Skin preparations

Abstract

To provide a novel skin external active ingredient that is composed of a natural product-derived component having excellent biological safety and effectiveness.SOLUTION: A skin external preparation contains extract from Crocus sativus (Iridaceae, Crocus) as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an external skin preparation containing a plant extract as an active ingredient. [Background technology]

[0002] It has been known that ultraviolet rays, chemical substances, allergens, etc. stimulate living cells, causing the alteration of their constituents and promoting the synthesis of cytokines and active oxygen. It is also known that the altered cellular components, cytokines, or active oxygen induce inflammation and abnormal deposition of melanin pigment in cells, causing age spots, freckles, melasma, etc.

[0003] Conventionally, various ultraviolet absorbers, antioxidants, and anti-inflammatory agents have been proposed for the purpose of preventing and improving skin health and aging, and cosmetics, foods, beverages, and medicines containing them have been put on the market. For example, ultraviolet absorbers such as cinnamic acid derivatives and benzophenone derivatives; antioxidants such as vitamin C, vitamin E, and catalase; and anti-inflammatory agents such as glycyrrhizic acid or its salts, allantoin, and tranexamic acid have been proposed, but there is a demand for safer antioxidants and anti-inflammatory agents derived from natural products.

[0004] Although plant extracts have been proposed as active ingredients that can be incorporated into cosmetics, there have been problems with efficacy and safety.

[0005] [Patent Document 1] Japanese Patent Publication No. 61-024522 [Patent Document 2] Japanese Patent Publication No. 04-305519 DISCLOSURE OF THEINVENTION [Problem to be solved by the invention]

[0006] In view of the above problems, the present inventors have conducted extensive research to find new active ingredients derived from natural products. As a result, they have newly discovered that an extract of saffron (Crocus sativus) of the genus Crocus in the family Iridaceae has the effect of suppressing gene expression of factors that contribute to the production of inflammatory substances (cytokines) induced by external stimuli such as ultraviolet rays, and is useful as an active ingredient in external skin preparations such as cosmetics. [Means for solving the problem]

[0007] The present invention is an external skin preparation containing, as an active ingredient, an extract of saffron (Crocus sativus) of the genus Crocus in the family Iridaceae. The present invention is a skin external preparation containing an extract of saffron (Crocus sativus) of the genus Crocus in the family Iridaceae as an active ingredient, and is characterized in that the extract contains Kaempferol 3-O-beta-Sophoroside. Effect of the Invention

[0008] The present invention provides an external preparation for the skin that contains an extract of saffron (Crocus sativus) of the genus Crocus in the family Iridaceae as an active ingredient, and can provide an external preparation for the skin that protects the skin from external stimuli such as ultraviolet rays based on the effects of the active ingredient, saffron extract. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] In the present invention, saffron (Crocus sativus) of the genus Crocus in the family Iridaceae is used as the extraction material for the plant extract. The whole plant, flowers, leaves, stems, stamens, pistils, and roots can be used as the extraction part, but the whole plant or flowers are preferred. When flowers are used, the stamens and / or pistils may be included.

[0010] The preparation of the plant extract containing the active ingredient of the present invention can be carried out by washing the plant to be extracted with water in advance, if necessary, to remove foreign matter, leaving it as it is or drying it, and further cutting or crushing it, if necessary, and contacting it with an extraction solvent according to a conventional method such as immersion, countercurrent extraction, steam distillation, etc. In addition, in the present invention, a supercritical extraction method may be adopted.

[0011] Examples of extraction solvents used in the extract treatment include water; lower alcohols such as methanol, ethanol, and propanol; polyhydric alcohols such as ethylene glycol, propylene glycol, 1,3-butylene glycol, and glycerin; esters such as ethyl acetate, butyl acetate, and methyl propionate; ketones such as acetone and methyl ethyl ketone; ethers such as ethyl ether and isopropyl ether; and hydrocarbon solvents such as n-hexane, toluene, and chloroform, and these may be used alone or in combination of two or more.

[0012] In preparing the extract, the pH of the extract is not particularly limited, but is generally preferably in the range of pH 3 to 9. The pH can be adjusted by adding an alkalinity regulator such as sodium hydroxide, sodium carbonate, or potassium hydroxide, or an acidity regulator such as citric acid, hydrochloric acid, phosphoric acid, or sulfuric acid to the above-mentioned extraction solvent.

[0013] The extraction conditions such as extraction temperature and time vary depending on the type and pH of the solvent used, or the size of the plant material, but for example, in the case of the immersion method using methanol or ethanol, or a mixed solvent of water and a lower alcohol or polyhydric alcohol as the extraction solvent, the extraction temperature is in the range of 0 to 80° C. The extraction time is preferably in the range of 1 hour to 7 days in the case of cold extraction at 4° C., and in the range of 1 hour to 3 days in the case of medium temperature extraction at about 40° C., and in the case of high temperature extraction at 70 to 80° C., it is preferably in the range of 1 hour to 24 hours. In the case of the immersion method, the bath ratio is generally 1 to 200 times, preferably 1 to 100 times, the amount of the solvent relative to the plant material by weight.

[0014] The extract according to the present invention is characterized by containing "Kaempferol 3-O-beta-Sophoroside" as an active ingredient.

[0015] On the other hand, the extract according to the present invention is preferably prepared to have a high concentration of Kaempferol 3-O-beta-Sophoroside by combining one or more of the following: activated carbon treatment, ion exchange resin treatment, synthetic adsorbent, silica gel, and recrystallization treatment. Activated carbon may be obtained by treating raw materials such as pine trees, bamboo, coconut shells, walnut shells, and other plant materials, as well as coal and petroleum materials, with these raw materials as raw materials, using a physical method such as a high-temperature carbonization method using gases such as steam, carbon dioxide, and air, or by treating the raw materials with chemicals such as zinc chloride, and then heating them to make them porous. Examples of ion exchange resins include weakly basic anion exchange resins, strongly acidic cation exchange resins, strongly basic ion exchange resins, and weakly acidic cation exchange resins. Examples of synthetic adsorbents include nonionic resins such as chelating resins, strongly acidic cation adsorption resins, styrene-divinylbenzene copolymers, and methacrylic acid ester polymers. By any of these treatments, compounds that cause coloring problems (pigments such as anthocyanins) and compounds that cause skin irritation are removed from the extract. Compounds that cause safety problems such as coloring and skin irritation (pigments such as anthocyanins) are removed. By the above treatments, it is preferable that the compounds are contained in the extract in a range of 0.01% to 0.1%.

[0016] In the present invention, topical skin preparations include cosmetics, quasi-drugs, and topical medicines, and examples thereof include milky lotions, creams, lotions, essences, packs, lipstick, foundations, liquid foundations, make-up press powders, blushers, face powders, facial cleansers, body shampoos, scalp and hair shampoos, hair conditioners, shampoos or tonics for hair growth and nourishment, cleansing cosmetics such as soaps, and bath additives, but the present invention is not limited to these.

[0017] In addition to the extract of the present invention, the skin topical preparation may contain, as necessary, ingredients used in skin topical preparations, such as oily ingredients, surfactants (synthetic or natural), moisturizers, thickeners, preservatives and disinfectants, powder ingredients, UV absorbers, antioxidants, colorants, fragrances, etc. Furthermore, as long as the effectiveness and characteristics of the extract, hydrolysate or fermented product of the present invention are not impaired, there is no problem in combining and blending 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 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] Further, as the emulsifier or emulsification aid, 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, water-soluble polysaccharides derived from soybeans, soybean-derived protein and polysaccharide complexes, lanolin or its derivatives, cholesterol, stevia derivatives (stevia enzyme fermentation products, etc.), silicates (aluminum, magnesium, etc.), carbonates (calcium, sodium, etc.), saponin and its derivatives, lecithin and its derivatives (hydrogenated 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.

[0021] Further, 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, ceramide, lactic acid, urea, higher fatty acid octyldodecyl, seaweed extract, Bletilla serrata root (white) extract, 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, extract of Brussels sieboldii, 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] Examples of physiologically active ingredients include placenta extract, mulberry bark extract, saxifrage extract, perilla extract, rice bran extract or hydrolysate thereof, white mustard extract or hydrolysate thereof, fermented white mustard, peony extract or hydrolysate thereof, lactic acid bacteria fermented rice, purple barley extract, lotus seed extract or hydrolysate thereof, fermented lotus seed product, Job's tears hydrolysate, fermented Job's tears seed product, fermented royal jelly product, sake lees extract or ceramide contained therein, fermented sake lees product, Pandanus amaryllifolius Roxb. extract, Arcangelicia flava Merrilli extract, and chamomile extract.In addition, coral grass extract, rice leaf extract or hydrolysate thereof, eggplant (lotus, long eggplant, Kamo eggplant, rice eggplant, etc.) extract or hydrolysate thereof, apricot fruit extract, seaweed extract such as Eucalyptus tschonoskii, extract of marine flowering plants such as Zostera marina, fermented soy milk, jellyfish water, rice extract or hydrolysate thereof, fermented rice extract, germinated rice extract or hydrolysate thereof, fermented germinated rice, black bean extract or hydrolysate thereof, brown sugar extract or fermented product thereof, extract of Damask rose flower, ylang-ylang flower extract, extract of bamboo shoot skin, linoleic acid and its derivatives or processed products (for example, liposomal linoleic acid, etc.) ), collagen and its derivatives derived from animals or fish, elastin and its derivatives, glycyrrhizic acid and its derivatives (dipotassium salt, etc.), t-cycloamino acid derivatives, vitamin A and its derivatives, allantoin, diisopropylamine dichloroacetate, gamma-amino-beta-hydroxybutyric acid, gentian extract, licorice extract, carrot extract, ginseng extract or its fermented product, camellia extract, red ginseng extract, Mitsuishi Kami extract, loofah extract, Ulva pertusa extract, peach extract, plum extract, peach kernel extract, kiwi extract, sunflower extract, Zizyphus joazeiro extract, pau d'arco bark extract, daylily extract or fermented product, hibiscus flower extract or fermented product, silverleaf extract, cherimoya extract, mango extract, mangosteen extract, funori extract, oolong tea extract, red fuki extract, sliver extract, sweet potato extract, Japanese pepper peel or seed coat extract or hydrolysate, 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, Houttuynia cordata extract, Banpeiyu extract, aloe extract, aloe vera extract, fig flower extract, apple extract, white asparagus extract, red clover extract, etc.

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

[0034] Preparation Example 1. Preparation of extract (1) Saffron flowers were dried, and 2000g of purified water was added to 100g of the dried product, and the mixture was soaked at 40℃ for 2 hours. This was filtered to obtain 1750g of a brown, transparent saffron flower crude extract. The saffron flower crude extract was subjected to a styrene-divinylbenzene copolymer, washed with purified water and a 20w / w% aqueous ethanol solution, and then eluted with 1,3-butylene glycol. This was filtered, and purified water was added to obtain 4700g of a yellow, transparent saffron flower extract (solid concentration: 0.10%).

[0035] Test Example 1. Quantitative analysis of Kaempferol 3-O-beta-Sophoroside <Quantitative method> 1 g of the extract of Preparation Example 1 was precisely weighed out, and the mobile phase was added to make exactly 10 mL to prepare a sample solution. 10 μL of this sample solution was taken and tested by liquid chromatography under the following test conditions. The peak area of ​​the obtained chromatogram was calculated, and the concentration (%) of Kaempferol 3-O-beta-Sophoroside was calculated from the calibration curve prepared in advance. Kaempferol 3-O-beta-Sophoroside concentration in this product (%) = C / sample amount (g) x 10 C: Kaempferol 3-O-beta-Sophoroside concentration (w / v%) calculated from the calibration curve <Creating a calibration curve> Accurately weigh out about 5 mg of Kaempferol 3-O-beta-Sophoroside, add the mobile phase to make exactly 50 mL, and use this as the standard stock solution. Accurately take 1.5 mL of the standard stock solution, add the mobile phase to make exactly 10 mL each, and use this as the standard solution. Take 10 μL of the standard stock solution and standard solution, and perform the test by liquid chromatography under the above test conditions. A calibration curve was created by plotting the peak area of ​​the obtained chromatogram on the vertical axis and the Kaempferol 3-O-beta-Sophoroside concentration (w / v%) on the vertical axis. <Test conditions> Detector: Ultraviolet spectrophotometer (measurement wavelength: 254 nm) Column: A stainless steel tube having an inner diameter of 4.6 mm and a length of 25 cm is packed with 5 μm octadecylsilylated silica gel for liquid chromatography. Column temperature: constant temperature around 35℃ Mobile phase: 0.05% phosphoric acid aqueous solution / acetonitrile mixture (17:3) Flow rate: 0.8 mL per minute (retention time of Kaempferol 3-O-beta-Sophoroside is approximately 17 minutes)

[0036] As a result of Test Example 1, it was confirmed that the extract according to the Production Example contained 0.043% Kaempferol 3-O-beta-Sophoroside. In the present invention, for example, by adjusting the extraction process of Production Example 1, it is preferable that 0.01% to 0.1% Kaempferol 3-O-beta-Sophoroside is contained. [Table 1] JPEG0007664595000001.jpg52135

[0037] Test Example 2: Evaluation of the effect of UV rays on the suppression of inflammation-related gene expression Normal human epidermal cells were grown at 6 × 10 in HuMedia KG2 (Kurabo) containing a growth additive. 4 The concentration was adjusted to cells / mL, 1 mL of the solution was inoculated into a 24-well plate, and cultured at 37°C under 5% CO2 and saturated water vapor. After one day of culture, a culture medium containing the extract of Production Example 1 (the extract of Production Example 1 was added so that the final concentration as a solution was 2% relative to the total volume of the culture medium) was added and cultured. As a comparative control, a test group (control group) was set up in which a culture medium containing only a 30% aqueous solution of 1,3-butylene glycol (30-BG) was added instead of the extract of Production Example 1 (the final concentration of 30% 1,3-butylene glycol (30-BG) relative to the total volume of the culture medium was adjusted to 2%). After another day of culture, a UV-B lamp (TL20W / 12RS, manufactured by Philips) was used to illuminate the bottom of the incubator at approximately 50 mJ / cm. 2The cells in the test group 24 hours after the ultraviolet irradiation were collected with 0.5 mL of Trizol reagent (Invitrogen). 100 μL of chloroform (Wako Pure Chemical Industries, Ltd.) was added to the collected cells, mixed and stirred, and centrifuged for 15 minutes at 12,500 rpm and 4 ° C. with a centrifuge (TOMY / MX-160), and 200 μL of the aqueous layer was collected. 250 μL of isopropanol (Wako Pure Chemical Industries, Ltd.) was added to the collected aqueous layer, mixed and stirred, and centrifuged for 10 minutes at 12,500 rpm and 4 ° C. to obtain a precipitate of total RNA. 1 mL of 75% ethanol was added to the total RNA, and after stirring and washing, the precipitate was collected by centrifugation at 12,500 rpm and 4 ° C. for 10 minutes. The collected total RNA was reverse transcribed using a specified kit [PrimeScript RT reagent Kit with gDNA Eraser (Perfect Real Time) (Takara Bio)] to synthesize cDNA. Using the synthesized cDNA as a sample, the expression of various genes and the expression of the internal standard substance ACTB gene were detected using Thermal Cycler Dice (registered trademark) Real Time System Single (manufactured by Takara Bio Inc.) and SYBR (registered trademark) Premix Ex TaqTM II (Tli RNaseH Plus) [manufactured by Takara Bio Inc.]. Here, ACTB (β-actine) is one of the housekeeping genes (a gene that is expressed at a constant amount in many tissues and cells, is constantly expressed, and is essential for the maintenance and proliferation of cells), and since its expression level is always constant, it is used as an internal standard in PCR experiments. The test results were obtained by comparing the expression level of each gene in each test group when the expression level of the ACTB gene was constant. In this test system, the expression level of each gene in the control group was set to 100, and the relative value of the expression level of the gene in the other test groups was obtained.

[0038] The results of Test Example 2 are shown in Table 2. [Table 2] JPEG0007664595000002.jpg82129

[0039] As shown in Table 2, it was confirmed that the present invention remarkably suppresses the expression of factors involved in skin inflammation [chemokine CXCL16, interleukin-8 (IL-8), interleukin-6 (IL-6), interleukin-1 (IL-1α), chemokine CCL2, and HMGB1 (high mobility group box 1)]. This suggests that the present invention has an effect of suppressing skin inflammation.

[0040] Test Example 3. Evaluation of the effect of suppressing HMGB1 (High Mobility Group Box 1) HMGB1 is a nuclear factor that promotes the transcription of NF-kB, which is secreted at the early stage of inflammation, and in this Test Example 3, the effect of suppressing the production of this factor was evaluated. On the first day, epidermal cells (NHEK) and fibroblasts (NB1RGB) were seeded in a 96-well plate, and on the second day, a medium containing the extract of Production Example 1 was added to each culture system. On the third day, each cell was irradiated with UVB (50 mJ / cm) from the bottom. 2 ) was irradiated. 24 hours after irradiation, the cultured cell supernatant was collected and reacted with an ELISA kit to measure absorbance (450 nm), and the amount of HMGB1 (ng / mL) was calculated from the calibration curve. The absorbance was measured when a control group was added with 30% 1,3-butylene glycol instead of the extract of Production Example 1 and when it was not irradiated with ultraviolet light, and a relative value was calculated when the amount of HMGB1 in the control group without ultraviolet light irradiation was set at 100.

[0041] The test results for epidermal cells in Test Example 3 are shown in Table 3. [Table 3] JPEG0007664595000003.jpg59148

[0042] The test results for fibroblasts in Test Example 3 are shown in Table 4. [Table 4] As shown in Tables 3 and 4, it was confirmed that the extract of Production Example 1 inhibits UV-induced HMGB1 production in epidermal cells and fibroblasts. This suggests an inhibitory effect on early inflammation by inhibiting the transcription of NF-kB.

[0043] Test Example 4: Evaluation of ICAM-1 (Intercellular Adhesion Molecule-1) Inhibitory Effect ICAM-1 is an adhesion factor present on vascular endothelial cells and involved in the migration of leukocytes, and in this Test Example 3, the effect of inhibiting the production of this factor was evaluated. On the first day, fibroblasts (NB1RGB) were seeded in a 96-well plate, and on the second day, a medium containing the extract of Production Example 1 was added. On the third day, UVA (20 mJ) was irradiated from the bottom. 24 hours after irradiation, the cultured cell supernatant was collected. Vascular endothelial cells (HUVEC) were seeded in a 96-well plate, and the following day, the above-recovered fibroblast cell supernatant was added and cultured. After 6 days of culture, the cells were fixed and blocked, and reacted with anti-ICAM-1 antibody as the primary antibody and fluorescently labeled secondary antibody in that order. After washing and removing the secondary antibody, the fluorescence intensity (Ex = 544 nm, Em = 590 nm) was measured. Furthermore, DNA was fluorescently stained with hoechst33342, and then the fluorescence intensity (Ex = 355 nm, Em = 460 nm) was measured to calculate the amount of ICAM-1 per DNA. A group containing 30% 1,3-butylene glycol instead of the extract of Production Example 1 was used as a control, and the fluorescence intensity was measured when exposed to ultraviolet light and when not exposed to ultraviolet light, and the relative value was calculated with the amount of ICAM-1 in the control without ultraviolet light exposure set at 100.

[0044] The evaluation results of Test Example 4 are shown in Table 5. [Table 5] JPEG0007664595000005.jpg60147

[0045] As shown in Table 5, it was confirmed that the extract of Production Example 1 inhibited the production of ICAM-1 induced by ultraviolet light, suggesting an effect of inhibiting the worsening of inflammation caused by leukocyte infiltration.

[0046] Formulation example 1. Lotion [Ingredients] Part Olive oil 1.0 Polyoxyethylene (5.5) Cetyl Alcohol 5.0 Butylparaben 0.1 Extract of Preparation Example 1 5.0 Ethanol 5.0 Glycerin 5.0 1,3-Butylene glycol 5.0 Potassium hydroxide (appropriate amount) Purified water (enough to make the total amount 100 parts) Fragrance (appropriate amount)

[0047] Formulation example 2: Emulsion [Ingredients] Part Liquid Paraffin 6.0 Hexalan 4.0 Jojoba oil 1.0 Polyoxyethylene (20) Sorbitan Monostearate 1.0 Lipophilic glyceryl stearate 1.0 Soy Lecithin 1.5 Extract of Preparation 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 Sodium hyaluronate 0.01 Purified water (enough to make the total amount 100 parts) Fragrance (appropriate amount)

[0048] Formulation example 3. Emulsion An emulsion was obtained in the same manner as in Formulation Example 3, except that, among the ingredients of Formulation Example 2, 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide were replaced with 3.0 parts of arbutin.

[0049] Formulation example 4. Emulsion An emulsion was obtained in the same manner as in Formulation Example 3, except that, among the ingredients of Formulation Example 2, 2.0 parts of tranexamic acid were used in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide.

[0050] Formulation example 5. Emulsion An emulsion was obtained in the same manner as in Formulation Example 3, except that of Formulation Example 2, 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide were replaced with 3.0 parts of nicotinamide.

[0051] Formulation example 6. Lotion [Ingredients] Part Extract of Preparation Example 1 10.0 Ethanol 10.0 Glycerin 3.0 1,3-Butylene glycol 2.0 Methylparaben 0.2 Citric acid 0.1 Sodium citrate 0.3 Carboxyvinyl polymer 0.1 Xanthan gum 0.1 Fragrance (appropriate amount) Potassium hydroxide (appropriate amount) Purified water (enough to make the total amount 100 parts)

[0052] Prescription example 7. Essence [Ingredients] Part Ethanol 2.0 Glycerin 5.0 1,3-Butylene glycol 5.0 Methylparaben 0.1 Hyaluronic acid 0.1 Extract of Preparation Example 1 5.0 Citric acid 0.3 Sodium citrate 0.6 Purified water (enough to make the total amount 100 parts)

[0053] Example 8. 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 Extract of Preparation Example 1 5.0 Sodium carboxymethylcellulose 0.2 Bentonite 0.5 Propylene glycol 4.0 Triethanolamine 1.1 Methylparaben 0.1 Purified water (enough to make the total amount 100 parts) Titanium dioxide 8.0 Talc 4.0 Color pigment (appropriate amount)

[0054] Formulation example 9. Body shampoo [Ingredients] Part Sodium N-lauroylmethylalanine 25.0 Potassium coconut oil fatty acid solution (40%) 26.0 Coconut oil fatty acid diethanolamide 3.0 Methylparaben 0.1 Extract of Preparation Example 1 5.0 1,3-Butylene glycol 2.0 Purified water (enough to make the total amount 100 parts)

[0055] Formulation example 10. Hair shampoo [Ingredients] Part Sodium N-cocoyl methyl taurate 10.0 Sodium polyoxyethylene (3) alkyl ether sulfate 20.0 Lauryl dimethylaminoacetate betaine 10.0 Coconut oil fatty acid diethanolamide 4.0 Methylparaben 0.1 Citric acid 0.1 Extract of Preparation Example 1 2.0 1,3-Butylene glycol 2.0 Purified water (enough to make the total amount 100 parts)

[0056] Example 11. Hair conditioner [Ingredients] Part Polyoxyethylene (10) hydrogenated castor oil 1.0 Distearyldimethylammonium chloride 1.5 Stearyltrimethylammonium chloride 2.0 Glyceryl 2-ethylhexanoate 1.0 Cetanol 3.2 Stearyl alcohol 1.0 Methylparaben 0.1 Extract of Preparation Example 1 2.0 1,3-Butylene glycol 5.0 Purified water (enough to make the total amount 100 parts)

Claims

1. A skin topical agent containing as its active ingredient an extract of saffron (Crocus sativus) petals (excluding the mixed solvent extract consisting of betaine, lactic acid and water) from the Crocus genus of the Iridaceae family.

2. A skin topical preparation characterized in that the petal extract according to claim 1 contains Kaempferol 3-O-beta-Sophoroside (excluding Kaempferol 3-O-lactyl-Sophoroside).

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