JP2004217618A - Laminin 5 production promoting agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain an agent having an action to activate the promotion of laminin 5 productivity in an epidermal keratinocyte of the skin. <P>SOLUTION: The laminin 5 production promoting agent contains an extract derived from soybean as an active component. The agent is useful mainly in a cosmetic field or a dermatological field. <P>COPYRIGHT: (C)2004,JPO&NCIPI

Description

  The present invention relates to the technical field of cosmetics or dermatology, and more specifically, to a laminin 5 production promoter.

  Although the skin structure has been studied using a microscope and an electron microscope for a long time, it was not clear what component contributed to the maintenance of the skin structure. However, Dr. Burgesson discovered laminin 5 in 1991 (see, for example, Non-Patent Document 1), and further discovered the existence of an autoimmune disease against laminin 5 and a serious disease deficient in laminin 5 in the gene. (See Non-Patent Documents 2 and 3, respectively.) In this disease, blister formation was observed at the boundary between the epidermis and the dermis, which revealed that laminin 5 was an essential component for adhesion between the two. Laminin 5 is a main component of a structure composed of various glycoproteins called proteoglycans called basement membrane located at the boundary between the epidermis and the dermis. Laminin 5 further has an activity of promoting the adhesion of epidermal cells, and functions to directly bind epidermal cells to the basement membrane (see Non-Patent Document 4). From these facts, it is predicted that laminin 5 plays an essential role in maintaining the entire structure of the skin by maintaining a normal basement membrane structure.

  It is known that it is important for epithelial cells to adhere to basement membrane components in maintaining their cell functions (see Non-Patent Document 5). That is, it is essential that the epidermal basal cells are properly bound and adhered to the basement membrane for normal skin function expression.

On the other hand, in the senile skin, it has been reported that the basement membrane overlaps (see Non-Patent Document 6), the thickness of the basement membrane and the decrease of the component IV collagen (see Non-Patent Document 7) change, and the structural change of the basement membrane. Is thought to contribute to the deterioration of skin function in the elderly. Therefore, identification and acquisition of a substance that induces the reconstruction of the basement membrane structure by enhancing the production of laminin 5 and improves the skin function has been expected.
Rousselle et al. Cell Biol. 114,567,1991 Fine et al. Am. Acad. Dermatol. , 24, 119, 1991 Aberdam et al., Nat. Genet. 6,299,1994 Schmidhauser et al., Proc. Natn. Acad. U. S. A. , 87, 9118, 1990 Schmidhauser et al., Proc. Natn. Acad. U. S. A. , 87, 9118, 1990 Lavker et al. Invest. Dermatol. , 73, 59, 1979 azquez, Maturitas, 25, 209, 1996

  The present inventors have studied a wide range of animal and plant sources in order to obtain a substance capable of enhancing the production of laminin 5 as described above. As a result, conventionally, a soybean-derived preparation for extracting soybean lecithin, which is used in the technical field where particularly high safety is required, including cosmetics, as a naturally obtained surfactant, or It has been found that a preparation derived from soybean for extracting soybean saponin significantly enhances the ability of human epidermal keratinocytes to produce laminin-5.

  Accordingly, the present invention provides a preparation of laminin 5 containing a soybean-derived preparation, particularly an extract, as an active ingredient. Thus, the provided laminin-5 producing agent can prevent the functional deterioration accompanying the structural change of the basement membrane of normal skin, and can activate the skin with the reduced function. In addition, it can be used as a reagent for detecting the ability to produce laminin 5 in skin, particularly human epidermal keratinocytes, such as a positive control.

<Specific embodiment of the invention>
The soybean-derived preparations according to the present invention include all those having an activity of enhancing (or improving) the ability to produce laminin 5 in human epidermal keratinocytes. Therefore, specifically, the soybean-derived preparation may be a crushed product of soybean itself, or an extract obtained by extracting the crushed product with a suitable solvent. Such extracts include, but are not limited to, those extracted using hydrated lower alcohols, lower alcohols, lower alkyl ketones, lower alkyl ethers, polyhydric alcohols and mixed solvents of two or more of these. Can be mentioned.

  Specific examples of the water-containing lower alcohol include methanol, ethanol, iso-propanol and n-propanol containing water of less than 50% by weight, preferably 5 to 40%. Specific examples of the lower alcohol include: Methanol substantially free of water, ethanol, iso-propanol, n-propanol, n-butanol, sec-butanol, n-pentane, n-hexane, and specific examples of the lower alkyl ketone include acetone , Ethyl methyl ketone, butyl methyl ketone, and iso-butyl methyl ketone.

  Specific examples of the di-lower alkyl ether include diethyl ether, di-iso-propyl ether, di-n-propyl ether, di-n-butyl ether, n-butyl methyl ether, and n-butyl ethyl ether. Specific examples of the lower alkyl carboxylic acid ester include ethyl acetate and butyl acetate.

  Further, specific examples of the polyhydric alcohol include 1,3-butanediol, propylene glycol, glycerol, diglycerol and the like.

  Furthermore, preparations from extracts extracted from soybeans using hydrocarbons such as cyclohexane, toluene and xylene, and halogenated hydrocarbons such as dichloromethane and dichloroethane as extraction solvents are also used as long as the objects of the present invention are met. can do.

  Extraction of the soybean or the soybean crushed product using the above-mentioned solvent is performed according to a method known per se, and the extract is used as it is, or after drying, after re-dissolving the dried product in an appropriate solvent, the laminin 5 described below is extracted. The preparation used in the present invention can be selected by measuring the production activity. The extraction method includes, if necessary, immersing the dried soybeans or the ground soybeans in any of the above-mentioned solvents, leaving them to stand, or using a shaker, and is usually performed by an operation commonly used for solid-liquid extraction. It can be carried out. The operation temperature for extraction can be appropriately selected depending on the solvent system to be used, but the extraction is preferably performed at a temperature of 5 ° C. to the boiling point of the solvent used, preferably up to 60 ° C. The optimal time for the preparation used in the present invention varies depending on the solvent system and temperature used, and the extraction time can be easily determined by those skilled in the art by measuring the laminin 5 production activity described below. Would.

  The above extraction operation can be preferably performed with 70 to 95% by weight of methanol or ethanol (water content: 30 to 5%, pH is adjusted to 3 to 8 as necessary), anhydrous methanol, anhydrous ethanol, acetone and diethyl After immersion at room temperature (15 to 30 ° C.) using ether, 1,3-butanediol, or propylene glycol, the intended product is extracted. The extract thus obtained can be used as it is or further diluted with ethanol, or the dried product can be used as it is, or the dried product can be redissolved in, for example, ethanol to prepare the preparation used in the present invention.

  The skin activator of the present invention contains the above preparation in an amount sufficient to enhance laminin 5 production in epidermal keratinocytes. Such a sufficient amount is not critical because it varies depending on the preparation method from soybeans, but is an extract extracted from a dry and ground soybean product with aqueous methanol or ethanol, or a mixed solvent of ethanol or acetone and diethyl ether. When used as a preparation, its dry matter content is from 0.0001 to 20% by weight per total composition weight. Among such sufficient amounts, those skilled in the art can determine the optimum amount for the individual soybean-derived preparation by repeating the measurement of the production activity of laminin 5 in the cultured epidermal keratinocytes described below. Just fine.

  The skin activator of the present invention can be in the form of an aqueous solution, oil solution, other solution, emulsion, cream, gel, suspension, microcapsule, powder, granule, capsule, solid, or the like. After being prepared in these forms by a method known per se, lotion preparations, emulsions, creams, ointments, plasters, haptics, aerosols, injections, internal preparations (tablets, powders, granules, pills) Preparations, syrups, troches and the like), suppositories and the like, which can be applied, pasted, sprayed, injected, drunk, and inserted into the body. Among these preparations, external preparations for the skin such as lotion preparations, emulsions, creams, ointments, plasters, cataplasms, aerosols and the like are considered to be preparations suitable for the purpose of the present invention. The external preparation for skin described here includes drugs, quasi-drugs, and cosmetics, and will be used in the same meaning hereinafter.

  The preparations described in the preceding paragraph include excipients and flavors commonly used in preparing these preparations, as well as oils and fats, surfactants, preservatives, sequestering agents, water-soluble polymers, A tackifier, a powder component, an ultraviolet ray protective agent, a humectant, a medicinal component, an antioxidant, a pH adjuster, a detergent, a desiccant, an emulsifier, and the like can be appropriately compounded. When these various components are incorporated into the skin activator of the present invention, it is possible within a range that does not impair the intended effects of the present invention.

  Examples of the fats and oils include liquid fats and oils, solid fats and oils, waxes, hydrocarbon oils, higher fatty acids, higher alcohols, synthetic ester oils, and silicones.

  That is, as the liquid fat, avocado oil, camellia oil, evening primrose oil, turtle oil, macadamia nut oil, corn oil, mink oil, olive oil, rapeseed oil, egg yolk oil, sesame oil, persic oil, wheat germ oil, sasanqua oil, castor oil , Linseed oil, safflower oil, cottonseed oil, eno oil, soybean oil, peanut oil, teaseed oil, kaya oil, rice bran oil, cinnamon oil, Japanese kiri oil, jojoba oil, germ oil, triglycerin, glycerin trioctanoate, bird Glycerin isopalmitate, etc., as solid oils and fats, such as cocoa butter, coconut oil, horse fat, hardened coconut oil, palm oil, tallow, sheep butter, hardened tallow, palm kernel oil, lard, beef bone oil, moku kernel oil Waxes such as beeswax, candelilla wax, cotton wax, carnauba wax, bayberry wax, wart, hardened oil, beef tallow, mokuro, hardened castor oil, etc. Wax, whale wax, montan wax, bran wax, lanolin, kapok wax, lanolin acetate, liquid lanolin, sugar cane wax, isopropyl lanolin fatty acid, hexyl laurate, reduced lanolin, jojoba wax, hard lanolin, shellac wax, POE lanolin alcohol ether, POE lanolin alcohol acetate And hydrocarbon oils such as POE cholesterol ether, lanolin fatty acid polyethylene glycol, and POE hydrogenated lanolin alcohol ether. Examples of liquid oils include liquid paraffin, ozokerite, squalene, pristane, paraffin, ceresin, squalene, petrolatum, and microcrystalline wax. .

  Higher fatty acids include, for example, lauric acid, myristic acid, palmitic acid, stearic acid, behenic (behenic) acid, oleic acid, 12-hydroxystearic acid, undecylenic acid, tallic acid, isostearic acid, linoleic acid, linoleic acid, Eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and the like.

  As higher alcohols, for example, linear alcohols such as lauryl alcohol, cetyl alcohol, stearyl alcohol, behenyl alcohol, myristyl alcohol, oleyl alcohol, and cetostearyl alcohol, monostearyl glycerin ether (bacyl alcohol), 2-decyltetradecinol, lanolin Examples include alcohols, cholesterol, phytosterols, hexyldecanol, isostearyl alcohol, branched-chain alcohols such as octyldodecanol, and the like.

  Synthetic ester oils include isopropyl myristate, cetyl octoate, octyl dodecyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, decyl oleate, hexyl decyl dimethyloctanoate, cetyl lactate, myristyl lactate Lanolin acetate, isocetyl stearate, isocetyl isostearate, cholesteryl 12-hydroxystearate, ethylene glycol di-2-ethylhexylate, dipentaerythritol fatty acid ester, N-alkyl glycol monoisostearate, neopentyl glycol dicaprate, malic acid Diisostearyl, glycerin di-2-heptylundecanoate, trimethylolpropane tri-2-ethylhexylate, triisostearyl Trimethylolpropane phosphate, pentaneerythritol tetra-2-ethylhexylate, glycerin tri-2-ethylhexylate, trimethylolpropane triisostearate, cetyl 2-ethylhexanoate, 2-ethylhexyl palmitate, glyceryl trimmyristate, triglycol -2-heptylundecanoic acid glyceride, castor oil fatty acid methyl ester, oleic oil, cetostearyl alcohol, acetoglyceride, 2-heptylundecyl palmitate, diisobutyl adipate, N-lauroyl-L-glutamic acid-2-octyldodecyl ester , Di-2-heptylundecyl adipate, ethyl laurate, di-2-ethylhexyl sebacate, 2-hexyldecyl myristate, 2-hexyl palmitate Sill, 2-hexyldecyl adipate, sebacic acid diisopropyl, 2-ethylhexyl succinate, ethyl acetate, butyl acetate, amyl acetate, and triethyl citrate.

  Examples of silicones include linear polysiloxanes such as dimethylpolysiloxane, methylphenylpolysiloxane, and methylhydrogenpolysiloxane, cyclic polysiloxanes such as decamethylpolysiloxane, dodecamethylpolysiloxane, and tetramethyltetrahydrogenpolysiloxane. Examples thereof include a silicone resin and a silicone rubber forming a three-dimensional network structure.

  Examples of the surfactant include an anionic surfactant, a cationic surfactant, an amphoteric surfactant, and a nonionic surfactant.

  Examples of the anionic surfactant include soap base, fatty acid soaps such as sodium laurate and sodium palmitate, higher alkyl sulfates such as sodium lauryl sulfate and potassium lauryl sulfate, POE lauryl sulfate triethanolamine, POE lauryl sulfate Higher fatty acid amide sulfonic acids such as alkyl ether sulfates such as sodium, N-acyl sarcosine acids such as sodium lauroyl sarcosine, N-myristoyl-N-methyltaurine sodium, coconut oil fatty acid methyltauride sodium and sodium laurylmethyltauride Salts, phosphate salts such as sodium POE oleyl ether phosphate, POE stearyl ether phosphoric acid, sodium di-2-ethylhexyl sulfosuccinate, monolauroylmo Sulfosuccinates such as sodium ethanolamide polyoxyethylene sodium sulfosuccinate and sodium lauryl polypropylene glycol sulfosuccinate; alkylbenzene sulfonates such as linear dodecylbenzenesulfonate, sodium lineardodecylbenzenesulfonate triethanolamine and linear dodecylbenzenesulfonate; N N-acyl glutamates such as monosodium lauroylglutamate, disodium N-stearoylglutamate, monosodium N-myristoyl-L-glutamate, higher fatty acid ester sulfates such as hydrogenated coconut oil fatty acid sodium glycerin sulfate, and funnel oils. Sulfated oil, POE alkyl ether carboxylic acid, POE alkyl allyl ether carboxylate, α-olefin sulfonate Higher fatty acid ester sulfonate, secondary alcohol sulfate, higher fatty acid alkylolamide sulfate, sodium lauroyl monoethanolamidosuccinate, N-palmitoyl aspartate ditriethanolamine, sodium caseinate and the like.

  Examples of the cationic surfactant include alkyltrimethylammonium salts such as stearyltrimethylammonium chloride and lauryltrimethylammonium chloride, distearyldimethylammonium dialkyldimethylammonium chloride, and poly (N, N'-dimethyl-3,5-methylene chloride). Alkylpyridinium salts such as piperidinium) and cetylpyridinium chloride, alkyl quaternary ammonium salts, alkyldimethylbenzylammonium salts, alkylisoquinolinium salts, dialkylmorphonium salts, POE alkylamines, alkylamine salts, polyamine fatty acid derivatives , Amyl alcohol fatty acid derivatives, benzalkonium chloride, benzethonium chloride and the like.

  Examples of the amphoteric surfactant include 2-undecyl-N, N, N- (hydroxyethylcarboxymethyl) -2-imidazoline sodium and 2-cocoyl-2-imitazolinium hydroxide-1-carboxyethyloxy2. Imidazoline amphoteric surfactants such as sodium salt, betaine surfactants such as 2-heptadecyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, betaine lauryldimethylaminoacetate, alkylbetaine, amidobetaine and sulfobetaine And the like.

  Examples of the lipophilic nonionic surfactant include sorbitan monooleate, sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan sesquioleate, sorbitan trioleate, and penta-2 Sorbitan fatty acid esters such as diglycerol sorbitan ethylhexylate, diglycerol sorbitan tetra-2-ethylhexylate, glycerin mono-cottonseed oil fatty acid, glyceryl monoerucate, glycerin sesquioleate, glyceryl monostearate, α, α'-pyrooleate Glycerin polyglycerin fatty acids such as glycerin glutamate and glyceryl monostearate, and propylene glycol fatty acids such as propylene glycol monostearate Esters, hydrogenated castor oil derivatives, glycerin alkyl ethers, polyoxyethylene / methyl polysiloxane copolymers, and the like.

  Examples of the hydrophilic nonionic surfactant include POE sorbitan fatty acid esters such as POE sorbitan monooleate, POE-sorbitan monostearate, POE-sorbitan monooleate, POE-sorbitan tetraoleate, and POE-sorbit monolau. POE-sorbit monooleate, POE-sorbit pentaoleate, POE-sorbit fatty acid esters such as POE-sorbit monostearate, POE-glycerin monostearate, POE-glycerin monoisostearate, POE-glycerin triisoate POE glycerin fatty acid esters such as stearate, POE monooleate, POE distearate, POE monodioleate, and POE fatty acid esters such as ethylene glycol cysteate POE alkyl ethers such as POE lauryl ether, POE oleyl ether, POE stearyl ether, POE behenyl ether, POE 2-octyl dodecyl ether, POE cholestanol ether, POE octyl phenyl ether, POE nonyl phenyl ether, POE dinonyl phenyl ether, etc. POE alkyl phenyl ethers, Pluronics such as Pluronic, POE POP cetyl ether, POE POP 2-decyl tetradecyl ether, POE POP monobutyl ether, POE POP hydrogenated lanolin, POE POP glycerin ether POE -POP alkyl ethers, tetra-POE such as tetronic, etc.-Condensation products of tetra-POP ethylenediamine, POE castor oil, POE cured oil POE hardened castor oil monoisostearate, POE hardened castor oil triisostearate, POE hardened castor oil monopyroglutamic acid monoisostearic acid diester, POE hardened castor oil maleic acid and other POE castor oil hardened castor oil derivatives, POE POE beeswax lanolin derivatives such as sorbit beeswax, alkanolamides such as coconut oil fatty acid diethanolamide, lauric acid monoethanolamide, fatty acid isopropanolamide, POE propylene glycol fatty acid ester, POE alkylamine, POE fatty acid amide, sucrose fatty acid ester, POE Nonylphenylformaldehyde condensate, alkylethoxydimethylamine oxide, trioleyl phosphoric acid and the like can be mentioned.

  Examples of the preservative include methyl paraben, ethyl paraben, butyl paraben and the like.

  Examples of the sequestering agent include edetic acid sodium salt, EDTA and the like.

  Examples of the water-soluble polymer include natural polymers, semi-synthetic polymers, synthetic polymers, and inorganic polymers.

  Natural water-soluble polymers include gum arabic, tragacanth gum, galactan, guar gum, carob gum, karaya gum, carrageenan, tamarind gum, xanthan gum, pectin, agar, quince seed (quince), alge colloid (cassow extract), starch (rice, corn) , Potato, wheat), glycyrrhizic acid and the like, plant-based polymers such as xanthan gum, dextran, succinoglucan and pullulan, and animal-based polymers such as collagen, casein, albumin and gelatin.

  Examples of semi-synthetic water-soluble polymers include starch-based polymers such as dextrin, carboxymethyl starch, and methylhydroxypropyl starch, methylcellulose, nitrocellulose, ethylcellulose, methylhydroxypropylcellulose, hydroxyethylcellulose, cellulose dimethyldialkyl sulfate (12 to 20). A) alginate polymers such as ammonium, hydroxypropylcellulose, sodium carboxymethylcellulose (CMC), crystalline cellulose and cellulose powder; and alginic acid polymers such as sodium alginate and propylene glycol alginate.

  Examples of the synthetic water-soluble polymer include vinyl polymers such as polyvinyl alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, carboxyvinyl polymer, and alkyl-modified carboxyvinyl polymer, and polyoxyethylene polymers such as polyethylene glycol 2000, 4000, and 6000. And polyoxyethylene-polyoxypropylene copolymer-based polymers, acrylic polymers such as sodium polyacrylate, polyethylene acrylate, and polyacrylamide, polyethyleneimine, and cationic polymers.

  Examples of the inorganic water-soluble polymer include bentonite, aluminum magnesium silicate, laponite, hectorite, and silicic anhydride.

  Examples of the powder component include talc, kaolin, mica, sericite, sericite, muscovite, phlogopite, synthetic mica, mica, biotite, lithia mica, vermiculite, magnesium carbonate, calcium carbonate, aluminum silicate, and silica silicate. Barium acid, calcium silicate, magnesium silicate, strontium silicate, metal tungstate, magnesium, silica, zeolite, barium sulfate, calcined calcium sulfate (calculated gypsum), calcium phosphate, fluoroapatite, hydroxyapatite, ceramic powder, metal soap (Zinc myristate, calcium palmitate, aluminum stearate), inorganic powders such as boron nitride, polyamide resin powder (nylon powder), polyethylene powder, polymethyl methacrylate powder, polystyrene powder, styrene and aluminum Organic powders such as copolymeric resin powder of lylic acid, benzoguanamine resin powder, polytetrafluoroethylene powder and cellulose powder, inorganic white pigments such as titanium dioxide and zinc oxide, inorganic oxides such as iron oxide (iron oxide) and iron titanate Red pigment, inorganic brown pigment such as γ-iron oxide, yellow iron oxide, inorganic yellow pigment such as loess, black iron oxide, carbon black, inorganic black pigment such as lower titanium oxide, mango violet, cobalt violet Inorganic purple pigments such as chromium oxide, chromium hydroxide, cobalt titanate, etc .; inorganic blue pigments such as ultramarine and navy blue; titanium oxide coated mica; titanium oxide coated bismuth oxychloride; titanium oxide coated talc , Colored titanium oxide coated mica, bismuth oxychloride, pearl pigments such as fish scale foil, aluminum powder, mosquito Metal powder pigments such as upper powder, Red No. 201, Red No. 202, Red No. 204, Red No. 205, Red No. 220, Red No. 226, Red No. 228, Red No. 405, Orange No. 203, Orange No. 204, Yellow No. 205 Yellow No. 401, Blue No. 404, Red No. 3, Red No. 104, Red No. 106, Red No. 227, Red No. 230, Red No. 401, Red No. 505, Orange No. 205, Yellow No. 4, Yellow No. 5, Yellow No. 202, Yellow No. 203, Green No. 3, Blue No. 1 and the like, organic pigments such as zirconium, barium or aluminum lake, chlorophyll, natural coloring matter such as β-carotene, coloring agents such as titanium yellow, carsamine, safflower red, And the like.

  The ultraviolet ray protective agent includes both an "ultraviolet ray absorber" which is a substance that chemically absorbs ultraviolet rays, and an "ultraviolet ray blocking agent" which is a substance which scatters and reflects ultraviolet rays by a physical action.

  That is, as long-wavelength ultraviolet (UVA) absorbers, anthranilic acid-based ultraviolet absorbers such as methylanthranilate and homomenthyl-N-acetylanthranilate, for example, 2,4-dihydroxybenzophenone, 2,2-dihydroxy-4- Methoxybenzophenone, 2,2'-dihydroxy-4,4'-dimethoxybenzophenone, 2,2 ', 4,4'-tetrahydroxybenzophenone, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4 '-Methylbenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid, 4-phenylbenzophenone, 2-ethylhexyl-4'-phenyl-benzophenone-2-carboxylate, 2-hydroxy-4-n-octoxy Benzophenone, Benzophenone ultraviolet absorbers such as -hydroxy-3-carboxybenzophenone, for example, 2,2'-hydroxy-5-methylphenylbenzotriazole, 2- (2'-hydroxy-5'-t-octylphenyl) benzotriazole, Benzotriazole-based ultraviolet absorbers such as-(2'-hydroxy-5'-methylphenyl) benzotriazole, for example, dianisoylmethane, 4-methoxy-4'-t-butyldibenzoylmethane and the like can be mentioned. Among these long-wavelength ultraviolet absorbers, 4-methoxy-4'-tert-butyldibenzoylmethane, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxybenzophenone derivatives, for example, 2-hydroxy-4- Methoxybenzophenone-5-sulfonate is a long-wavelength ultraviolet absorber excellent in safety and effectiveness, and is preferred.

  Also, as a medium wavelength ultraviolet (UVB) absorber, para-aminobenzoic acid (hereinafter, referred to as PABA), PABA monoglycerin ester, N, N-dipropoxy PABA ethyl ester, N, N-diethoxy PABA ethyl ester, N, N- Benzoic acid-based UV absorbers such as dimethyl PABA ethyl ester, N, N-dimethyl PABA butyl ester, N, N-dimethyl PABA amyl ester, dipropylene glycol salicylate, ethylene glycol salicylate, myristyl salicylate, methyl salicylate, amyl salicylate, menthyl Salicylic acid type ultraviolet absorbers such as salicylate, homomenthyl salicylate, octyl salicylate, phenyl salicylate, benzyl salicylate, p-isopropanol phenyl salicylate and the like Octylcinnamate, ethyl-4-isopropylcinnamate, methyl-2,5-diisopropylcinnamate, ethyl-2,4-diisopropylcinnamate, methyl-2,4-diisopropylcinnamate, propyl-p-methoxycinnamate, Isopropyl-p-methoxycinnamate, isoamyl-p-methoxycinnamate, octyl-p-methoxycinnamate (2-ethylhexyl-p-methoxycinnamate), 2-ethoxyethyl-p-methoxycinnamate, cyclohexyl-p- Methoxy cinnamate, ethyl-α-cyano-β-phenyl cinnamate, 2-ethylhexyl-α-cyano-β-phenyl cinnamate, glyceryl mono-2-ethylhexanoyl-diparamethoxy cinnamate, methoxycinnamate Cinnamic acid-based ultraviolet absorbers such as octyl, 3,4,5-trimethoxycinnamic acid-3-methyl-4- [methylbis (trimethylsiloxy) silyl] butyl and p-dimethoxycinnamic acid monoethyl ester; Camphor derivatives such as-(4'-methylbenzylidene) -d, 1-camphor, 3-benzylidene-d, 1-camphor, 5- (3,3-dimethyl-2-norbornylidene) -3-pentyn-2-one; Urocanic acid, urocanic acid ethyl ester, 2-phenyl-5-methylbenzoxazole, dibenzalazine and the like.

Further, examples of the ultraviolet blocking agent include titanium oxide (TiO ₂ ), talc (MgSiO ₂ ), carmine (FeO ₂ ), bentonite, kaolin, zinc oxide (ZnO) and the like.

  Examples of the humectant include polyethylene glycol, propylene glycol, glycerin, 1,3-butylene glycol, hexylene glycol, xylitol, sorbitol, maltitol, chondroitin sulfate, hyaluronic acid, mucoitin sulfate, caronic acid, atelocollagen, cholesteryl-12. -Hydroxystearate, sodium lactate, bile salts, dl-pyrrolidone carboxylate, short-chain soluble collagen, diglycerin (EO) PO adduct, Izayobara extract, Achillea millefolium extract, melilot extract, etc. it can.

Examples of the medicinal component include whitening agents such as arbutin, vitamin C and derivatives thereof, kojic acid placental extract, glutathione, and saxifrage extract, glycyrrhizic acid derivatives, glycyrrhetinic acid derivatives, salicylic acid derivatives, hinokitiol, zinc oxide, and allantoin. Agents, royal jelly, photosensitizers, cholesterol derivatives, activators such as calf blood extract, etc., nonylate valenylamide, nicotinic acid benzyl ester, nicotinic acid β-butoxyethyl ester, capsaicin, zingerone, canthalis tincture, itctamol, caffeine, Tannic acid, α-borneol, tocopherol nicotinate, inositol hexanicotinate, cyclandate, cinnarizine, trazoline, acetylcholine, verapamil, cepharanthin, γ-oryzanol and the like Line accelerators: antiseborrheic agents such as sulfur and thiantole, for a variety of purposes, oakaku extract, spinach extract, sicon extract, peony extract, assembly extract, birch extract, sage extract, loquat extract , Carrot extract, aloe extract, mallow extract, iris extract, grape extract, yokuinin extract, loofah extract, lily extract, saffron extract, senkyu extract, ginger extract, hypericum extract, Ononisu extract, rosemary extract, garlic extract, capsicum extract, citrus unshiu peel, angelica, etc., retinol, vitamin a such as retinol acetate, riboflavin, riboflavin butyrate, vitamin B ₂ such as flavin adenine nucleotide, pyridoxine hydrochloride , Pyridoxy Vitamin _{B 2} such as dioctanoate, L- ascorbic acid, L- ascorbic acid dipalmitate ester, L- ascorbic acid-2-sodium sulfate, L- ascorbic acid phosphoric acid ester, DL-alpha-tocopherol -L- ascorbate Vitamin Cs such as dipotassium phosphate diester, pantothenates such as calcium pantothenate, D-pantoenyl alcohol, pantothenyl ethyl ether, acetyl pantothenyl ethyl ether, vitamin Ds such as ergocalciferol and cholecalciferol, nicotinic acid Nicotinic acid such as nicotinamide, nicotinic acid amide, benzyl nicotinate, vitamin E such as α-tocopherol, tocopherol acetate, DL-α-tocopherol nicotinate, DL-α-tocopherol succinate, vitamin P, bioti Vitamins and the like, can be mentioned.

  The drug components can be widely mixed as long as the desired effects of the present invention are not impaired by the compounding.

  Since the composition of the present invention thus prepared has an activity to enhance the ability to produce laminin 5 in human epidermal keratinocytes, the composition prevents a decrease in the function of the skin, particularly accompanying the structural change of the basement membrane, It can promote skin activation.

Hereinafter, the present invention will be described more specifically with reference to specific examples.
Measurement test for laminin 5 production ability:
(1) Culture of epidermal keratinocytes Epidermal keratinocytes were isolated from human foreskin and cultured in an epidermal keratinocyte growth medium (KGM) having a low calcium concentration. Bovine pituitary extract and EGF were added to this medium. After culturing the cells with KGM until the fourth generation, the adherent cells were suspended by treatment with trypsi-EDTA, and the cell aggregates were removed by filtration to obtain a uniform cell suspension. The cells were collected by centrifugation and resuspended in DMEM-F12 (2: 1) -0.1% BSA to 8 × 10 ⁴ / ml. This cell suspension was added to 0.5 ml of the same medium containing twice the concentration of the drug in 0.5 ml. The culture was performed at 37 ° C. for 24 hours using a 24-well plate. At the end of the culture, the culture supernatant was transferred to an Eppendorf tube and centrifuged at 15000 rpm for 5 minutes. The supernatant was transferred to a new tube and stored at -20 ° C until the day of laminin-5 measurement. To solubilize laminin 5 bound in the cells and on the culture plastic, Tris-HCl buffer pH 7.4 containing various surfactants was added to each well and stored overnight at -20 ° C. The next day, it was sonicated and frozen again. On the next day, after redissolving, the mixture was centrifuged at 15000 rpm for 5 minutes, the supernatant was transferred to a tube, and stored at -20 ° C until the day of laminin-5 measurement.

(2) Measurement of Laminin 5 by Sandwich ELISA Laminin 5 present in the culture supernatant and the cell layer was measured by sandwich ELISA. A monoclonal antibody to laminin α3 chain of laminin 5, BM165, was bound to the solid layer of a 96-well ELISA plate. In order to measure by laminin 5 sandwiching, 6F12 which is a monoclonal antibody against laminin β3 chain was biotinylated (b-6F12) and used as another kind of antibody. In this method, only a heterotrimer (α3β3γ2) capable of exerting a function is measured, and a heterodimer (β3γ2) is not detected. A sample is added to each well previously filled with a 3% gelatin / phosphate buffer solution containing b-6F12. The final dilution ratio of the sample in the well was 1/4 for the culture solution and 1/10 for the cell layer. The antigen-antibody reaction was performed at 37 ° C. for 2 hours. After washing, avidin HRP (horseradish peroxidase) solution was added, and the reaction was further performed for 30 minutes to 1 hour. After washing, an ABTS solution as a substrate for HRP was added, and the absorbance at 405 nm was measured using an ELISA plate reader. The calibration curve was prepared in the range of 0 to 40 ng / ml.

  The amount of laminin 5 produced was indicated by the sum of the amount released in the medium and the amount remaining in the cell layer.

(3) Results The results are shown in Table 1 below.

  As shown in Table 1, fetal calf serum, which is known to activate cells, showed a marked increase in laminin 5 production at a concentration of 1.5% as compared to the control group.

  The soybean saponin fraction (saponin, manufactured by soybean: manufactured by Wako Pure Chemical Industries, Ltd.) also showed an effect of increasing the production of laminin 5 to a level comparable to fetal calf serum. The effect was almost comparable to fetal calf serum.

  The soybean lecithin fraction (lecithin, manufactured by soybean: manufactured by Wako Pure Chemical Industries, Ltd.), although slightly weaker than the soybean saponin fraction, also exhibited a concentration-dependent laminin 5 production promoting action.

Formulation Example 1: Cream polyoxyethylene (addition of 20 mol) cetyl alcohol ether 1.0
Methylphenyl polysiloxane (20cs) 2.0
Liquid paraffin 3.0
2-hydroxy-4-methoxybenzophenone 5.0
Soybean saponin (saponin, soybean: Wako Pure Chemical Industries, Ltd.) 0.2
Propylene glycol 5.0
Glycerin 2.0
Ethyl alcohol 15.0
Carboxyvinyl polymer 0.3
Hydroxypropyl cellulose 0.1
2-aminomethylpropanol 0.1
Preservatives qs perfume qs ion exchange water qs (production method)
Propylene glycol, glycerin, ethyl alcohol, carboxyvinyl polymer, hydroxypropylcellulose, and 2-aminomethylpropanol are added to ion-exchanged water and heated to 70 ° C. (aqueous phase).

  Mix methylphenylpolysiloxane, liquid paraffin, polyoxyethylene cetyl alcohol ether, preservative, 2-hydroxy-4-methoxybenzophenone, soybean saponin, and flavor to adjust to 70 ° C (oil phase).

The aqueous phase was gradually added to the oil phase for pre-emulsification, and the emulsified particles were homogenized using a homomixer, followed by degassing and cooling to obtain a cream.
Formulation Example 2: Cream polyoxyethylene (addition of 20 mol) cetyl alcohol ether 1.0
Methylphenyl polysiloxane (20cs) 2.0
Liquid paraffin 3.0
2-hydroxy-4-methoxybenzophenone 5.0
Soybean lecithin (lecithin, soybean: Wako Pure Chemical Industries, Ltd.) 0.2
Propylene glycol 5.0
Glycerin 2.0
Ethyl alcohol 15.0
Carboxyvinyl polymer 0.3
Hydroxypropyl cellulose 0.1
2-aminomethylpropanol 0.1
Preservatives qs perfume qs ion exchange water qs (production method)
Propylene glycol, glycerin, ethyl alcohol, carboxyvinyl polymer, hydroxypropylcellulose, and 2-aminomethylpropanol are added to ion-exchanged water and heated to 70 ° C. (aqueous phase).

  Mix methylphenylpolysiloxane, liquid paraffin, polyoxyethylene cetyl alcohol ether, preservative, 2-hydroxy-4-methoxybenzophenone, soybean lecithin, and flavor to adjust to 70 ° C (oil phase).

The aqueous phase was gradually added to the oil phase for pre-emulsification, and the emulsified particles were homogenized using a homomixer, followed by degassing and cooling to obtain a cream.

Claims (5)

  An agent for promoting the production of laminin 5, comprising soy extract as an active ingredient.   2. The production promoter according to claim 1, wherein laminin 5 is laminin 5 in human epidermal keratinocytes.   The production accelerator according to claim 1 or 2, wherein the soybean extract is extracted from the ground soybean using a hydrous lower alcohol.   The production enhancer according to claim 1 or 2, wherein the soybean extract is a soybean saponin fraction, a soybean lecithin fraction, or a mixture thereof. A method for externally applying a soybean extract to human skin to improve laminin 5 production in epidermal keratinocytes of the skin.