Composition for anti-aging or skin regeneration containing isoproterocurenol

Abstract

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

Description

Technical Field

[0001] The present invention relates to a composition for anti-aging or skin regeneration, and more particularly, to a composition for anti-aging or skin regeneration comprising isoprocurcumenol. Background Art

[0002] Skin aging is an inevitable phenomenon. Aging leads to a loss of skin elasticity, wrinkles, and symptoms such as hyperpigmentation 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 rays.

[0003] As the skin ages, the function of cells declines, and at the same time, the skin is affected by the external environment, such as ultraviolet rays. First, as aging progresses, the division rate of keratinocytes slows down. Although the normal skin regeneration cycle is about 28 days, this cycle increases with aging. Secondly, it is known that the number of fibroblasts present in the epidermis decreases with aging, and at the same time, their function 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, resulting in an increase in dermal atrophy.

[0004] However, even though aging cannot be completely prevented even at 100 years of age, there is an increasing desire and demand for substances that can improve skin aging to enhance beauty. Therefore, there is a need for the rapid development of substances that slow down or improve skin aging.

[0005] Epidermal growth factor (Epidermal growth factor, EGF) is a growth factor that regulates the growth and survival of epidermis or epidermal cells. EGF is also considered to be a factor related to cancer and has been used medically to treat wounds and burns. Recently, EGF has been widely used as a raw material for cosmetics, and many studies have been carried out on the cosmetic composition for preventing skin aging comprising EGF, such as Korean Patent No. 2093884. Therefore, it is necessary to study a small molecule substance of natural origin with a function similar to that of EGF and stable and that can be used to pass through the skin barrier. However, the biosynthesis, separation and purification of known EGF are expensive, and it is difficult to store EGF in an active state. In addition, there is a problem that EGF cannot easily pass through 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 those of EGF and are stable and can be used to pass through the skin barrier. Summary of the Invention

[0007] Technical issues

[0008] One aspect of the present disclosure is to provide a composition for anti-aging or skin regeneration, which comprises isoproterocurenol, 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 pass through the skin barrier.

[0009] Technical Solution

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

[0011] [Formula (1)]

[0012]

[0013] Beneficial effects

[0014] According to one aspect of the present disclosure, isoprotocurenol can be extracted from various plant species such as zedoaria. Composition according to the present invention has the advantage of not having side effect by using it. In addition, like EGF, isoprotocurenol is combined with EGFR to activate the signal relevant to the survival, growth and proliferation of cells, thereby increasing the resistance and viability of skin cells to external stimuli, and enhancing the regeneration of skin damaged due to external stimuli or aging. In addition, although isoprotocurenol has a function similar to that of EGF, unlike EGF, isoprotocurenol is stable and small in size, and therefore has the advantage of being able to move through the skin barrier to arrive at the skin basal layer to perform function. Therefore, isoprotocurenol of the present invention can be used in a variety of ways, for example, in medicine, quasi-drug, cosmetics, spice material, functional biomaterial and functional food material, use. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The results of confirming that, when a biosensor protein (EGFP-SH2) was expressed by intracellular injection of an EGFR activity measurement biosensor system, the protein was expressed by EGF and also by isoproterocurenol are shown.

[0016] Figure 2 Shown are the results of determining the cytotoxicity to HaCaT cells as keratinocytes depending on the concentration of isoproterocurenol.

[0017] Figure 3Shown are the results of confirming that ERK and AKT, which are EGFR isoforms, are activated by isoproterocurenol.

[0018] Figure 4 Shown are the results of determining the expression patterns of egrl, c-myc, c-jun, and c-fos genes increased by isoproterocurenol.

[0019] Figure 5 Shown are the results of determining the growth-promoting effect of keratinocytes by isoproterocurenol by CCK-8 assay.

[0020] FIG. 6 shows photographs (a) and graphs (b) showing the results of the proliferation and growth promoting effects of keratinocytes by isoproterocurenol determined by wound healing evaluation.

[0021] Figure 7 It was shown that the resistance of keratinocytes under conditions of cell damage caused by UVB is increased by isoproterocurenol. DETAILED DESCRIPTION

[0022] Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings. However, the embodiments of the present invention may be modified into various forms, and the scope of the present invention is not limited to the embodiments described below.

[0023] The inventors have completed the present invention by experimentally confirming that isoproterocurenol, 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 so as to easily move through the skin barrier.

[0024] The present invention relates to the use of isoproterocurenol represented by the following formula (1) or a salt thereof for preparing a composition for anti-aging or skin regeneration:

[0025] [Formula (1)]

[0026]

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

[0028] [Formula (1)]

[0029]

[0030] Isoproterocurenol can be obtained from extracts of various plant species such as Curcuma zedoaria, and has a function similar to that of epidermal growth factor (EGF) that activates signals related to cell survival, growth, and proliferation in cells, thereby increasing the resistance and viability of skin cells to external stimuli and enhancing the regeneration of skin damaged due to external stimuli or aging.

[0031] The composition of the present invention may contain isoprotocurenol or a salt thereof in an amount of 10 nM to 100 μM, or 1 μM to 100 μM, for example 5 μM to 100 μM, preferably 10 μM to 100 μM, and more preferably 25 μM to 50 μM. When the concentration of isoprotocurenol or a salt thereof is less than 1 μM, the effect of isoprotocurenol or a salt thereof may not be fully expressed, and when the concentration is greater than 100 μM, the cytotoxicity may be significantly increased.

[0032] "Skin regeneration" in the present invention may refer to promoting the growth of skin cells and enhancing their viability to prevent cell damage from external or internal stimuli, reducing skin wrinkles, and enhancing skin elasticity. Furthermore, skin regeneration includes improving or treating skin damage caused by the external environment, such as skin and tissue damage caused by burns. Thus, the compositions of the present invention can improve or treat burn wounds.

[0033] In addition, the isoprotocurenol or its salt of the present invention can be combined with EGFR to promote cell proliferation or growth, thereby activating anti-aging or skin regeneration. The cells that proliferate or grow by the isoprotocurenol or its salt of the present invention can be epidermal cells, preferably keratinocytes (which are cells that produce keratin), but the present invention is not limited thereto.

[0034] Keratin, a protein that is a major component of various tissues in animals, makes hair shiny and elastic and gives skin vitality and elasticity. In addition, keratin protein in the stratum corneum of the skin has strong resistance to chemicals, thereby performing a protective function against chemical irritation to the skin.

[0035] Therefore, the composition of the present invention can enhance skin elasticity or improve skin wrinkle problems.

[0036] At the same time, the composition of the present invention can enhance resistance to cell damage caused by external stimuli, thereby preventing or improving cell damage caused by external stimuli. The external stimulus can be chemical stimulation caused by cosmetics or other external agents, physical stimulation or ultraviolet light, preferably ultraviolet light, but the present invention is not limited thereto.

[0037] Furthermore, the composition of the present invention may be a parenteral composition or an oral composition, such as a pharmaceutical composition, a cosmetic composition, or a health functional food composition.

[0038] Specifically, when the composition of the present invention is a pharmaceutical composition, the composition may further comprise a pharmaceutically acceptable carrier, and the composition may further comprise a suitable carrier, template, and diluent commonly used in the preparation of pharmaceutical compositions. The composition may also be formulated and used in the form of oral preparations, external preparations, suppositories (e.g., powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols), and sterile injections according to conventional methods known in the art.

[0039] Pharmaceutical composition according to the present invention can be prepared and used with the form of external preparation (such as powder, granule, tablet, capsule, suspension, emulsion, syrup and aerosol) and sterile injection, and can preferably have the preparation of cream, gel, patch, spray, ointment, plaster, lotion, liniment, paste or dressing.The carrier, excipient and diluent that can be included in described composition can be for comprising the compound of at least one of following: lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.

[0040] The compositions can be prepared using diluents or excipients (for example, fillers, extenders, binding agents, 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 extract with at least one excipient (for example starch, calcium carbonate, sucrose, lactose and gelatin). Except simple excipients, lubricants, such as magnesium stearate talc, can also be used. Except the commonly used simple diluents such as water and liquid paraffin in this area, liquid preparations (such as suspensoids, solutions, emulsions and syrups) for oral use can also include various excipients (for example wetting agents, sweeteners, air fresheners and preservatives).

[0041] Preparations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensoids, emulsions, lyophilized agents, and suppositories. Non-aqueous solvents and suspensoids may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil and injectable esters such as ethyl oleate. Witepsol, polyethylene glycol, Tween 61, cocoa butter, laurin, glycerin gelatin, etc. may be used as the matrix for suppositories.

[0042] The pharmaceutical composition according to the present invention can be used in a pharmacologically effective amount. In the present invention, a "pharmacologically effective amount" refers to an amount that is sufficient to treat a disease in a reasonable benefit / risk ratio that is applicable to 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 of administration and the rate of excretion, the time period for treatment, factors such as the drugs used simultaneously, and other factors well known to the medical field. The pharmaceutical composition according to the present invention can be used as a single therapeutic agent or in combination with other therapeutic agents, can be used sequentially or simultaneously with conventional therapeutic agents, and can be a single administration or multiple administrations. In view of all the above factors, it is important to use the minimum amount that can obtain the maximum effect without causing side effects. Such an amount can be easily determined by those skilled in the art to which the present invention pertains.

[0043] The pharmaceutical composition of the present invention can be administered to a subject by various routes. The pharmaceutical composition of the present invention is determined according to various factors (e.g., the disease to be treated, the route of administration, the age, sex and weight of the patient, and the severity of the disease) and the type of medicine as the active ingredient.

[0044] Composition of the present invention can provide with the form of health functional food composition.In addition, for the purpose of skin regeneration (for example, anti-aging, strengthening skin elasticity and improving the problem of skin wrinkles), described composition can be added in food. Health functional food composition according to the present invention can be used as it is or can be used together with other food or food ingredients, and can suitably use according to ordinary method. The amount of active component to be mixed can suitably determine according to intended use such as prevention and health care or therapeutic treatment. When preparing food or beverage, based on the weight of total composition, isoproterocurenol or its salt can be added in the health functional food composition according to the present invention usually with greater than 0 % by weight and as many as 15 % by weight, preferably 1 % by weight to 10 % by weight or less amount. However, isoproterocurenol has no problem aspect stability, therefore can use as active component with the amount equal to or greater than above scope.

[0045] The type of food is not particularly limited, but examples of foods that may contain the material are meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, and dairy products (including ice cream), various soups, beverages, tea, drinks, vitamin complexes, etc., and may include all health functional foods in the conventional sense.

[0046] The same as conventional beverages, this beverage can comprise various flavoring agents or natural carbohydrates as other ingredients.Natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol.Sweeteners can comprise natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame.The amount of natural carbohydrates can be about 0.01g to 0.20g of the health functional food composition according to the present invention for every 100mL, preferably about 0.04g to 0.10g.

[0047] In addition, the health functional food composition according to the present invention can include various nutritional supplements, vitamins, electrolytes, flavorings, coloring agents, pectin acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerol, carbonating agents for carbonated beverages, etc. In addition, the health functional food composition according to the present invention can include the pulp for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used alone or in combination. The amount of these additives is not very important, but can be selected within the range of 0.01 weight portion to 0.20 weight portion of the health functional food composition according to the present invention per 100 weight portions.

[0048] The composition of the present invention can be provided in the form of a cosmetic composition. The cosmetic composition according to the present invention can be formulated in the following forms: a skin lotion, a skin softener, a skin firming agent, an astringent, a lotion, an emulsion lotion, a moisturizing lotion, a nourishing lotion, a massage cream, a nourishing cream, a moisturizing cream, a hand cream, a foundation, an essence, a nourishing essence, a peel-off mask, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body wash, or a body cleanser.

[0049] The cosmetic composition according to the present invention may additionally comprise a compound selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high-molecular peptides, high-molecular polysaccharides and sphingolipids.

[0050] Any water-soluble vitamin can be used as long as it can be added to the cosmetic composition. Preferably, vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinamide, folic acid, vitamin C, vitamin H, etc. can be used. Its salts (for example, thiamine hydrochloride and ascorbic acid sodium salt) or derivatives thereof (for example, ascorbic acid-2-phosphate sodium salt and ascorbic acid-2-phosphate magnesium salt) can also be used as the water-soluble vitamins in the present invention. Water-soluble vitamins can be obtained by conventional methods (for example, microbial transformation method, method for purification from microbial culture and enzymatic method or chemical synthesis method).

[0051] 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) and the like. Derivatives thereof (e.g., ascorbyl palmitate, ascorbyl stearate, ascorbyl dipalmitate, dl-α tocopheryl acetate, dl-α tocopheryl nicotinate, vitamin E, DL-panthenol, D-panthenol and pantothenyl ethyl ether) can also be used as the oil-soluble vitamins in the present invention. Available vitamins can be obtained by conventional methods (e.g., microbial transformation methods, methods for purification from microbial cultures and enzymatic methods or chemical synthesis methods).

[0052] Any high-molecular 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. The high-molecular peptide can be obtained by conventional methods such as purification from microbial culture media, enzymatic methods or chemical synthesis methods, or can be conventionally obtained by purification from natural products such as pig or bovine dermis or silk fibers.

[0053] 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 salt (e.g., sodium salt). For example, chondroitin sulfate or its salt can be conventionally obtained by purification from mammals or fish.

[0054] Any sphingolipid can be used as long as it is added to cosmetics, and preferably includes ceramide, phytosphingosine, glycosphingolipids, etc. Sphingolipids can be purified from mammals, fish, shellfish, yeast, or plants by conventional methods, or can be obtained by chemical synthesis.

[0055] In addition to the above-mentioned essential ingredients, the cosmetic composition of the present invention may further contain other ingredients generally added to cosmetics as needed.

[0056] Other ingredients that may be added include oil and fat components, moisturizers, emollients, surfactants, organic or inorganic pigments, organic powders, UV absorbers, preservatives, bactericides, antioxidants, plant extracts, pH regulators, alcohols, pigments, fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, etc.

[0057] The oil and fat components may include ester-based oils and fats, hydrocarbon-based oils and fats, silicone-based oils and fats, fluorine-based oils and fats, animal oils and fats, vegetable oils and fats, and the like.

[0058] Ester oils and fats may include tri-2-ethylhexanoin, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, butyl stearate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, diisopropyl adipate, isoalkyl neopentanoate, glyceryl tri(caprylate or caprate), trimethylolpropane tri(2-ethylhexanoate), trimethylolpropane tri(2-ethylhexanoate), Isostearate, Pentaerythritol Tetra(2-ethylhexanoate), Cetyl Octanoate, Decyl Laurate, Hexyl Laurate, Decyl Myristate, Myristyl Myristate, Cetyl Myristate, Stearyl Stearate, Decyl Oleate, Cetyl Ricinoleate, Isostearate Laurate, Isotridecyl Myristate, Isocetyl Palmitate, Octyl Stearate, Isocetyl Stearate, Isodecyl Oleate, Octyldodecyl Oleate, Octyldodecyl Linoleate, Isopropyl Isostearate, Cetearyl 2-Ethylhexanoate, Stearyl 2-Ethylhexanoate, Hexyl Isostearate, Ethylene Glycol Dioctanoate, Ethylene Glycol Dioleate , Propylene Glycol Dicaprate, Propylene Glycol Di(Capryl or Capric Acid), Propylene Glycol Dicaprylate, Neopentyl Glycol Dicaprate, Neopentyl Glycol Dicaprylate, Tricaprylin, Tridecanoic Acid, Triisopalmitin, Triisostearin, Octyldodecyl Neopentanoate, Isostearate Caprylate, Octyl Isononanoate, Hexyldecyl Neodecanoate, Octyldodecyl Neodecanoate, Isocetyl Isostearate, Isostearyl Isostearate, Octyldecyl Isostearate, Polyglyceryl Oleate, Polyglyceryl Isostearate, Triisohexadecyl Citrate, Triisoalkyl Citrate, Triisooctyl Citrate, Lauryl Lactate, Myristyl Lactate, Cetearyl Lactate Alkyl esters, octyldecyl 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, dihydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, isocetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate, isostearyl 12-stearoylhydroxystearate, etc.

[0059] The hydrocarbon-based oils and fats may include squalene, liquid paraffin, α-olefin oligomers, isoparaffins, ozokerite, paraffin, liquid isoparaffins, polybutene, microcrystalline wax, petrolatum, and the like.

[0060] Silicone-based oils and fats may include polymethylsilicone, tolylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane / methylhexadecyloxysiloxane copolymer, dimethylsiloxane / methylstearyloxysiloxane copolymer, alkyl-modified silicone oil, amino-modified silicone oil, and the like.

[0061] Fluorine-based oils and fats may include perfluoropolyethers and the like.

[0062] Animal or vegetable oils and fats can 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 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, tallow, horse oil, mink oil, tilapia oil, jojoba oil, candelilla wax, carnauba wax, liquid lanolin, hydrogenated castor oil, etc.

[0063] The wetting agent may include a water-soluble low molecular weight moisturizer, a fat-soluble molecular moisturizer, a water-soluble polymer, or an oil-soluble polymer.

[0064] Water-soluble low-molecular moisturizers can include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or greater), polypropylene glycol (polymerization degree n = 2 or greater), polyglycerol B (polymerization degree n = 2 or greater), lactic acid, lactate / ester, etc.

[0065] Fat-soluble low-molecular moisturizers may include cholesterol, cholesterol esters, and the like.

[0066] The water-soluble polymer may include carboxyvinyl polymer, polyaspartate, tragacanth gum, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitosan, chitosan, dextrin, and the like.

[0067] The fat-soluble polymer may include polyvinyl pyrrolidone / eicosene copolymer, polyvinyl pyrrolidone / hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, high molecular weight silicone, and the like.

[0068] Emollients may include long-chain acyl glutamic acid cholesterol esters, hydroxystearate cholesterol esters, 12-hydroxystearate, stearic acid, rosin acid, lanolin fatty acid cholesterol esters, and the like.

[0069] The surfactant may include nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants and the like.

[0070] The nonionic surfactant can include self-emulsifying monostearate glyceryl, propylene glycol fatty acid ester, glycerol fatty acid ester, polyglycerol fatty acid ester, fatty acid dehydrated sorbitol ester, polyoxyethylene (POE) dehydrated sorbitol fatty acid ester, POE dehydrated sorbitol fatty acid ester, POE glycerol fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hydrogenated castor oil, POE castor oil, POE / POP (polyoxyethylene / polyoxypropylene) copolymer, POE / POP alkyl ether, polyether-modified silicone, lauric acid alkanolamide, alkylamine oxide, hydrogenated soybean lecithin, etc.

[0071] Anionic surfactants can include fatty acid soaps, α-acyl sulfonates, alkyl sulfonates, alkyl allyl sulfonates, alkyl naphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, alkylamide phosphates, alkyl acylalkyl taurates, N-acyl amino acid salts, POE alkyl ether carboxylates, alkyl sulfosuccinates, sodium alkyl sulfoacetates, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphates, and the like.

[0072] Cationic surfactants may include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetearyltrimethylammonium chloride, distearyldimethylammonium chloride, octadecyldimethylbenzyl ammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, diethylaminoethyl stearate, dimethylaminopropyl stearate, quaternary ammonium salts of lanolin derivatives, and the like.

[0073] The amphoteric surfactant may include carboxybetaine type, amidobetaine type, sulfobetaine type, hydroxysulfobetaine type, amidosulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amidoamine type and the like.

[0074] The organic or inorganic pigment 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 complexes thereof); 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 thereof.

[0075] Organic powders may include metal soaps such as calcium stearate; alkyl phosphate metal salts (e.g., sodium zinc palmitate, zinc laurate soap, and calcium laurate); polyvalent metal salts of acylamino acids (e.g., calcium N-lauroyl-β-alanine, zinc N-lauroyl-β-alanine, and calcium N-lauroylglycine); 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-palmitoylhydrazide, N-α-paritoylornithine, 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-aminocaprylic acid and α-aminolauric acid; polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene / styrene copolymer, tetrafluoroethylene, and the like.

[0076] 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, butylphenyl salicylate, trimethylol salicylate, benzyl cinnamate, 2-ethoxyethyl p-methoxycinnamate, octyl p-methoxycinnamate, glyceryl mono-2-ethylhexane di-p-methoxycinnamate, isopropyl p-methoxycinnamate, diisopropyl / diisopropyl myristate. Cinnamate mixture, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzophenone, tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-triphenylamine-p-(carbonyl-2'-ethylhexyl-1'-oxy)-1,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, etc.

[0077] Bactericides may include hinokitiol, triclosan, triclosan, chlorhexidine gluconate, phenoxyethanol, resorcinol, isopropyl methylphenol, azulene, salicylic acid, zincphyllione, benzalkonium chloride, photosensitizer No. 301, sodium mononitroguaiacol, undecylenic acid, and the like.

[0078] Antioxidants may include butylated hydroxyanisole, propyl gallic acid, erythorbic acid, and the like.

[0079] The pH adjuster may include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, and the like.

[0080] The alcohol may include higher alcohols such as cetyl alcohol.

[0081] Furthermore, ingredients that can be added are not limited to the above components, and any of the above ingredients may be used within the range not impairing the objects and effects of the present invention, provided that they are used in an amount of preferably 0.01% by weight to 5% by weight, more preferably 0.01% by weight to 3% by weight, based on the total weight of the composition.

[0082] When the preparation 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, silicone, bentonite, silicon dioxide, talc, zinc oxide and the like.

[0083] When the preparation of the present invention is a powder or a spray, the carrier component may include lactose, talc, silicon dioxide, aluminum hydroxide, calcium silicate or polyamide powder, and in particular, when the preparation of the present invention is a spray, the carrier component may additionally contain a propellant (e.g., chlorofluorocarbon, propane / butane or dimethyl ether).

[0084] When the formulation of the present invention is a solution or emulsion, the carrier component may include a solvent, solvating agent or emulsifier, such as water, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, aliphatic glycerides, polyethylene glycol or sorbitan esters of fatty acids.

[0085] When the formulation of the present invention is a suspension, the carrier components may include a liquid diluent (e.g., water, ethanol or propylene glycol), a suspending agent (e.g., ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters), microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, tracanth, etc.

[0086] When the preparation of the present invention is a surfactant-containing cleanser, the carrier component may include fatty alcohol sulfates / esters, fatty alcohol ether sulfates / esters, sulfosuccinic acid monoesters, isethionates / esters, imidazolinium derivatives, methyl taurates, sarcosinates / esters, fatty acid amide ether sulfates / esters, alkylamide betaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linseed oil derivatives, ethoxylated glyceryl fatty acid esters, and the like.

[0087] Hereinafter, the present invention will be described in more detail through working examples.The following embodiments are merely examples to help understand the present invention, and the scope of the present invention is not limited thereto.

[0088] Embodiments of the invention

[0089] (Implementation Plan)

[0090] Experimental Example 1. Measurement of Epidermal Growth Factor Receptor (EGFR) Activity of Isoproterocurenol

[0091] Phosphorylation of tyrosine residues present in the cytoplasmic portion of EGFR, an EGF receptor on the cell membrane, occurs with EGFR activation. Furthermore, by utilizing the characteristic that EGFR is internalized into cells when its activity is maintained, this experiment used a cell-based biosensor system that can measure EGFR activity (see Korean Patent Publication No. 2019-0014340).

[0092] The cells of the biosensor system were treated with 10 ng / ml of EGF and 100 μM of isoproterocurenol (purchased from InterPharm), and the changes in the fluorescence signal were observed. After the cells were cultured in serum-free medium for 24 hours, they were treated with isoproterocurenol.

[0093] As a result, Figure 1 As shown, it was confirmed that EGFR was activated in cells treated with 100 μM of isoproterocurenol compared with the control group treated with only the solvent DMSO.

[0094] Experimental Example 2. Measurement of cytotoxicity depending on isoproterocurenol concentration

[0095] The cytotoxicity of isoproterocurenol was measured using HaCaT cells as keratinocytes.

[0096] When cells are treated with MTT tetrazolium When treated, this substrate is reduced to MTT formazan by mitochondrial enzymes, and the color changes to bluish-purple. That is, since the bluish-purple color becomes deeper as the number of living cells increases, the bluish-purple absorbance at a wavelength of 540 nm reflects the concentration of living cells.

[0097] Specifically, after culturing cells to fill each well of a 96-well plate, the cells were treated with isoprotocurenol at concentrations of 0 μM, 10 μM, 25 μM, 50 μM, and 100 μM and cultured at 37°C in a 5% CO2 incubator for 24 hours and 48 hours. Thereafter, cell viability was measured using MTT reagent. A control group not treated with isoprotocurenol (0 μM) was treated with the same amount of solvent DMSO.

[0098] As a result, Figure 2 As shown, it was determined that when cells were incubated with 100 μM isoproterocurenol for 48 hours, some cytotoxicity was exhibited, but the difference was only 10%. Therefore, it was determined that the cytotoxicity in keratinocytes was very low.

[0099] Experimental Example 3. ERK and AKT activity due to isoproterocurenol

[0100] It is known that the activities of extracellular signal-regulated kinase (ERK) and protein kinase B (AKT) transmit signals necessary for cell growth and survival through the activity of EGFR. In addition, in Experimental Example 1, it was observed that in cells of a biosensor system for measuring EGFR activity, isoproterocurenol produced a signal due to the activation of EGFR. Therefore, it was measured whether the activity of EGFR was induced by isoproterocurenol to the activity of ERK and AKT.

[0101] Specifically, HaCaT cells were cultured in serum-free medium and treated with 10 μM isoproterenol for 10 minutes, 30 minutes, and 60 minutes, respectively. Phosphorylated and activated ERK and AKT were then analyzed by Western blotting. Phosphorylation antibodies, which are markers for activated ERK and AKT, and a universal antibody for overall measurement were used.

[0102] As a result, Figure 3 As shown, it was confirmed that 10 μM isoproterocurenol fully induced the activities of ERK and AKT within 10 minutes. It was also confirmed that ERK and AKT were rapidly activated within 30 minutes.

[0103] Experimental Example 4. Changes in gene expression due to isoproterocurenol

[0104] The signals generated by EGFR induce the expression of genes important for cell growth and survival via important signal-mediating proteins (such as ERK and AKT). The egrl, c-myc, c-jun, and c-fos genes are regulated by EGFR and are well known as genes that promote cell growth and survival. In Experimental Example 2, HaCaT cells cultured in serum-free medium were treated with 10 μM isoproterenol (which fully induces ERK and AKT activity), and after 1 hour, the changes in gene expression were analyzed.

[0105] Specifically, Tri-reagent was used for mRNA extraction, poly-A primers and reverse transcriptase were used to generate cDNA by reverse transcription polymerase chain reaction (RT-PCR), and real-time quantitative PCR was used to measure the amount of each mRNA. The amount of each mRNA was corrected for the amount of β-actin (which is a constituent gene).

[0106] As a result, Figure 4 As shown, egr1, c-myc, c-jun, and c-fos mRNA were increased.

[0107] Experimental Example 5. Measurement of Keratinocyte Growth and Proliferation by Isoproterocurenol

[0108] Cell counting kit-8 (CCK-8) is a water-soluble tetrazolium The cell number is determined by the absorbance at 450 nm (which is the orange wavelength displayed by CCK-8) and the orange water-soluble formazan is formed by the dehydrogenase in the mitochondria of living cells. Thus, the absorbance at 450 nm (which is the orange wavelength displayed by CCK-8) reflects the number of cells and is therefore used to measure the growth and proliferation of cells. The growth promotion effect of HaCaT cells as keratinocytes was measured by using non-cytotoxic isoproterocurenol at a concentration of 10 μM or less (i.e., 1 nM, 10 nM, 100 nM, 1 μM and 10 μM) by CCK-8 assay.

[0109] Specifically, when 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 contained in the medium and treated with isoproterocurenol at concentrations of 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 at a concentration of 1 ng / ml was used. After culturing for 24 hours, the cells were treated with CCK8 and cell growth was analyzed by measuring absorbance at 450 nm.

[0110] As a result, Figure 5 As shown, it was determined that when cells were treated with isoproterocurenol at concentrations of 10 nM, 100 nM, 1 μM, and 10 μM, the growth and proliferation of cells were significantly increased compared to the negative control group (one-way ANOVA, *p<0.05).

[0111] Experimental Example 6. Cell Growth Promotion Effect of Isoproterocurenol (Wound Healing Evaluation)

[0112] The cell growth-promoting effect of isoproterocurenol was determined by wound healing assessment. Specifically, after HaCaT cells were fully cultured in a 12-well plate, a yellow tip was used to scrape off cells from a certain area to form a wound area. The culture medium was then replaced with serum-free medium and treated with 1 μM isoproterocurenol. The solvent DMSO was used as a negative control, and 1 ng / ml of EGF was used as a positive control.

[0113] As a result, Figure 6a and 6b As shown, it was determined that the wound area was filled faster when treated with isoproterocurenol during the 6-day culture period compared to the negative control group, and showed a more significant difference compared to the negative control group from the 3rd day (t-test, *p<0.05).

[0114] Experimental Example 7. Effect of Isocurenol on Anti-UVB Stimulation

[0115] Among ultraviolet wavelengths, UVB wavelengths corresponding to 280 nm to 315 nm are allowed to penetrate the epidermal layer where many keratinocytes are present to cause cell damage. Since cell damage caused by UVB also leads to apoptosis, it was determined whether isoproterocurenol exhibits resistance to cell damage caused by UVB (due to its promotion of growth and survival signal transmission of keratinocytes).

[0116] Specifically, 1×10 4 A 96-well plate of HaCaT cells was treated with 500 mJ / cm 2 The cells were irradiated with UVB and then cultured in a serum-free medium containing 1 μM isoproterocurenol or EGF. DMSO solvent was used as a negative control group, and cell viability was measured using the CCK-8 method used in Experimental Example 6.

[0117] As a result, Figure 7 As shown, it was determined that in the negative control group, the 2 1 μM of UVB induced apoptosis, so that only about 40% of the cells survived, whereas due to 1 μM of isoproterocurenol, about 60% of the cells survived, which showed a statistically significant value (t-test, *p<0.05).

[0118] Industrial Applicability

[0119] 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 the present disclosure as defined by the appended claims.

Claims

1. Use of an isoproterocurenol represented by the following formula (1) or a salt thereof for preparing a cosmetic composition for external use for skin regeneration: [Formula (1)] 2 . The method according to claim 1 , comprising isoprotocuronol or a salt thereof in an amount of 10 nM to 100 μM.

3. The use according to claim 1, for preventing or improving cell damage caused by external stimulation. The use according to claim 3 , wherein the external stimulus is ultraviolet rays.

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

6. The use according to claim 1, for improving skin wrinkle problems.

Images