in the composition is cross-linked, about 75 wt.% of heparin in the composition is cross-linked, or about 85 wt.% of heparin in the composition is cross-linked.

[0038] In some embodiments, the composition comprises heparan sulfate that is present from about 0.01 wt. % to about 85 wt. % of the composition, about 0.01 wt. % to about 70 wt. % of the composition, about 0.01 wt. % to about 60 wt. % of the composition, about 0.01 wt. % to about 40 wt. % of the composition, about 0.01 wt. % to about 20 wt. % of the composition, or about 0.01 wt. % to about 10 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 10 wt. %, about 20 wt. %, about 50 wt. %, about 75 wt. %, about 85 wt. %, and any individual amount or any ranges between any two of these values. Heparan sulfate may be obtained from any source, and may have molecular weight ranging from 3 Da to 30,000 Da. In some embodiments, heparan sulfate may be uncross-linked or cross-linked, or a mixture of both uncross-linked and cross-linked, and the cross-linking can be performed by any cross-linking agent disclosed herein. For example, about 10 wt.% of heparan sulfate in the composition is cross-linked, about 15 wt.% of heparan sulfate in the composition is cross-linked, about 25 wt.% of heparan sulfate in the composition is cross-linked, about 35 wt.% of heparan sulfate in the composition is cross-linked, about 45 wt.% of heparan sulfate in the composition is cross-linked, about 50 wt.% of heparan sulfate in the composition is cross-linked, about 75 wt.% of heparan sulfate in the composition is cross-linked, or about 85 wt.% of heparan sulfate in the composition is cross-linked.

[0039] In some embodiments, the composition comprises chondroitin sulfate that is present from about 0.01 wt. % to about 85 wt. % of the composition, about 0.01 wt. % to about 70 wt. % of the composition, about 0.01 wt. % to about 60 wt. % of the composition, about 0.01 wt. % to about 40 wt. % of the composition, about 0.01 wt. % to about 20 wt. % of the composition, or about 0.01 wt. % to about 10 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 10 wt. %, about 20 wt. %, about 50 wt. %, about 75 wt. %, about 85 wt. %, and any individual amount or any ranges between any two of these values. Chondroitin sulfate may be obtained from any source, and may have molecular weight ranging from 3000 Da to 50,000 Da. In some embodiments, the chondroitin sulfate may be uncross-linked or cross-linked, or a mixture of both uncross-linked and cross-linked, and the cross-linking can be performed by any cross-linking agent disclosed herein. For example, about 10 wt.% of chondroitin sulfate in the composition is cross-linked, about 15 wt.% of chondroitin sulfate in the composition is cross-linked, about 25 wt.% of chondroitin sulfate in the composition is cross-linked, about 35 wt.% of chondroitin sulfate in the composition is cross-linked, about 45 wt.% of chondroitin sulfate in the composition is cross-linked, about 50 wt.% of chondroitin sulfate in the composition is cross-linked, about 75 wt.% of chondroitin sulfate in the composition is cross-linked, or about 85 wt.% of chondroitin sulfate in the composition is cross-linked.

[0040] In some embodiments, the composition comprises dermatan sulfate that is present from about 0.01 wt. % to about 85 wt. % of the composition, about 0.01 wt. % to about 70 wt. % of the composition, about 0.01 wt. % to about 60 wt. % of the composition, about 0.01 wt. % to about 40 wt. % of the composition, about 0.01 wt. % to about 20 wt. % of the composition, or about 0.01 wt. % to about 10 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 10 wt. %, about 20 wt. %, about 50 wt. %, about 75 wt. %, about 85 wt. %, and any individual amount or any ranges between any two of these values. Dermatan sulfate may be obtained from any source, and may have molecular weight ranging from 3000 Da to 50,000 Da. In some embodiments, dermatan sulfate may be uncross-linked or cross- linked, or a mixture of both uncross-linked and cross-linked, and the cross-linking can be performed by any cross-linking agent disclosed herein. For example, about 10 wt.% of dermatan sulfate in the composition is cross-linked, about 15 wt.% of dermatan sulfate in the composition is cross-linked, about 25 wt.% of dermatan sulfate in the composition is cross-linked, about 35 wt.% of dermatan sulfate in the composition is cross-linked, about 45 wt.% of dermatan sulfate in the composition is cross-linked, about 50 wt.% of dermatan sulfate in the composition is cross-linked, about 75 wt.% of dermatan sulfate in the composition is cross-linked, or about 85 wt.% of dermatan sulfate in the composition is cross-linked.

[0041] In some embodiments, the composition comprises keratan sulfate that is present from about 0.01 wt. % to about 85 wt. % of the composition, about 0.01 wt. % to about 70 wt. % of the composition, about 0.01 wt. % to about 60 wt. % of the composition, about 0.01 wt. % to about 40 wt. % of the composition, about 0.01 wt. % to about 20 wt. % of the composition, or about 0.01 wt. % to about 10 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 10 wt. %, about 20 wt. %, about 50 wt. %, about 75 wt. %, about 85 wt. %, and any individual amount or any ranges between any two of these values. Keratan sulfate may be obtained from any source, and may have molecular weight ranging from 3000 Da to 50,000 Da. In some embodiments, keratan sulfate may be uncross-linked or cross-linked, or a mixture of both uncross-linked and cross-linked, and the cross-linking can be performed by any cross-linking agent disclosed herein. For example, about 10 wt.% of keratan sulfate in the composition is cross-linked, about 15 wt.% of keratan sulfate in the composition is cross-linked, about 25 wt.% of keratan sulfate in the composition is cross-linked, about 35 wt.% of keratan sulfate in the composition is cross-linked, about 45 wt.% of keratan sulfate in the composition is cross-linked, about 50 wt.% of keratan sulfate in the composition is cross-linked, about 75 wt.% of keratan sulfate in the composition is cross-linked, or about 85 wt.% of keratan sulfate in the composition is cross-linked.

[0042] In some embodiments, the composition comprises elastin (and fragments thereof) that is present from about 0.01 wt. % to about 85 wt. % of the composition, about 0.01 wt. % to about 70 wt. % of the composition, about 0.01 wt. % to about 60 wt. % of the composition, about 0.01 wt. % to about 40 wt. % of the composition, about 0.01 wt. % to about 20 wt. % of the composition, or about 0.01 wt.

% to about 10 wt. % of the composition. Specific examples include about 0.01 wt. %, about 0.1 wt. %, about 10 wt. %, about 20 wt. %, about 50 wt. %, about 75 wt. %, about 85 wt. %, and any individual amount or any ranges between any two of these values. Elastin may be obtained from any source, and may have molecular weight ranging from 3000 Da to 150,000 Da. In some embodiments, elastin (and fragments thereof) may be uncross-linked or cross-linked, or a mixture of both uncross-linked and cross-linked, and the cross-linking can be performed by any cross-linking agent disclosed herein. For example, about 10 wt.% of elastin in the composition is cross-linked, about 15 wt.% of elastin in the composition is cross-linked, about 25 wt.% of elastin in the composition is cross-linked, about 35 wt.% of elastin in the composition is cross-linked, about 45 wt.% of elastin in the composition is cross-linked, about 50 wt.% of elastin in the composition is cross-linked, about 75 wt.% of elastin in the composition is cross-linked, or about 85 wt.% of elastin in the composition is cross-linked. [0043] In some embodiments, the compositions do not contain dermal fillers such as chondroitin sulfate, dermatan sulfate, and keratin sulfate. In some embodiments, the compositions. In some embodiments, the composition comprises a polyphenolic compound and a dermal filler selected from hyaluronic acid, collagen, heparin, and heparan sulfate.

Additional Agents

[0044] In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, elastogenic peptides, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0045] In some embodiments, the composition comprises glucose present from about 2 wt. % to about 20 wt. % of the composition, about 2 wt. % to about 15 wt. % of the composition, about 2 wt. % to about 10 wt. % of the composition, about 2 wt. % to about 8 wt. % of the composition, about 2 wt. % to about 6 wt. % of the

composition, or about 2 wt. % to about 4 wt. % of the composition. Specific examples include about 2 wt. %, about 4 wt. %, about 5 wt. %, about 10 wt. %, about 12 wt. %, about 15 wt. %, about 20 wt. %, and any individual amount or any ranges between any two of these values. In some embodiments, the glucose is present in composition at a concentration from about 0.5 micromolar to about 100 micromolar, about 0.5

micromolar to about 75 micromolar, about 0.5 micromolar to about 50 micromolar, about 0.5 micromolar to about 20 micromolar, or about 0.5 micromolar to about 10 micromolar.

[0046] In some embodiments, the composition comprises a copper compound present from about 2 wt. % to about 20 wt. % of the composition, about 2 wt. % to about 15 wt. % of the composition, about 2 wt. % to about 10 wt. % of the composition, about 2 wt. % to about 8 wt. % of the composition, about 2 wt. % to about 6 wt. % of the composition, or about 2 wt. % to about 4 wt. % of the composition. Specific examples include about 2 wt. %, about 4 wt. %, about 5 wt. %, about 10 wt. %, about 12 wt. %, about 15 wt. %, about 20 wt. %, and any individual amount or any ranges between any two of these values. Non-limiting examples of copper compound include copper sulfate, copper phosphate, copper chloride, copper iodide, and copper oxide.

[0047] In some embodiments, the composition comprises a divalent

manganese compound present from about 2 wt. % to about 20 wt. % of the composition, about 2 wt. % to about 15 wt. % of the composition, about 2 wt. % to about 10 wt. % of the composition, about 2 wt. % to about 8 wt. % of the composition, about 2 wt. % to about 6 wt. % of the composition, or about 2 wt. % to about 4 wt. % of the composition. Specific examples include about 2 wt. %, about 4 wt. %, about 5 wt. %, about 10 wt. %, about 12 wt. %, about 15 wt. %, about 20 wt. %, and any individual amount or any ranges between any two of these values. In some embodiments, the divalent manganese compound is present in composition at a concentration from about 0.5 micromolar to about 100 micromolar, about 0.5 micromolar to about 75 micromolar, about 0.5 micromolar to about 50 micromolar, about 0.5 micromolar to about 20 micromolar, or about 0.5 micromolar to about 10 micromolar. Non-limiting examples of a divalent manganese compound include manganese ascorbate, manganese-PCA (manganese salt of L-pyrrolidone carboxylic acid), manganese chloride, manganese nitrate, manganese sulfate, manganese gluconate, and combinations thereof.

[0048] In some embodiments, the composition comprises a trivalent iron compound present from about 2 wt. % to about 20 wt. % of the composition, about 2 wt. % to about 15 wt. % of the composition, about 2 wt. % to about 10 wt. % of the composition, about 2 wt. % to about 8 wt. % of the composition, about 2 wt. % to about 6 wt. % of the composition, or about 2 wt. % to about 4 wt. % of the composition.

Specific examples include about 2 wt. %, about 4 wt. %, about 5 wt. %, about 10 wt. %, about 12 wt. %, about 15 wt. %, about 20 wt. %, and any individual amount or any ranges between any two of these values. In some embodiments, the trivalent iron compound is present in composition at a concentration from about 0.5 micromolar to about 100 micromolar, about 0.5 micromolar to about 75 micromolar, about 0.5 micromolar to about 50 micromolar, about 0.5 micromolar to about 20 micromolar, or about 0.5 micromolar to about 10 micromolar. Non-limiting examples of a trivalent iron compound include ferric ammonium citrate and ferric chloride.

[0049] In some embodiments, the composition comprises aldosterone present from about 2 wt. % to about 20 wt. % of the composition, about 2 wt. % to about 15 wt. % of the composition, about 2 wt. % to about 10 wt. % of the composition, about 2 wt. % to about 8 wt. % of the composition, about 2 wt. % to about 6 wt. % of the

composition, or about 2 wt. % to about 4 wt. % of the composition. Specific examples include about 2 wt. %, about 4 wt. %, about 5 wt. %, about 10 wt. %, about 12 wt. %, about 15 wt. %, about 20 wt. %, and any individual amount or any ranges between any two of these values. In some embodiments, the aldosterone is present in composition at a concentration from about 0.05 micromolar to about 10 micromolar, about 0.05 micromolar to about 5 micromolar, about 0.05 micromolar to about 2 micromolar, about 0.05 micromolar to about 1 micromolar, or about 0.05 micromolar to about 0.1 micromolar.

[0050] In some embodiments, the composition comprises sodium ascorbate present from about 2 wt. % to about 20 wt. % of the composition, about 2 wt. % to about 15 wt. % of the composition, about 2 wt. % to about 10 wt. % of the composition, about 2 wt. % to about 8 wt. % of the composition, about 2 wt. % to about 6 wt. % of the composition, or about 2 wt. % to about 4 wt. % of the composition. Specific examples include about 2 wt. %, about 4 wt. %, about 5 wt. %, about 10 wt. %, about 12 wt. %, about 15 wt. %, about 20 wt. %, and any individual amount or any ranges between any two of these values. In some embodiments, sodium ascorbate is present in composition at a concentration from about 0.5 micromolar to about 200 micromolar, about 0.5 micromolar to about 100 micromolar, about 0.5 micromolar to about 50 micromolar, about 0.5 micromolar to about 20 micromolar, or about 0.5 micromolar to about 10 micromolar.

[0051] In some embodiments, the composition comprises one or more elastogenic peptides. The term“elastogenic peptides” includes peptides that stimulate elastin protein synthesis. Non-limiting examples of elastogenic peptides include GAAPG (SEQ ID NO: 1), GWPG (SEQ ID NO: 2), GGGPG (SEQ ID NO: 3),

GLLPG (SEQ ID NO: 4), GIIPG (SEQ ID NO: 5), GSSPG (SEQ ID NO: 6), GTTPG (SEQ ID NO: 7), GCCPG (SEQ ID NO: 8), GMMPG (SEQ ID NO: 9), GFFPG (SEQ ID NO: 10), GYYPG (SEQ ID NO: 11), GWWPG (SEQ ID NO: 12), GDDPG (SEQ ID NO: 13), GNNPG (SEQ ID NO: 14), GEEPG (SEQ ID NO: 15), GQQPG (SEQ ID NO: 16), GRRPG (SEQ ID NO: 17), GHHPG (SEQ ID NO: 18), GKKPG (SEQ ID NO: 19), GPPPG (SEQ ID NO: 20), G3Hyp3HypPG (Glycine-3-hydroxyproline-3- hydroxyproline-Proline-Glycine) (SEQ ID NO: 21), G4Hyp4HypPG (Glycine-4- hydroxyproline-4-hydroxyproline-Proline-Glycine) (SEQ ID NO: 22), RRPEV (SEQ ID NO: 23), QPSQPGGY (SEQ ID NO: 24), PGGY (SEQ ID NO: 25), GPGV (SEQ ID NO: 26), KPGV (SEQ ID NO: 27), GPGL (SEQ ID NO: 28), EGSA (SEQ ID NO: 29), PGGF (SEQ ID NO: 30), GGGA (SEQ ID NO: 31), KPGKV (SEQ ID NO: 32), PGGV (SEQ ID NO: 33), KPKA (SEQ ID NO: 34), GPGGV (SEQ ID NO: 35), GPQA (SEQ ID NO: 36), GGPGI (SEQ ID NO: 37), PGPGA (SEQ ID NO: 38), GPGGV (SEQ ID NO: 39), GQPF (SEQ ID NO: 40), GGKPPKPF (SEQ ID NO: 41), GGQQPGL (SEQ ID NO: 42), GGPGI (SEQ ID NO: 43), VGVAPG (SEQ ID NO: 44), IGVAPG (SEQ ID NO: 45), PGGVLPG (SEQ ID NO: 46), VGVVPG (SEQ ID NO: 47), IGLGPGGV (SEQ ID NO: 48), VGAMPG (SEQ ID NO: 49), VGLSPG (SEQ ID NO: 50), IGAMPG (SEQ ID NO: 51), IGLSPG (SEQ ID NO: 52), GVAPGV (SEQ ID NO: 53), VAPGVG (SEQ ID NO: 54), APGVGV (SEQ ID NO: 55), PGVGVA (SEQ ID NO: 56),

GVGVAP (SEQ ID NO: 57), VGLMPG (SEQ ID NO: 58), VGVMPG (SEQ ID NO: 59), VGASPG (SEQ ID NO: 60), VGAAPG (SEQ ID NO: 61), IGLMPG (SEQ ID NO: 62), IGVMPG (SEQ ID NO: 63), IGASPG (SEQ ID NO: 64), IGAAPG (SEQ ID NO: 65), IGLSPG (SEQ ID NO: 66), IGLAPG (SEQ ID NO: 67), IGVSPG (SEQ ID NO:

68), VGLAPG (SEQ ID NO: 69), VGVSPG (SEQ ID NO: 70), and combinations thereof. In some embodiments, the peptides are acetylated at N-or C-terminus. In some embodiments, the elastogenic peptide is N-terminal acetylated IGVAPG (SEQ ID NO: 71). Additional examples of elastogenic peptides as described in U.S. Patent Nos.

7803522, 7666829, 7566693, 7723308, and are incorporated herein by reference.

[0052] In some embodiments, the elastogenic peptide may be of Formula I:

X 1 -X2-X3 -X4-X5 -X6, wherein XI is V or I; X2 is G or V; X3 is A, V, or L; X4 is M, S, A, or T; X5 is A or P; and X6 is G (SEQ ID NO: 72). In some embodiments, XI is V or I; X2 is G; X3 is A or L; X4 is M or S; X5 is P; and X6 is G.

[0053] In some embodiments, the elastogenic peptide may be of Formula II: X1-X2-X3-X4-X5-X6-Z-X1-X2-X3-X4-X5-X6, wherein Z is a linking moiety; XI is independently selected from V and I; X2 is G; X3 is independently selected from A and L; X4 is independently selected from M and S; X5 is P; and X6 is G (SEQ ID NOS: 73- linker-73). The linking moiety Z may be comprised of, for example, but not limited to, at least one of alanine or any other amino acid, a disulfide bond, a carbonyl moiety, a hydrocarbon moiety optionally substituted at one or more available carbon atoms with a lower alkyl substituent. In some embodiments, the linking moiety Z is a lysine residue or lysine amide, i.e., a lysine residue wherein the carboxyl group has been converted to an amide moiety -CONH. Non-limiting example of peptides of Formula II include V GAMPGA AA AAV GAMPG (SEQ ID NO: 74), VGLSPGAAAAAVGLSPG(SEQ ID NO: 75), V GY APGA A A A A V GY APG (SEQ ID NO: 76), IGAMPGAAAAAIGAMPG (SEQ ID NO: 77), IGLSPGAAAAAIGLSPG (SEQ ID NO: 78), and

IGVAPGAAAAAIGV APG (SEQ ID NO: 79).

[0054] In some embodiments, the elastogenic peptides disclosed herein may be modified peptides such that any amino acid residue of the elastogenic peptide may carry a substitution selected from acetyl, amide, carbonyl, imide, thiol, alkyl; alkenyl; alkynyl; cycloalkyl; sulfonyl; sulfinyl; silyl; a fatty acid; pyroglutamyl; isocyanate, alkyl carbonyl, alkyl, a cylcloalkyl, thioester, urea, carbamate, sulfonamide, alkylamine, aryl, alkylaryl, heteroaryl, alky heteroaryl; furazanyl, ftiryl, imidazolidinyl, imidazolyl, imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl, oxazolyl, piperazinyl, and

combinations thereof. In some embodiments the elastogenic peptides disclosed herein may be substituted with D-amino acids.

[0055] In some embodiments, the elastogenic peptides disclosed herein are acetylated. In some embodiments, the elastogenic peptides are acetylated at the N- terminus. [0056] In some embodiments, the elastogenic peptides may be elastin protein fragments. Elastin fragments may be obtained commercially or may be generated by protease digestion, such as using proteinase K or thermolysin. For example,

commercially available Elastin E91 preparation from Protein Preparations, Inc., St. Louis, MO, is a suitable elastin product to subject to digestion, having about 1,000 to about 60,000 Dalton molecular weight. Additionally, a series of digests available under the trade name ProK, and specifically ProK-60 and ProK-60P, which are elastin peptide mixtures derived from the proteolytic digestion of insoluble elastin derived from bovine neck ligaments can also be used.

[0057] The elastogenic peptides disclosed herein may be present from about 2 wt. % to about 60 wt. % of the composition, about 2 wt. % to about 50 wt. % of the composition, about 2 wt. % to about 40 wt. % of the composition, about 2 wt. % to about 30 wt. % of the composition, about 2 wt. % to about 20 wt. % of the composition, or about 2 wt. % to about 10 wt. % of the composition. Specific examples include about 2 wt. %, about 10 wt. %, about 20 wt. %, about 30 wt. %, about 40 wt. %, about 50 wt. %, about 60 wt. %, and any individual amount or any ranges between any two of these values. In some embodiments, the elastogenic peptides are present from about 1 mg/mL to about 100 mg/mL, about 1 mg/mL to about 75 mg/mL, about 1 mg/mL to about 50 mg/mL, about 1 mg/mL to about 25 mg/mL, or about 1 mg/mL to about 10 mg/mL. Specific examples include about 1 mg/mL, about 5 mg/mL, about 10 mg/mL, about 20 mg/mL, about 50 mg/mL, about 75 mg/mL, about 100 mg/mL, and any individual amount or any ranges between any two of these values.

[0058] The elastogenic peptides disclosed herein may be synthesized by suitable methods such as by exclusive solid phase techniques, by partial solid-phase techniques, by fragment condensation or by classical solution phase synthesis. For example, the techniques of exclusive solid-phase synthesis are set forth in the textbook “Solid Phase Peptide Synthesis”, J.M. Stewart and J.D. Young, Pierce Chem. Company, Rockford, Illinois, 1984 (2nd. ed.), and M. Bodanszky,“Principles of Peptide

Synthesis”, Springer Yerlag, 1984. [0059] In some embodiments, the compositions may include an analgesic selected from the group consisting of lidocaine, benzocaine, tetracaine, bupivacaine, ***e, etidocaine, flecainide, mepivacaine, pramoxine, prilocaine, procaine, chloroprocaine, oxyprocaine, proparacaine, ropivacaine, dyclonine, dibucaine, propoxycaine, chloroxylenol, cinchocaine, dexivacaine, diamocaine, hexylcaine, levobupivacaine, propoxycaine, pyrrocaine, risocaine, rodocaine, and derivatives and stereoisomers thereof. The analgesic may be present from about 2 wt. % to about 20 wt. % of the composition, about 2 wt. % to about 15 wt. % of the composition, about 2 wt. % to about 10 wt. % of the composition, about 2 wt. % to about 8 wt. % of the composition, about 2 wt. % to about 6 wt. % of the composition, or about 2 wt. % to about 4 wt. % of the composition. Specific examples include about 2 wt. %, about 4 wt. %, about 5 wt. %, about 10 wt. %, about 12 wt. %, about 15 wt. %, about 20 wt. %, and any individual amount or any ranges between any two of these values.

[0060] In some embodiments, the compositions optionally comprise analgesics. In some embodiments, the compositions are free of analgesics.

[0061] In some embodiments, the composition comprises tetracycline from about 2 wt. % to about 20 wt. % of the composition, about 2 wt. % to about 15 wt. % of the composition, about 2 wt. % to about 10 wt. % of the composition, about 2 wt. % to about 8 wt. % of the composition, about 2 wt. % to about 6 wt. % of the composition, or about 2 wt. % to about 4 wt. % of the composition. Specific examples include about 2 wt. %, about 4 wt. %, about 5 wt. %, about 10 wt. %, about 12 wt. %, about 15 wt. %, about 20 wt. %, and any individual amount or any ranges between any two of these values.

[0062] Other polyphenolic compounds, elastin peptides, dermal fillers, and additional agents are described in U.S. Patent Nos. 7,803,522; 7,566,693; 7,666,829; 7,723,308; 7,745,225; 8, 148,327; 8,618,084; 9,387, 194, and are incorporated herein by reference.

[0063] In some embodiments, the composition comprises an agent that inhibits collagen deposition or formation, present from about 2 wt. % to about 50 wt. % of the composition, about 2 wt. % to about 40 wt. % of the composition, about 2 wt. % to about 30 wt. % of the composition, about 2 wt. % to about 20 wt. % of the

composition, about 2 wt. % to about 10 wt. % of the composition, or about 2 wt. % to about 5 wt. % of the composition. Specific examples include about 2 wt. %, about 10 wt. %, about 15 wt. %, about 25 wt. %, about 30 wt. %, about 40 wt. %, about 50 wt. %, and any individual amount or any ranges between any two of these values. In some embodiments, the agent that inhibits collagen deposition or formation is present in composition at a concentration from about 0.5 micromolar to about 200 micromolar, about 0.5 micromolar to about 100 micromolar, about 0.5 micromolar to about 50 micromolar, about 0.5 micromolar to about 20 micromolar, or about 0.5 micromolar to about 10 micromolar. Non-limiting examples of an agent that inhibits collagen deposition or formation include spironolactone, eplerenone, canrenone, and

dimethyloxaloylglycine (DMOG).

[0064] In some embodiments, the composition comprises povidone-iodine from about 0.01 wt. % to about 10 wt. % of the composition, a polyphenolic compound from about 0.01 wt. % to about 10 wt. % of the composition, and one or more

elastogenic peptides from about 2 wt.% to about 60 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese

compound, a trivalent iron compound, aldosterone, an agent that inhibits collagen deposition or formation, sodium ascorbate, or combinations thereof.

[0065] In some embodiments, the composition comprises povidone-iodine from about 0.01 wt. % to about 5 wt. % of the composition, a polyphenolic compound from about 0.01 wt. % to about 5 wt. % of the composition, and one or more elastogenic peptides from about 2 wt.% to about 30 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, an agent that inhibits collagen deposition or formation, sodium ascorbate, or combinations thereof.

[0066] In some embodiments, the composition comprises povidone-iodine from about 0.01 wt. % to about 2.5 wt. % of the composition, a polyphenolic compound from about 0.01 wt. % to about 2.5 wt. % of the composition, and one or more elastogenic peptides from about 2 wt.% to about 15 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, an agent that inhibits collagen deposition or formation, sodium ascorbate, or combinations thereof.

[0067] In some embodiments, the composition comprises povidone-iodine from about 0.01 wt. % to about 1 wt. % of the composition, a polyphenolic compound from about 0.01 wt. % to about 1 wt. % of the composition, and one or more elastogenic peptides from about 2 wt.% to about 6 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, an agent that inhibits collagen deposition or formation, sodium ascorbate, or combinations thereof.

[0068] In some embodiments, the composition comprises a polyphenolic compound from about 0.01 wt. % to about 10 wt. % of the composition, and one or more dermal fillers from about 0.01 wt.% to about 85 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, elastogenic peptides, an analgesic, or combinations thereof.

[0069] In some embodiments, the composition comprises a polyphenolic compound from about 0.01 wt. % to about 5 wt. % of the composition, and one or more dermal fillers from about 2 wt.% to about 30 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, elastogenic peptides, an analgesic, or combinations thereof.

[0070] In some embodiments, the composition comprises a polyphenolic compound from about 0.01 wt. % to about 2.5 wt. % of the composition, and one or more dermal fillers from about 2 wt.% to about 15 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, elastogenic peptides, an analgesic, or combinations thereof.

[0071] In some embodiments, the composition comprises a polyphenolic compound from about 0.01 wt. % to about 1 wt. % of the composition, and one or more dermal fillers from about 2 wt.% to about 6 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, elastogenic peptides, an analgesic, or combinations thereof.

[0072] In some embodiments, the composition comprises a polyphenolic compound from about 0.01 wt. % to about 10 wt. % of the composition, and botulinum toxin from about 0.1 wt.% to about 25 wt. % of the composition. The composition may further comprise hyaluronic acid, elastin, collagen, elastogenic peptides, an analgesic, or combinations thereof.

[0073] In some embodiments, the composition comprises a polyphenolic compound from about 0.01 wt. % to about 5 wt. % of the composition, and botulinum toxin from about 2 wt.% to about 10 wt. % of the composition. The composition may further comprise hyaluronic acid, elastin, collagen, elastogenic peptides, an analgesic, or combinations thereof.

[0074] In some embodiments, the composition comprises a polyphenolic compound from about 0.01 wt. % to about 2.5 wt. % of the composition, and botulinum toxin from about 2 wt.% to about 5 wt. % of the composition. The composition may further comprise hyaluronic acid, elastin, collagen, elastogenic peptides, an analgesic, or combinations thereof.

[0075] In some embodiments, the composition comprises a polyphenolic compound from about 0.01 wt. % to about 1 wt. % of the composition, and botulinum toxin from about 1 wt.% to about 3 wt. % of the composition. The composition may further comprise hyaluronic acid, elastin, collagen, elastogenic peptides, an analgesic, or combinations thereof. [0076] In some embodiments, the composition comprises povidone-iodine from about 0.01 wt. % to about 10 wt. % of the composition, a polyphenolic compound from about 0.01 wt. % to about 10 wt. % of the composition, metformin from about 2 wt.% to about 20 wt. % of the composition, and glucose from about 2 wt.% to about 20 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, an agent that inhibits collagen deposition or formation, sodium ascorbate, or combinations thereof.

[0077] In some embodiments, the composition comprises povidone-iodine from about 0.01 wt. % to about 5 wt. % of the composition, a polyphenolic compound from about 0.01 wt. % to about 5 wt. % of the composition, metformin from about 2 wt.% to about 10 wt. % of the composition, and glucose from about 2 wt.% to about 10 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, an agent that inhibits collagen deposition or formation, sodium ascorbate, or combinations thereof.

[0078] In some embodiments, the composition comprises povidone-iodine from about 0.01 wt. % to about 2.5 wt. % of the composition, a polyphenolic compound from about 0.01 wt. % to about 2.5 wt. % of the composition, metformin from about 2 wt.% to about 5 wt. % of the composition, and glucose from about 2 wt.% to about 5 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, an agent that inhibits collagen deposition or formation, sodium ascorbate, or combinations thereof.

[0079] In some embodiments, the composition comprises povidone-iodine from about 0.01 wt. % to about 1 wt. % of the composition, a polyphenolic compound from about 0.01 wt. % to about 1 wt. % of the composition, metformin from about 2 wt.% to about 2 wt. % of the composition, and glucose from about 2 wt.% to about 2 wt. % of the composition. The composition may further comprise a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, an agent that inhibits collagen deposition or formation, sodium ascorbate, or combinations thereof. [0080] In some embodiments, the compositions comprise metformin and glucose, wherein metformin is present from about 2 wt.% to about 20 wt. % of the composition, and glucose is present from about 2 wt.% to about 20 wt. % of the composition.

[0081] In some embodiments, the compositions comprise metformin and an agent that inhibits collagen deposition or formation, wherein metformin is present from about 2 wt.% to about 20 wt. % of the composition, and the agent that inhibits collagen deposition or formation is present from about 2 wt.% to about 50 wt. % of the composition. Non-limiting examples of an agent that inhibits collagen deposition or formation include spironolactone, eplerenone, canrenone, and dimethyloxaloylglycine (DMOG). In some embodiments, the compositions comprise metformin and

spironolactone.

Other Excipients

[0082] In some embodiments, the compositions disclosed herein may further include a solvent, a chelating agent, an emulsifier, a viscous carrier, a thickener, a stabilizing agent, an antioxidant, a preservative, a fragrance, a buffer, a humectant, a surfactant, a diluent, a disintegrant, a filler, a binder, an emollient, a pigment, a cooling agent, and the like.

[0083] In some embodiments, the composition comprises a solvent such as, water, saline, ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediols and isomers thereof, glycerol, glycerol triacetate, pentaerythritol, sorbitol, mannitol, Transcutol®, dimethyl isosorbide, polyethylene glycol,

polypropylene glycol, polyvinylalcohol, hydroxypropyl methylcellulose, 2-pyrrolidone, 2-piperidone, e-caprolactam, N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N- alkylpiperidone, N-alkylcaprolactam, dimethylacetamide, polyvinylpyrrolidone, ethyl propionate, tributylcitrate, acetyl triethylcitrate, acetyl tributyl citrate, triethylcitrate, ethyl oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene glycol monoacetate, propylene glycol diacetate, e-caprolactone and isomers thereof, d- valerolactone and isomers thereof, b-butyrolactone and isomers thereof, and combinations thereof. [0084] In some embodiments, the composition comprises a chelating agent such as, tetrasodium 3 -hydroxy-2, 2'-iminodisuccinate (HIDS), ethylenediamine-N,N'- disuccinic acid (EDDS), iminodisuccinic acid, tetrasodium glutamate diacetate, nitrilotriacetic acid (NT A)), ethylenediamine tetraacetate (EDTA), and methylglycin diacetic acid (MGDA), and combinations thereof.

[0085] In some embodiments, the composition comprises emulsifiers, such as ceteth-20, laureth-3, glyceryl stearate, polyethylene glycol, macrogol cetostearyl ether, stearic acid, stearyl alcohol, polysorbate 60, Irish moss, Tween 80, sorbitol

monostearate. glycol esters, fatty acids, fatty alcohols, fatty acid glycol esters, fatty esters, fatty ethers, esters of glycerin, esters of propylene glycol, fatty acid esters of polyethylene glycol, fatty acid esters of polypropylene glycol, esters of sorbitol, esters of sorbitan anhydrides, carboxylic acid copolymers, esters and ethers of glucose, ethoxylated ethers, ethoxylated alcohols, alkyl phosphates, polyoxyethylene fatty ether phosphates, fatty acid amides, acyl lactylates, soaps, polyethylene glycol 20 sorbitan monolaurate (polysorbate 20), polyethylene glycol 5 soya sterol, steareth-2, steareth-20, steareth-2l , ceteareth-20, PPG-2 methyl glucose ether distearate, ceteth-lO, polysorbate 80, cetyl phosphate, potassium cetyl phosphate, diethanol amine cetyl phosphate, polysorbate 60, glyceryl stearate, PEG- 100 stearate, tragacanth gum, 10-30 alkyl acrylate crosspolymers, and mixtures thereof. In some embodiments, commercially available emulsifying wax may also be used. A typical emulsifying wax contains mixture of cetearyl alcohol, polysorbate 60, PEG- 150 Stearate, Steareth-20, and glyceryl stearate, and combinations thereof.

[0086] In some embodiments, the composition comprises preservatives, such as methylparaben and propylparaben and the salts thereof (e.g. sodium or potassium salts), potassium sorbate, sodium benzoate, diazolidinyi urea, phenoxyethanol, DMDM hydantoin, sorbic acid, benzyl alcohol, formaldehyde, triclosan.methylisothiazolinone, methylchloroisothiazolinone, caffeine, citric acid, benzoic acid, butylated

hydroxytoluene, propylene glycol, organic acids, esters of parahydroxybenzoic acid(methyl, ethyl, propyl and butyl esters of parahydroxy benzoic acid, and their sodium salts etc), chloform,chlorocresol, phenoxyethanol, quaternary ammonium compoundsand butylated hydroxyanisole, and combinations thereof.

[0087] In some embodiments, the composition comprises buffer or pH adjusting agents, such as triethanolamine, triethylamine, diethylmethylamine, ethyldimethylamine, isopropyldimethylamine.one or more adipic acids, glycines, citric acids, calcium hydroxides, magnesium aluminometasilicates, buffers, typically

Bronsted- Lowry and/or Lewis acids and/or bases, or any combinations thereof

[0088] In some embodiments, the composition comprises a viscous carrier. Examples of useful viscous carriers include, but are not limited to, carbomers, polyacrylic acid, cellulosic derivatives, polycarbophil, polyvinylpyrrolidone, gelatin, dextrin, polysaccharides, polyacrylamide, polyvinyl alcohol, polyvinyl acetate, derivatives thereof and mixtures thereof. The compositions disclosed herein can have a viscosity at 25° C of at least about 10 cps or at least about 100 cps or at least about 1000 cps, more preferably at least about 10,000 cps and still more preferably at least about 70,000 cps or more, for example up to about 200,000 cps or about 250,000 cps, or about 300,000 cps or more, at a shear rate of O. l/second.

[0089] In some embodiments, the composition comprises a thickener, such as PEG- 150 distearate, PEG-7 glyceryl cocoate, PEG-200 hydrogenated glyceryl palmitate, PEG- 120 methyl glucose dioleate, carboxymethylene polymer, carboxyvinyl polymer, acrylates, C10-C30 alkyl acrylate crosspolymers, isopropyl myristate, and combinations thereof. Other examples include xantham gum, acacia, alginic acid, bentonite, carbopols, carbomer 940, carbomer 941 , gelatin, carbomer copolymer, aluminum monostearat, dextrin, magnesium aluminum silicate, silicon dioxide, sodium alginate, triethanolamine, polyvinyl alcohol, pectin, methylcellulose, hydroxypropyl cellulose, aqueous thickening agents, such as neutral, anionic-cationic polymers, and combinations thereof.

[0090] In some embodiments, the composition comprises a fragrance, such as aromatic agents selected from the group, but are not limited to, menthol, lavender, mint, cinnamon, chocolate, vanillin, and fruit extracts such as cherry, orange, strawberry, and grape, fennel oil, eucalyptus oil, carnation oil, ginger oil, sweet orange oil, and combinations thereof.

[0091] In some embodiments, the compositions comprise one or more antioxidants, such as vitamin E and its derivatives, a-tocopherol, y-tocopherol, d- tocopherol, ascorbyl palmitate, propyl gallate (PG), octyl gallate, dodecyl gallate, butylated hydroxy anisole (BHA) and butylated hydroxy toluene (BHT), and D-a- tocopheryl polyethylene glycol 1000 succinate.

[0092] In some embodiments, the compositions comprise a stabilizing agent selected from albumin, gelatin, casein, starch, hydroxyethyl starch, monosaccharides (e.g., mannose, glucose, fructose and xylose), polysaccharides (e.g., lactose, maltose, sucrose, raffinose and dextran), mannitol, sorbitol, glycerol, and combinations thereof.

[0093] In some embodiments, the composition comprises a cooling agent selected from but not limited to menthol; an isomer of menthol, a menthol derivative; 4- Methyl-3-(l-pyrrolidinyl)-2[5H]-furanone; WS-23, Icilin, Icilin Unilever Analog, 5- methyl-4-( l-pyrrolidinyl)-3-[2H]-furanone; 4,5-dimethyl-3-( l-pyrrolidinyl)-2[5H]- furanone; isopulegol, 3-(l-menthoxy)propane-l,2-diol, 3-(l-menthoxy)-2- methylpropane-l,2-diol, p-menthane-2,3-diol, p-menthane-3,8-diol, 6-isopropyl-9- methyl-l,4-dioxas-piro[4,5]decane-2-methanol, menthyl succinate and its alkaline earth metal salts, trimethylcyclohexanol, N-ethyl-2-isopropyl-5-methylcyclohexanecarb- oxamide, Japanese mint (Mentha arvensis) oil, peppermint oil, menthone, menthone glycerol ketal, menthyl lactate, 3-(l-menthoxy)ethan-l-ol, 3-(l-menthoxy)propan-l-ol, 3-(l-menthoxy)butan-l-ol, l-menthylacetic acid N-ethylamide, l-menthyl-4- hydroxypentanoate, l-menthyl-3-hydroxybutyrate, N,2,3-trimethyl-2-(l-methylethyl)- butanamide, and spearmint oil, and combinations thereof.

[0094] Emollient or lubricating vehicles that help hydrate the skin can also be used in the compositions. In some embodiments, emollients can be selected from the group, but are not limited to, liquid vaseline. paraffinum liquidum, petrolatum, proplylene glycol, fatty acid esters, mineral oil including dimethicone, waxes including white wax, spermacetic wax, squalene, cetearyl alcohol, cetostearyl alcohol, stearyl alcohol, 2-Octyldodecanol, mineral oil USP, light mineral oil NF, liquid paraffin BP, light liquid paraffin BP, candellilla wax, sweet almond oil, apricot oil, emu oil, argan oil, glycerin, coconut oil, grape seed oil, honey, lanolinand, and combinations thereof

[0095] In some embodiments, the composition comprises surfactants, such as a polysorbate, polyoxyethylene (20) sorbitan monostearate, polyoxyethylene (20) sorbitan monooleate, a polyoxyethylene fatty acid ester, Myrj 45, Myrj 49, Myrj 52 and Myrj 59; a polyoxyethylene alkylyl ether, polyoxyethylene cetyl ether, polyoxyethylene palmityl ether, polyethylene oxide hexadecyl ether, polyethylene glycol cetyl ether, a sucrose ester, a partial ester of sorbitol, sorbitan monolaurate, sorbitan monolaurate a monoglyceride, a diglyceride, isoceteth-20, a sucrose ester, or selected from the group consisting of steareth 2, glyceryl monostearate/PEG 100 stearate, glyceryl stearate, Steareth-2l, peg 40 stearate, polysorbate 60, polysorbate 80, sorbitan stearate, laureth 4, Sorbitan monooleate, ceteareth 20, steareth 20, ceteth 20, macrogol cetostearyl ether, ceteth 2, PEG-30 dipolyhydroxystearate, sucrose distearate, polyoxyethylene (100) stearate, PEG 100 stearate, laureth 4, cetomacrogol ether, cetearyl alcohol, cetearyl glucoside, oleyl alcohol, steareth-2, diisopropyl adipate, capric/caprilic triglicerides, polysorbate 20, montanov 68 (cetearyl alcohol (and) cetearyl glucoside.), sharonmix 824(a liquid blend of methyl paraben, ethyl paraben and propyl parabenin

phenoxyethanol), Simusol 165 (glyceryl stearate and PEG- 100 stea rate), methyl glucose sequistearate, Peg 30 dipolyhydroxystearate, sucrose stearic acid esters, sorbitan laureth, sorbitan stearate, and combinations thereof

[0096] In some embodiments, the compositions may include other skin care agents, including, but not limited to, retinol, steroids, sunblock, salicylate, minocycline, antifungals, peptides, antibodies, lidocaine, and the like and combinations thereof In some embodiments, other skin care agents include N-acyl amino acid compounds including, for example, N-acyl phenylalanine, N-acyl tyrosine, and the like, their isomers, including their D and L isomers, salts, derivatives, and mixtures thereof An example of a suitable N-acyl amino acid is N-undecylenoyl-L-phenylalanine is commercially available under the tradename SEPI WHITE®. Other skin active agents include, but are not limited to, Lavandox, Thallasine 2, Argireline NP, Gatuline In- Tense and Gatuline Expression, Myoxinol LS 9736, Syn-ake, and Instensyl®, Sesaflash™, N- acetyl D-glucosamine, panthenol (for example, DL panthenol available from Alps Pharmaceutical Inc.), tocopheryl nicotinate, benzoyl peroxide, 3-hydroxy benzoic acid, flavonoids (for example, flavanone, chalcone), farnesol, phytantriol, glycolic acid, lactic acid, 4-hydroxy benzoic acid, acetyl salicylic acid, 2- hydroxybutanoic acid, 2-hydroxypentanoic acid, 2-hydroxyhexanoic acid, cis- retinoic acid, trans-retinoic acid, retinol, retinyl esters (for example, retinyl propionate), phytic acid, N-acetyl-L-cysteine, lipoic acid, tocopherol and its esters (for example, tocopheryl acetate: DL-a- tocopheryl acetate), azelaic acid, arachidonic acid, and others.

[0097] In some embodiments, the compositions may include skin lightening agents, such as ascorbic acid compounds, vitamin B3 compounds, azelaic acid, butyl hydroxyanisole, gallic acid and its derivatives, glycyrrhizinic acid, hydroquinone, kojic acid, arbutin, mulberry extract, and combinations thereof.

[0098] In some embodiments, the compositions may include sunblock agents, such as but not limited to para-aminobenzoic acid (PABA), PABA esters (glyceryl PABA, amyldimethyl PABA and octyldimethyl PABA), benzophenones (oxybenzone and sulisobenzone), cinnamates (octylmethoxy cinnamate and cinoxate), salicylates (homomethyl salicylate) anthranilates, TiO2, avobenzone, bemotrizinol, bisoctrizole, 3- (4-methylbenzylidene)-camphor, cinoxate, diethylamino hydroxybenzoyl hexyl benzoate, dioxybenzone, drometrizole trisiloxane, ecamsule, ethylhexyl triazone, homosalate, menthyl anthranilate, octocrylene, octyl salicylate, iscotrizinol, isopentenyl- 4-methoxycinnamate, octyl- dimethyl-p-aminobenzoic acid, octyl-methoxycinnamate, oxybenzone, polysilicone- 15, trolamine salicylate, and ZnO, and combinations thereof.

[0099] In some embodiments, the compositions may further include one or more additives selected from the group consisting of vitamins, cosmetic peptides, oil control agents, sensation modifying agents, skin lightening agents, hydrating

compositions, a sunblock agent, a compound that absorbs or reflects UV photons, other skin care agent and combinations thereof. [0100] In some embodiments, the compositions described herein may be formulated as a liquid. Liquid dosage forms for topical administration may include diluents such as, for example, alcohols, glycols, oils, water, and the like. Such compositions may also include wetting agents or emulsifiers. In embodiments, the compositions may be formulated as oil-in-water or water-in-oil emulsion. A cream can be a water-in-oil (w/o) emulsion in which an aqueous phase is dispersed in an oil phase, or an oil-in-water (o/w) emulsion in which an oil is dispersed within an aqueous base. An ointment generally refers to a more viscous oil-in-water cream. Traditional ointment bases (i.e. carrier) include hydrocarbons (petrolatum, beeswax, etc.) vegetable oils, fatty alcohols (cholesterol, lanoilin, wool alcohol, stearyl alcohol, etc.) or silicones. Insoluble solids such as starch, zinc oxide, calcium carbonate, or talc can also be used in ointments and creams. Gel forms of the compositions described above can be formed by the entrapment of large amounts of aqueous or aqueous-alcoholic liquids in a network of polymers or of colloidal solid particles. Such polymers or colloids (gelling or thickening agents) are typically present at concentrations of less than 10% w/w and include carboxymethyl cellulose, hydroxypropylmethyl cellulose, hydroxyethyl cellulose, methyl cellulose, sodium alginate, alginic acid, pectin, tragacanth, carrageen, agar, clays, aluminum silicate, carbomers, and the like.

[0101] In some embodiments, the compositions may be formulated in any formulation suitable for dermal, subdermal, transdermal, or subcutaneous

administration, including, but not limited to, a solution, fluid, solid, semi-solid, jelly, paste, gel, or hydrogel. In some embodiments, the compositions may be formulated in any formulation suitable for topical administration, including, but not limited to, a solution, fluid, emulsion, suspension, solid, semi-solid, jelly, paste, gel, hydrogel, ointment, lotion, emulsion, cream, foam, mousse, liquid, spray, suspension, dispersion, powder, aerosol, or transdermal patch.

[0102] In some embodiments, the compositions described above can be formulated as aerosols in which the composition is dissolved in a propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas, and a co-solvent such ethanol, acetone, hexadecyl alcohol, and the like and combinations thereof

[0103] In some embodiments, the compositions described herein may be formulated as a hydrogels. Hydrogels are typically prepared by cross-linking various monomers and/or polymers to provide a three-dimensional polymer network. Non limiting examples of polymers include, polyoxyethylene-polypropylene block copolymers, ionic poly saccharides, such as chitosan or sodium alginate, cellulose, and biodegradable polymers, such as poly-lactides (PLA) and poly-glycolides (PGA), butylene succinate (PBS), polyhydroxyalkanoate (PHA), polycaprolactone acid lactone (PCL), polyhydroxybutyrate (PHB), glycolic amyl (PHV), PHB and PHV copolymer (PHBY), and poly lactic acid (PLA)-polyethylene glycol (PEG) copolymers (PLEG).

[0104] In some embodiments, the compositions disclosed herein can be in the form of transdermal patches. The transdermal patches can be in any conventional form such as, for example, a strip, a gauze, a film, and the like. Patch material may be nonwoven or woven (e.g., gauze dressing). Layers may also be laminated during processing. It may be nonocclusive or occlusive, but the latter is preferred for backing layers. The patch is preferably hermetically sealed for storage (e.g., foil packaging). The patch can be held onto the skin and ingredients of the patch can be held together using various adhesives. For example, the transdermal patch can be in the form of a band-aid type device, or it may be packaged in a small metal or plastic“cup”, which is strapped onto the appropriate site using an adhesive, tape, or an outer fabric or leather strap, similar to that worn as part of a watch. The entire patch may be disposable or may be refillable.

[0105] A wide variety of methods may be used for preparing the compositions described above. Broadly speaking, the compositions may be prepared by combining together the components of the composition, as described herein, at a temperature and for a time sufficient to provide a dermal composition. For example, in some

embodiments, the compositions components of the compositions may be dissolved, suspended, dispersed or otherwise mixed in a selected carrier or vehicle, at an effective concentration such that the condition to be treated is relieved or ameliorated.

[0106] The present invention also provides kits comprising the components of the combinations of the disclosure in kit form. A kit may include one or more components including, but not limited to a polyphenolic compound, and one or more components selected from povidone-iodine, metformin, a dermal filler, and botulinum toxin as discussed herein, optionally in association with one or more additional agents, as discussed herein. In one embodiment, the kit includes a polyphenolic compound in a first container, and any of the components in a second container. In another

embodiment, the kit includes a polyphenolic compound and one or more components in a first container, and any of the additional agents in a second container. The kit can further include instructions for administration of the polyphenolic compound, components, and additional agents.

[0107] In some embodiments, the kit may comprise ampoules, disposable syringes, capsules, vials, tubes, or the like. In some embodiments, the kit may comprise a single dose container or multiple dose containers comprising the compositions of embodiments herein. In some embodiments, each dose container may contain one or more unit doses. In some embodiments, the kit may include an applicator. In one embodiment, the kit includes the composition in a gel or ointment, sold in a tube. In another embodiment, the composition is provided in a viscous liquid (such as

carboxylmethylcellulose, hydroxypropylmethycellulose, polyethylene glycol, glycerin, polyvinyl alcohol, or oil containing drops) for rubbing into the skin. The compositions may have preservatives or be preservative-free (for example, in a single-use container).

[0108] In some embodiments, the compositions of embodiments herein can be applied topically using an applicator device. In some embodiments, a kit further includes an applicator device. In some embodiments, the applicator device permits application of the composition to a target site on the skin while preventing the composition from contacting non-target site areas of the skin. In some embodiments, the applicator device may allow the composition to be applied without first applying the composition to one's lingers. In some embodiments, the applicator device may include gloves, sponges, spatulas, swabs, syringes without needles, adhesive patches, or a combination thereof. In some embodiments, use of spatulas or swabs, or the like may require the device to be inserted into a container containing the composition. In some embodiments, use of syringes or adhesive patches may be accomplished by filling the syringe or patch with the composition. The composition may be expelled from the syringes onto the person's skin and/or may be topically spread on the skin by the spatulas or swabs.

METHODS

[0109] Also disclosed herein are methods to treat a subject. In some embodiments, a method of ameliorating a skin condition in a subject in need thereof comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0110] In some embodiments, a method of improving skin texture or appearance in a subject in need thereof comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0111] In some embodiments, a method of treating scars and/or wrinkles on a subject’s skin comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some

embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0112] In some embodiments, a method of improving appearance or reducing fine lines and wrinkles on a subject’s skin comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0113] In some embodiments, a method of stimulating elastogenesis in a subject in need thereof comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some

embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0114] In some embodiments, a method of inducing elastin production at a tissue site comprising scar tissue, visible lines or wrinkles in a subject comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin. In some embodiments, the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

[0115] In some embodiments, a method of treating a skin condition in an individual in need thereof, the method comprising administering a composition disclosed herein into a dermal region of the individual, wherein the administration improves the skin condition. Skin conditions treated by the disclosed compositions include, without limitation, augmentations, reconstructions, diseases, disorders, defects, or imperfections of a body part, region or area. In one aspect, a skin condition treated by the disclosed compositions include, without limitation, a facial augmentation, a facial reconstruction, a facial disease, a facial disorder, a facial defect, or a facial imperfection. In one aspect, a skin condition treated by the disclosed compositions include, without limitation, skin dehydration, a lack of skin elasticity, skin roughness, a lack of skin tautness, a skin stretch line or mark, skin paleness, a dermal divot, a sunken check, a thin lip, a retro-orbital defect, a facial fold, frown lines, crow’s feet, blepharospasm, strabismus, cervical dystonia, hyperhidrosis, glabellar lines, facial neck lines, or a wrinkle.

[0116] In some embodiments, the polyphenolic compound, and each of the components disclosed herein (povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin), and the additional agents can be present in separate compositions and can be administered simultaneously or sequentially. For example, compositions of polyphenolic compound and povidone-iodine and compositions containing one or more elastogenic peptides may be administered simultaneously or sequentially. In some embodiments, povidone-iodine compositions are administered followed by administering elastogenic peptide compositions. In some embodiments, elastogenic peptide compositions are administered followed by povidone-iodine compositions.

[0117] The compositions disclosed herein can be administered by injection dermally, subdermally, transdermally, or subcutaneously. The compositions disclosed herein can also be administered in a variety of dosage forms such as direct topical application, transdermal application, percutaneous application, or application via a transdermal patch and the like.

[0118] In some embodiments, the composition may be administered as needed, once daily, once in two days, once in three days, twice a week, every other week, every other day, or the like for one or more dosing cycles. A dosing cycle may include administration for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, or about 10 weeks.

[0119] In some embodiments, the composition may be administered as needed, such as once a month, once in two months, once in three months, once in 6 months, once in 12 months, once in 24 months, or once in 36 months. In some embodiment, administration of a composition disclosed herein can treat a skin condition for, e.g., about 6 months to about 12 months, about 6 months to about 15 months, about 6 months to about 18 months, about 6 months to about 21 months, about 6 months to about 24 months, about 9 months to about 12 months, about 9 months to about 15 months, about 9 months to about 18 months, about 9 months to about 21 months, about 6 months to about 24 months, about 12 months to about 15 months, about 12 months to about 18 months, about 12 months to about 21 months, about 12 months to about 24 months, about 15 months to about 18 months, about 15 months to about 21 months, about 15 months to about 24 months, about 18 months to about 21 months, about 18 months to about 24 months, about 21 months to about 24 months, about 24 months to about 36 months, or about 30 months to about 48 months.

[0120] In some embodiments, the amount of the botulinum toxin administered can vary according to the particular facial area being treated. Typically, no less about 5 units of a botulinum toxin type A is administered per injection site (i.e. to each muscle portion injected), per patient treatment session. For a botulinum toxin type A, preferably no less than about 20 units the botulinum toxin type A are administered per injection site and for a botulinum toxin type B, preferably no less than about 200 units of the botulinum toxin type B are administered per injection site. Generally, the total amount of botulinum toxin suitable for administration to a patient according to the methods of the invention disclosed herein may be about 100 units, about 400 units or about 4000- 5000 units respectively, per treatment session.

[0121] In some embodiments, the composition is administered by microneedle injection. Microneedle is a hollow needle having an exposed height of between about 0 and 1 mm and a total length of between about 0.3 mm to about 2.5 mm. Preferably, the microneedle is a hollow needle having a length of less than about 2.5 mm. Most preferably, the microneedle is a hollow needle having a length of less than about 1.7 mm. The composition comprising therapeutic cells and extracellular matrix component are delivered into the skin to a depth of at least about 0.3 mm and no more than about 2.5 mm by the microneedle.

[0122] In some embodiments, the method may comprise administering botulinum toxin compositions to the frontalis, corrugator, procerus, occipitalis, temporalis, trapezius and cervical paraspinal muscles of the subject, wherein the botulinum neurotoxin is administered, to the frontalis at about eighty units divided among four sites of injection; to the corrugator at about forty units divided among two sites of injection; to the procerus at about twenty units to one site of injection; to the occipitalis at about one hundred twenty units divided among six sites of injection to about one hundred sixty units divided among eight sites of injection; to the temporalis at about one hundred sixty units divided among eight sites of injection up to two hundred units divided among ten sites of injection; to the trapezius at about one hundred twenty units divided among six sites of injection up to about two hundred units divided among ten sites of injection and to the cervical paraspinal muscles at about eighty units divided among four sites of injection; wherein the botulinum neurotoxin is injected at 31 to 39 injection sites.

[0123] Without wishing to be bound by theory, it is believed that presence of polyphenolic acid in the compositions prolongs the life of the dermal filler

(glycosaminoglycans, collagen, and elastin) and botulinum toxin. Therefore, the frequency of administration of the dermal filler or the botulinum toxin may be reduced. In some embodiments, the presence of polyphenolic acid, such as tannic acid in the composition increases the stability and life span of dermal filler by about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, or about 10 weeks. In some embodiments, the presence of polyphenolic acid, such as tannic acid increases the stability and life span of botulinum toxin by about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, or about 10 weeks.

[0124] It is also believed that stimulation of the deposition of elastin at the site of injury by composition of the invention will aid in promoting wound healing while limiting scarring. Initially, the stimulated deposition of elastin will hold the injured tissue together. The stimulation of elastin synthesis by a composition of the invention according to some embodiments will further act as a chemotactic attractant for fibroblasts, endothelial cells, and inflammatory cells, which will promote healing of the injured tissue. Elastin synthesis at the site of injury will also lessen scarring since scar tissue is devoid of elastin, and elastin is an important component of uninjured skin. The stimulation and secretion of elastin into the matrix will also generally provide a favorable environment for the cells that participate in the healing process, further accentuating the wound healing process.

[0125] The composition, according to some embodiments, may also stimulate the migration of fibroblasts into the treated area. According to another embodiment, a composition of the invention will interact with elastin receptors on the fibroblasts and stimulate the secretion of elastin (i.e., elastogenesis). In one embodiment, a composition of the invention is applied to the wound and maintained in contact with the wound for an extended period of time, i.e., during the entire healing process or until at least closure of the wound occurs by new tissue.

[0126] In some embodiments, the methods disclosed herein further comprise applying heat to a tissue site and topically applying the compositions disclosed herein. Without wishing to be bound by theory, it is believed that exposure to mild heat stimulates elastogenesis. For example, the heat applied to tissue site may comprise a temperature of about 39 °C to about 43 °C, including, for example, about 39 °C, about 39.5 °C, about 40 °C, about 40.5 °C, about 41 °C, about 42 °C, or about 43 °C. In some embodiments, the tissue site may comprise scar tissue, visible lines, wrinkles and combinations thereof. In embodiments, the tissue site may comprise connective tissue.

In embodiments, the tissue may include, dermal fibroblasts, smooth muscles cells, mouth tissue, hair follicles, and corneal tissue. In some embodiments, the tissue site is exposed to heat for about 5 minutes to about 1 hour. Non-limiting examples include about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes or about 1 hour.

[0127] In some embodiments, the methods disclosed herein further comprise applying cold temperature to a tissue site and topically applying the compositions disclosed herein. For example, the cold temperature applied to tissue site may comprise a temperature of about 4 °C to about 20 °C, including, for example, about 4 °C, about 6 °C, about 8 °C, about 10 °C, about 12 °C, about 14 °C, or about 20 °C. In some embodiments, the cold temperature may be applied for about 5 minutes to about 1 hour. Non- limiting examples include about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes or about 1 hour.

[0128] In some embodiments, the methods disclosed herein further comprise ablating a skin surface and topically applying the compositions disclosed herein.

Without wishing to be bound by theory, it is believed that ablation of skin surface enhances penetration of elatogenic peptides resulting in increased elastogenesis. The ablation methods disclosed herein may include ablation by electromagnetic radiation, laser, dermal abrasion, microdermal abrasion, chemical peel, ultrasound, heating, cooling, or by a microneedle. Skin surface include any part of the body, including but not limited to face, hands, legs, neck, abdominal area, eyes, nose, and chest. [0129] Abrasion of the outer layer or epidermis of the skin (dermal abrasion) is desirable to smooth or blend scars, blemishes, or other skin conditions that may be caused by, for example, acne, sun exposure, and aging. Standard techniques used to abrade the skin have generally been separated into two fields referred to as

dermabrasion and microdermabrasion. Both techniques remove portions of the epidermis called the stratum corneum, which the body interprets as a mild injury. The body then replaces the lost skin cells, resulting in a new outer layer of skin.

Additionally, despite the mild edema and erythema associated with the procedures, the skin looks and feels smoother because of the new outer layer of skin.

[0130] Microdermabrasion refers generally to a procedure in which the surface of the skin is removed due to mechanical rubbing by a handpiece emitting a stream of sand or grit. For example, a handpiece can be used to direct an air flow containing tiny crystals of aluminum oxide, sodium chloride, or sodium bicarbonate. The momentum of the grit tends to wear away two to three cell layers of the skin with each pass of the handpiece. Alternatively, new“crystal-free” microdermabrasion techniques utilize a diamond-tipped handpiece without a stream of grit.

[0131] In some embodiments, prior to administering the composition the tissue surface is ablated with electromagnetic radiation, for instance using a so-called fractional laser treatment. By way of example, such methods employ electromagnetic radiation (EMR) having one or more wavelengths of between approximately 1,850 to 100,000 nanometers and with pulse widths of between approximately 1 femtosecond (1 x 10 ¹⁵ s) to 10 milliseconds (10^c 10^-3 s) with fluence in the range of from

approximately 1 J/cm² to 300 J/cm². In other examples, the tissue is ablated with electromagnetic radiation having one or more wavelengths of between approximately 2,200 to 5,000 nanometers. In still other examples, the tissue is ablated with

electromagnetic radiation having one or more wavelengths of between approximately 190 to 320 nanometers with fluence in the range of from 1 J/cm² to 300 J/cm².

Optionally, conditions selected for ablating portions of the tissue minimize the coagulation zone of tissue damage, for instance by keeping the coagulation zone to a relatively small diameter surrounding the ablated void. [0132] Electromagnetic radiation (EMR), particularly in the form of laser light or other optical radiation, has been used in a variety of cosmetic and medical

applications, including uses in dermatology, dentistry, ophthalmology, gynecology, otorhinolaryngology and internal medicine. For most dermatological applications,

EMR treatment can be performed with a device that delivers the EMR to the surface of the targeted tissue(s). EMR treatment is typically designed to (a) deliver one or more particular wavelengths (or a particular continuous range of wavelengths) of energy to a tissue to induce a particular chemical reaction, (b) deliver energy to a tissue to cause an increase in temperature, or (c) deliver energy to a tissue to damage or destroy cellular or extracellular structures, such as for skin remodeling. Examples of devices that have been used to treat the skin during cosmetic procedures such as skin rejuvenation include the Palomar® LuxIR, the Palomar® 1540, 1440 and 2940 Fractional Handpieces, the Reliant Fraxel® SR Laser and similar devices by Lumenis, Alma Lasers, Sciton and many others.

[0133] In some embodiments, the compositions disclosed herein can be administered to the subject’s skin after microneedling. Microneedling involves penetrating the skin with tiny, solid, sterile, microneedles, and this creates

microchannels that, for a short period of time, allow molecules larger than 500 daltons to cross the stratum comeum. Any microneedling device known in the art may be used for this method. In some embodiments, the microneedling devices have an electric motor housed inside a handheld stylus and are attached to a disposable plastic cartridge that has an array of tiny microneedles. In some embodiments, the depth of penetration of the microneedle into the skin as well as the speed in which the motor reciprocally propels may be controlled to achieve the desired outcome.

[0134] All references cited herein are incorporated by reference to the same extent as if each individual publication, database entry (e.g. Genbank sequences or GenelD entries), patent application, or patent, was specifically and individually indicated to be incorporated by reference. This statement of incorporation by reference is intended by Applicants, pursuant to 37 C.F.R. § 1.57(b)(1), to relate to each and every individual publication, database entry (e.g. Genbank sequences or GenelD entries), patent application, or patent, each of which is clearly identified in compliance with 37 C.F.R. § 1.57(b)(2), even if such citation is not immediately adjacent to a dedicated statement of incorporation by reference. The inclusion of dedicated statements of incorporation by reference, if any, within the specification does not in any way weaken this general statement of incorporation by reference. Citation of the references herein is not intended as an admission that the reference is pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents.

[0135] The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and the accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0136] The foregoing written specification is considered to be sufficient to enable one skilled in the art to practice the invention. Various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims.

EXAMPLES

Example 1: Effect of tannic acid and povidone- iodine

[0137] Biological effects of povidone-iodine and tannic acid will be tested in cultures of dermal fibroblasts derived from punch biopsies of healthy skin. All fibroblasts will be isolated by allowing them to migrate out of skin explants and then will be passaged by trypsinization and maintained in alpha-minimum essential medium supplemented with 20 mM Hepes, 1% antibiotics and antimycotics, 1% L-Glutamate and 2% fetal bovine serum (FBS). Cells will be plated (50x 10⁵ cells/dish) to reach confluency and will be cultured for 7 days in the presence and absence of povidone- iodine (1% final concentration) and tannic acid (1 pg/mL), and combination of both. [0138] Cellular proliferation rates of control, povidone-iodine, tannic acid, and the combination of both povidone-iodine will be assessed at the end point by counting of trypsinized cells, by total DNA assay using the DNeasy Tissue System from Qiagen and by assessment of [³H] -thymidine incorporation, which will be added to all cultures (2 pCi/well) for the last 24 hours. Additionally, the immuno-detection of proliferating cells will performed with a Ki67 antigen specific fluorescence-labeled antibody.

[0139] Quantifying elastin fibers: The cell cultures maintained in the presence and absence of indicated reagents will be fixed in cold 100% methanol at 20°C (for detection of elastin) or in 4% paraformaldehyde at room temperature (for detection of collagen I). The multiple parallel cultures will be incubated with 10 mg/ml of polyclonal antibody to tropoelastin (Elastin Products, Owensville, MI), or polyclonal antibody to collagen type I (Chemicon, Temecula, CA). Cultures will be incubated with the respective fluorescein-conjugated goat anti- rabbit, goat anti-mouse, or rabbit anti-goat secondary antibodies. Nuclei will be counter stained with propidium iodide

(Sigma, Sigma, St. Louis, MO). All cultures will be examined with a Nikon Eclipse El 000 microscope attached to a cooled CCD camera (Qlmaging, Retiga EX) and analyzed with the computer-generated morphometric analysis system, in which the Image-Pro Plus software (Media Cybernetics, Silver Springs, MD) estimates the proportion of areas marked with green fluorescence, in relation to the entire ( 1 square mm) analyzed field.

[0140] Q-PCR: Confluent cultures of skin fibroblasts will be treated for 18 h with or without the reagents of interest for different periods of time. At the end of the treatment period, total RNA will be extracted from individual cultures using the RNeasy Mini Kit, and the one step PCR or quantitative real time PCR reactions will be set up with the RT-PCR Kit. The amounts of tropoelastin mRNA obtained from triplicate cultures will be normalized by the levels of l8s mRNA and then analyzed by the

Comparative Ct Method, using software from Applied Biosystems and normalized to the amounts of GAPDH mRNA or l8s mRNA. [0141] Western blots: At the end of the indicated experiments, 24 h-old separate cultures will be lysed lysed with NP-40 buffer containing a cocktail of broad- spectrum inhibitors of proteinases and phosphatases. The 50 mg aliquots of protein extract will be resolved by SDS-PAGE gel (4-12% gradient) in reducing conditions and analyzed by Western blot with antibodies. Initial blots will also re-probed with monoclonal anti-b-actin antibody (Cell Signaling Technology Inc., Danvers, MA) to confirm the equal protein loading. The degree of expression will be measured by densitometry.

[0142] Quantitative assays of insoluble elastin: Cultures of fibroblasts (plated in 35 mm culture dishes at 100,000 cells/dish) will be maintained for 72 h with 2 mCi of [¾- valine/ml (Amersham Biosciences Ltd. Oakville, Canada), in the presence and absence of the indicated treatments. At the end of each experiment, the levels of metabolically labeled NaOH- insoluble elastin present in individual cultures will be assayed and normalized per their DNA content.

Example 2: Effect of tannic acid and metformin

[0143] Biological effects of metformin and tannic acid will be tested in cultures of dermal fibroblasts derived from punch biopsies of healthy skin. All fibroblasts will be isolated by allowing them to migrate out of skin explants and then will be passaged by trypsinization and maintained in alpha-minimum essential medium supplemented with 20 mM Hepes, 1% antibiotics and antimycotics, 1% L-Glutamate and 2% fetal bovine serum (FBS). Cells will be plated (50x 10⁵ cells/dish) to reach confluency and will be cultured for 7 days in the presence and absence of metformin (1% final concentration), glucose, and tannic acid (1 pg/mL), and combination of all the three.

[0144] Cellular proliferation rates, quantification of elastin fibers,

quantification of elastin mRNA, and detection by Western blots will be performed as described in Example 1. Example 3: Effect of tannic acid on dermal fillers

[0145] Compositions will be prepared containing 1 wt.% tannic acid and dermal filler (fragments of hyaluronic acid, collagen, heparin, heparan sulfate). Control formulations containing dermal filler alone will also be prepared. The compositions will be injected to subject’s skin. Compositions containing tannic acid would have improved stability of dermal filler relative to comparable controls, and subjects would need less frequent administration of dermal filler.

Example 4: Topical application of tannic acid

[0146] Compositions of dermal filler (fragments of hyaluronic acid, collagen, heparin, heparan sulfate) will be injected to subject’s skin. Compositions containing 1% tannic acid will be topically applied at the injection site. Topical application of tannic acid will improve the stability of dermal filler relative to comparable controls, and subjects would need less frequent administration of dermal filler.

Example 5: Effect of tannic acid on botulinum toxin

[0147] Compositions will be prepared containing 1 wt.% tannic acid and botulinum toxin. Control formulations containing botulinum toxin alone will also be prepared. The compositions will be injected to subject’s skin. Compositions containing tannic acid would display improved stability of botulinum toxin relative to comparable controls, and subjects would need less frequent administration of botulinum toxin.

Example 6: Topical application of tannic acid

[0148] Compositions of botulinum toxin will be injected to subject’s skin. Compositions containing 1% tannic acid will be topically applied at the injection site. Topical application of tannic acid will improve the stability of botulinum toxin relative to comparable controls, and subjects would need less frequent administration of botulinum toxin.

Claims

CLAIMS What Is Claimed Is:

1. A composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin.

2. The composition of claim 1, wherein the dermal filler is selected from the group consisting of hyaluronic acid, collagen, heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, elastin, and a combination thereof.

3. The composition of claim 1, wherein the polyphenolic compound is selected from the group consisting of gallic acid, cinnamic acid, caffeic acid, ferulic acid, tannic acid, ellagic acid, coumoric acid, and derivatives thereof, and combinations thereof.

4. The composition of claim 1, further comprising one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

5. The composition of claim 1, wherein the polyphenolic compound is present from about 0.01 wt. % to about 10 wt. % of the composition, and the dermal filler is present from about 0.01 wt.% to about 85 wt. % of the composition.

6. The composition of claim 1, wherein the polyphenolic compound is present from about 0.01 wt. % to about 10 wt. % of the composition, and the botulinum toxin is present from about 0.1 wt.% to about 25 wt. % of the composition.

7. The composition of claim 4, wherein the povidone-iodine is present from about 0.01 wt. % to about 10 wt. % of the composition, the polyphenolic compound is present from about 0.01 wt. % to about 10 wt. % of the composition, and the elastogenic peptide is present from about 2 wt.% to about 60 wt. % of the composition.

8. The composition of claim 4, wherein the povidone-iodine is present from about 0.01 wt. % to about 10 wt. % of the composition, the polyphenolic compound is present from about 0.01 wt. % to about 10 wt. % of the composition, metformin is present from about 2 wt.% to about 20 wt. % of the composition, and glucose is present from about 2 wt.% to about 20 wt. % of the composition.

9. A method of stimulating elastogenesis in a subject in need thereof comprising administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin.

10. The method of claim 9, wherein the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

11. The method of claim 9, wherein the method further comprises ablating a skin surface and topically administering the composition.

12. A method of ameliorating a skin condition in a subject in need thereof comprising administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin.

13. The method of claim 12, wherein the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

14. The method of claim 12, wherein the method further comprises ablating a skin surface and topically administering the composition.

15. A method of improving skin texture or appearance in a subject in need thereof comprising administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin.

16. The method of claim 15, wherein the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

17. The method of claim 15, wherein the method further comprises ablating a skin surface and topically administering the composition.

18. A method of treating scars and/or wrinkles on a subject’s skin comprises administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin.

19. The method of claim 18, wherein the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

20. The method of claim 18, wherein the method further comprises ablating a skin surface and topically administering the composition.

21. A method of improving appearance or reducing fine lines and wrinkles on a subject’s skin comprising administering to the subject a composition comprising a polyphenolic compound, and one or more components selected from povidone-iodine, potassium iodide, metformin, a dermal filler, and botulinum toxin.

22. The method of claim 21, wherein the composition further comprises one or more additional agents selected from glucose, a copper compound, a divalent manganese compound, a trivalent iron compound, aldosterone, sodium ascorbate, an elastogenic peptide, an analgesic, tetracycline, and an agent that inhibits collagen deposition or formation.

23. The method of claim 21, wherein the method further comprises ablating a skin surface and topically administering the composition.

24. The method of claim 21, wherein the eleastogenic peptide is selected from SEQ ID NOs:

1 -79.