KR101238963B1 - Skin whitening composition comprising complex herbal extract - Google Patents

Abstract

The present invention relates to a composition containing a complex herbal extract as an active ingredient, specifically exhibits excellent tyrosinase inhibitory activity, and shows a melanin synthesis inhibitory activity in B16F10 melanoma cell experiments. It can be usefully used as an external skin pharmaceutical composition and cosmetic composition.

Description

Skin whitening composition containing complex herbal extract as active ingredient {Skin whitening composition comprising complex herbal extract}

The present invention relates to a composition for skin whitening containing a complex herbal extract as an active ingredient.

1, Ando, S., O. Ando, Y. Suemoto, and Mishima Y. 1993. Tyrosinase gene transcription and its control by melanogenic inhibitors. J Invest Dermatol 100, 150S-155S.

References 2 Aroca, P., K. Urabe, K. Kobayashi, K. Taskamoto, and V. J. Hearing. 1993. Melanin biosynthesis patterns of following hormonal stimulation. J. Biol Chem 268, 25650-25655.

[3] Carmichael, J., WG DeGraff, AF Gazdar, JD Minna, and JB Mitchell. 1987.Evaluation of a tetrazolium based semiautomated colorimetric assay: assessment of chemosen- sitivity testing. Cancer Res . 47 , 936-942.

[4] Chin, J. E., and N. C. Cho. 2005. Effects of Houttuynia cordata extracts on tyrosinse gene expression. J. Korean Soc Food Sci Nutr 34, 1284-1288. 7

[Reference 5] Choi, B.W., B. H. Lee, K. J. Kang, E. S. Lee and N. H. Lee. 1998. Screening of the tyrosinase inhibitors from marine algae and medicinal plants. Kor J Pharmacogn. 29. 237-242.

[Reference 6] Ge, R. Y., C. H. Zhou, and Y. C. She. 1983. Influences of stigma croci and semen persicae on function of ovary-uterus in pseudopregnant rats. J Tra Chinese Med 3, 23-26.

7 Hosoi, J., E. Abe, T. Suda and T. Kuroki. 1985.Regulation of melanin synthesis of B16 melanoma cells by 1 alpha, 25-dihydroxyvitamin D3 and retinoic acid. J. cancer. Res. 45, 1474-1478.

[Reference 8] Imokawa, G. and Y. Mishima. 1982. Loss of melanogenic properties in tyrosinase induced by glucosylation inhibitors within malignant melanoma cells. Cancer Res 42,1994-2002.

9 References Jimenez-Cervantes C., F. Solano, T. Kobayashi, K. Urabe, V. J. Hearing, J. Lozano, and C. Garcia-Borron. 1994. A new enzymatic function in the melanogenic pathway. J. Biol Chem 269, 17993-18001.

10. Kim, S. J., D. H. Kweon, and J. H. Lee. 2006. Investigation of antioxidative activity and stability of ethanol extracts of Licorice Root (Glychrrhiza glabra). Korean J Food Sci Technol 38, 584-588.

11 Kosuge, T., H. Ishida, and M. Ishii. 1985. Studies on active substances in the herbs used for oketsu in chinese medicine. Ⅱ. On the anticoagulative pronciple in persicae seme. J Chem Pham Bull 33, 1496-1498.

[12] Lee, P. J. 2009. Inhibitory effect of muscat bailey a seed extract on melanin production in α-MSH stimulated B16 cell. J. Kor Plant Res 22, 477-482.

13, Lee, Y. H., S. S. Park, S. W. Lee, S. H. Lee, K. H. Park, Y. J. Choi, and S. W. Gal. 2006.Whitening effect of mycelial cuture broth of paecilomyces japonica in the mixture of cucumber and grape extracts. J Life Science 16, 870-875.

14, Lin, C. B., L. Babiarz, F. Liebel, E. Roydon Price, M. Kizoulis, G. J. Gendi menico, D. E. Fisher, and M. Seiberg. 2002. Modulation of microphthal mia-associated transcription factor gene expression alters skin pigmentation. J Invest Dermatol 119, 1330-1340.

Reference 15 Michikawa, M., K.T. Lim, J. G. McLarnon and S. U. Kim. 1994. Oxygen radical-induced neurotoxicity in spinal cord neuron cultures. J. Neurosci Res. 37, 62-70.

[16] Mun, Y. J., J. Kim, N. Y. Lim, S. Y. Lee, S. Gwak, C. Y. Hwang, and W. H. Woo. 2002. Korean J Oriental Physiology & Pathology 16, 1230-1235.

17. Park, H. J., J. S. Lee, J. D. Lee, N. J. Kim, J. H. Pyo, J. M. Kang, I. H. Choe, S. Y. Kim, B. S. Shim, J. H. Lee, and Sabina Lim. 2005. The anti inflammatory effect of Cinnamomi Ramulus. J Korean Oriental Med 26, 140-151.

18. Park, S. M., G. W. Lee, and Y. H. Cho. 2008.Effects of Rheum undulatum extract on antioxidant activity and activity of matrix metalloproteinase-1 in human skin fibroblasts. J Life science 18, 1700-1704.

19 Paval, S. 1993. Dynamics of melanogenesis intermediates. J. Invest Dermatology 100, 162-165.

20. Sakamoto, S., H. Kudo, T. Kawasaki, K. Kuwa, N. Kasahara, S. Sassa, and R. Okamoto. 1988.Effects of a chinese herbal medicine, keishi-bukuryo-gan on the gonadal of rats. J Ethnophamacol 23, 151-158.

[21] Tomohiro, I. and F. Yukio. 2005 Hot water extracts from Adzuki Beans (vigna angularis) stimulate not only melanogensis in cultured mouse B16 melanoma cells but also pigmentation of hair color in C3H mice. BioSci. Biotechnol. Biochem. 69, 873-882.

22. Voegeli, R. 1996. Elastase and typtase determination on human skin surface. Cosmetic & Toiletries. 111, 51-58.

Modern people promote various skin aging phenomena such as blemishes, freckles and skin pigmentation due to various internal and external factors such as ultraviolet rays and stress (Voegeli, R. 1996. Elastase and typtase determination on human skin surface. Cosmetic & Toiletries. 111, 51-58.). Pigmentation of skin regulates the initial reaction of melanin from biosynthesis to tyrosinase enzyme and L-tyrosine such as DHICA oxidase (TRP-1) with DOPA (3,4-dihydroxyphenyla-lanine) from DOPA to DOPA quinone. (Aroca, P., et al. 1993. Melanin biosynthesis patterns of following hormonal stimulation. J. Biol Chem 268, 25650-25655 .; Jimenez-Cervantes C., et al. 1994. A new enzymatic function in the melanogenic pathway.J. Biol Chem 269, 17993-18001 .; Paval, S. 1993. Dynamics of melanogenesis intermediates.J. Invest Dermatology 100, 162-165.). Based on this, studies on natural products that may affect the inhibition of melanin biosynthesis by inhibiting the activity of tyrosinase enzymes are being actively conducted. As a result, the inhibitory effect of tyrosinase gene expression using Echochocho extract (Chin, JE, et al. 2005. Effects of Houttuynia cordata extracts on tyrosinse gene expression.J. Korean Soc Food Sci Nutr 34, 1284-1288.), Lee, PJ 2009. Inhibitory effect of muscat bailey a seed extract on melanin production in α-MSH stimulated B16 cell.J. Kor Plant Res 22, 477-482.) Research using natural products is being actively conducted. In addition, currently known antioxidants and whitening materials are representatives such as arbutin, Kojic acid, ascorbic acid, and plant extracts such as baekbaekpi, mulberry, licorice.

It is a complex prescription of natural medicines and consists of five kinds of Chinese herbal medicines such as rhubarb, gyeji, licorice, doin, and mango. Rhubarb is a perennial herbaceous plant belonging to the Medicinalaceae, which is used as a medicinal herb. Rhubarb is an external medicine used for burn treatment and dermatological diseases. It has excellent concentration-dependent antioxidant activity and has been reported to inhibit MMP-1 activity (Park, SM, et al. 2008. Effects of Rheum undulatum extract on antioxidant activity and activity of matrix metalloproteinase-1 in human skin fibrob). Cinnamon is dried sprigs of cinnamon tree, and contains chemicals such as cinnamaldehyde, coumarin, β-sitosterol, and protocatechuic acid. This addition has been reported to inhibit the production of reactive substances of NO and PGE ₂ produced by the concentration and to reduce gene expression such as iNOS, COX-2 [Park, HJ, et al. 2005. The anti inflammatory effect of Cinnamomi Ramulus. J Korean Oriental Med 26, 140-151.]. Peach Seed is a peach seed, a natural antioxidant, anti-aging and anti-cancer agent, which has been used for a long time in women's diseases and degenerative diseases, and has been reported for anti tumor promoter and anti-blood activity (Ge, RY, et al. 1983. Influences of stigma croci and semen persicae on function of ovary-uterus in pseudopregnant rats.J Tra Chinese Med 3, 23-26 .; Sakamoto, S., et al. 1988. Effects of a chinese herbal medicine, keishi-bukuryo-gan on the gonadal of rats.J Ethnophamacol 23, 151-158 .; Kosuge, T., et al. 1985. Studies on active substances in the herbs used for oketsu in chinese medicine.II.On the anticoagulative pronciple in persicae seme.J Chem Pham Bull 33, 1496-1498.). Licorice has a similar effect to steroid hormones, which are anti-inflammatory, detoxifying and antipyretic. Antioxidant effect (Kim, SJ, et al. 2006. Investigation of antioxidative activity and stability of ethanol extracts of Licorice Root (Glychrrhiza glabra) .Korean J Food Sci Technol 38, 584-588.) . It is an anti-irritant and has an effect of inhibiting melanin synthesis by acting as a tyrosinase inhibitor as well as a pigment removing effect (Mun, YJ, et al. 2002. Korean J Oriental Physiology & Pathology 16, 1230-1235.). It is known to be superior to kojic acid. Natrii sulfas are associated with stronger peristalsis by acting in the body and mechanically stimulating the mucous membranes in the digestive tract.

This study was conducted to study the expression of proteins such as tyrosinase and TRP-1, which are involved in melanin content and melanin biosynthesis, using B16F10 melanoma cells in order to utilize complex herbal extracts as whitening cosmetics based on the activity of constituents. It was.

The present inventors completed the present invention by confirming that the complex herbal extract has excellent tyrosinase inhibitory activity.

In order to achieve the above object, the present invention provides a whitening skin external pharmaceutical composition containing a complex herbal extract as an active ingredient.

The present invention also provides a cosmetic composition for whitening containing a complex herbal extract as an active ingredient.

Complex herbal extracts defined herein have a dry mixed weight ratio (%) of rhubarb, gyeji, licorice, cabbage and manganese 1 to 9: 1 to 6: 1 to 6: 1 to 30: 1 to 30, preferably 1 to 6: 1-4: 1-4: 1-20: 1-20.

As used herein, an “extract” is one or more selected from the group consisting of water, ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, butylene glycol, propylene glycol, hydrobutylene glycol, hydrous propylene glycol, hydrous glycerin Solvents, preferably water and ethanol, most preferably 60% to 80% ethanol soluble extract.

The complex herbal extract is characterized in that it contains 0.1 to 50% by weight based on the total weight of the skin external pharmaceutical composition.

The pharmaceutical composition comprises a cream, gel, patch, spray, ointment, warning, lotion, linen, pasta or cataplasma formulation.

In addition, the cosmetic composition includes a lotion, skin, lotion, nutrition lotion, nutrition cream, massage cream, essence, the formulation of the pack.

Hereinafter, the present invention will be described in detail.

The herbal extract of the present invention can be obtained as follows.

The mixed weight ratio (%) of rhubarb, cinnamon, licorice, cabbage and mango of the present invention is 1-9: 1-6: 1-6: 1-30: 1-30, preferably 1-6: 1-4: 1 to 4: 1 to 20: the first step of mixing in 1 to 20; Water, ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, butylene glycol, propylene glycol, hydrous butylene glycol in volumes of about 1 to 30 times, preferably about 1 to 15 times the weight of the mixture , At least one solvent selected from the group consisting of hydrous propylene glycol and hydrous glycerin, preferably water and ethanol, preferably water, about 10 ° C. to 100 ° C., preferably about 30 ° C. to 90 ° C., for about 1 hour to 5 hours. A second step of extracting preferably by reflux cooling using an extraction method such as hot water extraction, reflux cooling extraction, immersion extraction or pressure extraction for a time; The extract of the present invention may be obtained through the preparation method comprising the third step of centrifugation, vacuum filtration, concentration, and lyophilization of the extract 1 to 5 times.

In addition, rhubarb, cinnamon, forget-me-not, licorice, cabbage is a herb that has been used for a long time as a herbal medicine and edible, the herbal extract of the present invention extracted from them also has no problems such as toxicity and side effects, it is a non-irritant sample in skin patch test Proven, you can use it with confidence even for long time use.

The external skin pharmaceutical composition containing the complex herbal extract of the present invention is prepared as a pharmaceutical composition in the form of an external skin preparation of cream, gel, patch, spray, ointment, warning, lotion, linen, pasta or cataplasma. Can be used, but is not limited thereto.

The preferred dosage of the complex herbal extract of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the preferred effect, the composite herbal extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 10 mg / kg. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

Complex herbal extract of the present invention can be used in a variety of cosmetics and face wash having a whitening effect.

Products to which the composition can be added include, for example, cosmetics such as lotion, skin, lotion, nourishing lotion, nourishing cream, massage cream, essence, pack, etc., and cleansing, face wash, soap, treatment, essence, etc. have.

Cosmetics of the present invention comprises a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids and seaweed extract.

The water-soluble vitamins may be any compound that can be incorporated into cosmetics, but preferably vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H, and the like. Their salts (thiamine hydrochloride, sodium ascorbate salt, etc.) and derivatives (ascorbic acid-2-sodium phosphate salt, ascorbic acid-2-magnesium phosphate salt, etc.) may also be used in the water-soluble vitamins that can be used in the present invention. Included. The water-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification from microorganism culture, enzyme or chemical synthesis.

The oil-soluble vitamin may be any compound that can be incorporated into cosmetics, but preferably vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol), and the like. And derivatives thereof (ascorbic palmitate, ascorbic stearate, ascorbic acid dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherolvitamin E, dl-pantothenyl alcohol, D-pantothenyl alcohol, pantotenylethyl Ethers, etc.) are also included in the oil-soluble vitamins used in the present invention. Oil-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification of microorganism culture, enzyme or chemical synthesis.

The polymeric peptide may be any compound as long as it can be compounded in cosmetics, and examples thereof include collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, and keratin. Polymeric peptides can be purified and obtained by conventional methods such as purification from microbial cultures, enzymatic methods or chemical synthesis methods, or can be purified and used from natural products such as dermis and pig silk such as pigs and cattle.

The polymer polysaccharide may be any compound as long as it can be blended into cosmetics. Preferably, hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof (sodium salt, etc.) may be mentioned. For example, chondroitin sulfate or its salt, etc. can be normally purified from a mammal or fish.

The sphingolipid may be any compound as long as it can be blended into cosmetics. Preferably, ceramide, phytosphingosine, sphingosaccharide lipid, etc. may be mentioned. Sphingo lipids can usually be purified from mammals, fish, shellfish, yeasts or plants by conventional methods or obtained by chemical synthesis.

The seaweed extract may be any compound as long as it can be blended into cosmetics. Preferably, the seaweed extract may include brown algae extract, red algae extract, green algae extract, and the like. Also, calginine, arginic acid, sodium arginate, Potassium arginate and the like are also included in the seaweed extract used in the present invention. Seaweed extract can be obtained by purification from seaweed by conventional methods.

The cosmetic of the present invention may be blended with other essential ingredients, if necessary, in combination with the essential ingredients.

Other components that may be added include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, Blood circulation accelerators, cooling agents, restriction agents, purified water and the like.

Examples of the fat or oil component include ester fats, hydrocarbon fats, silicone fats, fluorine fats, animal fats, and vegetable fats and oils.

As ester fats and oils, glyceryl tri2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl mystinate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl acid isocetyl, isostyl acid isostearyl, isostaryl palmitate, octylate acid octyldodecyl, isostearic acid isetyl, diethyl sebacate, adipine Acid isopropyl, isoalkyl neopentane, tri (capryl, capric acid) glyceryl, trimethyl ethyl trimethylol propane, triisostearic acid trimethylol propane, tetra 2-ethylhexanoic penta erythritol Cetyl caprylate, lauric acid decyl, hexyl laurate, decyl myristin, myristin acid myristyl, myritic acid cetyl, stearyl stearate, decyl oleate, rininooleic acid , Isostearyl laurate, isotridecyl myristin, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecate oleate, octylate decyl oleate, octyl dodecyl linoleate, isopropyl isopropyl acid, 2 -Cetostearyl ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicapric acid, propylene glycol dicacapate, capric acid Propylene glycol, dicapric acid neopentyl glycol, dioctanoate neopentyl glycol, tricaprylic acid glyceryl, triundecyl glyceryl, triisopalmitinate glyceryl, triisostearate glyceryl, neopentane dodecyl Isostearyl octanoate, octyl isononate, hexyl decyl neodecanoate, octyl dodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, Sothete octylate, polyglycerol oleate, polyglycerine isostearate, triisocetyl citrate, triisoalkyl citrate, triisoctyl citrate, lauryl lactate, myritic lactate, octyl lactate, octyl lactate, tricitric acid Ethyl, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malic acid, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipicate, diisopropyl sebacinate, Dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, physteryl isostearate, phytic oleate, 12-Steloylhydroxystearate isocetyl, 12-Steloylhydroxystearate stearyl, 12-stealo Esters such as monohydroxystearic acid isostearyl; and the like.

Hydrocarbon-based fats and oils, such as squalene, a liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, a liquid isoparaffin, polybutene, microcrystal wax, and a vaseline, etc. are mentioned as a hydrocarbon-type fats and oils.

Examples of the silicone-based oils and fats include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane, methylcetyloxysiloxane copolymer, dimethylsiloxane and methylsteoxysiloxane copolymer, and alkyl. Modified silicone oil, amino modified silicone oil and the like.

Examples of the fluorine-based oil include perfluoropolyether and the like.

Animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, soybean oil, soybean oil, corn oil, rapeseed oil, almond oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil. , Cottonseed oil, Palm oil, Cucumber nut oil, Wheat germ oil, Rice germ oil, Shea butter, Walnut colostrum oil, Marker demia nut oil, Meadow home oil, Egg yolk oil, Uji, Horse oil, Mink oil, Orange rape oil, Jojoba oil And animal or plant fats and oils such as candeler wax, carnava wax, liquid lanolin and hardened castor oil.

Examples of the moisturizing agent include a water-soluble low molecular moisturizer, a fat-soluble molecular moisturizer, a water-soluble polymer, and a fat-soluble polymer.

Water-soluble low molecular humectants include serine, glutamine, sorbitol, mannitol, pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or more), polypropylene Glycol (polymerization degree n = 2 or more), polyglycerol B (polymerization degree n = 2 or more), lactic acid, lactic acid salt, etc. are mentioned.

Examples of the fat-soluble low molecular humectants include cholesterol and cholesterol esters.

Examples of the water-soluble polymer include carboxyvinyl polymer, polyasparaginate, tragacanth, xanthan gum, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, water soluble chitin, chitosan, and dextrin. Can be.

Examples of the fat-soluble polymers include polyvinylpyrrolidone-eicosene copolymers, polyvinylpyrrolidone-hexadecene copolymers, nitrocellulose, dextrin fatty acid esters, polymer silicones, and the like.

Examples of the emollient include long-chain acyl glutamic acid cholesteryl esters, hydroxy stearic acid cholesterol, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl esters, and the like.

As surfactant, nonionic surfactant, anionic surfactant, cationic surfactant, amphoteric surfactant, etc. are mentioned.

As the nonionic surfactant, self-emulsifying glycerin monostearate, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerol fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE Glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hardened castor oil, POE castor oil, POE / POP (polyoxyethylene / polyoxypropylene) copolymer, POE / POP alkyl ether, polyether modified silicone, lauric acid Alkanolamide, alkylamine oxide, hydrogenated soybean phospholipid, etc. are mentioned.

Anionic surfactants include fatty acid soaps, alpha-acyl sulfonates, alkyl sulfonates, alkyl allyl sulfonates, alkyl naphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkylphosphates, and alkylamides. Phosphates, alkyloylalkyltaurine salts, N-acylamino acid salts, POE alkyl ether carboxylate salts, alkyl sulfosuccinate salts, sodium alkyl sulfo acetates, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, and the like. have.

As cationic surfactant, alkyl trimethylammonium chloride, stearyl trimethyl ammonium chloride, stearyl trimethyl ammonium chloride, cetostearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, stearyl dimethyl benzyl ammonium bromide, behenyl trimethyl ammonium chloride, chloride Benzalkonium, diethylaminoethyl stearate, dimethylaminopropyl stearate, lanolin derivatives, quaternary ammonium salts, and the like.

Examples of amphoteric surfactants include the carboxybetaine type, the amide betain type, the sulfobetain type, the hydroxysulfobetain type, the amide sulfobetain type, the phosphobetaine type, the aminocarboxylate type, the imidazoline derivative type, and the amideamine type. An amphoteric surfactant etc. are mentioned.

Examples of the organic and inorganic pigments include inorganic pigments such as silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, Bengala, clay, bentonite, titanium mica, titanium oxide, bismuth chloride, zirconium oxide, magnesium oxide, Inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, chromium oxide, chromium oxide, chromium hydroxide, Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene, styrene copolymer, Organic pigments such as silk powder, cellulose, CI pigment yellow, CI pigment orange, and composite pigments of these inorganic pigments and organic pigments;

As organic powder, Metal soaps, such as a calcium stearate; Alkyl phosphate metal salts such as sodium cetylinate, zinc lauryl acid and calcium laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc, and N-lauroylglycine calcium; Amide sulfonic acid polyvalent metal salts, such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N-epsilon-lauroyl-L-lysine, N-epsilon-palmitolyzine, N-alpha-paratoylol nitin, N-alpha-lauroyl arginine, N-alpha-cured fatty acid acyl arginine -Acyl basic amino acid; N-acyl polypeptides, such as N-lauroyl glycyl glycine; Alpha-amino fatty acids such as alpha-aminocaprylic acid and alpha-aminolauric acid; Polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene-styrene copolymer, ethylene tetrafluoride and the like.

Examples of the ultraviolet absorber include paraaminobenzoic acid, ethyl paraaminobenzoate, amyl paraaminobenzoic acid, octyl paraaminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylic acid and benzyl cinnamic acid. , Paramethoxy cinnamic acid-2-ethoxyethyl, paramethoxy cinnamic acid octyl, diparamethoxy cinnamic acid mono-2-ethylhexaneglyceryl, paramethoxy cinnamic acid isopropyl, diisopropyl diisopropyl cinnamic acid ester mixture, wuro Canonic acid, ethyl urokanoate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and salts thereof, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone sodium sulfonate, dihydroxybenzophenone , Tetrahydroxybenzophenone, 4- tert -butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino- p- (carbo-2'-ethylhexyl-1'-oxy) -1 , 3,5-triazine, 2- (2-hi And the like can be mentioned hydroxy-5-methylphenyl) benzotriazole.

Examples of the disinfectant include hinokitiol, trichloroacid, trichlorohydroxydiphenyl ether, crohexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zinc filitione, benzalkonium chloride, No. 301, mononitro and eicol sodium, and undecylenic acid.

Examples of the antioxidant include butylhydroxyanisole, propyl gallic acid, and erythorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fmaric acid, sodium pmarate, succinic acid, sodium succinate, sodium hydroxide, sodium dihydrogen phosphate, and the like.

Examples of the alcohol include higher alcohols such as cetyl alcohol.

In addition, the compounding component which may be added other than this is not limited to this, Moreover, Although all said components can be mix | blended within the range which does not impair the objective and effect of this invention, Preferably it is 0.01-5% weight with respect to gross weight, More Preferably 0.01-3% by weight.

The cosmetic of the present invention may take the form of a solution, an emulsion, a viscous mixture or the like.

Components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the complex herbal extract as an active ingredient, for example, such as stabilizers, solubilizers, vitamins, pigments and flavorings Phosphorus aids and carriers.

The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, and includes, for example, milky lotion, cream, lotion, pack, foundation, lotion, essence, hair cosmetic, and the like.

Specifically, the cosmetic composition of the present invention can be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutrition cream, a moisturizing cream, a hand cream, Packs, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleansers.

When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

Complex herbal extract of the present invention shows excellent tyrosinase inhibitory activity and shows excellent melanin synthesis inhibitory activity in experiments using B16F10 melanoma cells can be usefully used as a composition for skin whitening.

1 is a diagram showing the cell viability of B16F10 melanoma cells treated with the extract of the herbal medicine,
2 is a diagram showing the melanin production inhibitory effect of the complex herbal extracts,
Figure 3 is a diagram showing the effect of inhibiting tyrosinase activity according to the addition of the compound herbal extract,
Figure 4 is a diagram showing the expression of tyrosinase protein and TRP-1 after treatment with 70% ethanol extract of the compound,
Figure 5 is a diagram showing the expression of tyrosinase protein and TRP-1 after treatment with the herbal extract extract.

Below, The invention is illustrated in detail by the following examples and experimental examples.

However, the following examples and experimental examples are illustrative of the present invention, and the content of the present invention is not limited by the following examples and experimental examples.

Example 1 Preparation of a Complex Herbal Water Extract

Ingredients necessary for the preparation of the complex herbal extracts were purchased from Human Hub Co., Ltd. Samples were prepared by mixing 30g rhubarb, 20g gage, 20g nettle, 100g licorice and 100g phosphorus, adding 10 times the amount of distilled water, extracting by reflux cooling at 85 ° C for 3 hours, separating supernatant and sediment three times, and extracting them repeatedly. 51.12 g of herbal extract (hereinafter referred to as “DO-W”) was obtained, filtered through Whatman No. 1 filter paper, concentrated, freeze-dried and stored in a refrigerator to use as a sample of this experiment.

Example 2. Preparation of Complex Herbal Ethanol Extract

Constituents necessary for the preparation of the compound herbal ethanol extract were purchased from Human Hub Co., Ltd. Samples were prepared by mixing 30g rhubarb, 20g gage, 20g nettle, 100g licorice and 100g phosphorus, and immersion extraction at room temperature for 24 hours by adding 70% ethanol in 10 times and separating supernatant and precipitate three times. After repeated extraction, 19.58 g of a compound herbal 70% ethanol extract (hereinafter referred to as “DO-EtOH”) was obtained, filtered through Whatman No. 1 filter paper, concentrated, lyophilized, and stored as a sample in the refrigerator. .

Reference Example 1. Cell Culture

B16F10 melanoma cells according to the method of Michikawa (Michikawa, M., KT Lim, JG McLarnon and SU Kim. 1994. Oxygen radical-induced neurotoxicity in spinal cord neuron cultures. J. Neurosci Res. 37, 62-70.) After diluting the 0.25% trypsin solution in the cultured cells, the cells were isolated, and then, in Dulbeco's modified eagle's medium (DMEM) medium, 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (penicillin / streptomycin) ( 100U / ml) was added and incubated in a 37 ° C., 5% CO ₂ incubator.

Experimental Example 1. Measurement of cell viability by MTT assay

Cell viability was measured according to a method such as Carmichael (Carmichael, J., DeGraff WG, Gazdar AF, Minna JD, Mitchell JB and 1987. Evaluation of a tetrazolium based semiautomated colorimetric assay:.. Assessment of chemosen- sitivity testing Cancer Res . 47 , 936-942.). Melanoma (B16F10) was dispensed 0.18ml in a 96 well plate at 0.6 ~ 0.8 × 10 ³ cells / well. Samples were prepared for each concentration, added 0.02ml, and incubated in 37 ° C. and 5% CO ₂ incubator for 24 hours. In the control group, the same amount of distilled water as that of the sample was added and the cells were cultured under the same conditions. After incubation for 4 hours with the addition of 0.02ml of MTT solution prepared at 5mg / ml concentration, the culture medium was removed, and 0.15ml of DMSO: EtOH (1: 1) was added to each well, followed by reaction at room temperature for 30 minutes, followed by ELISA reader. Absorbance was measured at 550 nm. Cytotoxicity measurement was expressed as the absorbance reduction rate of the sample solution addition group and no addition group, as shown in Equation 1 below.

In order to investigate the effects of the combined herbal extracts on the proliferation of B16F10 melanoma cells, the mixed herbal 70% ethanol extract and water extract were treated with 10, 25, 50, 75, 100, 250 μg / ml concentration and 48 hours after The proliferation of the cells was observed. As a result, toxicity was not observed at concentrations of 250 μg / ml or less when the cell proliferation rate of the control group was 100% (see FIG. 1).

Experimental Example 2 melanin measurement

The amount of melanin was modified by Hosoi et al. (Hosoi, J., et al. 1985. Regulation of melanin synthesis of B16 melanoma cells by 1 alpha, 25-dihydroxyvitamin D3 and retinoic acid.J. cancer.Res. 45, 1474-1478.). Cells were cultured, washed twice with PBS, and then centrifuged to form cell precipitates. 150 μl of a 1N NaOH solution added with 10% DMSO was added and dissolved for 1 hour at 60 ° C., and the absorbance was measured at 405 nm. The melanin amount was obtained from a standard straight line prepared using synthetic melanin, and the melanin amount of the experimental group was expressed as a percentage of the melanin amount of the control group.

As a result of measuring the melanin production inhibitory effect of the complex herbal extract (see Fig. 2) it could be observed that the concentration-dependent inhibition. In addition, it was observed that the experimental group treated with α-MSH and the sample at the same time was inhibited compared to the control treated with α-MSH alone. This suggests that the combination drug acts on melanin biosynthetic signaling pathway induced by α-MSH. In addition, 70% ethanol extract was superior to water extract. As with the results of Tyrosinase activity measurement, it was observed that it plays a role in inhibiting melanin production in a concentration-dependent manner. It was confirmed that the compound herbal extract is an excellent material as a natural whitening cosmetic material by showing superior activity than Arbutin, which is used as a whitening ingredient in whitening cosmetics.

Experimental Example 3. Measurement of Tyrosinase Activity

Tyrosinase activity was measured by a modification of Choi5 et al. (Choi, B.W., et al. 1998. Screening of the tyrosinase inhibitors from marine algae and medicinal plants.Kor J Pharmacogn. 29. 237-242.). Lysis buffer (1% triton X-100, 0.1M Sodium phosphate buffer, 50mM PMSF, pH 6.8) was added to the cells of each well. After the cells were destroyed on ice and centrifuged, only the supernatant was collected and used as an enzyme solution. Prepare the substrate by dissolving L-DOPA in 0.1M Sodium phosphate buffer (pH 6.8) at 2 mg / ml concentration, add 40 μl of enzyme solution to 160 μl of substrate, warm at 37 ℃ for 1 hour, and produce 490nm. Measured at The inhibition rate is expressed as in Equation 2 below.

As a result of measuring tyrosinase activity according to the addition of the complex herbal extract, it was confirmed that tyrosinase activity was inhibited as the treatment concentration of the sample was increased (see FIG. 3). Inhibition of 57% activity was observed at 100 μg / ml of the combined herbal 70% ethanol extract, but the water extract did not show good activity at the same concentration as the 70% ethanol extract. Lee, YH, et al. 2006.Whitening effect of mycelial cuture broth of paecilomyces japonica in the mixture of cucumber and grape extracts. J Life Science 16, 870-875. According to the results of the study, it can be seen that the inhibitory activity of tyrosinase activity of the 70% ethanol extract of the compound herbal 70% thyrosinase activity when the addition of 10% of the sample is inhibited. The action of tyrosinase is an enzyme that causes melanin production in melanosomes, which can cause problems such as skin aging and pigmentation (Tomohiro, I. et al. 2005 Hot water extracts from Adzuki Beans ( vigna angularis) stimulate not only melanogensis in cultured mouse B16 melanoma cells but also pigmentation of hair color in C3H mice.BioSci. Biotechnol. Biochem. 69, 873-882.). Therefore, the herbal extracts may be used as whitening cosmetics by inhibiting tyrosinase activity.

Experimental Example  4. Western blot Expression of whitening-related proteins

B16F10 melanoma cells were aliquoted to 2 × 10 ⁴ cells / well in a 60 mm tissue culture dish and incubated for 24 hours. After removing the medium, the complex herbal extracts were replaced with the concentration-treated medium, incubated for 48 hours, and washed with PBS. Lysis buffer 100ul was added to lyse the cells and centrifuged (12,000rpm, 4 ℃, 20min) to remove the cell membrane components. The protein obtained by centrifugation was quantified by bradford assay, 60ul of protein was electrophoresed using 10% SDS-PAGE, and then transferred to PVDF membrane to inhibit the nonspecific binding of the antibody and then at 30 ~ 40V for 2 hours or more. transferred. After transfer, soak in ponceau, check band, wash twice with TBST, take out, overnight with 5% skim milk and remove background. After washing three times, the primary antibody (1: 1000) was attached for 1 hour, and then the secondary antibody (1: 1000) was attached again, and transferred to a film using ECLkit (Milipore, Germany). Band density was confirmed by Gel doc (Amersham Pharmacia, England).

Tyrosinase activity and melanin biosynthesis of the multi-drug extracts were confirmed to be excellent materials for inhibition and analysis of the expression patterns of melanin biosynthesis-related proteins. 70-% ethanol extract of the multi-drugs on B16F10 melanoma cells (see Figure 4), water extract (See FIG. 5) After treating 50 and 75 100 μg / ml, 48 hours later, tyrosinase protein and TRP-1 expression were confirmed by wastern blotting. At this time, β-actin, a house keeping gene, which has little difference in expression level even under various conditions of cells, was used as a positive control. Thus, the herbal extracts were observed to inhibit the expression of tyrosinase (Tyrosinase) and TRP-1 (DHICA Oxidase) in a concentration-dependent manner. In the case of the 70% ethanol extract, the tyrosinase protein showed 50% inhibitory effect at 100 μg / ml. TRP-1 was confirmed that little inhibition compared to the control. In the case of water extract of the combined herbal, tyrosinase inhibition was excellent at 100 μg / ml and TRP-1 inhibition was better than 70% ethanol extract. Inhibition of expression of tyrosinase and TRP-1 inhibits the expression of tyrosinase and TRP-1, enzymes directly involved in melanin biosynthesis by inhibiting continuous ERK activation in the cell signaling pathway by α-MSH. It is considered to be.

Hereinafter, the formulation of the present invention is exemplified as a cream, massage cream, lotion, skin lotion, essence, packs, cleansing foam formulation, but is not limited to the formulation comprising the cosmetic composition of the present invention.

Formulation Example  1. Cream composition

The oil phase and the water phase are heated and mixed at 75 ° C., respectively, and then cooled to room temperature.

Formulation Example  2. Massage cream  Composition

The oil phase and the water phase are each dissolved by heating at 75 ° C., and then cooled to room temperature.

Formulation Example  3. Lotion Composition

The oil phase and the water phase are heated, mixed and emulsified at 75 ° C., respectively, and cooled to room temperature.

Formulation Example  4. Skin Lotion Composition

The aqueous phase and the ethanol phase are each prepared, mixed and filtered.

Formulation Example  5. Essence Composition

The aqueous phase and the ethanol phase are each prepared, mixed and filtered.

Formulation Example  6. Pack Composition

The aqueous and ethanol phases are dispersed, dissolved and mixed, and then cooled to room temperature.

Formulation Example  7. Cleansing Foam  Composition

The aqueous and oil phases are dispersed, dissolved and mixed, and then cooled and cooled to room temperature.

Claims (7)

delete delete delete delete delete The dry and mixed weight ratio (%) of rhubarb, gyeji, licorice, cabbage and mango are 1 ~ 9: 1 ~ 6: 1 ~ 6: 1 ~ 30: 1 ~ 30 water and ethanol solvent extract as an active ingredient Cosmetic composition for whitening containing. 7. The cosmetic composition according to claim 6, wherein the cosmetic composition is in the form of a lotion, skin, lotion, nutrition lotion, nutrition cream, massage cream, essence, pack.

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