Collagen synthesis enzyme gene expression promoter

Abstract

The present invention aims to provide a novel collagen synthesis gene expression promoter. [Solution] The features of the present invention for solving the above problems are as follows. 1. A collagen synthesis gene expression promoter containing lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) (deposit number NITE ABP-04138) and cherry blossom extract as active ingredients. 2. The collagen synthesis gene expression promoter described in 1. above, characterized in that the blending ratio of lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) (deposit number NITE ABP-04138) to cherry blossom extract is 1 / 0.1 to 1 / 100 in terms of blending ratio (the lactic acid bacteria / cherry blossom extract).

Description

Technical Field

[0001] The present invention relates to a novel promoter for collagen synthase gene expression.

Background Art

[0002] Conventionally, lactic acid bacteria derived from Prunus serrulata var. lannesiana have been known as lactic acid bacteria derived from cherry blossoms, and are known to have an action of promoting IL-12 production (Patent Document 1).

[0003] On the other hand, the skin is composed of the epidermis, basement membrane, and dermis. In particular, the dermis is formed by fibroblasts and extracellular matrix such as collagen, elastin, and hyaluronic acid. In young skin, fibroblasts are active, skin homeostasis is maintained, and moisturizing power and elasticity are maintained, so the skin is fresh and healthy. However, due to the effects of ultraviolet rays, dryness, stress, aging, etc., collagen is decomposed and deteriorated, and the growth of fibroblasts slows down, resulting in a decrease in moisturizing function and elasticity, and the occurrence of wrinkles, dullness, and rough skin. In particular, type I collagen is important for skin strength, and the COL1A1 gene is involved in its synthesis. It is considered that by promoting the mRNA expression of COL1A1, the synthesis of type I collagen can be improved, and skin aging can be prevented or improved (Patent Document 2).

[0004] So far, as those having an action of promoting the production of collagen, for example, starfruit leaf extract (Patent Document 3), kudzu leaf wrinkle extract (Patent Document 4), etc. are known. In addition, as substances that promote the expression of COL1A1 mRNA, trehalangelin (Patent Document 5), apple polyphenol that suppresses the decrease in COL1A1 mRNA expression by UVB (Patent Document 6), etc. are known.

[0005] ​​​​​​​​​​​​​ [Patent Document 4] Japanese Patent Publication No. 2003-146837 [Patent Document 5] Japanese Patent Publication No. 2015-024985 [Patent Document 6] Japanese Patent Publication No. 2016-053007 [Disclosure of the Invention] [Problems that the invention aims to solve]

[0006] Against this backdrop, the inventors discovered that combining cherry blossom extract with a low concentration of lactic acid bacteria derived from weeping cherry blossoms (Prunus pendula) has excellent collagen synthesis gene (COL1A1) expression promoting properties, thus completing the present invention. In other words, the present invention aims to provide a novel collagen synthesis gene expression promoter. [Means for solving the problem]

[0007] The features of the present invention for solving the above problems are as follows. 1. A collagen synthesis gene expression promoter containing lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) (deposit number NITE ABP-04138) and cherry blossom extract as active ingredients. 2. The collagen synthesis gene expression promoter described in 1. above, characterized in that the blending ratio of lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) (deposit number NITE ABP-04138) to cherry blossom extract is 1 / 0.1 to 1 / 100 in terms of blending ratio (the lactic acid bacteria / cherry blossom extract). [Effects of the Invention]

[0008] By combining the above-mentioned lactic acid bacteria with cherry blossom extract, it has the effect of promoting collagen synthesis gene expression, and as a result, the cherry blossom lactic acid bacteria have the effect of maintaining skin moisture and elasticity. Furthermore, by adjusting the blending ratio of the lactic acid bacteria to the cherry blossom extract to 1 / 0.1 to 1 / 100 (lactic acid bacteria / cherry blossom extract), it exhibits an even superior effect as a collagen synthesis gene expression promoter, thereby synergistically enhancing the lactic acid bacteria's ability to maintain skin moisture and elasticity. [Brief explanation of the drawing]

[0009] [Figure 1] This graph shows the effect of the above-mentioned combination of lactic acid bacteria and cherry blossom extract on collagen synthase gene expression in NHDF cells. [Figure 2] This is the phylogenetic tree of the lactic acid bacterium (Lacticaseibacillus paracasei sp. Shidare) used in this example. [Figure 3] This is an explanatory diagram showing the method for identifying the lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) in this example. [Modes for carrying out the invention]

[0010] The present invention will be described in detail below. This invention relates to lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) deposit number ( Deposit number NITE ABP-04138 The active ingredients are lactic acid bacteria (hereinafter referred to as "Sakura lactic acid bacteria") and cherry blossom extract. The above-mentioned cherry blossom lactic acid bacteria originate from weeping cherry trees (Prunus pendula) and can be isolated and identified by the methods described in the examples of this specification.

[0011] The cherry blossoms used in the above-mentioned cherry blossom extract refer to a general term for plants belonging to the genus Cerasus of the Rosaceae family, excluding plums, peaches, apricots, etc., and generally refer to those belonging to the subgenus Cerasus.

[0012] The cherry species used in the present invention is not particularly limited. For example, cherries belonging to groups such as the Prunus jamasakura group, Prunus pendula group, Prunus incisa group, Prunus serrulata group, Prunus grayana group, Prunus verecunda group, etc. can be used, and it is not further limited to these groups.

[0013] The part of the cherry used as the raw material of the present invention is not particularly limited, and examples include leaves, stems, trunks, flowers, roots, fruits, etc. However, it is preferable to use leaves or flowers.

[0014] Here, when extracting by polar solvent extraction, the polar solvent to be used is not particularly limited. For example, water, methanol, ethanol, isopropanol, acetone, 1,3 - butylene glycol, ethylene glycol, propylene glycol, glycerin, acetic acid, ethyl acetate, ether, hexane, etc. can be mentioned. Among these, water, methanol, and ethanol are preferable because the active ingredients can be efficiently extracted. These may be used alone or in combination of two or more.

[0015] When using water as the extraction solvent, the extraction is preferably carried out at an extraction temperature of 20 to 100°C, preferably about 40 to 70°C. This is because if the extraction temperature is too low, the active ingredients are difficult to be extracted, and if the extraction temperature is too high, the cyanide compounds contained in the cherry are likely to remain, and the active ingredients are likely to be decomposed, which is not preferable. The type of water for extraction is not particularly limited, and tap water, distilled water, mineral water, alkaline ion water, etc. can be used.

[0016] When using hydrous alcohol as the extraction solvent, the alcohol concentration is preferably 20 wt%, more preferably 25 wt% to 50 wt%, and even more preferably 25 wt% to 30 wt%. This is because when the alcohol concentration is less than 20 wt%, it is difficult to obtain a high extraction amount of active ingredients, and when the alcohol concentration exceeds 50 wt%, the yield decreases due to the influence of impurities, etc. Also, when the alcohol concentration is 30 wt% or more, the extraction temperature is preferably 0 to 95°C, preferably about 0 to 60°C. In addition, for hydrous ethanol extraction, in order to improve the content rate of the active ingredients, it is advisable to repeat at various concentrations.

[0017] Also, when extracting with a polar solvent, the extraction method is not particularly limited. For example, any method such as continuous extraction, immersion extraction, countercurrent extraction, etc. can be adopted, and any apparatus can be used at room temperature or under reflux heating. In addition, when performing extraction by the methods described above, only one of these may be performed, or these methods may be combined. Also, these extractions may be performed only once or may be performed two or more times.

[0018] As a specific method, the extraction raw material is put into a treatment tank filled with an extraction solvent, and the active ingredient is eluted while stirring. For example, when using water or hydrous alcohol as the extraction solvent, a polar solvent about 3 to 100 times the amount (weight ratio) of the extraction raw material is used, and extraction is performed for about 1 minute to 150 hours. After eluting the active ingredient in the solvent, filtration is performed to remove the extraction residue, thereby obtaining an extract. Thereafter, the extract is subjected to treatments such as dilution, concentration, drying, purification, etc. according to conventional methods to obtain an extract containing high-concentration caffeoyl glucose. Examples of the purification method include methods such as activated carbon treatment, resin adsorption treatment, silica gel treatment, ion exchange resin, liquid-liquid countercurrent distribution, membrane separation, etc.

[0019] Furthermore, when performing extraction by supercritical extraction, the supercritical fluid used at this time is not particularly limited, and examples include carbon dioxide and nitrogen, etc. In addition, only one of these may be used, or two or more types may be used in combination. Among these, carbon dioxide is particularly preferred. This is because the active ingredient can be extracted more easily. Also, the extraction method at this time may be performed by a known method. Thereafter, the extract is subjected to treatments such as dilution, concentration, drying, purification, etc. according to conventional methods to obtain an extract containing high-concentration caffeoyl glucose.

[0020] Furthermore, a commercially available product can be used as the above-mentioned cherry extract. In this case, for example, it is preferable to use "Cherry Flower Extract" manufactured by Oriza Yuka Co., Ltd. This is because it contains high-concentration caffeoyl glucose.

[0021] The ratio of the above-mentioned cherry blossom lactic acid bacteria to the cherry blossom extract is not particularly limited, but the ratio (cherry blossom lactic acid bacteria / cherry blossom extract) can be 1 / 0.1 to 1 / 100, preferably 1 / 1 to 1 / 20, and even more preferably 1 / 5 to 1 / 12.

[0022] The collagen synthesis gene expression promoter of the present invention can be used as an ingredient in various foods and beverages. Examples of foods and beverages include general foods such as confectionery (gum, candy, caramel, chocolate, cookies, snacks, jelly, gummies, tablets, etc.), noodles (soba, udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, and beverages (juice, coffee, tea, carbonated drinks, sports drinks, etc.), as well as health foods (tablets, capsules, etc.) and nutritional supplements (nutritional drinks, etc.). The agent of the present invention can be appropriately incorporated into these foods and beverages.

[0023] These foods and beverages can be formulated with various ingredients depending on their type. For example, they can contain food ingredients such as glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, gum arabic, carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, calcium salts, colorants, flavorings, and preservatives.

[0024] When applying the agent of the present invention to food and beverages, the amount added is preferably such that the total content of the active ingredient relative to the food and beverage is 1 to 20 wt%, since the main purpose is disease prevention and health maintenance.

[0025] The collagen synthesis gene expression promoter of the present invention may be used as a material for pharmaceuticals (including pharmaceuticals and quasi-drugs). It can be manufactured by appropriately blending the agent of the present invention with raw materials for pharmaceutical formulations. Examples of formulation ingredients that can be incorporated into the agent of the present invention include excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cocoa butter, hydrogenated vegetable oil, kaolin, talc, etc.), binders (distilled water, physiological saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrants (sodium alginate, agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, starch, lactose, acacia powder, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (quaternary ammonium base, sodium lauryl sulfate, etc.), adsorbents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), and lubricants (purified talc, stearate, polyethylene glycol, etc.).

[0026] The collagen synthesis gene expression promoter of the present invention can generally be administered orally in the form of tablets, pills, soft or hard capsules, granules, powders, granules, liquids, etc.

[0027] The dosage may vary depending on the method of administration, the patient's condition, and the patient's age, but typically, adults can be administered 0.5 to 5000 mg of the active ingredient per day, and children can usually be administered 0.5 to 3000 mg. The ratio of collagen synthesis gene expression promoters can be adjusted as appropriate depending on the dosage form, but generally, it is appropriate to use approximately 0.3 to 15.0 wt% when administered orally or via mucosal absorption, and approximately 0.01 to 10 wt% when administered parenterally. Note that the dosage will vary depending on various conditions, so a smaller amount than the above may be sufficient in some cases, and it may also be necessary to administer a larger amount than the above range.

[0028] The collagen synthesis gene expression promoter of the present invention can also be expected to produce the above-mentioned effects when used as a topical skin preparation (including cosmetics, pharmaceuticals, and quasi-drugs). Examples of skin topical preparations that may contain the collagen synthesis gene expression promoter of the present invention include lotions, soaps, facial cleansers, bath additives, creams, lotions, toners, colognes, shaving creams, shaving lotions, cosmetic oils, sunscreens, face powders, foundations, perfumes, face masks, nail creams, nail polish, nail polish removers, eyebrow pencils, blushes, eye creams, eyeshadows, mascaras, eyeliners, lipsticks, lip balms, shampoos, conditioners, hair dyes, dispersions, and cleansing agents. Examples of pharmaceuticals or quasi-drugs that may contain the collagen synthesis gene expression promoter of the present invention include ointments, creams, and topical solutions.

[0029] In addition to the collagen synthesis gene expression promoter of the present invention, the above-described form of topical skin preparation may contain, to the extent that it does not impair the above-described effect, ingredients commonly used in topical skin preparations such as cosmetics and quasi-drugs, oils, higher alcohols, fatty acids, UV absorbers, powders, pigments, surfactants, polyhydric alcohols / sugars, polymers, physiologically active ingredients, solvents, antioxidants, fragrances, preservatives, etc. Examples are listed below, but the present invention is not limited to these examples.

[0030] (1) Examples of oil content Ester-based oil phase components: Glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, butyl stearate, butyl myristate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, diethyl adipate Sopropyl, Isoarachil Neopentanoate, Caprylic / Capric Triglyceride, Trimethylolpropane Tri-2-ethylhexanoate, Trimethylolpropane Triisostearate, Pentaerythritol Tetra-2-ethylhexanoate, Cetyl Caprylate, Decyl Laurate, Hexyl Laurate, Decyl Myristate, Myristyl Myristate, Cetyl Myristate, Stearyl Stearate, Decyl Oleate, Cetyl Ricinoleate, Isostearyl Laurate, Isotridecyl Myristate, Isocetyl Myristate, Iso Tearyl, Isocetyl Palmitate, Isostearyl Palmitate, Octyl Stearate, Isocetyl Stearate, Isodecyl Oleate, Octyldodecyl Oleate, Octyldodecyl Linoleate, Isopropyl Isostearate, Cetostearyl 2-Ethylhexanoate, Stearyl 2-Ethylhexanoate, Hexyl Isostearate, Ethylene Glycol Dioctanoate, Ethylene Glycol Dioleate, Propylene Glycol Dicaprate, Di(Capryl / Capric Acid)Propylene Glycol, Propylene Glycol Dicaprylate, Dicaprin Neopentyl glycol acid, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerin oleate, polyglycerin isostearate, dipropyl carbonate,Dialkyl (C12-18) carbonate, triisocetyl citrate, triisoarachil citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, 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 Examples include ropil, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, isocetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate, and isostearyl 12-stearoylhydroxystearate. Hydrocarbon-based oil phase components include squalane, liquid paraffin, α-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, petrolatum, etc. Animal and vegetable oils and their hydrogenated oils, and naturally derived waxes: animal oils and their hydrogenated oils such as beef tallow, hydrogenated beef tallow, pork tallow, hydrogenated pork tallow, horse oil, hydrogenated horse oil, mink oil, orange roughy oil, fish oil, hydrogenated fish oil, egg yolk oil, avocado oil, almond oil, olive oil, cocoa butter, kiwi seed oil, apricot kernel oil, kukui nut oil, sesame oil, wheat germ oil, rice germ oil, rice bran oil, safflower oil, shea butter, soybean oil, evening primrose oil, perilla oil, tea seed oil, camellia oil, corn oil, rapeseed oil, Examples include hydrogenated rapeseed oil, palm kernel oil, hydrogenated palm kernel oil, palm oil, hydrogenated palm oil, peanut oil, hydrogenated peanut oil, castor oil, hydrogenated castor oil, sunflower oil, grape seed oil, jojoba oil, hydrogenated jojoba oil, macadamia nut oil, meadowfoam oil, cottonseed oil, hydrogenated cottonseed oil, coconut oil, hydrogenated coconut oil, and other vegetable oils and their hydrogenated oils, beeswax, high-acid value beeswax, lanolin, reduced lanolin, hydrogenated lanolin, liquid lanolin, carnauba wax, montan wax, and other waxes. Silicone-based oil phase components include dimethylpolysiloxane, methylphenylpolysiloxane, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, methylhydrogenpolysiloxane, polyether-modified organopolysiloxane, dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane-methylstearoxane copolymer, alkyl-modified organopolysiloxane, terminal-modified organopolysiloxane, amino-modified silicone oil, amino-modified organopolysiloxane, dimethiconol, silicone gel, acrylic silicone, trimethylsiloxysilicate, silicone RTV rubber, etc. Examples of fluorine-based oil phase components include perfluoropolyethers, fluorine-modified organopolysiloxanes, fluorinated pitch, fluorocarbons, fluoroalcohols, and fluoroalkyl / polyoxyalkylene copolymerized organopolysiloxanes.

[0031] (2) Examples of higher alcohols Examples include lauryl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, behenyl alcohol, 2-ethylhexanol, hexadecyl alcohol, and octyldodecanol.

[0032] (3) Examples of fatty acids Examples include caprylic acid, capric acid, undecylenic acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid, arachidonic acid, behenic acid, erucic acid, and 2-ethylhexanoic acid.

[0033] (4) Examples of UV absorbers Para-aminobenzoic acid, amyl para-aminobenzoate, ethyl dihydroxypropyl para-aminobenzoate, glyceryl para-aminobenzoate, ethyl para-aminobenzoate, octyl para-aminobenzoate, octyl dimethyl para-aminobenzoate, ethylene glycol salicylate, octyl salicylate, triethanolamine salicylate, phenyl salicylate, butylphenyl salicylate, benzyl salicylate, homomenthyl salicylate, benzyl cinnamate, octyl para-methoxycinnamate, 2-ethylhexyl para-methoxycinnamate, glyceryl mono-2-ethylhexanoate dipara-methoxycinnamate, isopropyl para-methoxycinnamate, diethanolamine para-methoxyhydrocinnamate salt, diisopropyl / diisopropyl cinnamic acid ester mixture, urocanic acid, ethyl urocanic acid, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, di Examples include hydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzophenone, dihydroxydimethoxybenzophenone, hydroxyoctoxybenzophenone, tetrahydroxybenzophenone, butylmethoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2-ethylhexyl-1-oxy)-1,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, methyl-O-aminobenzoate, 2-ethylhexyl-2-cyano-3,3-diphenyl acrylate, phenylbenzimidazole sulfate, 3-(4-methylbenzylidene)camphor, isopropyldibenzoylmethane, 4-(3,4-dimethoxyphenylmethylene)-2,5-dioxo-1-imidazolidinepropionate 2-ethylhexyl, and their polymer derivatives and silane derivatives.

[0034] (5) Examples of powders and pigments Dyes such as Red No. 104, Red No. 201, Yellow No. 4, Blue No. 1, Black No. 401, lake dyes such as Yellow No. 4 AL Lake, Yellow No. 203 BA Lake, nylon powder, silk powder, urethane powder, Teflon (registered trademark) powder, silicone powder, polymethyl methacrylate powder, cellulose powder, starch, silicone elastomer spherical powder, polymers such as polyethylene powder, yellow iron oxide, red iron oxide, black iron oxide, chromium oxide, carbon black, ultramarine, Prussian blue, zinc oxide, titanium dioxide, acid Examples include white pigments such as cerium oxide, extender pigments such as talc, mica, sericite, kaolin, and plate-like barium sulfate, pearl pigments such as titanium mica, metal salts such as barium sulfate, calcium carbonate, magnesium carbonate, aluminum silicate, and magnesium silicate, inorganic powders such as silica and alumina, metal soaps such as aluminum stearate, magnesium stearate, zinc palmitate, zinc myristate, magnesium myristate, zinc laurate, and zinc undecylenate, bentonite, smectite, and boron nitride. There are no particular restrictions on the shape (spherical, rod-shaped, needle-shaped, plate-shaped, irregular shape, flake-shaped, spindle-shaped, etc.) and particle size of these powders. These powders may or may not have been pre-treated by conventional surface treatments such as fluorine compound treatment, silicone treatment, silicone resin treatment, pendant treatment, silane coupling agent treatment, titanium coupling agent treatment, oil treatment, N-acylated lysine treatment, polyacrylic acid treatment, metal soap treatment, amino acid treatment, lecithin treatment, inorganic compound treatment, plasma treatment, mechanochemical treatment, etc.

[0035] (6) Examples of surfactants Anionic surfactants include fatty acid soaps, α-acyl sulfonates, alkyl sulfonates, alkylallyl sulfonates, alkylnaphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, alkylamide phosphates, alkylylalkyl taurates, N-acyl amino acid salts, POE alkyl ether carboxylates, alkyl sulfosuccinates, alkyl sulfoacetate sodium, acylated hydrolyzed collagen peptides, perfluoroalkyl phosphates, and the like. Cationic surfactants include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, beheninamidepropyldimethylhydroxypropylammonium chloride, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salts of lanolin derivatives, and the like. Amphoteric surfactants include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, and amideamine type. Nonionic surfactants include propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE glycerin fatty acid esters, POE alkyl ethers, POE fatty acid esters, POE hydrogenated castor oil, POE castor oil, POE-POP copolymers, POE-POP alkyl ethers, polyether-modified silicone lauric acid alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like. Examples of natural surfactants include lecithin, saponin, and sugar-based surfactants.

[0036] (7) Examples of polyhydric alcohols and sugars Examples include ethylene glycol, diethylene glycol, polyethylene glycol, propylene glycol, dipropylene glycol, polypropylene glycol, glycerin, diglycerin, polyglycerin, 3-methyl-1,3-butanediol, 1,3-butylene glycol, sorbitol, mannitol, raffinose, erythritol, glucose, sucrose, fructose, xylitol, lactose, maltose, maltitol, trehalose, alkylated trehalose, mixed isomerized sugars, sulfated trehalose, pullulan, etc. Chemically modified versions of these can also be used.

[0037] (8) Examples of polymers Acrylic acid ester / methacrylic acid ester copolymer (Plussize, manufactured by Go-O Chemical Co., Ltd.), vinyl acetate / crotonic acid copolymer (Resin 28-1310, manufactured by NSC Corporation), vinyl acetate / crotonic acid / vinyl neodecanate copolymer (28-2930, manufactured by NSC Corporation), methyl vinyl ether maleate half ester (Gantlets ES, manufactured by ISP Corporation), T-butyl acrylate / ethyl acrylate / methacrylic acid copolymer (Rubymer, manufactured by BASF Corporation), vinylpyrrolidone / vinyl acetate / vinyl propionate copolymer (Rubiscol VAP, manufactured by BASF Corporation), vinyl acetate β / crotonic acid copolymer (Rubicet CA, BASF), vinyl acetate / crotonic acid / vinylpyrrolidone copolymer (Rubicet CAP, BASF), vinylpyrrolidone / acrylate copolymer (Rubiflex, BASF), acrylate / acrylamide copolymer (Ultrahold, BASF), vinyl acetate / butyl maleate / isobornyl acrylate copolymer (Advantage, ISP), carboxyvinyl polymer (Carbopol, BFGoodrich), acrylic acid / alkyl methacrylate copolymer (Pemulene, BF Examples include anionic polymer compounds such as those manufactured by Goodrich, amphoteric polymer compounds such as the amphoteric acetate of dialkylaminoethyl methacrylate polymer (Yukaformer, manufactured by Mitsubishi Chemical Corporation), octylacrylamide acrylate / hydroxypropyl acrylate / butylaminoethyl methacrylate copolymer (AMPHOMER, manufactured by NSC), cationic polymer compounds such as the quaternary compound of vinylpyrrolidone / dimethylaminoethyl methacrylate (GAFQUAT, manufactured by ISP), methylvinylimidazolium chloride / vinylpyrrolidone copolymer (Rubicort, manufactured by BASF), and nonionic polymer compounds such as polyvinylpyrrolidone (Rubiscol K, manufactured by BASF), vinylpyrrolidone / vinyl acetate copolymer (Rubiscol VA, manufactured by BASF), vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer 937, manufactured by ISP), and vinylcaprolactam / vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer VC713, manufactured by ISP).Furthermore, naturally derived polymer compounds such as cellulose or its derivatives, keratin and collagen or their derivatives, calcium alginate, pullulan, agar, gelatin, tamarind seed polysaccharides, xanthan gum, carrageenan, high methoxyl pectin, low methoxyl pectin, guar gum, acacia gum, crystalline cellulose, arabinogalactan, karaya gum, tragacanth gum, alginic acid, albumin, casein, curdlan, gellan gum, and dextran can also be suitably used.

[0038] (9) Examples of bioactive ingredients Bioactive ingredients include substances that provide some kind of bioactivity to the skin when applied to the skin. Examples include whitening ingredients, immune stimulants, anti-aging agents, UV protection agents, slimming agents, skin tightening agents, antioxidants, hair growth agents, hair tonics, moisturizers, blood circulation promoters, antibacterial agents, disinfectants, drying agents, cooling agents, warming agents, vitamins, amino acids, wound healing promoters, irritation relievers, analgesics, cell activators, enzyme components, etc. Examples of suitable ingredients include Angelica keiskei extract, avocado extract, Hydrangea macrophylla extract, Althaea officinalis extract, Arnica montana extract, Aloe vera extract, apricot extract, apricot kernel extract, Ginkgo biloba extract, Fennel extract, Turmeric extract, Oolong tea extract, Rosa multiflora extract, Echinacea japonica leaf extract, Scutellaria baicalensis extract, Phellodendron amurense extract, Coptis japonica extract, Barley extract, Hypericum perforatum extract, Lamium album extract, Nasturtium officinale extract, Orange extract, Dried seawater, seaweed extract, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, chamomile extract, carrot extract, Artemisia capillaris extract, licorice extract, carrot extract, Arctium moniliforme extract, cinchona extract, cucumber extract, guanosine, gardenia extract, bamboo grass extract, Sophora flavescens extract, walnut extract, grapefruit extract, clematis extract, chlorella extract, mulberry extract, gentian extract, black tea extract, yeast extract, burdock extract, corn Rice bran fermentation extract, rice germ oil, comfrey extract, collagen, lingonberry extract, asarum extract, bupleurum extract, umbilical cord extract, salvia extract, soapwort extract, bamboo extract, hawthorn extract, sansho extract, shiitake mushroom extract, rehmannia extract, lithospermum extract, perilla extract, linden extract, meadowsweet extract, peony extract, calamus root extract, birch extract, horsetail extract, ivy extract, hawthorn extract, ce Elderflower extract, yarrow extract, peppermint extract, sage extract, mallow extract, Cnidium officinale extract, Swertia japonica extract, soybean extract, jujube extract, thyme extract, tea extract, clove extract, cogongrass extract, citrus peel extract, angelica extract, calendula extract, peach kernel extract, spruce extract, Houttuynia cordata extract, tomato extract, natto extract, carrot extract, garlic extract, wild rose extract, hibiscus extract,Examples of extracts include Ophiopogon japonicus extract, parsley extract, honey, witch hazel extract, Parietaria trifolia extract, Isodon japonicus extract, bisabolol, loquat extract, coltsfoot extract, butterbur extract, Poria cocos extract, butcher's broom extract, grape extract, propolis, loofah extract, safflower extract, peppermint extract, linden extract, peony extract, hop extract, pine extract, horse chestnut extract, skunk cabbage extract, soapberry extract, lemon balm extract, peach extract, cornflower extract, eucalyptus extract, saxifrage extract, coix seed extract, mugwort extract, lavender extract, apple extract, lettuce extract, lemon extract, astragalus extract, rose extract, rosemary extract, Roman chamomile extract, royal jelly extract, etc. Furthermore, it contains biopolymers such as deoxyribonucleic acid, mucopolysaccharides, sodium hyaluronate, sodium chondroitin sulfate, collagen, elastin, chitin, chitosan, and hydrolyzed eggshell membrane; moisturizing ingredients such as amino acids, hydrolyzed peptides, sodium lactate, urea, sodium pyrrolidone carboxylate, betaine, whey, and trimethylglycine; oily ingredients such as sphingolipids, ceramides, phytosphingosine, cholesterol, cholesterol derivatives, and phospholipids; ε-aminocaproic acid, glycyrrhizic acid, β-glycyrrhetinic acid, lysozyme chloride, and guaiazulene. Immunostimulants such as hydroxyl, hydrocholtisone, vitamins such as vitamin A, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, calcium pantothenate, biotin, nicotinamide, and vitamin C ester, active ingredients such as allantoin, diisopropylamine dichloroacetate, and 4-aminomethylcyclohexanecarboxylic acid, antioxidants such as tocopherol, carotenoids, flavonoids, tannins, lignans, and saponins, cell activators such as alpha-hydroxy acids and beta-hydroxy acids, and blood circulation promoters such as gamma-oryzanol and vitamin E derivatives. Wound-healing agents such as retinol and retinol derivatives, whitening agents such as arbutin, kojic acid, placenta extract, sulfur, ellagic acid, linoleic acid, tranexamic acid, and glutathione, cepharanthine, licorice extract, capsicum tincture, hinokitiol, garlic iodide extract, pyridoxine hydrochloride, DL-α-tocopherol, DL-α-tocopherol acetate, nicotinic acid, nicotinic acid derivatives, calcium pantothenate, D-pantothenyl alcohol, acetyl pantothenyl ethyl ether, biotin, allantoin, isopropylmethylphenol, estradiol All, ethinylestradiol, capronium chloride, benzalkonium chloride, diphenhydramine hydrochloride, tacanal, camphor, salicylic acid, vanillyl nonyl acid, vanillyl nonanoate, piroctone olamine, glyceryl pentadecanoate, L-menthol, mononitroguaiacol, resorcinol, gamma-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormones, cantharis tincture, cyclosporine, zinc pyrithione, hydrocoltisone, minoxidil, polyoxyethylene sorbitan monostearate, peppermint oil,Examples include hair growth products containing Sasanishiki rice extract.

[0039] (10) Examples of antioxidants Examples include sodium bisulfite, sodium sulfite, erythorbic acid, sodium erythorbate, dilauryl thiodipropionate, tocopherol, tol biguanide, nordihydroguaiaretinic acid, parahydroxyanisole, butylhydroxyanisole, dibutylhydroxytoluene, ascorbyl stearate, ascorbyl palmitate, octyl gallate, propyl gallate, carotenoids, flavonoids, tannins, lignans, saponins, and plant extracts with antioxidant effects such as apple extract and clove extract.

[0040] (11) Example of Be Examples include purified water, ethanol, lower alcohols, ethers, LPG, fluorocarbons, N-methylpyrrolidone, fluoroalcohols, volatile linear silicones, and next-generation chlorofluorocarbons (CFCs). [Examples]

[0041] The following describes examples of the present invention. These examples are provided to confirm the various actions and effects of the lactic acid bacteria obtained by the present invention, and the scope of the present invention is not limited to these products and manufacturing methods.

[0042] Examples 1. Methods for Isolating and Identifying Sakura Lactobacillus The isolation and identification methods for *Lactobacillus orientale* are as follows. (1) Method for isolating lactic acid bacteria Petals of weeping cherry blossoms (Prunus pendula) were collected aseptically (using tweezers or gloves). The petals were then placed in a culture medium similar to that used in production (mainly yeast extract) and incubated overnight at 30-37°C. The culture solution was then spread onto BCP-added plate count agar (a medium that turns yellow around the colony when acid is produced, allowing for the selection of acid-producing bacteria) and left to stand for 24 hours. Colonies with yellowed edges were then streaked on BCP-added plate count agar. Single colonies were then collected and incubated for 20 hours in a yeast extract-based liquid medium to confirm culturability. Selection was based on turbidity (OD660 4.0 or higher). Genomes were extracted from strains that met the turbidity criteria, and lactic acid bacteria were selected based on 16s rRNA (700-800 bp from the beginning). (2) Method for identifying isolated cherry blossom lactic acid bacteria A phylogenetic tree was constructed and identification was performed using the full-length 16s rRNA (1400-1500 bp). This phylogenetic tree is shown in Figure 2. Since this is L. casei, it is difficult to determine whether it is L. casei or L. paracasei from the 16s rRNA alone, so a specific primer described in the references was used for identification (Figure 3). As a result, it was identified as Lacticaseibacillus casei, named Lacticaseibacillus casei shidare, and the deposit procedure was carried out (deposit number NITE ABP-04138). (References):Microbiology (Reading). 2017 Jul;163(7):950-960.

[0043] 2. About cherry blossom extract In this example, cherry blossoms were extracted and filtered with aqueous ethanol containing ascorbic acid and malic acid, then concentrated and vacuum-dried. The café oil glucose in the above cherry blossom extract is 2.2 It was %.

[0044] Test example: Evaluation of the effect of promoting collagen synthesis enzyme gene (COL1A1) expression. 1. Experimental Method (1) Method for culturing human dermal fibroblast cells (NHDF cells) Human dermal fibroblasts purchased from Kurabo Industries Ltd. were cultured and used in the experiment. The culture medium used was Dulbecco's Modified Eagles Medium (containing 1000 mg / L glucose, DMEM) supplemented with 10 (v / v) % FBS (Fetal bovine serum), 100 units / mL penicillin G, and 100 μg / mL streptomycin. Cell culture was performed at 75 cm². 2 The procedure was carried out in a culture flask at 37°C in the presence of 5% CO2. For subculturing, the cultured cells were washed twice with PBS (-), then detached from the flask with a 0.25 w / v% Trypsin-1 mM EDTA·4Na solution containing phenol red, and used for the experiment. (2) Evaluation method Evaluation method for promoting collagen (COL1A1) synthase gene expression using NHDF cells NHDF cell suspension (1.6 × 10⁻⁶) 5 Cells (1 mL / well) were seeded into a 12-well plate and cultured for 24 hours. After culturing, the culture medium was changed to one containing the test substance. After 24 hours of culture, RNA was extracted from the cells according to standard procedures, and the expression level of the COL1A1 gene was confirmed by RT-PCR.

[0045] 2. Results and Effects of the Examples Figure 1 shows the effects of cherry blossom lactic acid bacteria, cherry blossom extract, and a combination of cherry blossom lactic acid bacteria and cherry blossom extract on collagen synthase gene expression. No significant promotion of collagen synthase gene expression was observed with the addition of 1 μg / ml of cherry blossom lactic acid bacteria in NHDF cells. Similarly, no significant promotion of collagen synthase gene expression was observed with the addition of 10 μg / ml of cherry blossom extract. However, a significant promotion of collagen synthase gene expression was observed when combining 1 μg / ml of cherry blossom lactic acid bacteria and 10 μg / ml of cherry blossom extract, which showed no significant difference between the two. These results confirm that combining cherry blossom lactic acid bacteria and cherry blossom extract, which showed no significant difference between them, promotes collagen synthase gene expression and has the effect of maintaining skin moisture and elasticity. Furthermore, the most effective combination was found when the ratio (cherry blossom lactic acid bacteria / cherry blossom extract) was 1 / 10.

[0046] The following are examples of formulations for the collagen synthase gene expression promoter of the present invention (hereinafter simply referred to as "the agent of the present invention"). Note that the following formulation examples are not intended to limit the present invention. Example formulation 1: Chewing gum Sugar 53.0 wt% Gum base 20.0 Glucose 10.0 Starch syrup 16.0 Fragrance 0.5 The agent of the present invention 0.5 100.0 wt%

[0047] Example formulation 2: Gummy Reduced starch syrup 40.0 wt% Granulated sugar 20.0 Glucose 20.0 Gelatin 4.7 Wednesday 9.68 Yuzu juice 4.0 Yuzu flavor 0.6 Dye 0.02 Agent of the present invention 1.0 100.0 wt%

[0048] Formula example 3: Candy Sugar 50.0 wt% Starch syrup 33.0 Wednesday 14.4 organic acid 2.0 Fragrance 0.2 The agent of the present invention 0.4 100.0 wt%

[0049] Example of formulation 4: Yogurt (hard / soft) Milk 41.5 wt% Skim milk powder 5.8 Sugar 8.0 Agar 0.15 Gelatin 0.1 Lactic acid bacteria 0.005 The agent of the present invention 0.4 fragrance trace amount water residue 100.0 wt%

[0050] Formulation example 5: Soft drink Fructose-glucose liquid sugar 30.0 wt% Emulsifier 0.5 The agent of the present invention 0.3 Fragrance (appropriate amount) Purified water remainder 100.0 wt%

[0051] Formulation example 6: Tablet confectionery Sugar 76.4 wt% Glucose 19.0 Sucrose fatty acid ester 0.2 The agent of the present invention 0.5 Purified water 3.9 100.0 wt%

[0052] Formulation Example 7: Soft Capsules Brown rice germ oil 47.0 wt% Yuzu seed oil 40.0 Emulsifier 12.0 Agent of the present invention 1.0 100.0 wt%

[0053] Formulation example 8: Tablets Lactose 54.0wt% Crystalline cellulose 30.0 Starch decomposition product 10.0 Glycerin fatty acid ester 5.0 Agent of the present invention 1.0 100.0 wt%

[0054] Formula example 9: Cosmetic cream Squalane 20.0 wt% Beeswax 5.0 Refined jojoba oil 5.0 Glycerin 5.0 Glycerin monostearate 2.0 Polyoxyethylene (20) sorbitan Monosterate 2.0 Agent of the present invention 2.0 Preservative (appropriate amount) Fragrance (appropriate amount) Purified water remainder 100.0 wt%

[0055] Formula example 10: Lotion Ethanol 5.0 wt% Glycerin 2.0 1,3-Butylene glycol 2.0 Polyethylene oleyl ether 0.5 Sodium citrate 0.1 Citric acid 0.1 The agent of the present invention 0.1 Purified water remainder 100.0 wt%

[0056] Formula Example 11: Body Gel Macadamia nut oil 2.0 wt% Octyldodecyl myristate 10.0 Methylphenylpolysiloxane 5.0 Behenyl alcohol 3.0 Stearic acid 3.0 Batyl alcohol 1.0 Glyceryl monostearate 1.0 Polyoxyethylene sorbitol tetraoleate 2.0 Hydrogenated soybean phospholipid 1.0 Ceramide 0.1 Retinyl palmitate 0.1 Preservative (appropriate amount) Centella asiatica extract 1.0 Agent of the present invention 1.0 1,3-Butylene glycol 5.0 Purified water remainder 100.0 wt%

[0057] Formulation example 12: Emulsion Squalane 4.0 wt% Vaseline 2.5 Cetanol 2.0 Glycerin 2.0 Lipophilic glyceryl monostearate 1.0 Stearic acid 1.0 L-arginine 1.0 The agent of the present invention 0.5 Potassium hydroxide 0.1 fragrance trace amount Purified water remainder 100.0 wt%

[0058] Formula Example 13: Bath additive (liquid) Propylene glycol 50.0 wt% Ethanol 20.0 Sodium sulfate 5.0 The agent of the present invention 0.5 Lanolin 0.5 Avocado oil 0.5 Dye 1.5 Fragrance 22.0 100.0 wt% [Industrial applicability]

[0059] Based on the above, the present invention can provide a novel collagen synthesis gene expression promoter.

Claims

1. A collagen synthesis gene expression promoter containing lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) (deposit number NITE ABP-04138) and cherry blossom extract as active ingredients.

2. The collagen synthesis gene expression promoter according to claim 1, characterized in that the blending ratio of lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) (deposit number NITE ABP-04138) to cherry blossom extract is 1 / 0.1 to 1 / 100 in terms of blending ratio (the lactic acid bacteria / cherry blossom extract).

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