KR20170020066A - Topical composition comprising the isolated compounds from Rubus coreanus seed showing skin whitening effect - Google Patents

Abstract

 The present invention relates to a composition containing phytic acid or a suavissimoside Fl compound as an active ingredient isolated from a bokbunja seed extract, and the compounds of the present invention are useful as a whitening agent, anti-wrinkle agent, anti-inflammatory agent And a cosmetic composition of the present invention.

Description

[0001] The present invention relates to a composition for external use containing a bokbunja seed separation compound as an active ingredient,

The present invention relates to an external-use composition containing a bramblycem splitting compound as an active ingredient.

[1] Voegeli, R. 1996. Elastase and typing determination on human skin surface. Cosmetic & Toiletries. 111, 51-58.

[Literature 2] Aroca, P., et al. 1993. Melanin biosynthesis patterns of following hormonal stimulation. J. Biol Chem 268, 25650-25655 .;

[Literature 3] Jimenez-Cervantes C., et al. 1994. A new enzymatic function in the melanogenic pathway. J. Biol Chem 269, 17993-18001 .;

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

[Literature 5] Chin, J. E., et al. 2005. Effects of Houttuynia cordata extracts on tyrosinase gene expression. J. Korean Soc Food Sci Nutr 34, 1284-1288.)

[Literature 6] 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.

[Literature 7] Kim Dae-Ho, Park Jin-Hong, Kim Jung-hwa, Kim Cheol-Hee, Yoo Jin-hyun, Kwon Min-cheol, Lee Hyun-yong,

[8] Kim Se-Hyun, Jung Heon Jung, Yong-Seok Chang, Young-Ki Park, and Sun Chang Chang, Exploring the Fruit Characteristics and Antioxidant Activity of the Berries Strawberry Clones, Journal of Korean Society of Agricultural Science 94 (1): 11-15, 2005

(Takshi Seto, Takashi Tanaka, Osamu Tanaka and Naohiro Naruhashi, 1984. β-Glucosyl Esters of 19 α-hydroxyursolic acid derivatives in leaves of Rubus species. Phytochemistry 23 (12), 2829-2834)

[10] Andrew T. Bruman, Ginger M. chateauneuf, Brian R. Boyd, Richard E. Brown, Pushpalatha P. N. Murthy 1999. Conformational inversion processes in phytic acid: NMR spectroscopic and molecular modeling studies. Tetrahedron Letters 40: 4489-4492),

[Literature 11] Blois, M.S. 1958. Antioxidant determinations by the use of a stable free radical, Nature, 181: 1199-1200.)

[Literature 12] Aoshima H, Tsunoue H, Koda H, Kiso Y. Aging of whiskey increases 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. J. Agric. Food Chem. 2004; 52 (16): 5240-5244;

[Document 13] Kim HK, Kim YE, Do JR, Lee YC, Lee BY. Antioxidative activity and physiological activity of some Korean medicinal plants. Korean J. Food Sci. Technol. 1995; 27 (1): 80-85.

[14] Kim, K.W., Ha, K.T., Park, C.S., Jin, U.H., Chang, H.W., Lee, C.S., Kim, C.H. Polygonum cuspidatum, compared with baicalin and berberine, inhibits inducible nitric oxide synthase and cyclooxygenase-2 gene expressions in RAW264.7 macrophages. Vascul. Pharmacol. 47: 99-107, 2007.)

[Document 15] Carmichael, J., WG DeGraff, AF Gazdar, JD Minna, and JB Mitchell. 1987. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47 , 936-942.

[Literature 16] Pavel S, Muskiet FA. Eumelanin (precursor) metabolites as markers for pigmented malignant melanoma, a preliminary report. Cancer Detect Prev. 1983; 6 (1-2): 311-316; 20.

[17] Hearing VJ, Jimenez M. Mammalian tyrosinase, the critical regulatory control point in melanocyte pigmentation. Int. J. Biochem. 1987; 19 (12): 1141.

[Literature 18] yagi A, Kanbara T, Morinobu N. Inhibition of mushroom-tyrosinase by aloe extract. Planta Med. Dec. 53 (6): 515-7.

[Literature 19] (15. Yang YM, MTT assay, Korean Cell Line workshop. 1998.

[Literature 20] Park JG, Kramer BS, Steinberg SM, Carmichael J, Collins JM, Minna JD, Gazdar AF. Chemosensitivity testing of human colorectal carcinoma cell lines using a tetrazolium-based colorimetric assay. Cancer Res. 1987; 47 (22): 5875-5879.

[Document 21] Ko JS. Dermatology. Soomoonsa Press. Seoul. Korea. 2000: 73.

[Document 22]. Prota G. Recent advances in the chemistry of melanogenesis in mammals. J. Invest Dermatol. 1980; 75 (1): 122-127.

Modern people promote skin aging phenomena such as spots, freckles and skin pigmentation by inducing various skin troubles due to various internal and external factors such as ultraviolet rays and stress (Voegeli, R. 1996. Elastase and typing determination on human skin surface. Cosmetic & Toiletries. 111, 51-58.). Pigmentation of the skin is known to regulate the initial reaction from DOPA to DOPA quinone with tyrosinase enzyme, L-tyrosine, such as DHICA oxidase (TRP-1), and DOPA (3,4-dihydroxyphenyla-lanine) in melanin pigment biosynthesis (Aroca, P., et al 1993. Melanin biosynthesis patterns of the 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.). On the basis of this, studies on the search for natural products that inhibit the activity of tyrosinase enzyme and affect the inhibition of melanin biosynthesis are actively conducted. As a result, the tyrosinase gene expression inhibitory effect of horsetail extract (Chin, JE, et al., 2005. Effects of Houttuynia cordata extracts on tyrosinase gene expression, J. Korean Soc Food Sci Nutr 34, 1284-1288) Inhibitory effect of α-MSH-stimulated melanin on the production of melanin in B16 cells (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.) Are being actively studied. Other known antioxidative and whitening raw materials are Arbutin, Kojic acid, Ascorbic acid, etc. Plant extracts such as bark, mackerel and licorice are widely known.

It is known that bokbunja has the efficacy of interpolating renal (ren liver kidney), nomic, diuretic, and treatment of dysentery, wells, and urinary frequency. Studies on the physiological activity of bokbunja strawberry showed that two kinds of flavan-3-ol, one kind of proanthocyanidin and one kind of ellagitanin were separated from the stem and four phenol compounds such as flavonoid were isolated from the leaves. And it is known that it promotes the growth of human B cells and T cell lines that play an important role in antibody production in the human immune system. On the other hand, research on the development of processed foods has been carried out with the development of liquor using bokbunja, and the preparation of dry surface using immature bokbunja powder (Kim Dae-ho, Park Jin-hong, Kim Jung-hwa, Kim Cheol-Hee, Yu-jin Hyun, Kwon Min-cheol, Lee Hyun-yong, (Korean J. Medicinal Crop Sci.), 13 (3): 81 ~ 86, 2005. Seo Hyun Kim, Jung Heon Jang, Yong Seok Park, Young Ki Kim, (1): 11-15, 2005)

Previous studies on the effects of bokbunja have been focused on fruit characteristics, antioxidant activity or development as food, and there is little research on the effects of bokbunja seeds. In addition, the bokbunja was discarded during the processing due to the foreign body that was bite to eat the fruit, and the efficacy of antioxidant, whitening, anti-aging was verified to verify the efficacy of bokbunjae.

Bokbunja berries contain free sugars of protocatechuic acid, gallic acid and glucose (glucose), and bokbunja seed contains various fatty acids and triterpenoid compounds. The triterpenoid compounds are widely distributed in the vegetable field, but the details of the physiological action and the like are not clearly understood. The triterpenoid compounds are mainly contained in berry strawberry, wild strawberry, and the like. tormentic acid, niga-ichigoside, kaji-ichigoside, 23-hydroxytormentic acid, suavissimoside, and the like.

Accordingly, the present inventors have found that the antioxidative effect of phytic acid and suavissimoside Fl, which are compounds isolated from bokbunja seed extract, by DPPH free radical scavenging activity, etc .; High cell viability; Tyrosinase inhibitory activity, Whitening activity such as; And the wrinkle-reducing effect through the inhibition of the elastase (elastase), etc., to confirm that the composition is useful as a composition for treating and preventing skin whitening and skin aging.

In order to achieve the above object, the present invention provides a dermatological pharmaceutical composition for treating and preventing whitening and skin aging comprising a compound isolated from bokbunja seed extract as an active ingredient.

The present invention also provides a cosmetic composition for improving and preventing whitening and skin aging comprising a compound isolated from bokbunja seed extract as an active ingredient.

As used herein, an "extract" refers to an extract of at least one selected from the group consisting of water, ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, butylene glycol, propylene glycol, hydrolyzed butylene glycol, Preferably a water or water and ethanol mixed solvent, most preferably 60% to 80% ethanol soluble extract.

As used herein, "skin aging" includes wrinkles, stains, muscle wounds, skin damage due to ultraviolet light, skin cancer, and preferably wrinkles or stains.

As used herein, a "compound isolated from bramble seed extract" includes phytic acid and suavissimoside Fl.

The above-mentioned compound is characterized in that it comprises 0.1 to 50% by weight based on the total weight of the dermatological pharmaceutical composition.

The pharmaceutical composition includes a cream, a gel, a patch, a spray, an ointment, an alarm, a lotion, a liniment, a pasta or a cataplasmal formulation.

In addition, the cosmetic composition includes formulations of lotion, skin, lotion, nutrition lotion, nutritional cream, massage cream, essence, and pack.

Such compounds as defined herein may be prepared in accordance with methods conventional in the art with pharmaceutically acceptable salts and their solvates.

Accordingly, the present invention is characterized in that said compounds comprise isomers, pharmaceutically acceptable salts or solvates thereof.

As the salt of the present invention, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. The acid addition salt is prepared by a conventional method, for example, by dissolving the compound in an excess amount of an acid aqueous solution and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The molar amount of the compound and the acid or alcohol (e.g., glycol monomethyl ether) in water may be heated and then the mixture may be evaporated to dryness, or the precipitated salt may be subjected to suction filtration.

As the free acid, organic acids and inorganic acids can be used. As the inorganic acids, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid and the like can be used. Examples of the organic acids include methanesulfonic acid, p- toluenesulfonic acid, acetic acid, trifluoroacetic acid, Citric acid, lactic acid, glycollic acid, gluconic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid and the like.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt in particular, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Hereinafter, the present invention will be described in detail.

The compounds of the present invention can be obtained as follows.

Butanol, acetone, ethyl acetate, hexane, butylene glycol, propylene glycol, water, ethanol, methanol, propanol, isobutanol, and the like which have a volume of about 1 to 30 times, preferably about 1 to 15 times, Preferably from about 10 DEG C to about 100 DEG C, preferably from about 60 DEG C to about 90 DEG C, with the addition of at least one solvent selected from the group consisting of propylene glycol, propylene glycol, propylene glycol, butylene glycol, For 5 to 5 hours, preferably by a reflux cooling extraction method such as hot water extraction, reflux cooling extraction, immersion extraction or pressure extraction; A second step of adding the crude extract obtained in the above step to water in an amount of about 0.05 to 50 times, preferably 0.5 to 5 times (v / w%) of the weight of the crude extract to prepare a suspension; A non-polar solvent selected from n-hexane, heptane, methylene chloride, chloroform or ethyl acetate in an amount of about 0.5 to 20 times, preferably 1 to 5 times (v / w) A third step of sequentially obtaining a non-polar solvent-soluble fraction and a polar solvent-soluble fraction by sequentially repeating the fractionation step with hexane or ethyl acetate solvent, butanol and water; The non-polar solvent-soluble fraction is purified by a column chromatography method such as a silica gel column, a Sephadex column or an ion exchange resin, or a purification method such as a retreating method, preferably a developing solvent (developing a mixed solvent of chloroform and water with increasing polarity) And the purification process using a recrystallization method is repeatedly carried out in a fourth step to isolate the compound of the present invention such as phytic acid and suavissimoside Fl of the present invention.

The present inventors have found that the antioxidative effect of phytic acid and suavissimoside Fl of the present invention, such as DPPH free radical scavenging activity, etc., High cell viability; Tyrosinase inhibitory activity, Whitening activity such as; And the wrinkle-reducing effect through the inhibition of the elastase (elastase), etc., were found to be useful as a composition for the treatment and prevention of whitening and skin aging.

In addition, the bokbunja has been used for herbal medicine and food for a long time, and the herbal extract of the present invention extracted from the herb has long been proved to be a non-irritant sample in the skin patch test and has no problems such as toxicity and side effects. Can be used.

The dermatological pharmaceutical composition containing the compound of the present invention can be used as a pharmaceutical composition in the form of external preparation for skin such as cream, gel, patch, spray, ointment, warning agent, lotion, liniment, pasta or cataplasm However, the present invention is not limited thereto.

The preferred dosage of the compound of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention is preferably administered at 0.0001 to 100 mg / kg per day, preferably 0.001 to 10 mg / kg per day. 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.

The compounds of the present invention can be used variously in cosmetics and cleansers having a whitening effect.

Examples of products to which the present composition can be added include cosmetics such as lotion, skin, lotion, nutrition lotion, nutritional cream, massage cream, essence, pack, cleansing, cleanser, soap, have.

The cosmetic composition of the present invention comprises a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.

The water-soluble vitamin is not particularly limited as long as it can be compounded in cosmetics. Preferably, vitamin B, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, And their salts (thiamine hydrochloride, sodium ascorbate, etc.) or derivatives (sodium ascorbic acid-2-phosphate, magnesium ascorbate-2-phosphate etc.) can also be added to water-soluble vitamins . The water-soluble vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzymatic method, or a chemical synthesis method.

Usable vitamins include vitamins such as vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol) , Derivatives thereof (such as palmitic acid ascorbin, stearic acid ascorbic acid, dipalmitic acid ascorbin, dl-alpha tocopherol acetic acid, dl-alpha tocopherol nicotinic acid vitamin E, dl-pantothenyl alcohol, D-pantothenyl alcohol, Ether, etc.) are also included in the usable vitamins used in the present invention. Usability Vitamins can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzyme or a chemical synthesis method.

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. The polymeric peptide can be obtained by a conventional method such as purification from a culture broth of a microorganism, an enzymatic method, or a chemical synthesis method, or it can be purified from natural products such as ducks such as pigs and cows and silk fiber of silkworms.

The polymeric polysaccharide may be any compound as long as it can be incorporated in cosmetics, and examples thereof include hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof (sodium salt, etc.). For example, chondroitin sulfate or a salt thereof can be usually purified from mammals or fish.

Sphingo lipids may be any as long as they can be incorporated into cosmetics, and preferable examples thereof include ceramides, phytosphingosine and sphingoglycolipids. Sphingoid lipids can be purified from ordinary mammals, fish, shellfish, yeast or plants by conventional methods or can be obtained by chemical synthesis.

The seaweed extract may be any of those which can be compounded in cosmetics. Preferably, the seaweed extract is selected from the group consisting of algae extract, red pepper extract, green algae extract and the like. Also, the algae extract may be colored guanine, arginic acid, Potassium alginate and the like are also included in the seaweed extract used in the present invention. Seaweed extract can be obtained from seaweed by a conventional method.

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

Examples of the compounding ingredients that may be added include organic solvents such as a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorbent, a preservative, a bactericide, an antioxidant, a plant extract, a pH adjuster, A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water.

Examples of the oil retaining component include ester-based oil retaining, hydrocarbon-based oil retaining, silicone-based oil retaining, fluoric oil retaining, animal retention and plant retention.

Examples of ester-based fats include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isostearyl isostearate, Butyl isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isosilyl myristate, isostearic acid isostearyl, isostearyl palmitate, octyldodecyl myristate, Trimethylol propane, triisostearic acid trimethylol propane, tetra 2-ethylhexanoic acid pentaerythritol tetra (2-ethylhexanoate) , Decyl caprylate, decyl laurate, hexyl laurate, myristate decyl, myristyl myristate, myristine monoethyl stearate, stearyl stearate, decyl oleate, ricinoleic acid tri , Isostearyl stearate, isostearyl stearate, isodecyl stearate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl isostearate, isopropyl stearate, isopropyl stearate, isopropyl stearate, -Hexyl stearate, stearyl ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di (capryl, capric acid) propylene glycol, Propyleneglycol propionate, propyleneglycol propionate, dicaproic acid neopentyl glycol, dioctanoic acid neopentyl glycol, tricarboxylic acid glyceryl, triunsaturated glyceryl, triisopalmitic acid glyceryl, triisostearic acid glyceryl, neopentanoic acid octyldodecyl Octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, Octyldecyl lactate, octyldecyl lactate, octyldecyl lactate, polyglycerin oleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisobutyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, But are not limited to, ethyl, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, But are not limited to, dioctyl sebacate, stearic acid cholesteryl, isostearic acid cholesteryl, hydroxystearic acid cholesteryl, oleic acid cholesteryl, oleic acid dihydrocholesteryl, isostearic acid pitostearyl, Stearoyl hydroxystearic acid isostearyl, 12-stearoyl stearyl hydroxystearate, 12-stearo And monohydroxystearic acid and esters such as sostearyl.

Examples of the hydrocarbon hydrocarbon-based fats include hydrocarbon fats and oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, floating isoparaffin, polybutene, microcrystalline wax and vaseline.

Examples of silicone based oils include polymethyl silicone, methylphenyl silicone, methyl cyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane-methylstarchoxysiloxane copolymer, alkyl Modified silicone oils, and amino-modified silicone oils.

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

Examples of animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rape oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, , Corn oil, palm oil, palm oil, cucumber nut oil, wheat germ oil, rice germ oil, shea butter, coltsfoot colostrum, marker daisy nut oil, mead home oil, egg oil, , Canned wax, carnauba wax, liquid lanolin, hardened castor oil, and the like.

Examples of the moisturizing agent include water-soluble low-molecular moisturizing agents, oil-soluble molecular moisturizing agents, water-soluble polymers, and oil-soluble polymers.

Examples of the water-soluble low-molecular moisturizing agent include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B Glycol (polymerization degree n = 2 or more), polyglycerin B (polymerization degree n = 2 or more), lactic acid, lactic acid salt and the like.

Examples of the lipid-soluble low-molecular moisturizing agent include cholesterol and cholesterol ester.

Examples of the water-soluble polymer include carboxyvinyl polymer, polyaspartic acid, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, dextrin, etc. .

Examples of the oil-soluble polymer include polyvinylpyrrolidone / eicosene copolymer, polyvinylpyrrolidone / hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and polymer silicone.

Examples of the emollients include long chain acyl glutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid and lanolin fatty acid cholesteryl ester.

Examples of the surfactant include nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants.

Examples of the nonionic surfactant include self emulsifying monostearate glycerin, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbit fatty acid ester, POE (Polyoxyethylene / polyoxypropylene) copolymer, POE.POP alkyl ether, polyether-modified silicone, polyether-modified silicone, polyoxyethylene-polyoxypropylene (POE) Alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like.

Examples of the anionic surfactant include fatty acid soap, alpha-acylsulfonate, alkylsulfonate, alkylarylsulfonate, alkylnaphthalenesulfonate, alkylsulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl ginseng salt, Alkylsulfosuccinic acid salts, acylated hydrolyzed collagen peptide salts, and perfluoroalkyl phosphoric acid esters, and the like can be mentioned. have.

Examples of the cationic surfactant include alkyl trimethyl ammonium chloride, stearyl trimethyl ammonium chloride, stearyl trimethyl ammonium bromide, cetostearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, stearyl dimethyl benzyl ammonium chloride, behenyl trimethyl ammonium chloride, Benzalkonium, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salts of lanolin derivatives, and the like.

Examples of the amphoteric surfactant include carboxybetaine type, amide betaine type, sulfobetaine type, hydroxysulfobetaine type, amidosulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type and amide amine type Amphoteric surfactants and the like.

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, But are not limited to, polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluororesin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, Silk powder, cellulose, CI Pigment Yellow, CI Pigment Orange, and composite pigments of inorganic pigments and organic pigments thereof.

As the organic powder, metallic soap such as calcium stearate; Metal salts of alkyl phosphates such as sodium zinc cetylate, zinc laurylate and calcium lauryl laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc and N-lauroylglycine calcium; Amidosulfonic acid multivalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; Such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylidene, N-alpha-paratyylnitine, N-alpha-lauroyl arginine, Acyl basic amino acids; N-acylpolypeptides such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-aminocaprylic acid, alpha-aminoaurauric acid, and the like; Polyethylene, polypropylene, nylon, polymethylmethacrylate, polystyrene, divinylbenzene-styrene copolymer, ethylene tetrafluoride, and the like.

Examples of ultraviolet absorbers include paraaminobenzoic acid, ethyl parnamobenzoate, amyl paranobenzoate, octyl paranobenzoate, ethyleneglycol salicylate, phenyl salicylate, benzyl salicylate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate, benzyl cinnamate , Octyl methoxycinnamate, dioctyl methoxycinnamate, mono-2-ethylhexane glyceryl dipyrromethoxycinnamate, isopropyl paratumoxycinnamate, diisopropyl-diisopropyl cinnamate ester mixture, Carninoic acid, ethyl urocanoate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and salts thereof, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzophenone , Tetrahydroxybenzophenone, 4- tert -butyl-4'-methoxydibenzoylmethane, 2,4,6-trianylino- p- (carbo-2'-ethylhexyl- , 3,5-triazine, 2- (2-H 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, gallic acid propyl, and eicosorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogenphosphate and the like.

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

In addition, any of the above components may be blended within the range not impairing the objects and effects of the present invention, but it is preferably 0.01 to 5% by weight, more preferably 0.01 to 5% by weight, Preferably 0.01 to 3% by weight.

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

The ingredients contained in the cosmetic composition of the present invention may contain, as an active ingredient, the ingredients conventionally used in cosmetic compositions in addition to the above-mentioned compounds, for example, conventional additives such as stabilizers, solubilizers, vitamins, And a carrier.

The cosmetic composition of the present invention can be prepared into any formulation conventionally produced in the art, and examples thereof include emulsions, creams, lotions, packs, foundations, lotions, essences, and hair cosmetics.

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 .

When 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. In the case of a spray, in particular, / 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.

Antioxidative effects such as DPPH free radical scavenging activity on phytic acid and suavissimoside Fl which are compounds isolated from the bokbunja seed extract of the present invention; High cell viability; Tyrosinase inhibitory activity, Whitening activity such as; And the wrinkle-reducing effect through the inhibition of the elastase (elastase), thereby being useful as a composition for treating and preventing whitening and skin aging.

FIG. 1 is a graph showing the electron donating ability of phytic acid and suavissimoside F1,
FIG. 2 is a graph showing the inhibitory activity of nitric oxide in Raw264.7 cells of phytic acid and suavissimoside F1,
FIG. 3 is a graph showing cell viability of B16F10 melanoma cells treated with phytic acid and suavissimoside F1,
FIG. 4 is a graph showing the cell viability of phytic acid and suavissimoside F1 in CCD-986sk cells,
FIG. 5 is a graph showing the tyrosinase inhibitory effect of phytic acid and suavissimoside F1,
FIG. 6 shows the effect of inhibiting elastase in phytic acid and suavissimoside F1 purified water,
FIG. 7 is a graph showing MMP-1 inhibitory activity of phytic acid and suavissimoside F1 in CCD-986sk cells,
FIG. 8 shows cell viability in phagocytic cells of phytic acid and suavissimoside F1 (Raw 267.7).

Below, The present invention will be described 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.

Reference Example 1. Reagents used for separation purification

RP-18 (12nm s-150um, YMC * GEL ODS-A, YMC Co., Japan) for silicagel (70-230 mesh, colum chromatography, merck, Darmstadt, Germany) Were used. The analytical instrument used was Shimadzu's LC-20AT pump, SPD-M20A PDA, ELSD HPLC, and nuclear magnetic resonance spectrum (JNM-ECA500, Japan). The instrument used for the physiological activity experiments was a UV / VIS spectrophotometer (Hitachi, Japan), a rotary vacuum evaporator (Tokyo, Rikakikai Co.), a centriefuge (Hitachi, Japan), a freeze drier _{, Japan), CO 2 incubator (} Hanbaek Scientific Co. Korea), pH meter (Metrohm, Switzerland), BOD incubator (Hanbaek Co. Korea), autoclave (Hanbaek Scientific Co. Korea), ELISA reader (Bio Rad, Japan) the Respectively.

Example 1 Isolation of Active Ingredients from Bokbunja Seed Extract

1-1. material

The brambled seeds used in this experiment( Rubus coreanus seed)Was purchased from Samdongbond Omija Farm in early December 2014 and used as an experimental material.

1-2. Sample extraction and purification

2 kg of fermented brambles was pulverized and immersed in 70% ethanol for 24 hours and filtered. This process was repeated twice, and then concentrated under reduced pressure at 50 ° C to obtain a viscous 70% ethanol concentrate. The concentrate was dissolved in 100 mL of methanol, 500 mL of water and 500 mL of hexane were added, and the liquid was extracted. 500 mL of ethyl acetate was further added to the aqueous layer and the mixture was subjected to liquid extraction.

The ethyl acetate fraction extract was concentrated and purified by silica gel open column chromatography. The column was loaded with a solvent system of hexane / ethyl acetate (9: 1, v / v), then hexane / ethyl acetate as mobile phase 9: 1 to 0:10 and again eluted with ethyl acetate / methanol 10: 0 to 2: v / v).

The obtained purified product was subjected to silica gel chromatography or LiChroperp RP-18 column chromatography and suavissimoside F1 (Formula 1), which exhibited the following physical properties in ethyl acetate / methanol (8: 2) 1984), ethyl acetate / methanol (7: 3 - 2: 1), ethyl acetate / methanol (2: 5: 5) fractions of the following formula (2), Andrew T. Bruman, Ginger M. chateauneuf, Brian R. Boyd, Richard E. Brown, Pushpalatha PNMurthy 1999 Tetrahedron Letters 40: 4489-4492), respectively, were obtained.

[Chemical Formula 1]

Suavissimoside F1 < / RTI >

- Molecular Weight : 680.8405

- Structural formula: C ₃₆ H ₅₆ O ₁₂

(2)

Pic acid

- Molecular Weight : 660.04

_- Structural formula: C ₆ H ₁₈ O ₂₄ P ₆

Experimental Example 1. Identification of antioxidant activity and anti-inflammatory effect

1-1. Reagent preparation

(DPPH, D9132), xanthine, xanthine oxidase, pyrogallol, hyaluronidase, hyaluronic acid, p- dimethylaminobenzaldehyde (D2004), sodium nitrite S5506) and griess reagent (G4410) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

1-2 Measurement of electron donating ability

In order to confirm the electron donating ability of the sample of the above example, the Blois method described in the literature was modified to perform the following experiment (Blois, MS 1958. Antioxidant determinations by the use of a stable free radical, Nature, 181: 1199- 1200.)

Electron donating ability (EDA) was measured by modifying Blois' method (9). 1 mL of 0.2 mM 1,1-diphenyl-2-picrylhydrazyl (DPPH) was added to 2 mL of each sample solution, The absorbance was then measured at 517 nm. The electron donating ability was expressed by the absorbance reduction rate of the sample solution addition group and the no addition group.

1,1 - diphenyl - 2 - picrylhydrazyl (DPPH), which is used for electron donating ability measurement, is a very stable free radical and is a purple compound showing characteristic absorption at 517 nm. DPPH is very stable in organic solvent such as alcohol and it is decolorized by proton-radical scavenger in antioxidant mechanism. Therefore, DPPH is widely used to search for antioxidant materials with various natural materials because it has an advantage of being able to easily observe antioxidant activity with naked eyes. In addition, free radicals are likely to cause aging by binding to lipids or proteins in the body. Phenolic compounds have a strong ability to reduce or offset free radicals, which can be used as a measure to inhibit free radical aging in the body (13) Aoshima H, Tsunoue H, Koda H, Kiso Y. Aging of whiskey increases 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. J. Agric. Food Chem 2004; 52 (16): 5240-5244; 14. Kim, J., Kim, Y., Do, JR, and Lee, Y., Antioxidative activity and physiological activity of some Korean medicinal plants, J. Food Sci. Technol. 1995; 27 (1): 80-85.).

The experimental results are shown as Figure 1. The results of measuring the electron donating ability of Phytic acid, suavissimoside F1. Phytic acid and suavissimoside F1 showed electron donating ability of more than 90% at a concentration of 1000 μg / ml. C, it was confirmed that the compound is excellent in antioxidant efficacy.

1-3. Anti-inflammatory effect measurement

In order to confirm the antiinflammatory effect of the sample of the above example, the method described in the literature was modified to carry out an experiment for the inhibition of nitrogen oxide (Nitric oxide) as follows (Kim, KW, Ha, KT, , Chang, HW, Lee, CS, Kim, CH Polygonum cuspidatum, compared with baicalin and berberine, inhibits inducible nitric oxide synthase and cyclooxygenase-2 gene expressions in RAW264.7 macrophages. Vascul. Pharmacol. 47: 99-107, 2007. )

Nitric oxide (NO) measurement was performed by measuring the amount of nitric oxide (NO) in the cell supernatant as nitrite and nitrate. Griess reagent (G4410 Sigma, USA), which is a safe form after nitrite reduction to nitrite, was used. In a 6 well plate, 2 × 10 ⁶ cells were washed twice with PBS when confluence was 80% After culturing for more than 12 hours using the medium, 10 μg / mL lipopolysacchride (L6636LPS) was added to all wells except for the control group. After 2 hours, the samples were treated by concentration. The amount of NO produced was measured by absorbance at 540 nm after collecting supernatant by time and reacting with griess reagent for 10 minutes.

Excess inflammatory factors such as Nitric oxide (NO) and Prostaglandin E2 (PGE2) are formed by inductive NO synthase (iNOS) and cyclooxygenase (COX) -2 in the body inflammation process. Among these, NO has various physiological functions such as body defense function, signal transduction function, neurotoxicity, and vasodilation. Nitric oxide synthase (NOS), which forms NO, is divided into three kinds of isoenzymes such as type Ⅰ, Ⅱ and Ⅲ according to physicochemical properties. Type Ⅰ (neuronal NOS, nNOS) and type Ⅲ (endothelial, eNOS) are classified into constitutive NOS because they are continuously present in the cells. In some cells, specific stimulants such as LPS, cytokides and bacterial toxins , And inductive NOS (iNOS) type III, which is expressed only when exposed (27). These NOS convert L-argine to L-citruLline and form NO. These NOS are absolutely highly expressed by iNOS, which plays an important pathological role. In general, the formation of NO plays an important role in killing bacteria or eliminating tumors, but the formation of excessive NO by pathologic causes causes inflammation and causes tissue damage, gene mutation and nerve damage.

In order to measure the degree of inhibition of NO production Raw264.7 cell Phytic acid, was also indicated as 2 the results of measuring the amount of NO produced was treated suavissimoside F1 at different concentrations to the cells. 1, 5, 10, and 25 μg / ml, the NO production was decreased in a concentration-dependent manner. In particular, at 25 μg / ml of the highest concentration, I could.

Experimental Example 2. 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.)

MTT screening for cell viability is performed using a 96-well plate. The test results can be easily read by using an ELISA reader (muLtiwell microplate reader), which is widely used as a cytotoxicity and cell proliferation detection method It is one of the methods. In the case of cancer cells, the yellow water soluble MTT tetrazolium is reduced to a purple colored water insoluble MTT formazan by the dehydrogenase action of mitochondria during metabolism. The absorbance of MTT formazan is maximized at wavelengths near 550 nm, reflecting the metabolically vigorous concentration of cells (Pavel S, Muskiet FA, Eumelanin (precursor) metabolites as markers for pigmented malignant melanoma, a preliminary report. Cancer Detect Prev., 1983; 6 (1-2): 311-316; 20. Hearing VJ, Jimenez M. Mammalian tyrosinase, the critical regulatory control point in melanocyte pigmentation, Int. J. Biochem. : 1141.).

2-1. Cell culture

The cells were cultured using DuLbeco's modified eagle's medium (SH30081.03 DMEM) supplemented with 10% fetal bovine serum (SH30243.31 FBS) and 1% penicillin / streptomycin (100 U / mL) The cells were subcultured in a 5% CO ₂ incubator at 37 ° C.

2-2. Cell lines and reagents used for measuring cell viability

Cell lines used for cell viability were purchased from the American Type Culture Collection (ATCC), melanoma cells B16F10 (CRL6475), fibroblast cells CCD-986sk (CRL-1947) and macrophage cells Raw 264.7 (TIB-71) The cell viability was measured by using DMEM (SH30081.03 DuLbecco's Modified Eagle Medium), fetal bovine serum (FBS), peni cillin / streptomycin, trypsin 250 and 0.4% trypan blue stain using Gibco BRL Co. (Grand Island, USA) and Haemacytometer 2-yl] -2,5-diphenyl-tetrazoliumbromide (M2128, MTT) was purchased from Sigma Chemical Co. (St. Louis, Mo., USA).

2-3. Measurement of cell viability by MTT assay

Each cell line [melanoma (B16F10), fibroblast (CCD-986sk), and macrophage (Raw 264.7) cells] was dispensed in a 96 well plate at a concentration of 0.6 to 8 × 10 ³ cells / well in an amount of 0.18 ml. mL, and incubated at 37 ° C in a 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 adding 0.02 mL of the MTT solution prepared at a concentration of 5 mg / mL for 4 hours, the culture solution was removed, and 0.15 mL of DMSO: Ethanol (1: 1) was added to each well and reacted at room temperature for 30 minutes. Absorbance was measured at 550 nm. The cell viability was measured by the absorbance reduction rate of the sample solution addition group and the no addition group.

2-4. Experiment result

As a result of the experiment, it was determined whether the production of nitric oxide was induced by lipopolysaccharide (LPS) or the decrease of cell popuLation due to cytotoxicity of the compound, as shown in FIG. 3. As a result, Phytic acid and suavissimoside F1 showed high cell proliferation at concentrations of 1, 5, 10 and 25 μg / ml compared with 50 and 100 μg / ml, respectively. It was confirmed that toxicity was observed at the concentration of μg / ml.

The survival rate of melanoma cells by phytic acid and suavissimoside F1 was shown in Fig. The cell viability was 80% in the α-MSH-treated group compared with the normal cell group. The phytic acid and suavissimoside F1 treatment showed higher survival rate at the concentration of 1, 5 μg / ml compared with the control group. / ml, it was confirmed that the cell viability was 80% or more.

Experimental Example 3. Whitening activity test

L-DOPA, porcine pancreas elastase (PPE), N -succinyl- (L-Ala) ₃ - p- nitrophenyl-alanine , collagenase and 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg were purchased from Sigma Chemical Co. (St. Louis, MO, USA). The procollagen type I C-peptide EIA kit (Takara-Bio Inc. Japan) was used for the measurement of collagen biosynthesis.

3-1. Tyrosinase inhibitory activity

In order to test the tyrosinase inhibitory activity of the samples obtained in the examples, Yagi et al. (Yagi A, Kanbara T, Morinobu N. Inhibition of mushroom-tyrosinase by aloe extract. Planta Med 1987 Dec; 53 (6): 515-7 ..)

To the reaction mixture, 0.2 mL of mushroom tyrosinase (T3824, 110 U / mL) was added to a mixture of 0.2 mL of the substrate solution in which 10 mM L-DOPA had been dissolved and 0.5 mL of the sample solution in 0.5 mL of 0.175 M sodium phosphate buffer (pH 6.8) And the DOPA chrome produced in the reaction solution was measured at 475 nm. The tyrosinase inhibitory activity was expressed as the absorbance reduction ratio of the sample solution and the non - added sample.

Melanin, a key determinant of skin tone, is biosynthesized in melanosomes in pigmented cells called melanocytes in the epidermal basal layer. The starting material for the synthesis of melanin is tyrosine, an amino acid. Tyrosine is oxidized by tyrosinase in melanocytes to L-3,4-dihydroxyl phenylalanine (DOPA) and DOPA quinone. It is known that DOPA quinone then becomes DOPA chrome, 5,6-dihydroxyindole, indole-5,6-quinone and then melanin by polymerization with indole-5,6-quinone (15 Yang YM, MTT assay, Korean Cell Line workshop 1998; 16. Park JG, Kramer BS, Steinberg SM, Carmichael J, Collins JM, Minna JD, Gazdar AF. Chemosensitivity testing of human colorectal carcinoma cell lines using a tetrazolium-based colorimetric assay Cancer Res 1987; 47 (22): 5875-5879; 17. Ko JS, Dermatology, Soomoonsa Press, Seoul, Korea, 2000: 73.). In addition, tyrosinase plays an important role in the formation of melanin in the skin. Tyrosine hydoxylase, which oxidizes tyrosine in melanosome to form DOPA, acts as a DOPA oxidase that oxidizes DOPA to form DOPA quinone. (Prota G. Recent advances in the chemistry of melanogenesis in mammals. J. Invest. Dermatol. 1980; 75 (1): 122-127.).

The experimental results, so the skin is shown in Figure 5 of mushroom tyrosinase inhibitory activity measurement results are derived in order to measure the tyrosinase-inhibiting activity that can effectively inhibit the biosynthesis of melanin in the polymer. Phytic acid and suavissimoside F1 were inhibited by tyrosinase, especially at the concentration of 1000 μg / ml.

Experimental Example 4. Effect of wrinkle-improving effect

4-1. Measurement of Elastase Inhibitory Activity

In order to test the elastase inhibitory activity of the samples obtained in the examples, Cannell et al. (12) were used to test the results as follows (12. Cannell RJP, Kellan SJ, Owsians AM, Walker JM. Results of a large scale screen of microalgae for the production of protease inhibitors (Planta Med. 1988; 54 (1): 10-14.).

The elastase present in the neutrophil granules of the human body is an enzyme that degrades elastin, a substrate protein important for maintaining skin elasticity in the dermis, and is a nonspecific hydrolytic enzyme capable of degrading collagen, another important substrate protein. Therefore, elastase inhibitors have been shown to improve skin wrinkles (21) Tsuji N, Moriwaki S, Suzuki Y, Takema Y, Imokawa G. The role of elastase secreted by fibroblasts in wrinkle formation: (2): 283-290). It is also one of the leukocyte granulase enzymes that decompose elastin in the body. The activity of the enzymes in abnormal tissues is extremely high, which is a direct cause of tissue destruction. 1980); 255 (4): 1301-1304; 24. DeWitt DL, Rollins TE (1986), pp. , Day JS, Gauger JA, Smith WL, Orientation of the active site, and antigenic determinants of prostaglandin endoperoxide synthase in the endoplasmic reticulum. J. Biol. Chem., 1981, 256 (20): 10375-10382.

N-succinyl- (L-Ala) ₃ - p- nitroanilide S4760, sigma was used as a substrate and the amount of p- nitroanilide produced from the substrate at 37 ° C for 20 minutes was measured at 445 nm. 0.5 mL of each test solution was added to 0.5 mL of each test solution. 0.5 mL of porcine pancreas elastase (2.5 U / mL E1250, Sigma) dissolved in 50 mM tris-HCl buffer (pH 8.6) N-succinyl- (L-Ala) ₃ - p- nitroanilide (0.5 mg / mL) dissolved in 50 mM tris-HCl buffer (pH 8.6) was added and the reaction was allowed to proceed for 20 minutes. Elastase inhibitory activity was expressed as the absorbance reduction ratio of the sample solution and the non - added sample.

As a result of measuring the elastase inhibitory activity associated with this wrinkle formation it is shown in FIG. Phytic acid and suavissimoside F1 showed elastase inhibitory activity in a dose dependent manner and 24% and 25% of elastase inhibitory activity at a concentration of 1000 μg / ml.

4-2. Measurement of inhibitory activity of MMP-1

In order to test the MMP-1 inhibitory activity of the samples obtained in the examples, the following methods were applied as described in the literature (Chung JH, Kang S, Varani J, Lin J, Fisher GJ, Voorhees JJ 2000. Decreased extracellular -signal-regulated kinase and increased stress-activated MAP kinase activities in aged human skin in vivo J. Invest. Dermatol. 115: 117-182 ..)

Cells are inoculated on a 96-well plate at a concentration of 1 × 10 ⁴ cells / well, and the samples are added to each well and cultured in a CO ₂ incubator for 24 hours. The cultures of the cells thus obtained are collected and used for the experiment. The amount of collagen biosynthesis in the cell culture medium was determined by measuring the amount of propeptide using a procollagen type-Ⅰ C peptide (PIP) EIA kit (MK101, Takara). MMP-1 was measured using an Abcam ELISA system (ab100603 GE Healthcare) do.

4-2. Fibroblast cell (CCD- 986sk ) Survival rate confirmation

Phytic acid, suavissimoside results confirm the viability of the fibroblast cells by F1 as shown in FIG. 7 Phytic acid, suavissimoside F1 due to UVB irradiation cell viability is degraded CCD-986sk cell 1, at 5, 10 μg / ml concentration of 100 % Cell growth rate. Therefore, in this study, the inhibition rate was measured at a cell concentration of 10 μg / mL or less to measure MMP-1 expression.

4- 3. MMP -1 inhibition activity measurement

Reconstitution of matrix proteins is involved in various biological reactions, in particular MMPs play an important role in the degradation of substrate proteins. Among them, MMP-1 is involved in intestitial collagenase, and the increase of MMP-1 in skin exposed to ultraviolet light causes skin aging by degrading collagen composing skin and causing deficiency of substrate protein (25,26 ) Thus, in this study, MMP-1 inhibition activity of phytic acid and suavissimoside F1 in CCD-986sk cells was measured.

As a result, as shown in FIG. 8, the UVB-treated control group showed higher MMP-1 expression than the normal cell group, and treated with phytic acid and suavissimoside F1 showed 20% inhibition of MMP-1 .

Phytic acid, and suavissimoside F1 were tested for antioxidant activity, whitening, wrinkle improvement and anti - inflammatory effects. The antioxidative activity of phytic acid, suavissimoside F1 was measured and the electron donating ability was measured to be 90% or more at a concentration of 1000 μg / ml. In order to confirm the whitening effect, tyrosinase inhibitory activity of Phytic acid and suavissimoside F1 purified water was more than 40%. In the case of phytic acid, suavissimoside F1 purified water, Inhibition activity of more than 20% at 1,000 μg / ml and more than 20% inhibition activity of MMP-1 at a concentration of 5 μg / ml. In order to measure the anti-inflammatory activity, Raw264.7 cells were treated with LPS to determine the expression pattern of Nitric oxide and its associated iNOS protein. As a result, it was confirmed that Phytic acid and suavissimoside F1 inhibited more than 40% I could. Phytic acid and suavissimoside F1 were added to a mask pack to confirm the efficacy of cosmetic addition. From the above results, it was confirmed that the whitening effect and the wrinkle improving effect of the purified water were confirmed, and the possibility of functional cosmetic material was confirmed.

Hereinafter, formulations of cream, massage cream, lotion, skin lotion, essence, pack, and cleansing foam are exemplified as the formulation examples of the present invention, but the formulations including the cosmetic composition of the present invention are not limited thereto.

Formulation Example  1. Cream composition

The oil phase and water phase are heated to 75 ° C and cooled to room temperature.

Formulation Example  2. Massage Cream  Composition

The oil phase and water phase are mixed by heating at 75 DEG C and then cooled to room temperature.

Formulation Example  3. lotion composition

The oil phase and water phase are mixed and emulsified by heating at 75 ° C and then cooled to room temperature.

Formulation Example  4. Skin lotion composition

The water phase and the ethanol phase are respectively prepared and mixed and then filtered.

Formulation Example  5. Essence composition

The water phase and the ethanol phase are respectively prepared and mixed and then filtered.

Formulation Example  6. Pack composition

The water phase and the ethanol phase are dispersively dissolved and mixed, and then cooled to room temperature.

Formulation Example  7. Cleansing Foam  Composition

The water phase and the oil phase are dispersed and dissolved, mixed and sieved, and then cooled to room temperature.

Claims (4)

A pharmaceutical composition for external application for the treatment and prevention of whitening and skin aging comprising phytic acid or suavissimoside Fl compound isolated from bokbunja seed extract as an active ingredient. The dermatological pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is a cream, a gel, a patch, a spray, an ointment, a warning agent, a lotion, a liniment, a pasta or a cataplasma. A cosmetic composition for improving and preventing whitening and skin aging comprising phytic acid or suavissimoside Fl compound as an active ingredient isolated from bokbunja seed extract. The cosmetic composition according to claim 3, wherein the cosmetic composition is a formulation of lotion, skin, lotion, nutrition lotion, nutritional cream, massage cream, essence, pack.

Images