Composition for anti-aging or skin regeneration comprising ocainine

Abstract

The present invention relates to a composition for anti-aging or skin regeneration containing ocainine, or use of ocainine or a salt thereof, and particularly, to a composition for anti-aging or skin regeneration containing ocainine represented by the following chemical formula (1) or a salt thereof as an active ingredient, and use of ocainine or a salt thereof.

Description

Technical Field

[0001] This invention relates to compositions for anti-aging or skin regeneration, and more particularly, to compositions for anti-aging or skin regeneration containing oxanin. Background Technology

[0002] Skin aging is an inevitable phenomenon. Aging leads to a loss of skin elasticity, wrinkles, and symptoms such as pigmentation and epidermal atrophy. Skin aging is mainly divided into physiological aging (which indicates changes in the function, structure, and shape of the skin) and aging caused by external stimuli such as ultraviolet radiation.

[0003] As skin ages, cell function declines, and simultaneously, the skin is affected by external environmental factors such as ultraviolet radiation. First, with aging, the rate of keratinocyte division slows down. While the normal skin regeneration cycle is approximately 28 days, this cycle lengthens with aging. Second, the number of fibroblasts present in the epidermis is known to decrease with aging, and their function also declines. Fibroblasts are cells that produce collagen and elastin fibers to provide elasticity to the skin. However, when the number of fibroblasts decreases and their function declines, dermal atrophy occurs. Finally, the expression of matrix proteases increases with aging. Therefore, collagen and elastin fibers are broken down by matrix proteases, leading to increased dermal atrophy.

[0004] However, even at the age of 100, aging cannot be completely prevented, and the expectation and demand for substances that can improve skin aging to enhance beauty are increasing. Therefore, there is a need to rapidly develop substances that slow down or improve skin aging.

[0005] Epidermal growth factor (EGF) is a growth factor that regulates the growth and survival of the epidermis or epidermal cells. EGF is also considered a cancer-related factor and has been used medically to treat wounds and burns. Recently, EGF has been widely used as a raw material in cosmetics, and numerous studies have been conducted on cosmetic compositions containing EGF for preventing skin aging, such as Korean Patent No. 2093884. Therefore, there is a need to research small molecule substances of natural origin that have similar functions to EGF, are stable, and can be used to cross the skin barrier. However, the biosynthesis, isolation, and purification of EGF are known to be expensive, and it is difficult to store EGF in its active state. Furthermore, there is a problem that EGF cannot easily cross the skin barrier due to its large molecular weight.

[0006] Therefore, there is a need to study small molecules of natural origin that have functions similar to EGF, are stable, and can be used to cross the skin barrier. Summary of the Invention

[0007] Technical issues

[0008] One aspect of this disclosure is to provide compositions for anti-aging or skin regeneration, said compositions comprising oxaline (a small molecule substance of natural origin) or a salt thereof as an active ingredient to provide a composition for anti-aging or skin regeneration that has a function similar to that of EGF and is stable and can be used to cross the skin barrier.

[0009] Technical solution

[0010] According to one aspect of this disclosure, the present invention provides a composition for anti-aging or skin regeneration, comprising oxaline or a salt thereof represented by formula (1) as an active ingredient:

[0011] [Equation (1)]

[0012]

[0013] Beneficial effects

[0014] According to one aspect of this disclosure, oxcarbazone can be extracted from various plant species, such as Bidens pilosa. The compositions according to the invention have the advantage of not having side effects when used. Furthermore, like EGF, oxcarbazone binds to EGFR to activate signals associated with cell survival, growth, and proliferation, thereby increasing the resistance and viability of skin cells to external stimuli and enhancing skin regeneration damaged by external stimuli or aging. Moreover, while oxcarbazone has functions similar to EGF, unlike EGF, oxcarbazone is stable and small in size, and therefore has the advantage of being able to move across the skin barrier to reach the basal layer of the skin to perform its functions. Therefore, the oxcarbazone of the present invention can be used in a variety of ways, such as in pharmaceuticals, quasi-pharmaceuticals, cosmetics, fragrance materials, and biofunctional materials. Attached Figure Description

[0015] Figure 1 The results show the results of determining the expression of the biosensor protein (EGFP-SH2) via EGF and also via oxcarpine when the biosensor system for measuring EGFR activity via intracellular injection is used.

[0016] Figure 2 The results show the cytotoxicity of HaCaT cells, which are keratinocytes, determined by CCK-8 assay.

[0017] Figure 3 The results show that ERK and AKT, as EGFR subtypes, are activated by oxcanin.

[0018] Figure 4The results show the results of determining the expression patterns of the egrl, c-myc, c-jun, and c-fos genes increased by oxcanin.

[0019] Figure 5 The results show the results of determining the growth-promoting effect of oxalanine on keratinocytes by CCK-8 assay.

[0020] Figure 6 shows photographs (a) and (b) illustrating the results of determining the proliferative and growth-promoting effects of oxaline on keratinocytes by wound healing assay.

[0021] Figure 7 The study showed that keratinocyte resistance increased due to oxaline under conditions of UVB-induced cell damage. Detailed Implementation

[0022] Preferred embodiments of the invention will be described below with reference to the accompanying drawings. However, embodiments of the invention can be modified to have various forms, and the scope of the invention is not limited to the embodiments described below.

[0023] The inventors completed this invention by experimentally determining that oxaline, a small molecule substance of natural origin, has a function similar to that of epidermal growth factor (EGF), and is stable and small in size, thus easily moving across the skin barrier.

[0024] This invention relates to the use of oxcarpine or a salt thereof, represented by formula (1), in the preparation of compositions for anti-aging or skin regeneration:

[0025] [Equation (1)]

[0026]

[0027] Specifically, the present invention provides a composition for anti-aging or skin regeneration, comprising oxcanin or a salt thereof represented by formula (1) as an active ingredient:

[0028] [Equation (1)]

[0029]

[0030] Oxycarnin, or 2′,3,3′,4,4′-pentahydroxychalcone, can be obtained from extracts of various plant species, such as Bidens pilosa, and has a function similar to that of epidermal growth factor (EGF), which activates signals in cells related to cell survival, growth, and proliferation. This increases the resistance and viability of skin cells to external stimuli and enhances the regeneration of skin damaged by external stimuli or aging.

[0031] The compositions of the present invention may contain oxcarnitine or a salt thereof in amounts of 10 nM to 100 μM, or 0.1 μM to 100 μM, for example 5 μM to 100 μM, preferably 1 μM to 50 μM, and more preferably 1 μM to 10 μM. When the concentration of oxcarnitine or a salt thereof is less than 0.1 μM, the effects of oxcarnitine or a salt thereof may not be adequately expressed, and when the concentration is greater than 100 μM, cytotoxicity may increase significantly.

[0032] In this invention, "skin regeneration" can refer to promoting skin cell growth and enhancing viability to prevent cell damage from external or internal stimuli, reducing wrinkles, and improving skin elasticity. Furthermore, skin regeneration includes improving or treating skin damage caused by external environmental factors, such as skin and tissue damage caused by burns. Therefore, the compositions of this invention can improve or treat burn wounds.

[0033] Furthermore, the oxcarbazone or its salt of the present invention can bind to EGFR to promote cell proliferation or growth, thereby activating anti-aging or skin regeneration. The cells proliferating or growing through the oxcarbazone or its salt of the present invention can be epidermal cells, preferably keratinocytes (cells that produce keratin), but the present invention is not limited thereto.

[0034] Keratin, a protein that forms the main component of various tissues in animals, gives hair its luster and elasticity, and endows skin with vitality and elasticity. Furthermore, keratin proteins in the stratum corneum of the skin are highly resistant to chemicals, thus performing a protective function against chemical irritants to the skin.

[0035] Therefore, the compositions of the present invention can enhance skin elasticity or improve the problem of skin wrinkles.

[0036] Furthermore, the compositions of the present invention can enhance resistance to cell damage caused by external stimuli, thereby preventing or improving cell damage caused by external stimuli. External stimuli can be chemical stimuli caused by cosmetics or other topical agents, physical stimuli, or ultraviolet light, preferably ultraviolet light, but the present invention is not limited thereto.

[0037] Furthermore, the compositions of the present invention can be parenteral or oral compositions, such as pharmaceutical or cosmetic compositions.

[0038] Specifically, when the compositions of the present invention are pharmaceutical compositions, the compositions may additionally contain a pharmaceutically acceptable carrier, and the compositions may additionally contain suitable carriers, templates, and diluents commonly used in the preparation of pharmaceutical compositions. The compositions can also be formulated and used in the form of oral formulations, topical formulations, suppositories (e.g., powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols) and sterile injections according to methods conventionally known in the art.

[0039] The pharmaceutical compositions according to the invention can be formulated and used in the form of external preparations (e.g., powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols) and sterile injections, and are preferably in the form of creams, gels, patches, sprays, ointments, plasters, lotions, liniments, pastes, or poultices. The carriers, excipients, and diluents that may be included in the compositions can be compounds comprising at least one of the following: lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil.

[0040] The composition can be prepared using diluents or excipients (e.g., fillers, extenders, binders, wetting agents, disintegrants, and surfactants). For example, solids for oral administration include tablets, pills, powders, granules, capsules, etc., and such solids can be prepared by mixing an extract with at least one excipient (e.g., starch, calcium carbonate, sucrose, lactose, and gelatin). In addition to simple excipients, lubricants such as magnesium stearate talc can also be used. In addition to simple diluents commonly used in the art, such as water and liquid paraffin, liquid formulations for oral administration (e.g., suspensions, solutions, emulsions, and syrups) can also contain a variety of excipients (e.g., wetting agents, sweeteners, air fresheners, and preservatives).

[0041] Formulations intended for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Wipedesol, polyethylene glycol, Tween 61, cocoa butter, lauryl alcohol, glycerin gelatin, etc., may be used as bases for suppositories.

[0042] The pharmaceutical compositions according to the invention can be administered at a pharmacologically effective amount. In this invention, a "pharmacologically effective amount" refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio suitable for medical treatment. The effective dose level can be determined based on the type and severity of the patient's disease, the activity of the drug, the sensitivity to the drug, the duration and route of administration and the rate of excretion, the duration of treatment, factors such as concurrently used drugs, and other factors known in the medical field. The pharmaceutical compositions according to the invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, can be administered sequentially or concurrently with conventional therapeutic agents, and can be administered once or multiple times. In view of all the above factors, it is important to administer the minimum amount that yields the maximum effect without causing side effects. Such an amount can be readily determined by those skilled in the art to which this invention pertains.

[0043] The pharmaceutical compositions of the present invention can be administered to a subject via a variety of routes. The pharmaceutical compositions of the present invention are determined based on various factors (e.g., the disease to be treated, the route of administration, the patient's age, sex, and weight, and the severity of the disease) and the type of drug as the active ingredient.

[0044] The compositions of the present invention can be provided in the form of cosmetic compositions. Formulations of the cosmetic compositions according to the present invention can be in the following forms: skin cleansers, skin softeners, astringents, lotions, emulsions, moisturizing lotions, nourishing lotions, massage creams, nourishing creams, moisturizing creams, hand creams, foundations, serums, nourishing serums, peel-off masks, soaps, cleansing foams, cleansing washes, cleansing creams, body washes, or body cleansers.

[0045] The cosmetic compositions according to the present invention may additionally comprise compounds selected from water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, and sphingolipids.

[0046] Any water-soluble vitamin can be used, as long as it can be added to a cosmetic composition. Preferably, vitamins B1, B2, B6, pyridoxine, pyridoxine hydrochloride, B12, pantothenic acid, niacin, niacinamide, folic acid, vitamin C, and vitamin H can be used. Salts of these vitamins (e.g., thiamine hydrochloride and sodium ascorbate) or derivatives thereof (e.g., sodium ascorbate-2-phosphate and magnesium ascorbate-2-phosphate) can also be used as the water-soluble vitamins in this invention. Water-soluble vitamins can be obtained by conventional methods (e.g., microbial transformation, purification from microbial cultures, and enzymatic or chemical synthesis).

[0047] Any available vitamin can be used, as long as it can be added to cosmetics, and preferably includes vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (e.g., d1-α-tocopherol, d-α-tocopherol, and d-α-tocopherol), etc. Derivatives thereof (e.g., ascorbate palmitate, ascorbate stearate, ascorbate dipalmitate, dl-α-tocopherol acetate, dl-α-tocopherol nicotinate, vitamin E, DL-panthenol, D-panthenol, and panthenyl ether) can also be used as oil-soluble vitamins in this invention. Available vitamins can be obtained by conventional methods (e.g., microbial transformation, purification from microbial cultures, and enzymatic or chemical synthesis).

[0048] Any high molecular weight peptide can be used, as long as it can be added to cosmetics, and preferably includes collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin, etc. High molecular weight peptides can be obtained by conventional methods such as purification from microbial culture media, enzymatic methods or chemical synthesis, or can be conventionally obtained by purification from natural products such as dermal hides of pigs or cattle or silk fibers.

[0049] Any high molecular weight polysaccharide can be used, as long as it can be added to cosmetics, and preferably includes hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or its salts (e.g., sodium salts). For example, chondroitin sulfate or its salts can be routinely obtained by purification from mammals or fish.

[0050] Any sphingolipid can be used, as long as it is added to cosmetics, and preferably includes ceramides, phytosphingosine, glycosphingolipids, etc. Sphingolipids can be purified from mammals, fish, shellfish, selenium-enriched yeasts, or plants by conventional methods, or obtained by chemical synthesis.

[0051] In addition to the essential ingredients mentioned above, the cosmetic compositions of the present invention may also contain other ingredients commonly added to cosmetics, as needed.

[0052] Other ingredients that can be added include oils and fats, moisturizers, emollients, surfactants, organic or inorganic pigments, organic powders, UV absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation enhancers, cooling agents, antiperspirants, purified water, etc.

[0053] Oil and fat components may include ester-based oils and fats, hydrocarbon-based oils and fats, organosilicon-based oils and fats, fluorine-based oils and fats, animal oils and fats, vegetable oils and fats, etc.

[0054] Ester oils and fats may include glyceryl tri-2-ethylhexanoate, hexadecyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isochetyl isostearate, butyl stearate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isochetyl myristate, isostearyl myristate, isostearyl palmitate, octyl dodecyl myristate, isochetyl isostearate, diethyl sebacate, diisopropyl adipate, isoalkyl neopentanoate, triglycerides (octanoate or decanoate), trimethylolpropane tris(2-ethylhexanoate), trimethylolpropane tris(2-ethylhexanoate), trimethylolpropane tris(2-ethylhexanoate) Isostearate, Pentaerythritol tetra(2-ethylhexanoate), hexadecyl octanoate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, hexadecyl myristate, stearate, decyl oleate, hexadecyl castor oil, isostearate laurate, isotriadecyl myristate, isochetyl palmitate, octyl stearate, isochetyl stearate, isodecanyl oleate, octyl dodecyl oleate, octyl dodecyl linoleate, isopropyl isostearate, cetearyl 2-ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate Propylene glycol didecanoate, propylene glycol di(octanol or decanoic acid), propylene glycol dicaprylate, neopentyl glycol didecanoate, neopentyl glycol dicaprylate, tricaprylic acid glyceride, tridecanoic acid glyceride, triisopalmitoyl glyceride, triisostearate glyceride, octyl dodecyl neopentanoate, octyl isostearate, octyl isononanoate, hexyl decyl neodecanoate, isochetyl isostearate, isostearate, octyl decyl isostearate, polyglycerol oleate, polyglycerol isostearate, triisohexadecyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, hexadecyl lactate Alkyl esters, octyl decyl lactate, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di(2-ethylhexyl) succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, cholesterol stearate, cholesterol isostearate, cholesterol hydroxystearate, cholesterol oleate, dihydrocholesterol oleate, phytosterol isostearate, phytosterol oleate, isochetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate, isostearyl 12-stearoylhydroxystearate, etc.

[0055] Hydrocarbon-based oils and fats can include squalene, liquid paraffin, α-olefin oligomers, isoalkanes, ceresin, paraffin, liquid isoalkanes, polybutene, microcrystalline wax, petrolatum, etc.

[0056] Silicone-based oils and fats may include polymethylsiloxane, toluene silicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecylcyclosiloxane, dimethylsiloxane / methylhexadecyloxysiloxane copolymer, dimethylsiloxane / methylstearyloxysiloxane copolymer, alkyl-modified silicone oil, amino-modified silicone oil, etc.

[0057] Fluorine-based oils and fats can include perfluoropolyethers, etc.

[0058] Animal or vegetable oils and fats may include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, safflower oil, soybean oil, corn oil, rapeseed oil, almond oil, palm kernel oil, palm oil, castor oil, sunflower seed oil, grapeseed oil, cottonseed oil, coconut oil, macadamia nut oil, wheat germ oil, rice germ oil, shea butter, evening primrose oil, macadamia nut oil, meadowfoam seed oil, egg yolk oil, beef tallow, horse oil, mink oil, tilapia oil, jojoba oil, candelilla wax, carnauba wax, liquid lanolin, hydrogenated castor oil, etc.

[0059] Wetting agents may include water-soluble low-molecular-weight moisturizers, lipid-soluble molecular moisturizers, water-soluble polymers, or oil-soluble polymers.

[0060] Water-soluble low-molecular-weight moisturizers may include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-carboxylate, glycerin, propylene glycol, 1,3-butanediol, ethylene glycol, polyethylene glycol B (degree of polymerization n=2 or greater), polypropylene glycol (degree of polymerization n=2 or greater), polyglycerol B (degree of polymerization n=2 or greater), lactic acid, lactate / ester, etc.

[0061] Lipid-soluble low-molecular-weight moisturizers can include cholesterol, cholesterol esters, etc.

[0062] Water-soluble polymers may include carboxyvinyl polymers, polyaspartate salts / esters, astragalus gum, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitosan, chitin, dextrin, etc.

[0063] Fat-soluble polymers may include polyvinylpyrrolidone / eicosene copolymer, polyvinylpyrrolidone / hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and high molecular weight organosilicon.

[0064] Emollients may include long-chain acylglutamic acid cholesterol ester, hydroxystearic acid cholesterol ester, 12-hydroxystearate, stearic acid, rosin acid, lanolin fatty acid cholesterol ester, etc.

[0065] Surfactants can include nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, etc.

[0066] Nonionic surfactants may include self-emulsifying glyceryl monostearate, propylene glycol fatty acid esters, glyceryl fatty acid esters, polyglycerol fatty acid esters, fatty acid dehydrated sorbitol esters, polyoxyethylene (POE) dehydrated sorbitol fatty acid esters, POE dehydrated sorbitol fatty acid esters, POE glyceryl fatty acid esters, POE alkyl ethers, POE fatty acid esters, POE hydrogenated castor oil, POE castor oil, POE / POP (polyoxyethylene / polyoxypropylene) copolymer, POE / POP alkyl ethers, polyether-modified organosilicon, lauric acid alkanolamide, alkylamine oxides, hydrogenated soybean lecithin, etc.

[0067] Anionic surfactants can include fatty acid soaps, α-acylsulfonates, alkylsulfonates, alkylallyl sulfonates, alkylnaphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, alkylamide phosphates, alkyl acyl alkyl taurines, N-acyl amino acid salts, POE alkyl ether carboxylates, alkyl sulfosuccinates, sodium alkyl sulfoacetate, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphates, etc.

[0068] Cationic surfactants may include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetearyltrimethylammonium chloride, distearate dimethylammonium chloride, octadecyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, diethylaminoethyl stearate, dimethylaminopropyl stearate, and quaternary ammonium salts of lanolin derivatives.

[0069] Amphoteric surfactants can include carboxybetaine type, amide betaine type, sulfobetaine type, hydroxysulfonyl ammonium salt type, amide sulfobetaine type, phosphoryl betaine type, aminocarboxylate type, imidazoline derivative type, amide amine type, etc.

[0070] Organic or inorganic pigments may include inorganic pigments (e.g., silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, ferric oxide, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, chromium oxide, chromium hydroxide, smithsonite and its complexes); organic pigments (e.g., polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenolic resin, fluororesin, silicone resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene / styrene copolymer, silk powder, cellulose, CI pigment yellow and CI pigment orange); and composite pigments of inorganic and organic pigments.

[0071] Organic powders may include metal soaps such as calcium stearate; alkyl phosphate metal salts (e.g., sodium zinc palmitate, zinc lauryl soap, and calcium laurylate); polyvalent metal salts of acyl amino acids (e.g., calcium N-lauroyl-β-alanine, zinc N-lauroyl-β-alanine, and calcium N-lauroyl glycine); polyvalent metal salts of amide sulfonic acids (e.g., calcium N-lauroyl-taurate and calcium N-palmitoyl-taurate); acyl basic amino acids (e.g., N-ε-lauroyl-L-lysine, N-ε-palmitoylhydrazine, N-α-paritolylornithine, N-α-lauroylarginine, and N-α-hardened tallow fatty acid acylarginine); N-acyl polypeptides such as N-lauroylglycylglycine; α-amino fatty acids such as α-aminooctanoic acid and α-aminolauric acid; polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene / styrene copolymer, tetrafluoroethylene, etc.

[0072] UV absorbers may include p-aminobenzoic acid, ethyl p-aminobenzoate, amyl p-aminobenzoate, octyl p-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butyl phenyl salicylate, trimethylcyclohexyl salicylate, benzyl cinnamate, 2-ethoxyethyl p-methoxycinnamate, octyl p-methoxycinnamate, glyceryl mono-2-ethylhexane di-p-methoxycinnamate, isopropyl p-methoxycinnamate, diisopropyl / diisopropyl cinnamate. Mixtures of cinnamic acid esters, ethyl urethane, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzophenone, tetrahydroxybenzophenone, 4-tert-butyl-4′-methoxybenzoylmethane, 2,4,6-triphenylamine-p-(carbonyl-2′-ethylhexyl-1′-oxy)-1,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, etc.

[0073] Fungicides may include physalisol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcinol, isopropyl methylphenol, azulene, salicylic acid, zincyllione, benzalkonium chloride, photosensitizer No. 301, sodium mononitroguaiacol, undecenoic acid, etc.

[0074] Antioxidants can include butylated hydroxyanisole, propyl gallic acid, isoascorbic acid, etc.

[0075] pH adjusters can include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, etc.

[0076] Alcohols can include higher alcohols such as cetyl alcohol.

[0077] Furthermore, the ingredients that can be added are not limited to the above components, and any of the above components can be used within the scope of not impairing the purpose and effect of the present invention, provided that they are used in an amount of preferably 0.01% to 5% by weight, more preferably 0.01% to 3% by weight, based on the total weight of the composition.

[0078] When the formulation of the present invention is a lotion, paste, cream or gel, the carrier component may include animal fiber, plant fiber, wax, paraffin, starch, tracanth, cellulose derivatives, polyethylene glycol, organosilicon, bentonite, silica, talc, zinc oxide, etc.

[0079] When the formulation of the present invention is a powder or a spray, the carrier component may include lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder, and in particular, when the formulation of the present invention is a spray, the carrier component may additionally include a propellant (e.g. chlorofluorocarbon, propane / butane or dimethyl ether).

[0080] When the formulation of the present invention is a solution or emulsion, the carrier component may contain a solvent, solvator or emulsifier, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylethylene glycol oil, glycerol aliphatic esters, polyethylene glycol or dehydrated sorbitol esters of fatty acids.

[0081] When the formulation of the present invention is a suspension, the carrier component may include a liquid diluent (e.g., water, ethanol or propylene glycol), a suspending agent (e.g., ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene dehydrated sorbitol ester), microcrystalline cellulose, aluminum hydroxide, bentonite, agar, tracanth, etc.

[0082] When the formulation of the present invention is a detergent containing a surfactant, the carrier component may include fatty alcohol sulfate / ester, fatty alcohol ether sulfate / ester, sulfosuccinate monoester, hydroxyethanesulfonate / ester, imidazoline. Derivatives, methyl taurine, sarcosine salts / esters, fatty acid amide ether sulfates / esters, alkyl amide betaine, fatty alcohols, fatty acid glycerides, fatty acyl diethanolamine, vegetable oils, linoleic acid derivatives, ethoxylated glycerol fatty acid esters, etc.

[0083] The invention will be described in more detail below through working examples. The following embodiments are merely examples to help understand the invention, and the scope of the invention is not limited thereto.

[0084] Invention Embodiments

[0085] (Implementation Plan)

[0086] Experimental Example 1. Measurement of epidermal growth factor receptor (EGFR) activity of oxcarbamate

[0087] Phosphorylation of tyrosine residues in the cytoplasmic portion of EGFR, which is an EGF receptor for the cell membrane, occurs in conjunction with EGFR activation. Furthermore, by utilizing the characteristic that EGFR is internalized into the cell when EGFR activity is maintained, this experiment uses a cell-based biosensor system capable of measuring EGFR activity (see Korean Patent Publication No. 2019-0014340).

[0088] Cells of the biosensor system were treated with 10 ng / ml EGF and 100 μM oxcarbazine (purchased from InterPharm), and changes in fluorescence signal were observed. Cells were treated with oxcarbazine after being cultured in serum-free medium for 24 hours.

[0089] As a result, such Figure 1 As shown, EGFR was activated in cells treated with 100 μM oxcarbazine compared to the control group treated with DMSO alone.

[0090] Experimental Example 2. Measurement of cytotoxicity depending on oxcarbazine concentration

[0091] The cytotoxicity of oxacin was measured using HaCaT cells as keratinocytes.

[0092] When cells are used as a yellow, water-soluble substrate, MTT tetrazolium During treatment, this substrate is reduced to MTT formazan by mitochondrial enzymes, and the color changes to blue-violet. In other words, the blue-violet color becomes deeper as the number of live cells increases, so the absorbance of blue-violet at 540 nm reflects the concentration of live cells.

[0093] Specifically, after culturing cells to fill each well of a 96-well plate, the cells were treated with 0 μM, 10 μM, 25 μM, 50 μM, and 100 μM oxcarbamate and cultured at 37°C in a 5% CO2 incubator for 24 and 48 hours. Cell viability was then measured using the MTT assay. The control group, untreated with 0 μM oxcarbamate, was treated with the same amount of DMSO.

[0094] As a result, such Figure 2 As shown, treatment with 25 μM and 50 μM oxcarbazine revealed some cytotoxicity, while culturing cells with 100 μM oxcarbazine for 48 hours showed 50% to 60% toxicity. Therefore, subsequent experiments were conducted using oxcarbazine concentrations up to 10 μM.

[0095] Experimental Example 3. Due to the ERK and AKT activities of oxcarbamate

[0096] It is known that the activities of extracellular signal-regulated kinases (ERK) and protein kinase B (AKT) transmit signals essential for cell growth and survival through the activity of EGFR. Furthermore, in Experiment 1, it was observed that oxcarnitine generates a signal due to EGFR activation in cells of a biosensor system used to measure EGFR activity. Therefore, the study measured whether EGFR activity is induced by oxcarnitine to the activities of ERK and AKT.

[0097] Specifically, HaCaT cells were cultured in serum-free medium and treated with 10 μM oxcarbazine for 10 min, 30 min, and 60 min, respectively. Subsequently, phosphorylated and activated ERK and AKT were analyzed by Western blotting. Phosphorylated antibodies serving as markers of activated ERK and AKT, and a universal antibody for measuring all of them were used as antibodies.

[0098] As a result, such Figure 3 As shown, 10 μM oxcarbazine was determined to adequately induce the activity of ERK and AKT within 10 minutes. ERK and AKT were also determined to be rapidly activated within 30 minutes.

[0099] Experimental Example 4. Due to changes in oxcarpine gene expression

[0100] Signals generated by EGFR induce the expression of genes important for cell growth and survival via key signal-mediated proteins (such as ERK and AKT). The egrl, c-myc, c-jun, and c-fos genes are regulated by EGFR and are known to promote cell growth and survival. In Experiment 2, HaCaT cells cultured in serum-free medium were treated with 10 μM oxcarbamate (which sufficiently induces ERK and AKT activity), and changes in gene expression were analyzed after 1 hour.

[0101] Specifically, Tri-reagent was used for mRNA extraction, and cDNA was generated via reverse transcription polymerase chain reaction (RT-PCR) using poly-A primers and reverse transcriptase. Real-time quantitative PCR was used to measure the amount of each mRNA. The amount of each mRNA was corrected using the amount of β-actin (a constituent gene).

[0102] As a result, such Figure 4 As shown, egrl, c-myc, c-jun, and c-fos mRNA levels increased.

[0103] Experimental Example 5. Measurement of keratinocyte growth and proliferation using oxaline

[0104] Cell Counting Kit-8 (CCK-8) is a water-soluble tetrazolium compound called WST-8. The salt reagent is used to form an orange, water-soluble formazan via dehydrogenases in the mitochondria of living cells. The absorbance at 450 nm (the orange wavelength indicated by CCK-8) reflects the number of cells and is therefore used to measure cell growth and proliferation.

[0105] Specifically, in 1×10 4 After culturing HaCaT cells in 96-well plates for 24 hours, the cells were replaced with serum-free medium to remove growth factors and treated with non-cytotoxic oxcarbazin at concentrations of 10 μM or lower (i.e., 1 nM, 10 nM, 100 nM, 1 μM, and 10 μM). As a negative control, the same amount of solvent DMSO was used, and as a positive control, EGF was used at a concentration of 1 ng / ml. After 24 hours of culture, cells were treated with CCK8, and cell growth was analyzed by measuring absorbance at 450 nm.

[0106] As a result, such Figure 5 As shown, treatment with 100 nM, 1 μM and 10 μM oxcarbazine significantly increased cell growth and proliferation compared to the negative control group (one-way ANOVA, *p<0.05).

[0107] Experimental Example 6. Cell growth promoting effect of oxcarbamate (wound healing evaluation)

[0108] The cell growth-promoting effect of oxcarbazin was determined by a wound healing assay. Specifically, after HaCaT cells were adequately cultured in 12-well plates, a certain area of ​​cells was scraped off using a yellow-tipped plate to create a wound area, the medium was replaced with serum-free medium, and the cells were treated with 1 μM oxcarbazin. DMSO was used as a negative control, and 1 ng / ml EGF was used as a positive control.

[0109] As a result, such Figure 6a and 6b As shown, when treated with oxcarbazin during the 6-day culture period, the wound area was filled more quickly compared to the negative control group, and the difference was more significant from day 3 onwards compared to the negative control group (t-test, *p<0.05).

[0110] Experimental Example 7. The efficacy of oxcarbazin against ultraviolet (UVB) stimulation.

[0111] In the ultraviolet wavelength range, UVB wavelengths corresponding to 280 nm to 315 nm were used to penetrate the epidermal layer, in which numerous keratinocytes reside, to induce cell damage. Since UVB-induced cell damage also leads to apoptosis, it was determined whether oxaline exhibited resistance to UVB-induced cell damage (due to its promotion of keratinocyte growth and the transmission of survival signals).

[0112] Specifically, 1×10⁻⁶ cells will be cultivated within them. 4 A 96-well plate containing HaCaT cells was treated with 500 mJ / cm². 2 Cells were irradiated with UVB and then cultured in serum-free medium containing 1 μM oxcarbamate or EGF. DMSO was used as a negative control, and cell viability was measured using the CCK-8 assay used in Experiment 5.

[0113] As a result, such Figure 7 As shown, it was determined that in the negative control group, due to 500 mJ / cm 2 UVB induces apoptosis, resulting in only about 40% of cells surviving, while 1 μM oxcarbazine results in about 60% cell survival, which is statistically significant (t-test, *p<0.05).

[0114] Industrial applicability

[0115] While exemplary embodiments have been shown and described above, it will be apparent to those skilled in the art that modifications and variations may be made without departing from the scope of this disclosure as defined by the appended claims.

Claims

1. Use of oxcarpine or a salt thereof, represented by formula (1), in the preparation of compositions in the form of topical formulations for skin regeneration: [Equation (1)] 2. The use according to claim 1, wherein the composition comprises oxcarbamate or a salt thereof in an amount of 10 nM to 100 μM.

3. The use according to claim 1, for improving or treating skin and tissue damage caused by burns.

4. The use according to claim 1, for preventing or improving cell damage from external stimuli.

5. The use according to claim 4, wherein the external stimulus is ultraviolet light.

6. The use according to claim 1, for enhancing skin elasticity.

7. The use according to any one of claims 1 to 6, wherein the composition is a pharmaceutical composition.

8. The use according to any one of claims 1 to 2 and 4 to 6, wherein the composition is a cosmetic composition.

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