Topical skin preparations

Abstract

To provide a novel active ingredient that can be blended in a skin external preparation, and exhibits skin-whitening effect, anti-aging effect and ultraviolet damage recovery effect.SOLUTION: A novel active ingredient of a skin external preparation is extract from carrot of Panax (Araliaceae), and contains ginsenoside Rg3 and Rg5.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to an extract of Panax ginseng C.A. Meyer belonging to the genus Panax of the Araliaceae family, which can be incorporated into external preparations for the skin (including the scalp).

Background Art

[0002] Conventionally, techniques for using Panax ginseng or its extract as an active ingredient in external skin preparations are known as described in Patent Documents 1 to 6, for example. However, conventional extracts of Panax ginseng had problems in terms of effectiveness and safety.

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Patent Document 6

Disclosure of the Invention

Problems to be Solved by the Invention

[0003] As a result of intensive research to solve the above problems, the present inventors newly found that an extract obtained from Panax ginseng C.A. Meyer belonging to the genus Panax of the Araliaceae family and containing at least ginsenoside Rg3 and Rg5 has a whitening effect, an anti-aging effect, and / or a repair effect on cell damage caused by ultraviolet rays, and is useful as an active ingredient in external skin preparations. Also, it was newly found that ginsenoside Rg5 is useful as a whitening component.

Means for Solving the Problems

[0004] The present invention relates to a topical skin whitening agent containing a ginseng extract, which is obtained from Panax ginseng (Panax CA Meyer), a plant belonging to the genus Panax in the family Araliaceae, and contains ginsenoside Rg3 and ginsenoside Rg5, as an active ingredient. The present invention relates to an anti-aging topical skin preparation containing an extract of Panax ginseng (Panax ginseng CA Meyer), which belongs to the genus Panax in the family Araliaceae, and which contains ginsenoside Rg3 and ginsenoside Rg5 as an active ingredient. The present invention relates to a topical skin preparation for repairing cell damage caused by ultraviolet light, which contains a ginseng extract obtained from Panax ginseng (Panax CA Meyer), a plant belonging to the genus Panax in the family Araliaceae, and which contains ginsenoside Rg3 and ginsenoside Rg5 as an active ingredient. The present invention is a skin whitening agent comprising ginsenoside Rg5. [Effects of the Invention]

[0005] According to the present invention, a ginseng extract containing ginsenoside Rg3 and ginsenoside Rg5 provides a topical skin preparation for whitening, anti-aging, or repairing cell damage caused by ultraviolet rays, due to its whitening effect, anti-aging effect, and UV damage repair effect. Furthermore, ginsenoside Rg5 can be provided as a topical skin whitening ingredient. [Brief explanation of the drawing]

[0006] [Figure 1] This figure shows the results of a skin whitening effect evaluation test. [Figure 2] This figure shows the results of a skin whitening effect evaluation test. [Figure 3] This figure shows the results of a skin whitening effect evaluation test. [Figure 4] This figure shows the results of a skin whitening effect evaluation test. [Figure 5] This figure shows the results of a cell repair test after UV irradiation. [Figure 6] This figure shows the evaluation results of intracellular glutathione levels after UV irradiation. [Figure 7] This figure shows the evaluation results of intracellular glutathione levels after UV irradiation. [Figure 8] This figure shows the results of the fibroblast activation test. [Best Mode for Carrying Out the Invention]

[0007] The Panax ginseng (Panax ginseng CA Meyer), belonging to the genus Panax in the family Araliaceae, is used in this invention. The roots and fibrous roots are particularly preferred as the extractant parts of the Panax ginseng.

[0008] In the present invention, a ginseng extract containing ginsenosides Rg3 and Rg5 is obtained by subjecting ginseng roots (particularly fibrous roots are preferred) to a treatment such as heating. In the present invention, Rg3 is preferably present in an amount of 20 ppm or more, more preferably 40 ppm or more, and even more preferably 60 ppm or more. Ginsenoside Rg5 is preferably present in the extract in an amount of 10 ppm or more, more preferably 15 ppm or more, and even more preferably 25 ppm or more.

[0009] Furthermore, in the extract according to the present invention, the content ratio of ginsenoside Rg3 to ginsenoside Rg5 is preferably 2:1 to 4:1 by weight.

[0010] The carrot extract containing the above ginsenoside Rg3 and ginsenoside Rg5 can be obtained, for example, by heat treatment. As the heating temperature conditions, heat at 100°C to 121°C for 1 hour or more, and after heating, extract the carrot with water, a lower alcohol, a polyhydric alcohol or a mixed solvent thereof to obtain a carrot extract containing an amount of ginsenoside Rg3 and Rg5 that exhibits a whitening effect, an anti-aging effect and a repair effect on cell damage caused by ultraviolet rays. Also, as described later, from the viewpoint of the whitening effect, it is preferable that the amounts of ginsenoside Rg1 and Rb1 in the extract are smaller. It is preferably at least less than the amount of ginsenoside Rg3.

[0011] As the above extraction solvent, from the viewpoints of extraction of active ingredients and safety, a mixed solvent of water and a polyhydric alcohol (ethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin, etc.) is preferable. The mixing ratio of water and the polyhydric alcohol is preferably 70:30 to 10:90.

[0012] The extract obtained by the above method may be formulated into a skin external preparation as a liquid extract or in a dry powder state.

[0013] The extract or its dried product according to the present invention can be formulated into a skin external preparation (cosmetics, quasi-drugs, external pharmaceuticals). Examples of skin external preparations include, for example, emulsions, creams, lotions, essences, packs, lipsticks, foundations, liquid foundations, makeup press powders, blushers, face powders, facial cleansers, body shampoos, scalp and hair shampoos, hair conditioners, hair growth and hair nourishing shampoos or tonics, soaps and other cleansing cosmetics, and furthermore, bath agents, etc., but the present invention is not limited thereto.

[0014] The amount of extract or concentrate or dried product thereof according to the present invention is in the range of 0.0001 to 10.0% by weight (solids by weight, the same applies hereinafter) in the case of basic cosmetics, in the range of 0.0001 to 5.0% by weight in the case of makeup cosmetics, and in the range of 0.0001 to 20.0% by weight in the case of cleansing cosmetics.

[0015] When the extract according to the present invention is used in a topical skin preparation, ingredients that can be incorporated into topical skin preparations, such as oily components, surfactants (synthetic and natural), emulsifiers, moisturizers, thickeners, preservatives / bactericides, defoaming agents, powder components, antioxidants, chelating agents, pH adjusters, pigments, fragrances, etc., can be appropriately added as needed. Furthermore, there is no problem in combining it with other physiologically active ingredients as long as the effectiveness and characteristics of the ginseng extract or its concentrate or dried product according to the present invention are not impaired.

[0016] Here, oily components include, for example, lotus oil, rose oil, rosehip oil, olive oil, jojoba oil, castor oil, soybean oil, rice oil, rice bran oil, rice germ oil, coconut oil, chamomile oil, Roman chamomile oil, palm oil, cocoa oil, meadowfoam oil, bergamot oil, rosehip oil, coffee arabic seed oil, lavender oil, shea butter, geranium oil, pine seed oil, tea tree oil, avocado oil, macadamia nut oil, vanilla oil, plant-derived oils such as squalane; animal-derived oils such as mink oil and turtle oil; beeswax, carnauba wax, rice wax. Examples include waxes such as custard and lanolin; hydrocarbons such as liquid paraffin, petrolatum, paraffin wax, and squalane; fatty acids such as myristic acid, palmitic acid, stearic acid, oleic acid, isostearic acid, and eicosenoic acid; higher alcohols such as lauryl alcohol, cetanol, and stearyl alcohol; synthetic esters and synthetic triglycerides such as isopropyl myristate, isopropyl palmitate, butyl oleate, cetyl isooctanoate, 2-ethylhexylglyceride, and higher fatty acid octyldodecyl (octyldodecyl stearate, etc.).

[0017] Furthermore, examples of surfactants 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 alkylphenyl ether sulfates, polyoxyethylene alkyl ether phosphates, and α-sulfonated fatty acid alkyl ethers. Anionic surfactants such as telates and polyoxyethylene alkylphenyl ether phosphates; cationic 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-dialkylmorphonium salts, and polyethylene polyamine fatty acid amide salts; and amphoteric surfactants such as N,N-dimethyl-N-alkyl-N-carboxymethylammonium betaine, N,N,N-trialkyl-N-alkyleneammonium carboxybetaine, and N-acylamidopropyl-N′,N′-dimethyl-N′-β-hydroxypropylammonium sulfobetaine can be used.

[0018] Furthermore, emulsifiers or emulsifying aids may include stevia derivatives such as enzyme-treated stevia, saponins or their derivatives, casein or its salts (such as sodium), sugar-protein complexes, sucrose or its esters, lactose, water-soluble polysaccharides derived from soybeans, complexes of soybean-derived proteins and polysaccharides, lanolin or its derivatives, cholesterol, stevia derivatives (such as enzyme-treated stevia), silicates (such as aluminum and magnesium), carbonates (such as calcium and sodium), saponins and their derivatives, lecithin and its derivatives (such as hydrogenated lecithin), lysolecithin or its derivatives, lactic acid bacteria-fermented rice, lactic acid bacteria-fermented germinated rice, lactic acid bacteria-fermented grains (such as wheat, beans, and other grains).

[0019] Furthermore, examples of humectants include glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, sorbitol, xylitol, sodium pyrrolidone carboxylate, 2-methacryloyloxyethyl phosphorylcholine / butyl methacrylate copolymer solution, as well as sugars such as trehalose, Tremella fuciformis polysaccharide, 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, sphingoglycolipids, ceramides (human-type ceramides, yuzu ceramides, etc.), lactic acid, urea, higher fatty acid octyldodecyl, seaweed extract, Bletilla striata root extract, various amino acids and their derivatives.

[0020] Furthermore, examples of thickening agents include components derived from brown algae, green algae, or red algae such as alginic acid, agar, carrageenan, and fucoidan; Bletilla striata root extract; polysaccharides such as 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; 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.

[0021] Furthermore, preservatives and disinfectants include, for example, urea; parahydroxybenzoic acid esters such as methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, and butyl parahydroxybenzoate; phenoxyethanol, dichlorophene, hexachlorophene, chlorhexidine hydrochloride, benzalkonium chloride, salicylic acid, ethanol, undecylenic acid, phenols, jamal (imidazodinylurea), polyphosphate, propanediol, 1,2-pentanediol, various essential oils, bark distillates, radish ferment filtrate, and plant-derived ethanol or 1,3-butylene glycol from sugarcane, etc.

[0022] In addition, powder components include, for example, sericite, titanium dioxide, talc, kaolin, bentonite, zinc oxide, magnesium carbonate, magnesium oxide, zirconium oxide, barium sulfate, anhydrous silicic acid, mica, nylon powder, polyethylene powder, silk powder, cellulose-based powders, grain powders (rice, wheat, corn, millet, etc.), and legume powders (soybeans, adzuki beans, etc.).

[0023] Examples of UV 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, and aloe extract.

[0024] Examples of defoaming agents include ethanol, isopropanol, disiloxane, dimethylpolycyclosane, dimethicone silica, trisiloxane, silylated silica, dimethicone, trimethylsiloxysilicate, and DPG isobornyl ether.

[0025] Examples of antioxidants include butylhydroxyanisole, butylhydroxytoluene, propyl gallate, astaxanthin, extract of Callicarpa japonica, extract of Bletilla striata root, extract of Paeonia lactiflora, vitamin E and its derivatives (e.g., vitamin E nicotinate, vitamin E linoleate, etc.).

[0026] Examples of chelating agents include trisodium ethylenediamine hydroxyethyl triacetate, EDTA or its salts, gluconic acid, phytic acid, sodium polyphosphate, sodium metaphosphate, and tetrasodium hydroxyethanediphosphonate.

[0027] Examples of pH adjusting agents 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.

[0028] Furthermore, 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, potassium 4-methoxysalicylate, magnolignan (5,5'-dipropyl-biphenyl-2,2'-diol), hydroxybenzoic acid and its derivatives, vitamin E and its derivatives, α-hydroxy acids, and AMP (adenosine monophosphate, adenosine monophosphate). These may be used individually or in combination.

[0029] Furthermore, 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 sugar derivatives such as kojic acid glucoside. Ascorbic acid derivatives include, for example, L-ascorbic acid-2-phosphate sodium, L-ascorbic acid-2-phosphate magnesium, L-ascorbic acid-2-sulfate sodium, and L-ascorbic acid-2-sulfate magnesium. Ascorbic acid sugar derivatives such as ester salts, L-ascorbic acid-2-glucoside, L-ascorbic acid-5-glucoside, ascorbyl tocopheryl maleate, ascorbyl tocopheryl potassium phosphate, myristyl 3-glyceryl ascorbate, caprylyl 2-glyceryl ascorbate, 6-position acylated products of these ascorbic acid sugar derivatives (acyl groups include hexanoyl, octanoyl, and decanoyl groups), L-ascorbic acid tetraisopalmitate, L-ascorbic acid tetralaurate, etc. Examples of hydroquinone derivatives include tetra-rubic acid fatty acid esters, 3-O-ethyl ascorbic acid, sodium L-ascorbic acid-2-phosphate-6-O-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 as hydroquinone derivatives, arbutin (hydroquinone-β-D-glucopyrano Examples of tranexamic acid derivatives include α-arbutin (hydroquinone-α-D-glucopyranoside), tranexamic acid esters (e.g., lauryl tranexamic acid ester, hexadecyl tranexamic acid ester, cetyl tranexamic acid ester or their salts), amide forms of tranexamic acid (e.g., methyl tranexamic acid amide), resorcinol derivatives include, for example, 4-n-butylresorcinol, 4-isoamylresorcinol, and 2,5-dihydroxybenzoic acid derivatives include, for example, 2,Examples of nicotinic acid derivatives include 5-diacetoxybenzoic acid, 2-acetoxy-5-hydroxybenzoic acid, and 2-hydroxy-5-propionyloxybenzoic acid. Examples of nicotinic acid derivatives include nicotinamide and benzyl nicotinate. Examples of α-hydroxy acids include lactic acid, malic acid, succinic acid, citric acid, and α-hydroxyoctanoic acid.

[0030] Furthermore, the physiologically active ingredients include, for example, placenta extract, mulberry bark extract, saxifrage extract, perilla extract, rice bran extract or its hydrolysate, white mustard extract or its hydrolysate, white mustard ferment, peony extract or its hydrolysate, lactic acid bacteria extract, bifidobacteria extract, beautyberry extract, lotus seed extract or its hydrolysate, lotus seed ferment, Job's tears hydrolysate, Job's tears seed ferment, royal jelly ferment, soybean sprout extract, sake lees extract or ceramide contained therein, sake lees ferment, Pandanus amaryllis extract, and Arcangeli. Shea flava extract, calendula extract, chamomile extract, coral grass extract, rice leaf extract or its hydrolysate, eggplant (lotus, long eggplant, Kamo eggplant, American eggplant, etc.) extract or its hydrolysate, apricot fruit extract, seaweed extract such as Eriocaulon sieboldii, marine flowering plant extract such as Zostera margaritacea, soy milk ferment, jellyfish water, rice extract or its hydrolysate, rice fermentation extract, germinated rice extract or its hydrolysate, germinated rice ferment, black bean extract or its hydrolysate, brown sugar extract or its ferment, Damask rose flower extract, ylang-ylang flower extract, bamboo shoot peel extract Linoleic acid and its derivatives or processed products (e.g., liposomal linoleic acid), collagen and its derivatives derived from animals or fish, elastin and its derivatives, proteoglycans derived from animals or fish, glycyrrhizic acid and its derivatives (e.g., dipotassium salt), t-cycloamino acid derivatives, vitamin A and its derivatives, allantoin, diisopropylamine dichloroacetate, γ-amino-β-hydroxybutyric acid, gentian extract, licorice extract, ginseng extract, Panax ginseng (white ginseng) extract, fermented Panax ginseng, camellia extract, Mitsuishikon Bu extract, loofah extract, sea lettuce extract, peach extract, plum extract, peach kernel extract, kiwi extract, sunflower extract, juazeiro extract, pau d'arco bark extract, daylily extract or ferment, hibiscus flower extract or ferment, rhododendron extract, cherimoya extract, mango extract, mangosteen extract, sea lettuce extract, oolong tea extract, red bell pepper extract, orchid extract, sweet potato extract, sansho pepper peel or seed coat extract or hydrolysate thereof, safflower extract, Casablanca lily extract, sweet potato extract or ferment thereof, guava leaf extract,These include Artemisia capillaris extract, clove extract, olive leaf extract, horse chestnut extract, ginger extract, vanilla extract, Houttuynia cordata extract, pomelo extract, aloe extract, aloe vera extract, tulip extract, centella asiatica extract, fig flower extract, apple extract, white asparagus extract, kudzu extract, Codonopsis pilosula extract, or their hydrolysates.

[0031] Next, the present invention will be described in more detail with reference to manufacturing examples, test examples, and embodiments, but the present invention is not limited thereto. In the following, all parts refer to parts by weight, and all percentages refer to weight percent.

[0032] Manufacturing Example 1. Preparation of Panax Ginseng Extract (1) Ten grams of ginseng root hairs were immersed in 40 grams of purified water and heated at 100°C to 120°C for 1 hour. The heated root hairs were extracted with 50% butylene glycol to obtain 86 grams of ginseng extract (solid content concentration 1.7%).

[0033] Manufacturing Example 2. Preparation of Panax Ginseng Extract (2) Ten grams of ginseng root hairs were immersed in 40 grams of purified water and heated at 100°C to 120°C for 2 hours. The heated root hairs were extracted with 50% butylene glycol to obtain 80 grams of ginseng extract (solid content concentration 2.0%).

[0034] Manufacturing Example 3. Preparation of Panax Ginseng Extract (3) Ten grams of ginseng root hairs were immersed in 40 grams of purified water and heated at 100°C to 120°C for 3 hours. The heated root hairs were extracted with 50% butylene glycol to obtain 90 grams of ginseng extract (solid content concentration 2.3%).

[0035] Manufacturing Example 4. Preparation of Panax Ginseng Extract (4) 100g of ginseng root hairs were immersed in 500g of purified water and heated at 100°C to 115°C for 2 hours. After heating, the root hairs were immersed in purified water to obtain 1000g of ginseng extract (solid content concentration 2.0%).

[0036] Manufacturing Example 5. Preparation of Panax Ginseng Extract (5) 20g of ginseng root hairs were immersed in 80g of purified water and heated at 100°C to 115°C for 4 hours. The heated root hairs were extracted with 75% butylene glycol to obtain 219g of ginseng extract (solid content concentration 1.8%).

[0037] Comparative manufacturing example 1. Preparation of Panax ginseng extract 10 g of ginseng root hairs were immersed in 40 g of purified water and extracted at 40°C to obtain 105 g of ginseng extract (solid content concentration 1.45%).

[0038] Test Example 1. Quantitative Test Method for Ginsenosides Rg3 and Rg5 10 μL of each of the preparations from Preparations 1-5 and Comparative Preparation 1 were taken and tested by liquid chromatography under the following test conditions. The peak areas of the resulting chromatograms were determined, and the concentrations of ginsenosides Rg3 and Rg5 were calculated from a pre-prepared calibration curve. (1) Test conditions Detector: UV absorbance spectrophotometer (measurement wavelength: 203 nm) Column: A stainless steel tube with an inner diameter of 4.6 mm and a length of 15 cm was used, packed with 5 μm octadecylsilylated silica gel for liquid chromatography. Column temperature: 30℃ Mobile phase A: Acetonitrile Mobile phase B: Water Mobile phase delivery: The concentration gradient was controlled by changing the mixing ratio of mobile phase A and mobile phase B as shown in Table 1. [Table 1] JPEG0007836035000001.jpg43153(2) Creation of a calibration curve First, 5 mg of ginsenoside Rg3 was weighed out and 90% acetonitrile aqueous solution was added to make a standard stock solution of 25 mL. Next, 2.5 mL and 5.0 mL of the ginsenoside Rg3 standard stock solution were accurately taken, and mobile phase was added to each to make exactly 10 mL, which was then used as the standard solution. Then, 10 μL of the standard stock solution and 10 μL of the standard solution were taken and tested by liquid chromatography under the aforementioned test conditions. A calibration curve was created by plotting the peak area of ​​the obtained chromatogram on the x-axis and the concentration of ginsenoside Rg3 (w / v ppm) on the y-axis. For Rg5, 5 mg was weighed out and 90% acetonitrile aqueous solution was added to make a standard stock solution of 25 mL. Next, 1.0 mL, 2.5 mL, and 5.0 mL of the ginsenoside Rg5 standard stock solution were accurately taken, and mobile phase was added to each to make exactly 10 mL, which was then used as the standard solution. Then, 10 μL of the standard solution was taken and tested by liquid chromatography under the aforementioned test conditions. A calibration curve was created by plotting the peak area of ​​the obtained chromatogram on the x-axis and the concentration of ginsenoside Rg5 (w / v ppm) on the y-axis.

[0039] Table 2 shows the measurement results of ginsenosides Rg3 and Rg5 contained in the extracts of Production Examples 1-5 and the extract of Comparative Production Example 1. [Table 2] JPEG0007836035000002.jpg55126

[0040] Test Example 2. Melanin Production Inhibition Evaluation Test (1) Culture of B16 cells B16 cells (mouse melanoma cells) were cultured in DMEM (Dulbecco's modified Eagle's medium (Sigma-Aldrich, St Louis, MO, USA)) containing 10% FBS (fetal bovine serum), 100 units / ml penicillin, and 100 μg / ml streptomycin (Wako, Osaka, Japan) at 5% CO2 and a temperature of 37°C. (2) Test solution treatment Using the same culture method as in (1) above, 5 × 10⁶ cells were cultured in a 6-well multiplate. 4B16 cells were seeded at a concentration of 1 cell / well. 24 hours later, a mixed solution of 3-isobutyl-1-methylxanthine (IBMX) as a melanin inducer, the test solution, and 2 mL of 10% FBS, 100 units / ml penicillin, and 100 μg / ml streptomycin-containing DMEM was prepared in each well, replacing the culture medium. The cells were then cultured at 5% CO2 and 37°C. The test solutions prepared were 1.0% (concentration as solution) of extracts from Production Examples 1-3 and their comparative control (extract from Comparative Production Example 1), and 0.5% (concentration as solution) of extracts from Production Examples 4 and 5 and their comparative control (extract from Comparative Production Example 1). Furthermore, a control (-IBMX) was set up when the culture medium was cultured without IBMX, and instead of the test solution, 0.75% or 0.5% 1,3-butylene glycol (BG) was added. In addition, a control (+IBMX) was set up when IBMX was added, and 0.75% or 0.5% BG was added instead of the test solution. (3) Measurement of melanin content After 72 hours of sample processing, the culture medium was removed, and each well was washed with 2 mL of PBS. Then, 50 μL of 1% TritonX-100 (SIGMA) and 250 μL of 1N NaOH were added in that order, mixed thoroughly, and collected in 1.5 mL tubes. The collected tubes were incubated at 98°C for 30 minutes to prepare cell lysates. The lysates were thoroughly mixed, and 80 μL / well was added to a 96-well multiplate. The absorbance at 405 nm was measured using a microplate reader (ARVO). TM X3). (4) Measurement of protein content The cell lysate prepared in (3) above was used with the Pierce BCA Protein Assay Kit (Thermo Scientific) according to the manual. The absorbance at 560 nm was measured using a microplate reader (ARVO). TM X3). (5) Evaluation of melanin production Melanin and protein levels were calculated using the method described above, and melanin production was evaluated from the melanin / protein ratio. The evaluation values ​​were calculated as relative values ​​with the control (-IBMX) set to 100.

[0041] The results of Test Example 2 are shown in Figures 1 and 2. As shown in Figures 1 and 2, it was confirmed that the extracts from Production Examples 1 to 5 exhibited significantly superior melanin production inhibitory effects compared to the extract from Comparative Production Example 1.

[0042] Test Example 3. Whitening evaluation test of ginsenosides Rg3, Rg5, Rb1, and Rg1. As test solutions, solutions of ginsenosides Rg3, Rg5, Rb1, and Rg1 were used instead of the production examples 1-5 and comparative production example 1 of Test Example 2. Dimethyl sulfoxide (DMSO) was used as the solvent to dissolve IBMX. A control (-IBMX) was set up when the culture medium was cultured with DMSO instead of ginsenosides, without adding IBMX. Furthermore, a control (+IBMX) was set up when the culture medium was cultured with IBMX and DMSO instead of the test solution. The final concentration of DMSO was adjusted to 0.1%.

[0043] The results of Test Example 3 are shown in Figures 3 and 4. As shown in Figure 3, Rg3 and Rg5 exerted a melanin production inhibitory effect at low concentrations, while as shown in Figure 4, Rb1 and Rg1 were confirmed to promote melanin production.

[0044] Test Example 4: Evaluation Test of Cell Damage Repair Caused by Ultraviolet Radiation (1)Cell (HaCaT) culture HaCaT cells (human keratinocyte cells) were cultured in DMEM (Dulbecco's modified Eagle's medium (Sigma-Aldrich, St Louis, MO, USA)) containing 5% FBS (fetal bovine serum), 100 units / mL penicillin, and 100 μg / mL streptomycin (Wako, Osaka, Japan) at 5% CO2 and a temperature of 37°C. (2) Test solution treatment Using the same culture method as in (1) above, 1 × 10⁶ cells were cultured in a 6-well multiplate. 5 HaCaT cells were seeded at a concentration of 1 cell / well. 24 hours later, a mixed solution of the test solution and 2 mL of 10% FBS, 100 units / ml penicillin, and 100 μg / mL streptomycin-containing DMEM was prepared and added to each well, replacing the culture medium. Cells were then cultured at 5% CO2 and 37°C. For the evaluation of cell viability repair, preparations 1-3 and their comparison (extract from comparative preparation 1) were prepared to a concentration of 0.5% (solution concentration). For the evaluation of intracellular glutathione production promotion, extracts from preparations 1, 2, and 5 and their comparison (extract from comparative preparation 1) were prepared to a concentration of 1.0% (solution concentration). A control was set where cells were cultured in a culture medium with 0.25% or 1.0% BG added instead of the test solution. (3) UV-B treatment (HaCaT cells) Five hours after treatment with the test solution, the samples were exposed to UV-B (60 mJ) using a UVB irradiator (TL20W / 12RS lamp, Dermaray 200; Muranaka). UV intensity was measured using an ultraviolet radiation sensor (UVX-31; TGK). After UV-B exposure, the samples were cultured at 5% CO2 and 37°C. A control group (UV-) that was not irradiated with UV-B was set up for comparison. (4) Cell viability evaluation (HaCaT cells) The procedure was performed using the Cell Counting Kit-8 (CCK8; Dojindo) according to the manual. The following modifications were made from the manual: After 24 hours of UV-B exposure, the culture medium was removed, and the CCK8 solution was added to each well. The mixture was incubated at 5% CO2 and 37°C for 1 hour. The CCK8 solution was collected in a 96-well plate, and the absorbance at 450 nm was measured using a microplate reader (ARVO). TM X3). The evaluation value was calculated as a relative value with the control (UV-) without UV-B irradiation set to 100. (5-1) Glutathione (GSH) measurement (HaCaT cells) After 24 hours of UV-B exposure, the culture medium was removed, and each well was washed with 2 mL of PBS. 300 μL of 0.05% Trypsin-EDTA (Gibco) was added, and the mixture was incubated at 5% CO2, 37°C, for 5 minutes. 300 μL of DMEM was added, and the mixture was collected in a 1.5 mL tube. 500 μL of PBS was then added to each well, and the mixture was collected in the same tube. Centrifuge (2000 rpm, 3 minutes) and the supernatant was removed. 1 mL of PBS was added, and the mixture was centrifuged (2000 rpm, 3 minutes) and the supernatant was removed. 250 μL of 0.5% Triton-X100 was added, and the mixture was thoroughly vortexed and sonicated. Centrifuged (13200 rpm, 10 minutes) and the supernatant was collected. The collected supernatant was subjected to GSSG / GSH Quantification using the Dojindo Quantification Kit according to the manual. The absorbance at 405 nm was measured using a microplate reader (ARVO TM X3). (5-2) Protein level measurement (HaCaT cells) The supernatant collected in the "Glutathione Measurement" described above was analyzed using the Pierce BCA Protein Assay Kit (Thermo Scientific) according to the manual. The absorbance at 560 nm was measured using a microplate reader (ARVO). TM X3). (5-3) Glutathione level evaluation (HaCaT cells) Glutathione (GSH) and protein levels were calculated using the method described above, and glutathione levels were evaluated from the glutathione / protein ratio. The evaluation values ​​were calculated as relative values ​​with the control group (UV-) without UV-B irradiation set to 100.

[0045] The results of Test Example 4 are shown in Figures 5 to 7. As shown in Figure 5, the extract according to the present invention was confirmed to exhibit a remarkable effect in repairing cells damaged by ultraviolet light compared to Comparative Production Example 1. Furthermore, as shown in Figures 6 and 7, the extract according to the present invention was also confirmed to have a remarkable effect in restoring the amount of intracellular glutathione that has decreased due to ultraviolet irradiation, compared to Comparative Production Example 1.

[0046] Test Example 5. Evaluation Test of Fibroblast Activation Effect Human dermal-derived fibroblasts NB1RGB were placed in 1 × 10⁶ wells of Eagle Minimum Essential Medium containing 0.5% NCS in a 96-well microplate. 4 Cells were seeded per well and pre-cultured for 1 day under conditions of 37°C and 5.0% CO2. Then, extracts from production examples 4 and 5 and comparative production example 1 were added to the culture medium as sample solutions to a concentration of 0.5% (as solution), and cultured for a further 3 days under the same conditions. Next, the medium was removed, 0.03% MTT was added, and the culture was held at 37°C for 1 hour. The resulting formazan was extracted with isopropanol, and the MTT value was measured at a wavelength of 570-630 nm using a microplate reader (Model 680, Bio-Rad). The same procedure was performed for the control case (no sample added) where PBS(-) was added instead of the sample solution. The relative value of the MTT value with each sample added to the MTT value obtained here was calculated and expressed as the fibroblast MTT activity rate (%). In addition, to confirm that the test system was functioning correctly, the same test was performed when 100 mM glucose was added as a positive control instead of the sample solution.

[0047] The results of Test Example 5 are shown in Figure 8. As shown in Figure 8, it was confirmed that the extract according to the present invention exhibits a significant fibroblast-activating effect compared to Comparative Production Example 1.

[0048] Example prescription 1. Lotion [Ingredients] Part Lavender oil 1.0 Glyceryl caprylate 3.0 Polyglyceryl-10 Laurate 3.0 Methylparaben 0.1 Extract from Production Example 1 2.0 White Ginseng Extract 2.0 Ginseng Fermented Extract 2.0 Sweet potato fermented extract 1.0 Guava leaf extract 1.0 Water-soluble collagen 1.0 Hydrolyzed collagen 1.0 Water eggplant extract 1.0 Strawberry flower extract 1.0 Glycerin 2.0 1,3-Butylene glycol 5.0 Sodium citrate 0.2 Purified water, in an amount that makes the total volume 100 parts.

[0049] Prescription example 2. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 2.0 parts of the extract from Production Example 2 were used instead of the extract from Production Example 1 contained in Formulation Example 1.

[0050] Prescription example 3. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 2.0 parts of the extract from Production Example 3 were used instead of the extract from Production Example 1 contained in Formulation Example 1.

[0051] Prescription example 4. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 2.0 parts of the extract from Production Example 4 were used instead of the extract from Production Example 1 contained in Formulation Example 1.

[0052] Prescription example 5. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 2.0 parts of the extract from Production Example 5 were used instead of the extract from Production Example 1 contained in Formulation Example 1.

[0053] Prescription example 6. Emulsion [Ingredients] Part Squalane 5.0 Hexaran 3.0 Rosehip oil 1.0 Camellia oil 1.5 Polyglyceryl laurate 1.0 Lipophilic glyceryl stearate 1.0 Bisabolol 1.0 Behenyl alcohol 3.0 Hydrogenated soy lecithin 1.5 Extract from manufacturing example 5 2.0 White Ginseng Extract 2.0 Ginseng Fermented Extract 2.0 L-ascorbic acid-2-glucoside 2.0 Sweet potato fermented extract 1.0 Alpinia zerumbet leaf extract 1.0 Guava leaf extract 1.0 Licorice extract 1.0 Codonopsis pilosula hydrolyzed extract 1.0 Water-soluble collagen 1.0 Hydrolyzed collagen 1.0 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, in an amount that makes the total volume 100 parts.

[0054] Prescription example 7. Cream [Ingredients] Part Olive oil 5.0 Jojoba oil 5.0 Meadowfoam oil 5.0 Squalane 5.0 Behenyl alcohol 2.0 Palmitic acid 2.5 Lactic acid fermented rice 2.0 Hydrogenated lecithin 0.5 Glyceryl stearylate 3.0 Extract from Production Example 1 2.0 White Ginseng Extract 2.0 Ginseng Fermented Extract 2.0 Sweet potato fermented extract 1.0 Alpinia zerumbet leaf extract 1.0 Guava leaf extract 1.0 Hydrolyzed rice extract 2.0 Water-soluble collagen 1.0 Hydrolyzed collagen 1.0 Xanthan gum 1.0 Purified water, in an amount that makes the total volume 100 parts.

[0055] Prescription example 8. Cream The cream was obtained in the same manner as in Formula Example 1, except that 2.0 parts of the extract from Production Example 2 were used instead of the extract from Production Example 1 included in Formula Example 7.

[0056] Prescription example 9. Cream The cream was obtained in the same manner as in Formulation Example 1, except that 2.0 parts of the extract from Production Example 3 were used instead of the extract from Production Example 1 included in Formulation Example 7.

[0057] Prescription example 10. Cream The cream was obtained in the same manner as in Formulation Example 11, except that 2.0 parts of the extract from Production Example 4 were used instead of the extract from Production Example 1 included in Formulation Example 7.

[0058] Prescription Example 11: Facial Cleanser [Ingredients] Part Coconut oil 2.0 Stearyl stearylate 2.0 Glyceryl caprylate 2.0 Extract from Production Example 1 2.0 White Ginseng Extract 2.0 Ginseng Fermented Extract 2.0 Water-soluble collagen 1.0 Glycerin 2.0 Fig bark extract 1.0 Job's tears seed extract 1.0 Clove extract 1.0

Claims

1. A melanin production inhibitor in pigment cells, containing a ginseng extract obtained from Panax ginseng (Panax CA Meyer), a plant belonging to the genus Panax in the family Araliaceae, and containing ginsenoside Rg3 and ginsenoside Rg5 as the active ingredient.

2. An inhibitor of the decrease in glutathione in epidermal cells caused by ultraviolet light, containing an extract of Panax ginseng (Panax CA Meyer), which belongs to the genus Panax in the family Araliaceae, and which contains ginsenoside Rg3 and ginsenoside Rg5, as the active ingredient.

3. A fibroblast activator containing a ginseng extract as an active ingredient, obtained from Panax ginseng (Panax CA Meyer), a plant belonging to the genus Panax in the family Araliaceae, and containing ginsenoside Rg3 and ginsenoside Rg5.

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