KR100550818B1 - Composition comprising the extract of Phellinus linteus or compounds isolated therefrom having tyrosinase inhibitory activity - Google Patents

Abstract

The present invention relates to a whitening cosmetic composition or a composition for preventing browning of food containing a compound that can be separated or synthesized from a situation mushroom non-polar solvent soluble extract or situation mushroom extract having tyrosinase inhibitory activity. Compounds that can be isolated or synthesized from solvent-soluble extracts or mushroom extracts act on tyrosinase, an enzyme that plays a key role in melanin biosynthesis, and effectively inhibits the enzyme activity. Thus, cosmetic compositions or foods for preventing skin aging and whitening It can be usefully used as a browning prevention composition.

Situation mushroom, extract, tyrosinase inhibition, compound, whitening, cosmetic composition, food, browning prevention composition.

Description

Composition comprising the extract of Phellinus linteus or compounds isolated there from having tyrosinase inhibitory activity}             

1 is a diagram showing the tyrosinase inhibitory activity of the crude extract and each solvent fraction of the situation mushroom,

FIG. 2A is a diagram of a Rainweber-Burk plot of tyrosinase and L-tyrosine with and without protocatechualdehyde addition,

FIG. 2B is a diagram of the Rainweber-Burk plot of tyrosinase and L-tyrosine with and without 5-hydroxymethyl-2-peraldehyde. FIG.

The present invention relates to a whitening cosmetic composition or a composition for preventing browning of foods containing compounds which can be separated from or synthesized from a situation mushroom nonpolar solvent-soluble extract or a situation mushroom extract.

Tyrosinase (EC 1.14.18.1) is a polyphenol oxidase (PPO) (Mayer AM, Phytochemistry , 26 , pp 11-20, 1987; Whitaker JR et al., In Food Enzymes, Structure and Mechanism , pp271-307, 1995), a copper-containing enzyme found in various animals, plants, and microorganisms, including humans. Among the amino acids, tyrosine is oxidized by tyrosinase to convert DOPA (3,4-dihydroxyphenylalanine). Dopaquinone is converted into 5,6-dihydroxyindole, which is then converted into 5,6-dihydroxyindole, which is polymerized to bind with protein to form melanin pigment. Melanin pigment, which is produced by melanocytes in the body skin, is a phenolic polymer that has a complex form of black pigment and protein, and is involved in the pigmentation of skin, hair, and eyes. In addition to aesthetic problems such as freckles, it also promotes skin aging and causes skin cancer (Hearing VJ et al., FASEB J. , 5 , pp2902-2909, 1991). As such, the enzymatic oxidation that forms melanin in L-tyrosine is of great importance because its end product, melanin, has a variety of functions, as well as changes in melanin biosynthesis cause different disease states.

In addition to the actions described above, tyrosinase is involved in the browning of fruits, vegetables and crustaceans, and has been applied in the development of drugs to control pests as an important enzyme involved in the shelling of insects (Andersen SO, Anul. Rev. Entomol. , 24 , pp 29-61, 1979). Tyrosinase has also been shown to play a central role in the neurotoxicity of dopamine, as well as to contribute to neurodegeneration associated with Parkinson's disease (Xu Y. et al., Mol Brain Res. , 45 , pp 159-162, 1997). In addition, many researches are being conducted on anticipation of an effect as a whitening cosmetic using a tyrosinase inhibitor or a therapeutic agent such as hyperpigmentation, and a whitening agent for preventing melanin overproduction of epithelial cell layers using various compounds isolated from plants. There are attempts to develop cosmetics and pharmaceutical products.

Korean Patent Application No. 2000-0073136 discloses a tyrosinase inhibitor comprising Eoseongcho extract and a whitening cosmetic containing the same, and Korean Patent Registration No. 190992 discloses a composition for skin whitening containing astronomical extract and tyrosinase inhibitor. have.

However, whitening agents and browning inhibitors that have been researched and developed so far have various disadvantages such as poor safety such as cytotoxicity and mutagenicity, low activity and poor skin permeability.

Therefore, there is a need for further studies on components having anti-tyrosinase activity that is safe and effective in practical use. In particular, the safety of tyrosinase inhibitors should be considered first because they are generally used without any standard when used in food or cosmetics.

On the other hand, inhibitors of enzyme reactions include competitive inhibitors and non-competitive inhibitors. Competitive inhibitors are substances that prevent the substrate from binding to the free enzyme, and are competitive and mutually exclusive because they have the same binding site for the enzyme. Competitive inhibitors are originally thought to be non-metabolizable derivatives of the substrate or another substrate of the enzyme or the reaction product. Typical noncompetitive inhibitors do not affect the binding of the substrate, and the substrate also does not affect the binding of the inhibitor. Substrate ( S ) and Inhibitor ( I ) is reversible and binds to other sites randomly and independently. That is, I binds to enzyme ( E ) or ES and S binds to E or EI . In other words, it is thought that a non-competitive inhibitor binds to and inhibits the catalytic site of the enzyme (Segel IH, In Biochemical calculations , p246, 1976). Nerya O. et al ., J. Agric. Food Chem. , 51 , pp 1201-1207 , 2003, are one of the structural features of noncompetitive inhibitors, with the presence of resorcinol moieties. Competitive inhibitors have the ability to chelate copper in enzymes.

Phellinus is native to the mulberry tree trunk as a wood-clay mushroom (Phellinus linteus) belonging to the mud mushrooms in (Phellinus) of pine scales mushrooms and (Hymenochaetaceae) and is known widely as is because both yellow conditions (桑黃) except hat surface . Fruiting hot water extract of P. linteus has been the subject of many studies as it has been known to have a retarding effect on cancers of the digestive system (Ikekawa, T., et al ., Gann , 59 , pp155-157, 1968). , Inhibitory effect of polysaccharides on tumor growth and metastasis (Sasaki, T., et al ., Chem. Pharm. Bull ., 19 , pp 881-826, 1971; Han, SB, et al ., Immunopharmacol , 41 , pp157 164, 1999), antitumor activity expected by preventing changes in gap junctional intercellular communication (GJIC) in gap binding by hydrogen peroxide (H ₂ O ₂ ) (Cho, JH, et al., Carcinogenesis , 23 , pp1163-1471, 2002), antimutagenic activity that induces the activity of quinone oxidoreductase and glutathione S-transferase and increases glutathione levels to prevent cancer (Shon, YH and Nam, KS, J. Ethnopharmacol. , 77 , pp 103-109, 2001). Oh et al. Revealed that the hydrothermal extract of S. mushrooms enhanced B-lymphocyte function (Oh, GT, et al. , Arch. Pharm. Res. , 15 , pp 379-381, 1992), and that this was due to polysaccharides (Song, KS et al ., Chem. Pharm. Bull. , 43 , pp2105-2108, 1995), Lee et al . (Lee, JH et al ., Chem. Pharm. Bull. , 44 , pp1093-1095, 1996) are acidic. Reported the properties of carbohydrate-peptide binding of heteroglycopeptides, and polysaccharides stimulate humoral and cellular immune responses (Kim, HM, et al ., Int. J. Immunopharmac. , 18 (5) , pp295-303, 1996). Various biological activities of Felinus linteus have been reported. In artificial cultivated mushrooms, it has been reported to have anti-cancer effects by activating beta-glucan (β-glucan) biological immune capacity (Rhee, YK, et al ., J. Food Sci. Technol. , 32 , pp 477-480, 2000).

Mushrooms are generally classified into high molecular polysaccharides and low molecular weight materials. Substances that exhibit pharmacological activity, such as anticancer and immune enhancing effects, of polymer polysaccharides of mushrooms, including situation mushrooms, are known as beta-glucan polysaccharides having a specific structure. These are all beta-1, 3-glucan (β-1,3-glucan) as a backbone, beta-1, 6, glucan (β-1,6-glucan) side by side, X-ray and NMR Has been reported to be poorly soluble in water having a triple helix structure (Chihara, G., et al ., Cancer Res. , 30 , pp 2776-2781, 1970). It is also known that the presence of steric structures is important for the expression of beta-glucan's antitumor activity in the relationship between structure and anticancer activity (Deslandes, Y. et al ., Macromolecules , 13 , pp1466-1471, 1980). In addition, when analyzing polysaccharides having anti-tumor activity, it is reported that glucose is mainly composed of galactose, mannose, arabinose, etc. (Kim) , YS, et al ., Arch. Pharm. Res. , 17 , pp 337-342, 1994). The contents of the protein polysaccharides of the fruiting body and the culture mycelium of the woody mud mushrooms were similar to those of the common polysaccharides. It appears to be different depending on the degree of polymerization and the binding mode of the main chain (Lee, JW, et al ., Kor. J. Mycol. , 27 , pp 424-429, 1999; Lee, JW and Bang, K., Food Ind. Nut., 6 , pp 25-33, 2001). However, it is difficult to rule out the role of the low molecular weight material of the fruiting body is necessary to study.

In addition, as the extract of the situation mushroom is known to prevent skin aging and whitening effect, the Korean cosmetics company has filed a patent and developed a product (Pacific, Korean Patent Application No. 10-1996-57166, Dec. 1998. 04 Registration; Hanbul Cosmetics, Korea Patent Application No. 10-1996-69542, Apr. 08, 1999; Nadri Cosmetics, Korea Patent Application No. 10-2001-10171, 2002. 04. 09 Registration), for the active ingredient Not known at all.                         

Therefore, the present inventors confirmed that the ethyl acetate soluble fraction of the crude mushroom crude extract strongly inhibited the activity of tyrosinase in mushrooms and inhibited the oxidation of L-tyrosinase, and at the same time separated and identified the active ingredients to complete the present invention. .

In order to achieve the above object, the present invention is a compound having a tyrosinase inhibitory activity, provides a whitening cosmetic composition or a composition for preventing browning of food containing a compound that can be separated or synthesized from a soluble mushroom non-polar solvent soluble extract or a mushroom mushroom extract do.

In order to achieve the above object, the present invention provides a whitening cosmetic composition or food browning prevention composition containing a Phellinus linteus non-polar solvent soluble extract.

The situation mushroom may use a cap, edge or fruiting body, preferably fruiting body.

The situation mushroom non-polar solvent soluble extract is a non-polar solvent selected from methylene dichloride (CH ₂ Cl ₂ ), ethyl acetate, n -butanol, and preferably extracted with ethyl acetate.

In addition, the present invention is protocatechualdehyde (Protocatechualdehyde: Compound 1), hydroxymethyl-2-peraldehyde (5-Hydroxymethyl-2-furaldehyde: Compound 2), succinic acid (Succnic acid: Compound 3), fumaric acid (compound 4), 9-Methyl soya-carebroside II (compound 5) and 9-methyl soya-cerbroside II hexaacetate (9- Methyl soya-cerebroside II hexaacetate: Provides a cosmetic composition for whitening containing a combination of a group selected from at least one compound group of compound 6).

In addition, it comprises a composition for preventing browning of food containing one or more compounds selected from the compounds 1 to 6.

The situation mushroom extract is a crude extract or a nonpolar solvent soluble extract, and includes water, a lower alcohol such as ethanol, methanol, butanol, or a mixed solvent thereof, preferably an extract soluble in methanol, and the nonpolar solvent soluble extract is dichloride. Non-polar solvent selected from methylene (CH ₂ Cl ₂ ), ethyl acetate, n -butanol, preferably extracted with ethyl acetate.

Hereinafter, the present invention will be described in detail.

Compounds that can be separated or synthesized from the situation mushroom nonpolar solvent-soluble extract or the situation mushroom extract of the present invention can be obtained as follows.

The crude mushroom extract of the present invention is pulverized after pulverizing the fruit mushroom body after pulverization, powder, water of about 2 to 20 times the dry weight of the situation mushroom, preferably about 2 to 5 times the volume of water, methanol, ethanol And a lower solvent such as butanol or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10, preferably about 0.5 hours to an extraction temperature of 20 to 100 캜, preferably 50 to 100 캜 with methanol. Extraction method such as hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction for 2 days, preferably 1 hour to 1 day, preferably by hydrothermal extraction, 1 to 5 times, preferably 3 times continuous extraction The resulting filtrate was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure at 20 to 100 ° C., preferably at 20 to 70 ° C., using a vacuum rotary concentrator to obtain crude mushroom extracts available in water, lower alcohols or mixed solvents thereof. I can get it.

The crude mushroom extract is suspended in water and then extracted using a solvent in the order of methylene dichloride, ethyl acetate, n -butanol to obtain a soluble extract of the situation mushroom nonpolar solvent of the present invention, preferably an ethyl acetate soluble fraction. More specifically, methane dichloride: water: methanol is mixed with crude mushroom extract, that is, methanol extract, in a ratio, preferably 10: 9: 1, to prepare methylene dichloride soluble fraction and water soluble fraction. may be obtained, again by extracting the soluble fraction in ethyl acetate and to yield the ethyl acetate-soluble fraction and soluble fraction, n in the soluble fraction-butanol-soluble fraction and soluble fraction-n was added to butanol Can be obtained.

By measuring the tyrosinase inhibitory activity of these non-polar solvent soluble fractions, silica gel column chromatography (SiO ₂ column chromatography) can be performed on the fraction having the most excellent activity, preferably ethyl acetate soluble fraction, and methylene dichloride. In a mixed solvent of methanol, preferably methylene dichloride, and a 9: 1 mixed solvent of methanol, column chromatography was carried out at a constant volume per hour, preferably at a rate of 1000 ml, to obtain 12 fractions from F1 to F12. Can be.

The F3 fraction showing the best tyrosinase inhibitory activity among the 12 fractions was searched for, and again subjected to silica gel column chromatography, and a mixed solvent of hexane: ethyl acetate, preferably hexane: ethyl acetate 3: 2 Column chromatography was performed at a constant volume per hour, preferably 1000 mL, with a mixed solvent of to obtain eight fractions of F3-1 to F3-8, in the fractions of F3-2 and F3-4. Compounds 1 and 2 can be isolated and identified respectively.

In addition, the F8 fraction, which shows the next strong tyrosinase inhibitory activity, can be separated by ethyl acetate again by silica gel column chromatography to obtain 9 fractions from F8-1 to F8-9, and Sephadex from the F8-4 fraction. Compounds 3 and 4 can be isolated and identified by LH-20 column chromatography, and F5-5 can be isolated and identified by Sephadex LH-20 column chromatography.

In addition, the compound 5 may be acetylated by adding a suitable solvent, preferably acetic anhydride (Ac ₂ O) / pyridine for 12 to 48 hours to acetylate the compound 6 to be obtained.

The present invention provides a method for separating and purifying the compounds of 1 to 5, which are separated from the soluble mushroom non-polar solvent soluble extract or the lean mushroom extract. Also provided is a method of partially compounding from compound 5 to yield compound 6.

The present invention provides a whitening cosmetic composition or a composition for preventing browning of food containing compounds that can be separated from or synthesized from a situation mushroom non-polar solvent soluble extract or situation mushroom extract obtained by the above method.

Tyrosinase, an enzyme that is essential for melanin biosynthesis, is investigated in order to investigate the efficacy as a whitening cosmetic composition or food browning prevention composition of compounds that can be isolated or synthesized from the situation mushroom non-polar solvent soluble extract or situation mushroom extract obtained above As a result of measuring the inhibitory activity, it was confirmed that the compounds capable of separating or synthesizing from the situation mushroom non-polar solvent soluble extract or the situation mushroom extract according to the present invention effectively inhibited tyrosinase activity.

The present invention provides a cosmetic composition useful for skin whitening wherein the amount of the compound that can be separated or synthesized from the nonpolar solvent-soluble extract or the situation mushroom extract has 0.01 to 30% by weight, preferably 0.01 to 5.0% by weight of the total cosmetic composition. In addition to cleansing, face wash, soaps, shampoos, rinses and treatments, it provides a new use of cosmetic compositions useful for skin whitening.

The composition of the present invention can be used in a variety of cosmetics, face wash and shampoo for the whitening effect. Examples of products to which the composition can be added include cosmetics such as various skins, lotions, creams, essences, nutrients, lotions, patches, and packs, and soaps, cleansing, body cleansers, face washes, and essences. .                     

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 vitamin may be any compound that can be incorporated into cosmetics, but preferably vitamin B ₁ , vitamin B ₂ , vitamin B ₆ , pyridoxine, pyridoxine, vitamin B ₁₂ , pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H and the like, and salts thereof (thiamine hydrochloride, sodium ascorbate salt) and derivatives (ascorbic acid-2-sodium phosphate salt, ascorbic acid-2-magnesium phosphate salt, etc.) can also be used in the present invention. It is included in water soluble vitamins. 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 formulated in cosmetics, but preferably vitamin A, carotene, vitamin D ₂ , vitamin D ₃ , and vitamin E (d1-α-tocopherol, d-α-tocopherol, d-δ-tocopherol) And derivatives thereof (ascorbic palmitate, ascorbic stearate, ascorbic acid dipalmitine, dl-α-tocopherol acetate, dl-α-tocopherolvitamin E, DL-pantothenyl alcohol, D- Pantothenyl alcohol, Pantothenyl ethyl ether, etc.) are contained in the oil-soluble vitamin used by this invention. Oil-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification of microorganism culture, enzyme or chemical synthesis.

The polymer peptide may be any compound as long as it can be incorporated into cosmetics. Preferably, collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin and the like can be given. 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, pit spingosine, sphingolipid lipid and the like can be given. 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, brown seaweed extract, red flush extract, green algae extract, and the like, and carrageenan, arginic acid, sodium arginate and argin purified from these seaweed extracts are preferred. Potassium ginate 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.

In addition to the said essential component, you may mix | blend the cosmetics of this invention with the other components normally mix | blended with cosmetics as needed.                     

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 di (capryl capric acid) Propylene glycol caprylate, neopentyl glycol dicapric acid, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl tritridecyl, glyceryl triisopalmitinate, glyceryl triisostearate, octyldode neopentane Sil, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isstearyl isostearate, Isostearic acid octyldecyl, polyglycerol oleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myritic lactic acid, cetyl lactate, octyl lactate, trit citrate Ethyl, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipic acid, 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-steal One hydroxy stearic acid and the like esters such as cetearyl source.

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

Examples of the silicone-based oils and fats include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane and methylcetyloxysiloxane copolymer, dimethylsiloxane and methylsteoxysiloxane copolymer, Alkyl modified silicone oil, amino modified silicone oil, etc. are mentioned.

Perfluoro polyether etc. are mentioned as fluorine-based fats and oils.

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 (polymerization degree n = 2 or more), polypropylene glycol (Polymerization degree n = 2 or more), polyglycerol (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.

Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerol fatty acid esters, sorbitan fatty acid esters, POE (polyoxyethylene) sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE Glycerin Fatty Acid Ester, POE Alkyl Ether, POE Fatty Acid Ester, POE Cured Castor Oil, POE Castor Oil, POE · POP (Polyoxyethylene Polyoxypropylene) Copolymer, POE / POP Alkyl Ether, Polyether Modified Silicone, Laurin Acid Alkanolamide, alkylamine oxide, hydrogenated soybean phospholipid, etc. are mentioned.

As anionic surfactant, fatty acid soap, (alpha)-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl phosphorus salt, alkylamide Phosphates, alkyloylalkyltaurine salts, N-acylamino acid salts, POE alkyl ether carboxylates, alkyl sulfosuccinates, sodium alkyl sulfoacetates, 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.

Organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium film mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide Inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, calamine, carbon black and composites thereof; 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-β-alanine calcium, N-lauroyl-β-alanine zinc, and N-lauroylglycine calcium; Amide sulfonic acid polyvalent metal salts, such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N-acyl basic amino acids, such as N (epsilon) -lauroyl-L- lysine, N (epsilon) -palmitolysine, N (alpha)-partoylol nitin, N (alpha)-lauroyl arginine, and N (alpha) -cured fatty acid acyl arginine; N-acyl polypeptides, such as N-lauroyl glycyl glycine; α-amino fatty acids such as α-aminocaprylic acid and α-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 salicylate and cinnamic acid benzyl , Paramethoxy cinnamic acid 2-ethoxyethyl, paramethoxy cinnamic acid octyl, diparamethoxy cinnamic acid mono 2-ethylhexane glyceryl, paramethoxy cinnamic acid isopropyl, diisopropyl diisopropyl cinnamic acid ester mixture, urokanoic acid Ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and salts thereof, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenonedisulfonate, dihydroxybenzophenone, tetra Hydroxybenzophenone, 4- tert -butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino- p- (carbo-2'-ethylhexyl-1'-oxy) -1,3 , 5-triazine, 2- (2-hydric When the like can be mentioned 5-methylphenyl) benzotriazole.

As fungicides, hinokithiol, trichloric acid, trichlorohydroxydiphenyl ether, chlorohexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, ginphylthione, benzalkonium chloride, Photosensitizer No. 301, sodium mononitroguicol, undecylenic acid, etc. are mentioned.

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 it is 0.01-3 weight% compounding.

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

Examples of the form of the cosmetic composition are not particularly limited, and examples thereof include emulsions, creams, lotions, patches, packs, foundations, lotions, essences, soaps, and the like.

Specific examples of the cosmetic of the present invention face wash, face wash foam, cleansing cream, cleansing milk, cleansing lotion, massage cream, cold cream, moisturizing cream, emulsion, lotion, patch, pack, aftershave cream, sunburn prevention cream, suntan oil , Soap, body shampoo, press powder, lu spout, eye shadow, hand cream, lipstick and the like.

The cosmetic of the present invention is selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extracts in addition to compounds that can be separated or synthesized from essential mushrooms, non-polar solvent-soluble extracts or situation mushroom extracts, which are essential ingredients Ingredients (such as compounding ingredients which may be added in addition to those exemplified above as required) may be added to known methods, for example, to the methods described in, for example, "Dermal Application Formulation Manual", Matsumoto Michio Supervision Edition (issued by Seishi, 1985) It can obtain by preparing accordingly.

In addition, the present invention provides a whitening cosmetic additive containing compounds that can be separated from or synthesized from a situation mushroom nonpolar solvent-soluble extract or a situation mushroom extract.

The cosmetic additives can be used in the production process or finished products such as existing cosmetics and cleansing agents, and can be used, and has the function of skin whitening.

It can also be used in skins, creams, lotions, essences, nutrients, cosmetics, patches, massage packs, systemic cleansers, soaps, cleansing agents, face washes, beauty veins and other care preparations.

Situation mushroom non-polar solvent soluble extract of the present invention or compounds that can be separated or synthesized from the situation mushroom extract can be used safely in long-term use for the purpose of prevention because there is little toxicity and side effects.

In another aspect, the present invention provides a composition for preventing browning of food as an active ingredient, which can be separated or synthesized from the situation mushroom non-polar solvent soluble extract or the situation mushroom extract obtained by the production method.

In addition, compounds that can be separated or synthesized from the situation mushroom non-polar solvent soluble extract or the situation mushroom extract of the present invention can be used as a food additive for preventing browning of food.

At this time, the food additive may be added to the food by immersing, spraying or mixing the food, the ratio of such additives is generally selected from the range of 0.01% to 10%.

The foods are fruits, vegetables, dried or cut products of fruit or vegetables, fruit juices, vegetable juices, mixed juices thereof, chips, noodles, livestock processed foods, fish processed foods, dairy products, fermented milk foods, cereal foods, microorganisms. It includes any one or more of fermented foods, bakery, condiments, meat processing, acidic beverages, licorice, herbs.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following Examples and Experimental Examples are merely 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 crude mushroom crude extract

The situation mushrooms used in the present invention were tested by receiving lyophilized powder from Joze Pharmaceutical Co., Ltd., and the samples were stored in the laboratory of the Pukyong University (No. 20020304). After lyophilization, 4.5 kg of the powder obtained by lyophilization was added with 10 liters of methanol, and reflux-cooled was extracted three times at a predetermined time interval (12 h, 6 h, 3 h) at 70 ° C. After filtration under reduced pressure, the filtrate extract was removed from methanol at 40 ° C. with a vacuum rotary concentrator to obtain 838 g of crude mushroom extract as a residue, and 2 g of the extract was used as a sample for enzyme activity detection.

Example 2. Preparation of Situation Mushroom Methylene Dichloride Soluble Fraction

4 l of a mixed solvent of methylene dichloride: water: methanol = 10: 9: 1 was added to 838 g of the situation mushroom crude extract obtained in Example 1, followed by mixing three to four times to obtain 2 l of water-soluble fraction and dichloride. After obtaining 2 L of methylene soluble fraction, methylene dichloride soluble fraction was dried to obtain 101.5 g of dry powder of methylene dichloride soluble fraction, which was used as a sample, and 2 g of this was used as a sample for enzyme activity screening.

Example 3. Preparation of Situation Mushroom Ethyl Acetate Soluble Fraction

2 L of ethyl acetate was added to 2 L of the water-soluble fraction obtained in Example 2, followed by mixing three to four times to obtain 2 L of the water-soluble fraction and 2 L of the ethyl acetate soluble fraction. The ethyl acetate soluble fraction was dried. 36.7 g of a dry powder of an ethyl acetate soluble fraction was obtained and used as a sample, and 2 g of this was used as a sample for enzyme activity screening.

Example 4 Situation Mushroom ｎ Preparation of Butanol Soluble Fraction

2 L of butanol was added to 2 L of the water-soluble fraction obtained in Example 3, followed by mixing 3 to 4 times to obtain 2 L of the water-soluble fraction and 2 L of the butanol-soluble fraction, followed by drying the water-soluble fraction and the butanol-soluble fraction. 244 g of the dry powder of the water-soluble fraction and 366 g of the dry powder of the butanol-soluble fraction were obtained and used as a sample, 2 g of which was used as a sample for enzyme activity screening.

Example 5. Isolation of Compounds

Silica gel column chromatography was performed on 36.7 g of the situation mushroom ethyl acetate soluble fraction of Example 3, which showed the tyrosinase inhibitory activity of each of the solvent soluble fractions and showed the most excellent activity. At this time, a mixed solvent of methylene dichloride: methanol 9: 1 was used as a developing solvent, and the fractionation was performed at 1000 ml / hour using Kiesel gel 60 (Kiesel gel 60, 230-400 mash) as a fixed phase, After dividing into 12 lower fractions up to F12, the F3 fraction (20.76 g) showing the best tyrosinase inhibitory activity of each fraction was measured, and silica gel column chromatography was performed. At this time, a mixed solvent of hexane: ethyl acetate 3: 2 was used as a developing solvent, and the fractionation was performed by 1000 ml / hour using Fiesel gel 60 (Kiesel gel 60, 230-400 mesh) as a fixed phase. From 1 to 8 subfractions of F3-8, 376.8 mg of protocatechualdehyde (Compound 1) of Formula 1 and 5-hydroxymethyl-2-perform of Formula 2 from F3-2 and F3-4 Each 1.74 g of aldehyde (Compound 2) was isolated.                     

Next, F8 fraction (6.33 g) showing strong tyrosinase inhibitory activity was subjected to silica gel column chromatography using ethyl acetate as a developing solvent, and divided into nine subfractions from F8-1 to F8-9. Sepadex LH-20 column chromatography was performed to separate 47 mg of succinic acid (Compound 3) and 36 mg of fumaric acid (Compound 4) of Formula 3, respectively, and F8-5. A precipitate formed by Sephadex LH-20 column chromatography (49 mg) was purified by a C ₁₈ sep-pak cartridge to obtain 9-methyl-soya-cerebroside II (Compound 5) ) 18 mg was obtained.

In addition, the reaction solution containing 0.3 ml of acetic anhydride (Ac ₂ O) and 0.2 ml of pyrimidine in Compound 5 (18 mg) was allowed to stand at room temperature for 24 hours, and then the reaction solution was evaporated with nitrogen gas. 14.9 mg of 9-methyl-soya-cerebroside II hexaacetate (compound 6) of 6 was obtained.

Analysis of each compound by various analyzers such as NMR and MS is as follows.

(1) Compound 1: Protocatechualdehyde                     

Form of matter: Yellowish crystals.

EI-MS m / z (R. int): 138 (92) [M ⁺ ], 137 (100) [MH], 109 (73), 81 (65), 63 (33), 53 (34).

IR Ｖ _max (KBr) cm ⁻¹ : 3222, 3329, 1652, 1596, 1535, 1445, 1297, 1165.

¹ H-NMR (CD ₃ OD): 9.68 ( CH 0), 7.30 (dd, J = 1.70, 9.48 Hz), 7.28 (d, J = 1.41 Hz), 6.90 (d, J = 8.1 Hz).

¹³ C-NMR (CD ₃ OD): 193.8 ( C HO), 154.5 (C-4), 148.0 (C-3), 131.6 (C-1), 127.2 (C-6), 117.0 (C-5) , 116.2 (C-2).

(2) Compound 2: 5-hydroxymethyl-2-peraldehyde                     

Form of material: Yellow oily phase.

EI-MS m / z: 126 [M ⁺ ].

IR ｖ _max (KBr) cm ⁻¹ : 3380 (OH), 2848, 1655 (C═O).

¹ H-NMR (CD ₃ OD): 9.52 (-CHO), 7.37 (1 H, d, J = 3.5 Hz, H-3), 6.67 ( ₁ H, d, J = 3.5 Hz, H-4), 4.60 (2H, s, CH ₂ OH).

¹³ C-NMR (CD ₃ OD) δ: 180.2 (-CHO), 164.0 (C-5), 154.7 (C-2), 125.6 (C-3), 111.7 (C-4), 58.4 (CH ₂ OH ).

(3) Compound 3: Succinic Acid                     

Form of material: White crystalline form.

Molecular formula of material: C ₄ H ₆ O ₄ .

¹ H-NMR (MeOH- d ₄ , 400 MHz) δ: 2.55 (4H, s, CH ₂ ),

¹³ C-NMR (MeOH- d ₄ , 100 MHz) δ: 30.6 (CH ₂ ), 176.9 (COOH).

(4) Compound 4: Fumaric Acid

Form of material: White crystalline form.

Molecular formula of material: C ₄ H ₄ O ₄ .

¹ H-NMR (MeOH- d ₄ , 400 MHz) δ: 6.75 (2H, s, CH).

¹³ C-NMR (MeOH- d ₄ , 100 MHz) δ: 135.9 (CH), 168.8 (COOH)

(5) Compound 5: 9-methyl soya-cerebroside II

Appearance of substance: white powder,

Molecular formula of substance: C ₄₁ H ₇₇ NO _9,

Negative-FABMS m / z 726.5 [M-H] < + >,

Positive-FABMS m / z 750.4 [M + Na] ⁺ , 710.5 [MH ₂ O + H] ⁺ , 548.4 [M-glucose + H] ⁺

HR-FABMS m / z: 750.5498 [M + Na] ⁺ (calculated for C ₄₁ H ₇₇ NO ₉ Na, m / z 750.5496, Δ +0.2 mmu).

IR Ｖ _max (KBr) cm ⁻¹ : 3406 (OH), 2923, 2853, 1655, 1546 (amide), 1466, 1083 (glycocidic CO), 878, 720 [(CH ₂ ) n], 637.

¹ H-NMR (pyrimidine- d ₅ , 400 MHz) δ: 8.36 (1H, d, J = 8.7, NH), 6.00 (1H, dd, J = 15.4, 5.9 Hz, H-4), 5.92 (1H , dt, J = 15.5, 5.2 Hz, H-5), 5.24 (1H, t, J = 7.0 Hz, H-8), 4.91 (1H, d, J = 7.8 Hz, H-1 ''), 4.80 ( 1H, m, H-2), 4.75 (1H, m, H-3), 4.71 (1H, dd, J = 10.2, 3.9 Hz, H-1a), 4.56 (1H, m, H-2 ′), 4,50 (1H, dd, J = 10.2, 2.5 Hz, H-6 ″ a), 4.34 (1H, dd, J = 11.7, 5.2 Hz, H-6 ″ b), 4.23 (1H, dd, J = 10.2, 3.8 Hz, H-1b), 4.21 (2H, m, H-3 ″, 4 ″), 4.02 (1H, t, J = 8.0 Hz, H-2 ″), 3.89 (1H, m, H− 5 ″), 2.14 (2H, t, J = 4.5 Hz, H-6), 1.99 (3H, t, J = 7.5 Hz, H-10, 3′b), 1.78 (1H, m, H-4 ′ b), 1.69 (1H, m, H-4'a), 1.60 (3H, s, H-19), 1.39 (2H, m, H-7), 1.26-1.23 [s, (CH2) n], 1.21 (2H, m, H-15 ') 0.84 (6H, t, J = 6.6 Hz, H-18, 16').

¹³ C-NMR (pyrimidine- d ₅ , 100 MHz) δ: 136.0 (C-9), 132.3 (C-5), 131.8 (C-4), 124.1 (C-8), 105.6 (C-1 ″ ), 78.5 (C-5 ″), 78.4 (C-3 ″), 75.0 (C-2 ″), 72.4 (C-2 '), 72.3 (C-3), 71.4 (C-4 ″), 70.1 (C-1), 62.6 (C-6 ″), 54.5 (C-2), 39.9 (C-10), 35.6 (C-3 '), 33.0 (C-6), 32.0 (C-16, 14 ') 29.9 ~ 29.5 (C-11 ~ 15, 5' ~ 13 '), 28.1 (C-7), 25.8 (C-4'), 22.9 (C-17, 15 '), 16.0 (C-19) , 14.2 (C-18, 16 ')

(6) Compound 6: 9-methyl soya-cerebroside II hexaacetate

¹ H-NMR (CDCl ₃ , 400 MHz) δ: 6.33 (1H, d, J = 9.1 Hz, NH), 5.82 (1H, d, J = 15.1 Hz, H-4), 5.39 (1H, dd, J = 15.2, 7.4 Hz, H-5), 5.31 (1H, t, J = 6.9 Hz), 5.18 (1H, t, J = 9.5 Hz), 5.15 (1H, dd, J = 7.3, 4.9 Hz), 5.07 (2H, t, J = 9.7 Hz), 4.95 (1H, dd, J = 8.0, 9.5 Hz), 4.47 (1H, dd, J = 8.0 Hz, H-1 ''), 4.30 (1H, m), 4.23 (1H, dd, J = 12.3, 4.5 Hz), 4.13 (1H, dd, J = 12.4, 2.3 Hz), 3.93 (1H, dd, J = 10.3, 3.9 Hz), 3.69 (1H, ddd, J = 10.0 , 4.5, 2.4 Hz), 3.61 (1H, dd, J = 10.4, 4.5 Hz), 2,05 (3H, t, J = 3.1), 2.17 (COC H ₃ ), 2.10 (COC H ₃ ), 2.03 ( COC H ₃ ), 2.02 (2 × COC H ₃ ), 2.00 (COC H ₃ ), 1.94 (2H, t, J = 7.2 Hz), 1.79 (2H, m), 1.57 (3H, s, C H ₃ ) , 1.25 [(C H ₂ ) n], 0.87 (6H, t, J = 6.8 Hz, 2 × C H ₃ )

¹³ C-NMR (CDC _l3 , 100 MHz) δ: 170.6, 170.2, 169.8, 169.7, 169.6, 169.4, 169.2 (all Me C O, NH C O), 136.8, 136.2, 124.5, 122.9, 100.5, 73.9, 73.1 , 72.7, 71.9, 71.2, 68.2, 67.2, 61.8, 50.6, 39.7, 32.6, 31.9, 31.9, 31.8, 29.7 ~ 29.3, 28.0, 27.4, 24.7, 22.7, 21.0 (CO C H ₃ ), 20.9 (CO C H ₃ ), 20.7 (CO C H ₃ ), 20.6 (3 × CO C H ₃ ), 16.0, 14.1, 14.1

Experimental Example 1. The Tyrosinase Inhibitory Effect of Crude Mushroom Extract and Solvent Fraction

Tyrosinase inhibition effect of crude mushroom extract and each solvent fraction was measured by slightly modifying the method of No. et al. (No, JK, et al. , Life Science , 65 , pp241-246, 1999) using L-tyrosine as a substrate . .

First, 170 μl of a reaction mixture obtained by mixing 1 mM L-tyrosine (Jansen Chimikasa), 50 mM potassium phosphate buffer (pH 6.5) and distilled water at a ratio of 10: 10: 9 was obtained in Example 1. A crude mushroom extract (methanol extract) and 10 µl of each solvent fraction obtained in Examples 2 to 4 and 20 µl of an aqueous solution of mushroom tyrosinase (1000 units / ml) were added to prepare a 96-well microparticle. The plate was placed in a microplate. After the reaction solution was incubated at 25 ° C. for 30 minutes, the amount of dopachrome produced was measured at a wavelength of 490 nm in a microplate reader (Versa max, Moleccular Devices, Calif., USA).

The active unit (U) of this enzyme was determined by the amount of enzyme that produced 1 μole of dopachrome per minute under the conditions mentioned above. The tyrosinase inhibitory activity% was calculated by the following equation. In the following equation, the absorbance of the sample is the absorbance at 490 nm without the inhibitor, the standard absorbance at 490 nm without the inhibitor and the control absorbance at 490 nm without the inhibitor and without the enzyme. Absorbance, and the absorption coefficient of dopachrome at 490 nm is 3.3 x 10 ³ . Inhibitory activity was expressed as the mean of 50% inhibitory concentration of three values obtained by interpolation of the concentration-inhibition curve. Meanwhile, Kojisan (Sigma) was used as a control.

% Inhibition = {1- (sample absorbance-controlled absorbance) / standard absorbance} × 100

As a result of measuring the tyrosinase inhibitory activity of the methanol extracts of the situation mushrooms and their respective solvent fractions, the IC ₅₀ value was 21.95 ㎍ / ml in ethyl acetate soluble fractions as shown in FIG. Tyrosinase inhibitory activity was shown in the order of methylene soluble fraction> n -butanol soluble fraction> methanol soluble fraction> water soluble fraction (see Table 1).

Therefore, the highest activity ethyl acetate soluble fractions were rescanned for tyrosinase inhibitory activity in the 12 lower fractions (F1 to F12) obtained by silica gel column chromatography, among which Compounds 1 and 2 were found to be the most active F3. , And compounds 3 to 5 were isolated and identified at F8, respectively.

sample IC ₅₀ ^a (μg / ml)   Raw Mushroom Extract (Methanol Extract) 846.3   Situation Mushroom Methylene Dichloride Soluble Fraction 339.2   Situation Mushroom Ethyl Acetate Soluble Fraction 21.9   Situation Mushroom Butanol Soluble Fraction 769.8   Situation Mushroom Number Soluble Fraction > 1000   Koji-san (control) 3.1 ^a inhibitory activity was expressed as the mean of 50% inhibitory concentrations of three values obtained by interpolation of concentration-inhibition curves.

Experimental Example 2 Tyrosinase Inhibitory Effect of Compounds Isolated from Situation Mushroom

Tyrosinase inhibitory activity of the five compounds obtained in Example 5 was performed in the same manner as in Experimental Example 1.

As a result, as can be seen in Table 2, the tyrosinase inhibitory activity of protocatechualdehyde (1) and 5-hydroxymethyl-2-peraldehyde (5-HMF) (2) was IC ₅₀ values of 0.4 and 90.8 μg. / Ml, protocatechualdehyde showed about 10 times better activity than the positive control kojic acid (3.1 μg / ml).

sample IC ₅₀ ^a (μg / ml)   Protocatechualdehyde (1) 0.4   5-hydroxymethyl-2-peraldehyde (2) 90.8   Succinic acid (3) > 500   Fumal (4) > 500   9-Methyl Soya-Serebroside II (5) > 500   Koji-san (control) 3.1 ^a inhibitory activity was expressed as the mean of 50% inhibitory concentrations of three values obtained by interpolation of concentration-inhibition curves.

Experimental Example 3. Kinetic analysis of the compounds isolated from the situation mushroom

Kinetic analysis of protocatechualdehyde (1) and 5-hydroxymethyl-2-peraldehyde (5-HMF) (2) in the compound obtained in Example 5 was measured by the following method.

First, the reaction solution was prepared in mushroom tyrosinase dissolved in 50 mM potassium phosphate buffer in four different concentrations from 0.5 mM to 2 mM using L-tyrosine as a substrate, followed by protocatechualdehyde (1) and 5-hydride. Roxymethyl-2-peraldehyde (5-HMF) (2) was added to the reaction solution at various concentrations, respectively, and the dopachrome production was measured at 490 nm. (Michaelis constant) of the tyrosinase Km, K _i (inhibition constant) of Vmax (maximal velocity) and inhibitors line Weber-Burke plot was calculated by (Lineweaver-Burk plots).

The K _i values of protocatechualdehyde (1) and 5-hydroxymethyl-2-peraldehyde (5-HMF) ( 2 ) are 1.1 × 10 ⁻⁶ M and 1.4 × 10 ⁻³ M, which are not comparable to competitive inhibitors. Each appeared as a competitive inhibitor (see FIGS. 2A and 2B).

Hereinafter, a formulation example of the cosmetic composition will be described, but the present invention is only intended to be described in detail and not intended to limit it.

Soap manufacturers

Situation Mushroom Ethyl Acetate Soluble Fraction of Example 3 ... 1-30 (%)

Maintenance ...

Sodium hydroxide ...............

Sodium Chloride .........................................

Spices Quantity

The whole amount was 100 with purified water, and soap was prepared by the above blending ratio.

Lotion Manufacturing 1

Situation Mushroom Ethyl Acetate Soluble Fraction of Example 3 ...... 3.00 (%)

L-ascorbic acid-2-phosphate magnesium salt ......... 1.00

Water Soluble Collagen (1% Aqueous Solution) ............ 1.00

Sodium Citrate ... 0.10

Citric Acid ........................................... 0.05

Licorice Extract ......... 0.20

1,3-butylene glycol ... 3.00

The whole amount was 100 with purified water, and a lotion was prepared by the above blending ratio.

Lotion Manufacturing 2

Situation Mushroom Ethyl Acetate Soluble Fraction of Example 3. 3.00 (%)

L-ascorbic acid-2-phosphate magnesium salt ......... 1.00

Water Soluble Collagen (1% Solution) ....................... 1.00

Sodium Citrate ... 0.10

Citric Acid ........................................... 0.05

1,3-butylene glycol ... 3.00

The whole amount was 100 with purified water, and a lotion was prepared at the above compounding ratio.

Lotion Manufacturing 3

Situation Mushroom Ethyl Acetate Soluble Fraction of Example 3 ..... 4.00 (%)

Brown algae extract ......................................... 1.00

Sodium Citrate ... 0.10

Citric Acid ........................................... 0.05

1,3-butylene glycol ... 3.00

The whole amount was 100 with purified water, and a lotion was prepared at the above blending ratio (%).

Cream manufacturers

Situation Mushroom Ethyl Acetate Soluble Fraction of Example 3 ........ 1.00 (%)

Polyethylene Glycol Monostearate ............... 2.00                     

Self-emulsifying glycerin monostearate ......................... 5.00

Cetyl Alcohol ... 4.00

Squalene ......................................... 6.00

Glyceryl tri2-ethylhexanoate ......... 6.00

Sphingose Sugar Lipids ......................................... 1.00

1.3-Butylene Glycol ... 7.00

The whole amount was 100 with purified water, and a cream was prepared at the above compounding ratio.

Pack manufacturer

Situation Mushroom Ethyl Acetate Soluble Fraction of Example 3 ........ 5.00 (%)

Polyvinyl Alcohol ... 13.00

L-ascorbic acid-2-phosphate magnesium salt ......... 1.00

Lauroylhydroxyproline ............... 1.00

Water Soluble Collagen (1% Aqueous Solution) ............... 2.00

1,3-butylene glycol ... 3.00

Ethanol ......................................... 5.00

The whole amount was 100 with purified water, and a pack was prepared at the above compounding ratio.

Essence

Situation Mushroom Ethyl Acetate Soluble Fraction of Example 3 ... 2.00 (%)                     

Hydroxyethylene Cellulose (2% Aqueous Solution) ... 12.00

Xanthan gum (2% aqueous solution) ....................... 2.00

1,3-butylene glycol ......... 6.00

Dark glycerin ... 4.00

Sodium Hyaluronate (1% Aqueous Solution) ......................... 5.00

The whole amount was 100 with purified water, and a cosmetic liquid was prepared at the blending ratio (%).

Compounds that can be isolated or synthesized from the soluble mushroom non-polar solvent soluble extract or the soluble mushroom extract of the present invention effectively inhibit tyrosinase activity, an enzyme that is essential for melanin biosynthesis, and thus can be used as a browning prevention composition or a composition for preventing browning, such as food Can be.

Claims (11)

Whitening cosmetic composition containing Phellinus linteus ethyl acetate soluble extract. delete delete Situation Mushroom Ethyl Acetate Separated or Synthesized from Soluble Extracts One of 5-hydroxymethyl-2-peraldehyde, succinic acid, fumaric acid, 9-methyl soya-cerebroside II and 9-methyl soya-cerebroside II hexaacetate A cosmetic composition for whitening containing a combination of one group selected from the above compound group. delete The cosmetic composition according to claim 1 or 4, wherein the composition is used in cosmetics such as skins, lotions, creams, essences, nutrients, lotions, patches and packs. The cosmetic composition according to claim 1 or 4, wherein the composition is used as a soap, cleansing, whole body cleanser, face wash, and cosmetic liquid. Situation Mushroom Ethyl Acetate Separated or Synthesized from Soluble Extracts One of 5-hydroxymethyl-2-peraldehyde, succinic acid, fumaric acid, 9-methyl soya-cerebroside II and 9-methyl soya-cerebroside II hexaacetate Cosmetic additives for skin whitening containing a combination of the group selected from the above compound group. The method of claim 8, wherein the additives are skins, creams, lotions, essences, nutrients, cosmetics, patches, massage packs, systemic cleansers, soaps, cleansing agents, face washes, cosmetics and other care preparations Cosmetic Additives Usable For. delete delete

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