JP7406782B2 - Skin external preparations - Google Patents

Description

NPMD NITE AP-01947

The present invention relates to yeast or processed products thereof that can be incorporated into external preparations for the skin (including the scalp).

Conventionally, ingredients derived from natural products have been researched and developed as active ingredients to be incorporated into external skin preparations. However, when these natural product-derived ingredients are used as active ingredients in external skin preparations, there are problems in terms of effectiveness, stability, etc.

As a result of intensive research to solve the above problems, the present inventors found that among yeasts of the genus Saccharomyces, yeast derived from plants of the genus Lilium in the family Liliaceae has the effect of improving rough skin, moisturizing effect, and whitening effect. It has been newly discovered that it has an antioxidant effect, an effect to improve wrinkles and sagging, and an effect to improve pigmentation such as age spots and freckles, and is useful as an active ingredient in external skin preparations.

Conventionally, the use of yeast (eg, Saccharomyces cerevisiae) as an active ingredient in external preparations for skin has been known, for example, from Patent Documents 1 and 2. However, yeast derived from plants of the genus Lilium in the Liliaceae family or its processed products have the effect of improving rough skin, moisturizing effect, whitening effect, antioxidant effect, wrinkle and sagging effect, and pigmentation such as spots and freckles. It had an improving effect and was not known to be used as an active ingredient in external preparations for the skin.
Japanese Patent Application Publication No. 08-163983 JP2009-179642

The present invention is an external skin preparation containing a yeast extract derived from a plant of the genus Lilium of the family Liliaceae, or a concentrate or dry powder thereof as an active ingredient.

The present invention is an external skin preparation containing a yeast extract derived from a plant of the genus Lili, or a concentrate or dry powder thereof as an active ingredient. Or, due to the cell proliferation promoting effect of the dry powder, it is possible to provide an external skin preparation for improving rough skin, moisturizing, whitening, antioxidant, wrinkles, sagging, and pigmentation such as spots and freckles. .

The present invention will be explained in detail below. The yeast used in the present invention is a yeast derived from a plant of the genus Lilium in the family Liliaceae. As the plant of the genus Lilium, Lilium japonicum is preferred. Examples of the yeast derived from the Japanese summer lily include the yeast derived from the Japanese Japanese Patent Application Publication No. 2016-086763.

A yeast extract, or a concentrate or dried product thereof, can be prepared as follows. For example, methods for producing yeast extracts include cultivating yeast in a culture solution, hydrolysis of cultured yeast to solubilize cell components with acid or alkali, and utilizing proteolytic enzymes within the yeast cells. It can be obtained by an autolysis method, an enzymatic method using an enzyme agent such as a proteolytic enzyme, or a combination of these methods.

The carbon source for culturing yeast is not particularly limited, and examples of the carbon source include glucose, fructose, lactose, raffinose, and the like. Further, in addition to the carbon source, a nitrogen source may be added, and examples of the nitrogen source include amino acids, peptone, and the like.

The culture temperature of yeast is in the range of 25°C to 40°C, preferably in the range of 28°C to 35°C. Further, the pH is in the range of 3.0 to 8.0, preferably in the range of 3.5 to 7.0.

A yeast extract concentrate can be obtained by concentrating a yeast culture solution, or a solution obtained by a hydrolysis method, a yeast autolysis method, or an enzymatic method, using a vacuum concentrator or the like.

Further, a dried yeast extract can be obtained by drying a yeast culture solution or a concentrated solution thereof by a vacuum drying method, a spray drying method, or the like.

Further, the dried yeast extract may be a dried product to which excipients are added for the purpose of chemical stability and low hygroscopicity. As excipients, saccharides such as starch, glucose, crystalline cellulose, lactose, and dextrin, and sugar alcohols such as sorbitol, xylitol, erythritol, mannitol, maltitol, and lactitol can be used; Any substance may be used as long as it can be dried and powdered.

The yeast extract or its concentrate or dried product according to the present invention can be incorporated into external skin preparations (cosmetics, quasi-drugs, external pharmaceuticals). Examples of external skin preparations include emulsions, creams, lotions, essences, packs, lipsticks, foundations, liquid foundations, makeup press powders, blushers, whitening powders, face washes, body shampoos, scalp and hair shampoos, and hair conditioners. , shampoos or tonics for hair growth, hair nourishing, cleaning cosmetics such as soaps, bath salts, etc., but the present invention is not limited thereto.

In the case of basic cosmetics, the amount of the yeast extract or its concentrate or dried product according to the present invention is 0.002 to 5.0% by weight (solid content, the same applies hereinafter) in terms of solid content. In the case of makeup cosmetics, the range is from 0.002 to 5.0% by weight, and in the case of cleansing cosmetics, the range is from 0.002 to 20.0% by weight. In the case of hair cosmetics, the solid content ranges from 0.0001 to 10.0% by weight.

When the yeast extract or its concentrate or dried product according to the present invention is used in a skin external preparation, ingredients that can be incorporated into the skin external preparation, such as oily components, surfactants (synthetic and natural), emulsifiers, Moisturizers, thickeners, preservatives/bactericidal agents, antifoaming agents, powder components, antioxidants, chelating agents, pH adjusters, pigments, fragrances, and the like can be blended as appropriate. Further, there is no problem in blending the yeast extract in combination with other physiologically active ingredients as long as the effectiveness and characteristics of the yeast extract or its concentrate or dried product according to the present invention are not impaired.

Here, examples of oily ingredients include lotus oil, olive oil, jojoba oil, castor oil, soybean oil, rice oil, rice bran oil, rice germ oil, coconut oil, chamomile oil, palm oil, cacao oil, meadowfoam oil, and bergamot oil. Plant-derived oils and fats such as oil, rosehip oil, Arabic coffee seed oil, orchid oil, shea butter, tea tree oil, avocado oil, macadamia nut oil, vanilla oil, and plant-derived squalane; Animal oils such as mink oil and turtle oil Fats and oils derived from waxes such as beeswax, carnauba wax, rice wax, and lanolin; Hydrocarbons such as liquid paraffin, petrolatum, paraffin wax, and squalane; myristic acid, palmitic acid, stearic acid, oleic acid, isostearic acid, and eicosenoic acid Fatty acids such as lauryl alcohol, cetanol, stearyl alcohol and other higher alcohols; isopropyl myristate, isopropyl palmitate, butyl oleate, cetyl isooctanoate, 2-ethylhexylglyceride, higher fatty acid octyldodecyl (octyldodecyl stearate, etc.) and synthetic triglycerides.

In addition, examples of surfactants include polyoxyethylene alkyl ether, polyoxyethylene fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyethylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, polyoxyethylene glycerin fatty acid ester, Nonionic surfactants such as oxyethylene hydrogenated castor oil, polyoxyethylene sorbitol fatty acid ester; fatty acid salts, alkyl sulfates, alkylbenzene sulfonates, polyoxyethylene alkyl ether sulfates, polyoxyethylene fatty amine sulfates, polyoxy Anionic surfactants such as ethylene alkyl phenyl ether sulfates, polyoxyethylene alkyl ether phosphates, α-sulfonated fatty acid alkyl ester salts, polyoxyethylene alkyl phenyl ether phosphates; quaternary ammonium salts, primary to secondary Tertiary fatty amine salts, trialkylbenzylammonium salts, alkylpyridinium salts, 2-alkyl-1-alkyl-1-hydroxyethylimidazolinium salts, N,N-dialkylmorphonium salts, polyethylene polyamine fatty acid amide salts, etc. Cationic surfactant; N,N-dimethyl-N-alkyl-N-carboxymethylammoniobetaine, N,N,N-trialkyl-N-alkyleneammoniocarboxybetaine, N-acylamidopropyl-N',N Ampholytic surfactants such as '-dimethyl-N'-β-hydroxypropylammoniosulfobetaine and the like can be used.

In addition, as emulsifiers or emulsification aids, 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, soybean-derived Water-soluble polysaccharide, complex of soybean-derived protein and polysaccharide, lanolin or its derivatives, cholesterol, stevia derivatives (stevia enzyme-treated products, etc.), silicates (aluminum, magnesium, etc.), carbonates (calcium, sodium, etc.), saponin and its derivatives, lecithin and its derivatives (hydrogenated lecithin, hydroxylated lecithin, etc.), lactic acid bacteria-fermented rice, lactic acid bacteria-fermented germinated rice, lactic acid bacteria-fermented grains (wheat, beans, millet, etc.), etc. can also be blended.

In addition, as a moisturizing agent, for example, glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, sorbitol, xylitol, sodium pyrrolidone carboxylate, 2-methacryloyloxyethylphosphorylcholine/butyl methacrylate copolymer liquid In addition, saccharides such as trehalose, mucopolysaccharides (e.g., hyaluronic acid and its derivatives, chondroitin and its derivatives, heparin and its derivatives, etc.), tuberose polysaccharide, elastin and its derivatives, collagen and its derivatives, NMF-related Substances, hydrolyzed conchiolin, hydrolyzed silk, sphingomonas culture, glycosphingolipid, ceramide (human ceramide, yuzu ceramide, etc.), lactic acid, urea, higher fatty acid octyldodecyl, seaweed extract, silane root extract , various amino acids and their derivatives.

Thickeners include, for example, ingredients derived from brown algae, green algae, or red algae such as alginic acid, agar, carrageenan, and fucoidan; Silane root extract; pectin, locust bean gum, aloe polysaccharide, and Alcaligenes-producing polysaccharide. Gums such as xanthan gum, tragacanth gum, roasted bean gum, and guar gum; Cellulose derivatives such as carboxymethylcellulose and hydroxyethylcellulose; polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, mastic resin, acrylic acid/methacrylic acid copolymer Examples include synthetic polymers such as polymers; hyaluronic acid and its derivatives; polyglutamic acid and its derivatives.

Preservatives and disinfectants include, for example, urea; paraoxybenzoic acid esters such as methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, and butyl paraoxybenzoate; phenoxyethanol, dichlorophene, hexachlorophene, and chlorhexidine hydrochloride. , benzalkonium chloride, salicylic acid, ethanol, undecylenic acid, phenols, jamal (imidazodeinyl urea), polyphosphoric acid, propanediol, 1,2-pentanediol, various essential oils, bark dry distillate, fermented radish liquid, sugarcane These include plant-derived ethanol or 1,3-butylene glycol.

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, and silk. There are powders, cellulose powders, grains (rice, wheat, corn, millet, etc.) powders, legumes (soybeans, adzuki beans, etc.) powders, etc.

Examples of ultraviolet absorbers include ethyl para-aminobenzoate, ethylhexyl para-dimethylaminobenzoate, amyl salicylate and its derivatives, 2-ethylhexyl para-methoxycinnamate, 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, aloe extract etc.

Examples of antifoaming agents include ethanol, isopropanol, disiloxane, dimethylpolycyclosane, dimethicone silica, trisiloxane, silylated silica, dimethicone, trimethylsiloxysilicate, and DPG isobornyl ether.

Antioxidants include, for example, butylated hydroxyanisole, butylated hydroxytoluene, propyl gallate, astaxanthin, extract of Murasakibu, silane root extract, peony extract, vitamin E and its derivatives (for example, vitamin E nicotinate). , vitamin E linoleate, etc.).

Examples of the chelating agent include trisodium ethylenediaminehydroxyethyltriacetate, edetic acid or its salts, gluconic acid, phytic acid, sodium polyphosphate, sodium metaphosphate, and tetrasodium hydroxyethane diphosphonate.

Examples of the pH adjuster include citric acid or its salts, lactic acid or its salts, glycolic acid, succinic acid, hydrochloric acid, monoethanolamine, diethanolamine, triethanolamine, sodium hydroxide, potassium hydroxide, and the like.

In addition, whitening agents include t-cycloamino acid derivatives, kojic acid and its derivatives, ascorbic acid and its derivatives, hydroquinone and 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, AMP (adenosine mono phosphate, adenosine monophosphate), and these may be blended alone or in combination.

The above-mentioned kojic acid derivatives include, for example, kojic acid esters such as kojic acid monobutyrate, kojic acid monocaprate, kojic acid monopalmitate, and kojic acid dibutyrate, kojic acid ethers, and kojic acid sugars such as kojic acid glucoside. Ascorbic acid derivatives include, for example, sodium L-ascorbic acid-2-phosphate, magnesium L-ascorbic acid-2-phosphate, sodium L-ascorbic acid-2-sulfate, and L-ascorbic acid. -2-Ascorbic acid ester salts such as magnesium sulfate, L-ascorbic acid-2-glucoside, L-ascorbic acid-5-glucoside, ascorbyl tocopheryl maleate, ascorbyl tocopheryl potassium phosphate, myristyl 3-glyceryl ascorbic acid , ascorbic acid sugar derivatives such as caprylyl 2-glyceryl ascorbic acid, 6-position acylated products of these ascorbic acid sugar derivatives (acyl group is hexanoyl group, octanoyl group, decanoyl group, etc.), L-ascorbic acid tetraisopalmitate ester, L-ascorbic acid tetra-fatty acid esters such as L-ascorbic acid tetralaurate, 3-O-ethyl ascorbic acid, sodium L-ascorbic acid-2-phosphate-6-O-palmitate, glyceryl ascorbic acid or its like. 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 Examples of hydroquinone derivatives include arbutin (hydroquinone-β-D-glucopyranoside) and α-arbutin (hydroquinone-α-D-glucopyranoside); examples of tranexamic acid derivatives include tranexamic acid esters (e.g. lauryl tranexamic acid). ester, tranexamic acid hexadecyl ester, tranexamic acid cetyl ester or salts thereof), amides of tranexamic acid (for example, tranexamic acid methylamide), etc., and resorcinol derivatives include, for example, 4-n-butylresorcinol, 4-n-butylresorcinol, Examples of 2,5-dihydroxybenzoic acid derivatives include isoamylresorcinol, 2,5-diacetoxybenzoic acid, 2-acetoxy-5-hydroxybenzoic acid, 2-hydroxy-5-propionyloxybenzoic acid, and nicotinic acid. Examples of 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.

In addition, the physiologically active ingredients include, for example, placenta extract, sagebrush extract, saxifrage extract, perilla extract, rice bran extract or its hydrolyzate, white mustard extract or its hydrolyzate, and white mustard fermented product. , peony extract or its hydrolyzate, lactic acid bacteria extract, bifidobacterium extract, purple extract, lotus seed extract or its hydrolyzate, lotus seed ferment, coix seed hydrolyzate, coix seed ferment, royal jelly fermentation products, sake lees extract or ceramides contained therein, fermented sake lees, Pandanus amaryllifolius extract, Arcangelicia flava extract, Calendula extract, Chamomile extract, Coral grass extract, Rice leaf extract extracts or hydrolysates thereof, extracts of eggplants (lotus, long eggplants, Kamo eggplants, rice eggplants, etc.) or hydrolysates thereof, extracts of apricot fruits, extracts of seaweeds such as Katamen Kirincho, marine flowers such as eelgrass Plant extracts, fermented soy milk, jellyfish water, rice extracts or hydrolysates thereof, fermented rice extracts, germinated rice extracts or hydrolysates thereof, fermented germinated rice products, black soybean extracts or hydrolysates thereof, Brown sugar extract or its fermented product, damask rose flower extract, ylang-ylang flower extract, bamboo shoot peel 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-hydroxy Butyric acid, gentian extract, licorice extract, carrot extract, panax ginseng extract or its fermented product, camellia extract, red ginseng extract, honeysuckle extract, loofah extract, Ulva extract, peach extract, plum extract , peach kernel extract, kiwi extract, sunflower extract, juazeiro extract, pau d'arco bark extract, daylily extract or fermented product, hibiscus flower extract or fermented product, silverwort extract, cherimoya extract, mango extract, mangosteen extract, funori extract, oolong tea extract, Benifuki extract, silane extract, sweet potato extract, extract of Japanese pepper peel or seed coat or its hydrolyzate, safflower flower extract, Casablanca extract, Sweet potato extract or fermented product thereof, guava leaf extract, olive leaf extract, horse chestnut extract, ginger extract, vanilla extract, Houtyami extract, banbaiyu extract, aloe extract, aloe vera extract, fig Examples include flower extract, apple extract, white asparagus extract, cacao extract, ginseng extract, or a hydrolyzate thereof.

Next, the present invention will be explained in more detail with reference to production examples, test examples, and examples, but the present invention is not limited thereto. In addition, in the following, all parts mean parts by weight, and all percentages mean weight %.

Manufacturing example 1. Preparation of yeast extract (1)
To 2700 g of sterilized GP liquid medium, 300 g of a preculture of yeast (Saccharomyces cerevisiae) derived from Sasayuri, which had been previously cultured in the same medium, was added, and cultured at 30°C for 20 hours with aeration and stirring. After heat sterilization, the pH was adjusted to 8.5 with an aqueous sodium hydroxide solution, and the mixture was heat-treated at 90°C for 3 hours with stirring. After adjusting the pH of this liquid, it was filtered to obtain 1604 g of yeast culture extract (solid content concentration 1.12%).

Production example 2. Preparation of yeast extract (2)
The yeast culture extract prepared by the method shown in Production Example 1 was concentrated to obtain 17 g of yeast culture extract powder. After adding a small amount of water and dissolving it, a small amount of ethanol and 833 g of D-mannitol were added, and the mixture was vacuum-dried with stirring to obtain 850 g of D-mannitol-shaped yeast culture extract powder.

Production example 3. Preparation of yeast extract (3)
To 2,700 g of sterilized GP liquid medium, 300 g of a preculture of yeast (Saccharomyces cerevisiae) derived from Sasayuri, which had been previously cultured in the same medium, was added, and cultured at 30°C for 20 hours with aeration and stirring. After heat sterilization, the pH was adjusted to 2.5 with an aqueous hydrochloric acid solution, and heat treatment was performed at 90°C for 3 hours with stirring. After adjusting the pH of this liquid, it was filtered to obtain 1219 g of yeast culture extract (solid content concentration 1.08%).

Manufacturing example 4. Adjustment of yeast culture solution (4)
To 2,700 g of sterilized GP liquid medium, 300 g of a preculture of yeast (Saccharomyces cerevisiae) derived from Sasayuri, which had been previously cultured in the same medium, was added, and cultured at 30°C for 20 hours with aeration and stirring. After heat sterilization, heat treatment was performed at 90°C for 3 hours. After adjusting the pH of this liquid, it was filtered to obtain 1536 g of yeast culture extract (solid content concentration 0.60%).

Test example 1. Evaluation test for the effect of promoting collagen synthesis in fibroblasts Human dermis-derived fibroblasts NB1RGB were seeded at 1× ¹⁰ cells/well in a 96-well microplate containing Eagle's minimum essential medium containing 0.5% NCS, and incubated at 37°C at 5.0% After pre-culture for 1 day under CO ₂ conditions, Production Example 1 was added to the medium at a concentration of 1.0% and 2.0% (as a solution), and cultured for an additional 5 days under the same conditions. Next, the medium was removed, the cells were fixed with cold methanol and cold ethanol, and then stained with a saturated picric acid aqueous solution containing 0.1% Sirius Red. After washing with purified water, extraction was performed with a 1:1 solution of 0.1% NaOH:methanol, and the amount of collagen was measured at a wavelength of 540 nm using a microplate reader (Model 680, manufactured by Bio-Rad). Perform the same operation as above for the sample without addition (control) in which PBS(-) was added instead of Production Example 1, and calculate the relative value of the collagen amount when each sample was added to the collagen amount obtained here. This was determined and taken as the fibroblast collagen synthesis rate (%). In addition, in order to confirm whether the test system was functioning normally, a similar test was conducted in the case where 1 mM ascorbic acid phosphate magnesium salt was added as a positive control instead of Production Example 1.

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

As shown in Table 1, it was confirmed that the present invention has an extremely excellent collagen synthesis promoting effect. This suggests that the present invention has the effect of improving wrinkles and sagging.

Test example 2. Fibroblast Hyaluronic Acid Synthesis Promotion Evaluation Human dermal-derived fibroblasts NB1RGB were seeded at 1× ¹⁰ cells/well in a 96-well microplate containing Eagle's minimum essential medium containing 0.5% NCS, and incubated at 37°C and 5.0% CO _{2 .} After pre-culture for 1 day under the following conditions, Production Example 1 was added to the medium at a concentration of 1.0% and 2.0% (as a solution), and cultured for an additional 5 days under the same conditions. Thereafter, the supernatant was collected from each well, and the amount of hyaluronic acid in the supernatant was quantified using a hyaluronic acid measurement kit (Quantikine (registered trademark) ELISA Hyaluronan Immunoassay, R&D SYSTEMS, USA). Using a control group in which PBS(-) was added instead of Production Example 1, the relative value of the amount of hyaluronic acid synthesized in each sample added group to the amount of hyaluronic acid synthesized in the control group was calculated, and the hyaluronic acid synthesis promotion effect (%) was calculated. I asked for To confirm that the test was conducted correctly, we also set up a group in which 10mM of N-acetyl-D-glucosamine was added as a positive control.

The results of Test Example 2 are shown in Table 2.
[Table 2]

As shown in Table 2, it was confirmed that the present invention has an extremely excellent effect of promoting hyaluronic acid synthesis. This suggests that the present invention has the effect of improving wrinkles and sagging.

Test example 3. Intrafibroblast ROS suppression test Human dermal-derived fibroblasts (NB1RGB) were prepared at 2 × ¹⁰ cells/mL in Eagle's minimum essential medium containing 10% NCS, and 100 μL was seeded in a 96-well microplate. % carbon dioxide gas and saturated water vapor at 37°C. After 24 hours, a culture solution containing Production Example 1 was added to the culture solution at a final concentration of 0.5%, 1%, or 2%, and cultured. In addition, a test group in which a culture medium containing PBS(-) was added to the final concentration of 2% was set as a control group and used as a comparison group. After 48 hours, the supernatant of the test area was removed and 2',7'-dichlorodihydrofluorescein diacetate (DCFH-2DA) was incorporated into the cells using a fluorescence plate reader (Fluoroscan Ascent, manufactured by Thermo Labsystems). The fluorescence intensity (excitation wavelength: 485 nm, absorption wavelength: 538 nm) was measured, and this was taken as the amount of oxidative damage. After the measurement, the amount of protein in the cell lysate was measured, and the amount of oxidative damage per amount of protein was calculated.

The results of Test Example 3 are shown in Table 3.
[Table 3]

As shown in Table 3, it was confirmed that the present invention has an extremely excellent effect of suppressing intracellular oxidative damage. This suggests that the present invention has an antioxidant effect.

Test example 4. Evaluation test for suppressing effect on neutrophil migration activity Human dermis-derived fibroblasts NB1RGB were seeded at 9 × ¹⁰ cells/well in a 24-well microplate containing Eagle's minimum essential medium containing 10% NCS, and incubated at 37°C at 5.0% After pre-culturing for one day under CO ₂ conditions, Production Example 1 was added to the medium at a concentration of 1.0% (as a solution) and further cultured. The next day, 5 J of ultraviolet A waves were irradiated using an ultraviolet lamp (model number SLUV-6, manufactured by As One Corporation), and the culture supernatant 24 hours after the end of irradiation was used as a chemokine-inducing solution. At the same time, a non-ultraviolet irradiation area was set up for comparison, and the collected culture supernatant was used as a chemokine non-induced solution. 500 μL of the collected culture solution was added to the receptor side of Transwell (Corning, model number 3415). Human leukemia cell line HL60 cells were cultured in RPMI medium containing 1.25% DMSO and 10% FBS for 5 days to induce differentiation into neutrophil-like cells, adjusted to 1 x ¹⁰ cells/mL, and cultured in Transwell. 300 μL was seeded on the insert side. Two hours after seeding the differentiated HL60 cells, cells that migrated to the receptor side were obtained. In addition, by adding 200 μL of 0.5mM EDTA/PBS(-) solution to a separate well from seeding, moving the cell culture insert after migration, and leaving it for 30 minutes, the cells remaining on the bottom of the cell culture insert can be removed. I got it. After the migrated cells were combined and 2',7'-dichlorodihydrofluorescein diacetate was incorporated, the fluorescence intensity (excitation wavelength 485 nm, absorption wavelength 538 nm) was measured using a fluorescence plate reader (Fluoroscan Ascent, manufactured by Thermo Labsystems). The obtained fluorescence intensity was expressed as a relative value when the chemokine-uninduced control was set as 100.

The results of Test Example 4 are shown in Table 4.
[Table 4]

As shown in Table 4, it was confirmed that the present invention has a significantly superior effect of suppressing neutrophil migration activity. This suggests that the present invention has the effect of improving wrinkles and sagging.

Test example 5. IL-8 secretion suppression test Human dermal-derived fibroblasts NB1RGB were seeded at 9 × ¹⁰ cells/well in a 24-well microplate containing Eagle's minimum essential medium containing 10% NCS, and the conditions were 37°C and 5.0% CO _2. After pre-culture for 1 day, Production Example 1 was added to the medium at a concentration of 1.0% (as a solution) and further cultured. The next day, the cells were irradiated with 5 J of ultraviolet A waves using an ultraviolet lamp (As One, model number SLUV-6), and the culture supernatant 24 hours after the end of irradiation was used as an IL-8 induction solution. At the same time, a non-ultraviolet irradiation area was set up for comparison, and the collected culture supernatant was used as an IL-8 non-induced solution. The amount of IL-8 in the collected culture solution was measured using an IL-8 measurement kit (manufactured by Boster Biological Technology). The amount of IL-8 obtained was expressed as a relative value when the uninduced control was set as 100.

The results of Test Example 5 are shown in Table 5.
[Table 5]

As shown in Table 5, it was confirmed that the present invention has an extremely excellent effect of suppressing the secretion of IL-8 (a factor involved in skin inflammation). This suggests that the present invention has the effect of suppressing skin inflammation.

Test example 6. Evaluation test for suppressing effect on melanin synthesis Cultured B16 mouse melanoma cells B16-F10 were seeded at 2.4 x ¹⁰ cells/well in a 24-well microplate, and incubated in RPMI1640 medium containing 10% FBS at 37°C and 5.0% _CO2. After pre-culturing for 1 day, a solution prepared by diluting Production Example 1 to a final concentration of 2.0% (as a solution) and a theophylline-containing medium adjusted to a final concentration of 1 mM were added to RPMI1640 medium containing 10% FBS. and cultured under the same conditions for 2 days. Next, the culture medium was removed, and 200 μL of 1N NaOH/10% dimethyl sulfoxide solution was added per well, sealed, and incubated at 50° C. for 2 hours to lyse the cells. 100 μL of this solution was transferred to another 96-well microplate, and the melanin value was measured at a wavelength of 490 nm using a microplate reader (Model 680, manufactured by Bio-Rad). On the other hand, transfer 5 μL of the same cell lysis solution to another 96-well microplate, add 200 μL of Dye Reagent Concentrate (Bio-Rad) diluted 5 times with purified water, and transfer to a microplate reader (Model 680, Bio-Rad). absorbance at a wavelength of 570 nm was measured using Separately, a known amount of bovine serum albumin (manufactured by Sigma) was serially diluted and the amount of protein equivalent to albumin was measured from a standard curve obtained by performing the same procedure. The obtained absorbance was divided by the amount of protein to determine the amount of melanin per protein. The same procedure as above was performed for the sample without addition (control) in which PBS(-) was added instead of Production Example 1, and the amount of melanin per protein obtained here was compared to the amount of melanin per protein when each sample was added. The relative value of the amount of melanin was determined and defined as the melanin synthesis rate (%). For comparison, a similar test was also conducted in the case where 2 mM kojic acid was added instead of Production Example 1 (positive control).

The results of Test Example 6 are shown in Table 6.
[Table 6]

As shown in Table 6, it was confirmed that the present invention has an extremely excellent melanin synthesis inhibiting effect. This suggests that the present invention is effective in preventing and improving pigmentation such as spots and freckles.

Prescription example 1. Lotion [Ingredients] Part Jojoba oil 1.0
Polyoxyethylene (5.5) Cetyl Alcohol 5.0
Methylparaben 0.1
Yeast extract of Production Example 1 2.0
Glycerin 5.0
1,3-butylene glycol 5.0
Sodium citrate 0.2
Purified water Amount that makes the total amount 100 parts

Prescription example 2. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 2.0 parts of the yeast extract of Production Example 3 was used in place of the yeast extract of Production Example 1 contained in Formulation Example 1.

Prescription example 3. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 2.0 parts of the yeast extract in Production Example 4 was used in place of the yeast extract in Production Example 1 contained in Formulation Example 1.

Prescription example 4. 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
Yeast 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
Carboxymethyl cellulose 0.3
Xanthan gum 0.2
Sodium hyaluronate 0.01
Purified water Amount that makes the total amount 100 parts

Prescription example 5. Emulsion A milky lotion was obtained in the same manner as in Formulation Example 4, except that 3.0 parts of arbutin was used in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide among the ingredients in Formulation Example 4. Ta.

Prescription example 6. Emulsion A milky lotion was prepared in the same manner as in Formulation Example 4, except that 5.0 parts of nicotinic acid amide was used in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide in the ingredients of Formulation Example 4. I got it.

Prescription example 7. Emulsion A milky lotion was prepared in the same manner as in Formulation Example 5, except that 2.0 parts of tranexamic acid was used in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide in the ingredients of Formulation Example 4. Obtained.

Prescription example 8. Emulsion Same as Prescription Example 4 except that 2.0 parts of tranexamic acid cetyl ester hydrochloride is used in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide in the ingredients of Prescription Example 4. A milky lotion was obtained.

Prescription example 9. Emulsion [Ingredients] Part Squalane 3.0
Cetyl alcohol 1.0
Polyethylene glycol monostearate 0.2
Dipropylene glycol 1.0
Cetyl isooctanoate 1.0
Dimethylpolycycloxane 1.0
Yeast extract of Production Example 1 2.0
Lactic acid bacteria fermented rice 2.0
Seaweed extract 1.0
Raffinose 2.0
1,3-butylene glycol 2.0
L-ascorbic acid-2-glucoside 2.0
Potassium hydroxide 0.5
Acetylated sodium hyaluronate 0.2
Guar gum 3.0
Ceramide 1.0
Purified water Amount that makes the total amount 100 parts

Prescription example 10. Emulsion A milky lotion was obtained in the same manner as in Formulation Example 9, except that 3.0 parts of arbutin was used in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide among the ingredients in Formulation Example 9. Ta.

Prescription example 11. Emulsion A milky lotion was prepared in the same manner as in Formulation Example 9 except that 5.0 parts of nicotinic acid amide was used in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide in the ingredients of Formulation Example 9. I got it.

Prescription example 12. Emulsion A milky lotion was prepared in the same manner as in Formulation Example 10, except that 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide were replaced with 2.0 parts of tranexamic acid in the ingredients of Formulation Example 9. Obtained.

Prescription example 13. Cream [Ingredients] Part Olive oil 5.0
Jojoba oil 5.0
Squalane 5.0
Behenyl alcohol 2.0
Palmitic acid 2.5
Yeast extract powder of Production Example 2 2.0
Lactic acid bacteria fermented rice 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 Amount that makes the total amount 100 parts

Prescription example 14. Cream [Ingredients] Part Polyoxyethylene sorbitan monostearate 1.5
Cetyl alcohol 3.0
Glyceryl monostearate 1.0
Squalane 5.0
Olive oil 5.0
Dipropylene glycol 3.0
Yeast extract of Production Example 3 2.0
Gentiana extract 1.0
Licorice extract 2.0
Citric acid 0.2
Methylparaben 0.2
Purified water Amount that makes the total amount 100 parts

Example 15. Liquid foundation [Ingredients] Part Stearic acid 2.4
Propylene glycol monostearate 2.0
Cetostearyl alcohol 0.2
liquid lanolin 2.0
Liquid paraffin 3.0
Isopropyl myristate 8.5
Propylparaben 0.05
Yeast extract powder of Production Example 2 5.0
Sodium carboxymethylcellulose 0.2
Bentonite 0.5
Propylene glycol 4.0
Triethanolamine 1.1
Methylparaben 0.1
Titanium oxide 8.0
Talc 4.0
Coloring pigment Appropriate amount Purified water Amount to make the total amount 100 parts

Prescription example 16. Body shampoo [Ingredients] Part N-lauroylmethylalanine sodium 25.0
Coconut oil fatty acid potassium solution (40%) 26.0
Coconut oil fatty acid diethanolamide 3.0
Methylparaben 0.1
Yeast extract powder of Production Example 2 5.0
1,3-butylene glycol 2.0
Purified water Amount that makes the total amount 100 parts

Prescription example 17. Hair shampoo [Ingredients] Part N-coconut oil fatty acid methyltaurine sodium 10.0
Polyoxyethylene (3) alkyl ether sodium sulfate 20.0
Lauryldimethylaminoacetic acid betaine 10.0
Coconut oil fatty acid diethanolamide 4.0
Methylparaben 0.1
Citric acid 0.1
Yeast extract of Production Example 1 2.0
1,3-butylene glycol 2.0
Purified water Amount that makes the total amount 100 parts

Example 18. Hair conditioner [Ingredients] Part Polyoxyethylene (10) Hydrogenated castor oil 1.0
Distearyldimethylammonium chloride 1.5
Stearyltrimethylammonium chloride 2.0
Glyceryl 2-ethylhexanoate 1.0
Setanol 3.0
Stearyl alcohol 1.0
Methylparaben 0.1
Yeast extract powder of Production Example 2 2.0
1,3-butylene glycol 5.0
Purified water Amount that makes the total amount 100 parts

Claims (3)

A skin treatment for improving rough skin containing an extract of yeast (Patent Microorganism Depositary Center accession number NITE AP-01947) derived from Lilium japonicum of the Liliaceae family, or its concentrate or dry powder as an active ingredient. Topical preparation. An anti-aging product containing an extract of yeast (Patent Microorganism Depositary Center accession number NITE AP-01947) derived from Lilium japonicum of the Liliaceae family, or its concentrate or dry powder as an active ingredient. External skin preparation. A skin whitening product containing an extract of yeast (Patent Microorganism Depositary Center accession number NITE AP-01947) derived from Lilium japonicum of the Liliaceae family (Liliaceae), or its concentrate or dry powder as an active ingredient. Topical preparation.