Lactic acid bacterium and skin external preparation

Abstract

To provide a component derived from natural sources and its applications in a skin external preparation (such as quasi-drugs or cosmetics).SOLUTION: The present invention provides a lactic acid bacterium as "Lactococcus lactis" derived from the flowers of Prunus x yedoensis (deposit number: NBRC115206), and a skin external preparation (quasi-drug or cosmetic) comprising a component derived from the lactic acid bacterium.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to lactic acid bacteria or extracts thereof that can be incorporated into external preparations for skin. [Background technology]

[0002] In the past, research and development has been conducted on ingredients derived from natural products as active ingredients to be incorporated into topical skin preparations. However, when using these ingredients derived from natural products as active ingredients in topical skin preparations, there were issues not only regarding their effectiveness and stability, but also regarding ensuring the stability of the materials.

[0003] In view of the above-mentioned problems, microbial-derived components have attracted attention as components derived from natural products. For example, techniques for using lactic acid bacteria-derived components as active ingredients in foods or cosmetics have been disclosed (Patent Documents 1 to 6). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent Publication No. 2000-264811 [Patent Document 2] Patent Publication No. 2006-028047 [Patent Document 3] Patent Publication No. 2007-070305 [Patent Document 4] Patent Publication No. 2011-168520 [Patent Document 5] Patent Publication No. 2022-122490 [Patent Document 6] Patent Publication No. 2023-040559 DISCLOSURE OF THEINVENTION [Problem to be solved by the invention]

[0005] As a result of intensive research aimed at solving the above problems, the present inventors have newly discovered that lactic acid bacteria derived from cherry blossoms (Lactococcus lactis, accession number: NBRC115206) have an effect of activating epidermal cells and are useful as an active ingredient in topical skin preparations. [Means for solving the problem]

[0006] The present invention relates to an external skin preparation containing, as an active ingredient, lactic acid bacteria (Lactococcus lactis, accession number: NBRC115206) derived from cherry blossoms, an extract of said lactic acid bacteria, or a concentrate or dry powder of the lactic acid bacteria extract. Effect of the Invention

[0007] The present invention relates to an external skin preparation containing, as an active ingredient, lactic acid bacteria derived from cherry blossoms (Lactococcus lactis, deposit number: NBRC115206), an extract of the lactic acid bacteria, or a concentrate or dry powder of the lactic acid bacteria extract. According to the present invention, by incorporating the active ingredient, lactic acid bacteria extract, or a concentrate or dry powder thereof, it is possible to provide an external skin preparation that exhibits an epidermal cell activation effect. BEST MODE FOR CARRYING OUT THEINVENTION

[0008] The present invention will be described in detail below. The lactic acid bacteria used in the present invention are Lactococcus lactis, derived from Somei-Yoshino cherry blossoms (Deposit No. NBRC115206).

[0009] The lactic acid bacteria extract, or a concentrate or dried product thereof, can be prepared as follows: For example, the lactic acid bacteria extract can be produced by a method of culturing lactic acid bacteria in a culture medium, a hydrolysis method in which the cultured lactic acid bacteria are solubilized with an acid or alkali to solubilize the bacterial components, an autolysis method using protease or the like present in the lactic acid bacteria cells, an enzymatic method using an enzyme such as a protease, or a combination of these methods.

[0010] The carbon source used in culturing lactic acid bacteria is not particularly limited, and examples of the carbon source include glucose, fructose, lactose, raffinose, etc. In addition to the carbon source, a nitrogen source may be added, and examples of the nitrogen source include amino acids and peptones.

[0011] The culture temperature for the lactic acid bacteria is in the range of 20° C. to 40° C., and preferably in the range of 28° C. to 37° C. The pH is in the range of 3.0 to 8.5, and preferably in the range of 3.5 to 7.5.

[0012] A concentrate of a lactic acid bacteria extract can be obtained by concentrating a culture solution of lactic acid bacteria, or a solution obtained by a hydrolysis method, an autolysis method of lactic acid bacteria, or an enzymatic method, using a vacuum concentrator or the like.

[0013] Furthermore, a dried product of a lactic acid bacteria extract can be obtained by drying a culture solution of lactic acid bacteria or a concentrated solution thereof by a vacuum drying method, a spray drying method or the like.

[0014] The dried product of the lactic acid bacteria extract may be prepared by adding an excipient for the purpose of chemical stability and low hygroscopicity. The excipient may be a sugar such as starch, glucose, crystalline cellulose, lactose, or dextrin, or a sugar alcohol such as sorbitol, xylitol, erythritol, mannitol, maltitol, or lactitol, but any substance may be used as long as it can be mixed with the lactic acid bacteria extract to form a dried powder.

[0015] The lactic acid bacteria extract or its concentrate or dry matter according to the present invention can be incorporated into external skin preparations (cosmetics, quasi-drugs, external medicines). Examples of external skin preparations include milky lotions, creams, lotions, essences, packs, lipsticks, foundations, liquid foundations, make-up press powders, blushers, face powders, face washes, body shampoos, scalp and hair shampoos, hair conditioners, shampoos or tonics for hair growth and nourishment, cleansing cosmetics such as soaps, and bath agents, but the present invention is not limited thereto.

[0016] The amount of the lactic acid bacteria extract or its concentrate or dry matter according to the present invention is in the range of 0.002 to 5.0% by weight (% by weight of solids, the same applies below) in the case of basic cosmetics, 0.002 to 5.0% by weight in the case of makeup cosmetics, 0.002 to 20.0% by weight in the case of cleansing cosmetics, and 0.0001 to 10.0% by weight in the case of hair cosmetics.

[0017] When the lactic acid bacteria extract or its concentrate or dried product according to the present invention is used in an external preparation for skin, it may be appropriately blended with ingredients that can be blended in external preparations for skin, such as oily ingredients, surfactants (synthetic or natural), emulsifiers, moisturizing agents, thickeners, preservatives / bactericides, antifoaming agents, powder ingredients, antioxidants, chelating agents, pH adjusters, colorants, fragrances, etc. Furthermore, as long as the effectiveness and characteristics of the lactic acid bacteria extract or its concentrate or dried product according to the present invention are not impaired, it may be blended in combination with other physiologically active ingredients.

[0018] Examples of oily components include plant-derived oils such as lotus oil, olive oil, jojoba oil, castor oil, soybean oil, rice oil, rice bran oil, rice germ oil, coconut oil, chamomile oil, palm oil, cocoa oil, meadowfoam oil, bergamot oil, rosehip oil, coffee tree seed oil, lambender oil, shea butter, tea tree oil, avocado oil, macadamia nut oil, vanilla oil, and plant-derived squalane; animal-derived oils such as mink oil and turtle oil; waxes such as beeswax, carnauba wax, rice wax, and lanolin; liquid paraffin, Examples of the fatty acids include petrolatum, paraffin wax, squalane, etc.; myristic acid, palmitic acid, stearic acid, oleic acid, isostearic acid, eicosenoic acid, etc.; higher alcohols such as lauryl alcohol, cetanol, stearyl alcohol, etc.; synthetic esters and synthetic triglycerides such as isopropyl myristate, isopropyl palmitate, butyl oleate, cetyl isooctanoate, 2-ethylhexyl glyceride, and higher fatty acid octyldodecyl (e.g., octyldodecyl stearate), etc.

[0019] Examples of the surfactant include nonionic surfactants such as polyoxyethylene alkyl ethers, polyoxyethylene fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyethylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, polyoxyethylene glycerin fatty acid esters, polyoxyethylene hydrogenated castor oil, and polyoxyethylene sorbitol fatty acid esters; fatty acid salts, alkyl sulfates, alkylbenzene sulfonates, polyoxyethylene alkyl ether sulfates, polyoxyethylene fatty amine sulfates, polyoxyethylene alkyl phenyl ether sulfates, polyoxyethylene alkyl ether phosphates, and α-sulfonated fatty acid alkyl esters. Examples of usable surfactants include anionic surfactants such as quaternary ammonium salts, primary to tertiary fatty amine salts, trialkylbenzylammonium salts, alkylpyridinium salts, 2-alkyl-1-alkyl-1-hydroxyethylimidazolinium salts, N,N-dialkylmorpholinium salts, and polyethylenepolyamine fatty acid amide salts; and amphoteric surfactants such as N,N-dimethyl-N-alkyl-N-carboxymethylammoniobetaine, N,N,N-trialkyl-N-alkyleneammoniocarboxybetaine, and N-acylamidopropyl-N',N'-dimethyl-N'-β-hydroxypropylammoniosulfobetaine.

[0020] In addition, the emulsifier or emulsification aid may also be blended with stevia derivatives such as enzyme-treated stevia, saponin or its derivatives, casein or its salts (sodium, etc.), sugar and protein complexes, sucrose or its esters, lactose, soy-derived water-soluble polysaccharides, soy-derived protein and polysaccharide complexes, lanolin or its derivatives, cholesterol, stevia derivatives (enzyme-treated stevia, etc.), silicates (aluminum, magnesium, etc.), carbonates (calcium, sodium, etc.), saponin and its derivatives, lecithin and its derivatives (hydrogenated lecithin, hydroxide lecithin, etc.), lactic acid bacteria-fermented rice, lactic acid bacteria-fermented germinated rice, lactic acid bacteria-fermented grains (wheat, beans, millet, etc.), etc.

[0021] Examples of moisturizing agents include glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, sorbitol, xylitol, sodium pyrrolidone carboxylate, 2-methacryloyloxyethyl phosphorylcholine-butyl methacrylate copolymer liquid, and further examples include sugars such as trehalose, mucopolysaccharides (e.g., hyaluronic acid and its derivatives, chondroitin and its derivatives, heparin and its derivatives, etc.), tuberose polysaccharides, elastin and its derivatives, collagen and its derivatives, NMF-related substances, hydrolyzed conchiolin protein, hydrolyzed silk, Sphingomonas culture, sphingoglycolipids, ceramides (human ceramide, yuzu ceramide, etc.), lactic acid, urea, higher fatty acid octyldodecyl, seaweed extract, Bletilla serrata root (white) extract, and various amino acids and their derivatives.

[0022] Further, examples of thickening agents include components derived from brown algae, green algae, or red algae, such as alginic acid, agar, carrageenan, and fucoidan; Bletilla serrata root (white) extract; polysaccharides such as pectin, locust bean gum, aloe polysaccharide, and Alcaligenes polysaccharide; gums such as xanthan gum, tragacanth gum, roast bean gum, and guar gum; cellulose derivatives such as carboxymethyl cellulose and hydroxyethyl cellulose; synthetic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, mastic resin, and acrylic acid / methacrylic acid copolymer; hyaluronic acid and its derivatives; and polyglutamic acid and its derivatives.

[0023] Examples of preservatives and disinfectants include urea; paraoxybenzoic acid esters such as methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, and butyl paraoxybenzoate; phenoxyethanol, dichlorophene, hexachlorophene, chlorhexidine hydrochloride, benzalkonium chloride, salicylic acid, ethanol, undecylenic acid, phenols, jamal (imidazolidinyl urea), polyphosphoric acid, propanediol, 1,2-pentanediol, various essential oils, bark distillate, radish fermentation liquid, and ethanol or 1,3-butylene glycol derived from plants such as sugar cane.

[0024] Examples of powder components include sericite, titanium oxide, talc, kaolin, bentonite, zinc oxide, magnesium carbonate, magnesium oxide, zirconium oxide, barium sulfate, silicic anhydride, mica, nylon powder, polyethylene powder, silk powder, cellulose-based powder, powder of grains (rice, wheat, corn, millet, etc.), powder of beans (soybeans, adzuki beans, etc.), and the like.

[0025] Examples of ultraviolet absorbers include ethyl paraaminobenzoate, ethylhexyl paradimethylaminobenzoate, amyl salicylate and its derivatives, 2-ethylhexyl paramethoxycinnamate, octyl cinnamate, oxybenzone, 2,4-dihydroxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonate, 4-tert-butyl-4-methoxybenzoylmethane, 2-(2-hydroxy-5-methylphenyl)benzotriazole, urocanic acid, ethyl urocanate, and aloe extract.

[0026] Examples of the antifoaming agent include ethanol, isopropanol, disiloxane, dimethylpolycyclohexane, dimethicone silicate, trisiloxane, silylated silica, dimethicone, trimethylsiloxysilicate, and DPG isobornyl ether.

[0027] Examples of antioxidants include butylhydroxyanisole, butylhydroxytoluene, propyl gallate, astaxanthin, extract of Brussels sprouts, extract of Bletilla serrata root, extract of peony root, vitamin E and its derivatives (e.g., vitamin E nicotinate, vitamin E linoleate, etc.).

[0028] Examples of the chelating agent include trisodium ethylenediaminehydroxyethyltriacetate, edetic acid or its salts, gluconic acid, phytic acid, sodium polyphosphate, sodium metaphosphate, and tetrasodium hydroxyethanediphosphonate.

[0029] Examples of pH adjusters include citric acid or salts thereof, lactic acid or salts thereof, glycolic acid, succinic acid, hydrochloric acid, monoethanolamine, diethanolamine, triethanolamine, sodium hydroxide, and potassium hydroxide.

[0030] Further, examples of whitening agents include t-cycloamino acid derivatives, kojic acid and its derivatives, ascorbic acid and its derivatives, hydroquinone or its derivatives, ellagic acid and its derivatives, nicotinic acid and its derivatives, resorcinol derivatives, tranexamic acid and its derivatives, 4-methoxysalicylic acid potassium salt, magnolignan (5,5'-dipropyl-biphenyl-2,2'-diol), hydroxybenzoic acid and its derivatives, vitamin E and its derivatives, α-hydroxy acid, and AMP (adenosine monophosphate), which may be blended alone or in combination.

[0031] Examples of the kojic acid derivatives include kojic acid esters such as kojic acid monobutyrate, kojic acid monocaprate, kojic acid monopalmitate, and kojic acid dibutyrate, kojic acid ethers, and kojic acid sugar derivatives such as kojic acid glucoside. Examples of the ascorbic acid derivatives include ascorbic acid derivatives such as sodium L-ascorbic acid 2-phosphate, magnesium L-ascorbic acid 2-phosphate, sodium L-ascorbic acid 2-sulfate, and magnesium L-ascorbic acid 2-sulfate. Ascorbic acid sugar derivatives such as ester salts, L-ascorbic acid-2-glucoside, L-ascorbic acid-5-glucoside, ascorbyl tocopheryl maleate, ascorbyl tocopheryl phosphate K, myristyl 3-glyceryl ascorbate, caprylyl 2-glyceryl ascorbate, etc., 6-acylated products of these ascorbic acid sugar derivatives (the acyl group is a hexanoyl group, an octanoyl group, a decanoyl group, etc.), L-ascorbic acid tetraisopalmitate, L-ascorbic acid tetralaurate, etc. Ascorbic acid tetra fatty acid esters, 3-O-ethyl ascorbic acid, L-ascorbic acid-2-phosphate-6-O-sodium palmitate, glyceryl ascorbic acid or its acylated derivatives, ascorbic acid glycerin derivatives such as bisglyceryl ascorbic acid, aminopropyl L-ascorbic acid phosphate, hyaluronic acid derivatives of L-ascorbic acid, 3-OD lactose-L-ascorbic acid, isostearyl ascorbyl phosphate, etc., and hydroquinone derivatives include arbutin (hydroquinone-β-D-glucopyranoside). amide), α-arbutin (hydroquinone-α-D-glucopyranoside), etc.; tranexamic acid derivatives include tranexamic acid esters (e.g., tranexamic acid lauryl ester, tranexamic acid hexadecyl ester, tranexamic acid cetyl ester or a salt thereof), tranexamic acid amides (e.g., tranexamic acid methylamide), etc.; resorcinol derivatives include, for example, 4-n-butylresorcinol, 4-isoamylresorcinol, etc.; 2,5-dihydroxybenzoic acid derivatives include, for example, 2,Examples of nicotinic acid derivatives include nicotinamide and benzyl nicotinate, and examples of α-hydroxy acids include lactic acid, malic acid, succinic acid, citric acid, and α-hydroxyoctanoic acid.

[0032] Further, examples of physiologically active ingredients include placenta extract, FK706, linoleic acid and its derivatives or processed products (e.g., liposomal linoleic acid, etc.), animal- or fish-derived collagen and its derivatives, elastin and its derivatives, glycyrrhizic acid and its derivatives (dipotassium salt, etc.), t-cycloamino acid derivatives, vitamin A and its derivatives, allantoin, diisopropylamine dichloroacetate, γ-amino-β-hydroxybutyric acid, gentian extract, and licorice extract.

[0033] The present invention will now be described in more detail with reference to Production Examples, Test Examples and Examples, but the present invention is not limited thereto. In the following, all parts mean parts by weight, and all % means % by weight.

[0034] Manufacturing Example 1. Preparation of lactic acid bacteria extract (1) A liquid medium containing glucose and peptone was prepared and sterilized, and 18 g of a preculture solution of lactic acid bacteria (Lactococcus lactis NBRC115206) derived from Somei-Yoshino flowers that had been cultured in GYP liquid medium was added to 900 g of the sterilized liquid medium, and the mixture was left to stand for 6 days at 30°C. This lactic acid bacteria culture solution was heat sterilized and filtered to obtain 749 g of lactic acid bacteria culture solution (solid concentration 1.10%).

[0035] Test Example 1. Epidermal cell activation effect test Human epidermal cells (NHEK) were cultured at 8 × 10 in a 96-well microplate containing HuMedia KG2 medium (Kurabo). 3The cells were seeded at 100 cells / well, pre-cultured for 1 day under conditions of 37°C and 5.0% CO2, and then HuMedia KB2 medium containing the extract of Production Example 1 (2.0%) as a sample solution was added, and the cells were cultured under the same conditions for another 2 days. After culturing for 2 days, the supernatant was discarded, and the cells were washed once with PBS(-), and 100μL / well of hoechst33342 reagent diluted 100 times with PBS(-) was added, and the cells were incubated at 37°C for 1 hour to fluorescently stain the DNA. Then, the fluorescence intensity [excitation: 355nm, emission: 460nm: fluorescence microplate reader (Fluoroscan Ascent, Thermo Fisher Scientific)) was measured to determine the amount of DNA. The same operation was performed on a section where 30% 1,3-butylene glycol was added instead of the sample solution of Production Example 1, and the relative value of the fluorescence intensity of each sample-added section to the fluorescence intensity (DNA amount) obtained here was determined, and the cell proliferation rate (%) was obtained.

[0036] The results of Test Example 1 are shown in Table 1. [Table 1] JPEG2025025767000001.jpg34122

[0037] As shown in Table 1, it was confirmed that the lactic acid bacteria extract according to the present invention has a significantly superior epidermal cell activation effect. This suggests that the lactic acid bacteria extract according to the present invention, when incorporated as an active ingredient in a skin topical agent, can activate epidermal cells and normalize epidermal turnover, leading to moisturized skin with a strong barrier function.

[0038] Formulation example 1. Lotion [Ingredients] Part Jojoba oil 1.0 Polyoxyethylene (5.5) Cetyl Alcohol 5.0 Methylparaben 0.1 Lactic acid bacteria extract of Production Example 1 2.0 Glycerin 5.0 1,3-Butylene glycol 5.0 Sodium citrate 0.2 Purified water (enough to make the total amount 100 parts)

[0039] Formulation example 2: Lotion A lotion was prepared with the same composition as that of Formulation Example 1, except that the lactic acid bacteria extract of Production Example 2 was used in place of the lactic acid bacteria extract of Production Example 1.

[0040] Formulation example 3. Emulsion [Ingredients] Part Squalane 5.0 Hexalan 3.0 Jojoba oil 1.0 Camellia oil 1.5 Polyoxyethylene (20) Sorbitan Monostearate 1.0 Lipophilic glyceryl stearate 1.0 Hydrogenated soy lecithin 1.5 Lactic acid bacteria extract of Production Example 1 3.0 L-Ascorbic acid-2-glucoside 2.0 Potassium hydroxide 0.5 Glycerin 3.0 1,3-Butylene glycol 2.0 Carboxymethylcellulose 0.3 Xanthan gum 0.2 Sodium hyaluronate 0.01 Purified water (enough to make the total amount 100 parts)

[0041] Formulation example 4. Emulsion An emulsion was prepared with the same composition as that of Formulation Example 3, except that the lactic acid bacteria extract of Production Example 2 was used in place of the lactic acid bacteria extract of Production Example 1.

[0042] Formulation example 5. Cream [Ingredients] Part Olive oil 5.0 Jojoba oil 5.0 Squalane 5.0 Behenyl Alcohol 2.0 Palmitic acid 2.5 Lactic acid bacteria extract of Production Example 1 2.0 Hydrogenated Lecithin 0.5 Carboxyvinyl polymer 0.3 Sodium alginate 0.2 Seaweed extract 2.0 Methylparaben 0.15 Ethylparaben 0.03 Purified water (enough to make the total amount 100 parts)

[0043] Formulation example 6. Cream A cream was prepared with the same composition as in Formulation Example 5, except that the lactic acid bacteria extract of Production Example 2 was used in place of the lactic acid bacteria extract of Production Example 1.

Claims

1. Lactococcus lactis (deposition number: NBRC115206) is a lactic acid bacterium derived from Somei-Yoshino cherry blossoms.

2. An epidermal cell activator containing the lactic acid bacteria according to claim 1.

3. A skin topical preparation containing the lactic acid bacteria according to claim 1.

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