KR101179589B1 - Composition comprising an extract of Magnolia officinalis Rehd. et Wils. or 4-O-methylhonokiol compound isolated therefrom for skin whitening - Google Patents

Abstract

The invention Magnolia (Magnolia officinalis Rehd . meat Wils . The present invention relates to a whitening composition containing an extract or a compound separated therefrom as an active ingredient. Specifically, the thick extract of the present invention and 4-O-methylhonokiol isolated therefrom are tyrosinase. Since the inhibitory effect and the inhibitory effect of melanin biosynthesis in B16 melanoma cells can be usefully used as an external skin pharmaceutical composition and cosmetic composition having a skin whitening effect.

Thick Extract, 4-O-Methyl Honokiol, Whitening

Description

A composition for skin whitening containing hydroxya extract or 4-O-methyl-honokiol separated therefrom as an active ingredient {Composition comprising an extract of Magnolia officinalis Rehd. et Wils. or 4-O-methylhonokiol compound isolated therefrom for skin whitening}

      The present invention relates to a skin whitening composition containing a thick extract extract or a compound separated therefrom as an active ingredient exhibiting a skin whitening effect.

Human skin color is determined by the amount of melanin, keratin and hemoglobin, of which melanin is the most decisive factor. Melanin is synthesized in melanocytes, the pigment cells in the basal layer of the skin, and transferred to the surrounding keratinocytes to give a human skin color (Kubo M. et al., Fragrance) . Journal , 8 , p 48, 1995).

When melanin is abnormally produced by external environmental factors, it may cause pigmentation such as blemishes and freckles, and may cause skin cancer. In particular, when skin cells are stimulated by ultraviolet rays, a signal for melanin production is generated by tyrosinase, an important enzyme in melanin synthesis, and these have a direct or indirect effect on melanocytes, leading to the production of melanin pigment (Seiji). M et al., Ann . NY Acad . Sci ., 100 , p497, 1963). Such melonin pigment production can be inhibited by using tyrosinase substrate analogs. Recently, melanin is produced by tyrosinase as well as tyrosinase related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2, Dopachrome tautomerase) as an important enzyme involved in the biosynthesis of melanin. It has been newly discovered that developing a substance that can inhibit these enzymes may be a way to prevent skin pigmentation (Marmol V. et al., FEBS Letter , 327 , p 307, 1993; Winder AJ et al., Pigment Cell Research , 7 , p305, 1994).

      Melanin synthesis inhibitors have been applied to the treatment of skin diseases, skin whitening agents, food browning inhibitors, etc. in medicine, cosmetics, food, etc. Recently, the demand is rapidly increasing, and as a result, various whitening agents have been developed and used, but various problems are raised. .

     Looking at the example of the prior invention published as a whitening agent, the method which can show a whitening effect by the use of ascorbic acid (Japanese Unexamined-Japanese-Patent No. 4-9320) is proposed. However, since ascorbic acid itself is a problem of phase stability in the prescription system, its active ingredient activity decreases with time, and various methods such as stabilization with capsules and liposomes have recently been proposed. have. Hydroquinone (Japanese Patent Laid-Open No. 6-192062) is used locally for the treatment of hyperpigmentation such as blemishes, freckles, spots and hyperpigmentation in pregnancy. Although these compounds show potent melanogenesis inhibitory activity, they are only allowed in some countries due to side effects such as degeneration or lethality of pigment cells and impaired cell function. have. Kojic acid (Japanese Patent Laid-Open Publication No. 56 ~ 7710) has excellent whitening effect due to its excellent inhibitory ability of tyrosinase, the only enzyme involved in the transformation of tyrosine into dopa and dopachrome. It is becoming. Arbutin (Japanese Patent Laid-Open No. Heisei 4 ~ 9315) has been proven to inhibit tyrosinase as well as koji acid.However, arbutin itself is hydroquinone after the sugar is separated by skin enzymes when it is applied to cosmetics with sugar attached to the hydroquinone. Skin irritation may be a concern, and there are problems such as low inhibitory activity, discoloration during use, and instability of the substance itself.

      Accordingly, there is an urgent need for a new whitening agent that is safe without side effects and can have a complex effect in actual clinical trials, rather than a substance that is involved in only one element.

Magnolia officinalis Rehd . et Wils.) has been used as a health agent for a long time in Korea, Japan, and China, and leafy leaves have been used for food in Korea and Japan (Information, Shinmingyo, Hyangjeomsa, Yeonglimsa, p.469, 1998). , magnolia, the alpha, beta-u death Mall (α, β-eudesmol), Magda nolol (magnolol), hono kiol (honokiol), also known as the given include obovatol (obovatol) (pharmacognosy Research, modern pharmacognosy , Hakchangsa, p.567, 2003). Currently used in Korea, the Chinese pepper is Magnolia which belongs to Magnolia. officinalis Rehd. et Wils ., Magnolia officinalis var. Magnolia, a deciduous tree belonging to biloba and Magnoliaceae. obovata ) and the Korean evergreen tree ( Machilus ), an evergreen tree belonging to the family Lauraceae. thunbergii , Machilus thunbergii var. obovata ), and depending on where it is used, it is used as a substitute for thick or thick pepper (Lee SR et al., Kor. J. Pharmacogn ., 17 , p199, 1986). Recently, a study using a thick gourd in Korea has been filed a patent for diabetic inhibitors (Jin-Suk Kim, Korea Patent Application No. 2003-0039241) utilizing the efficacy of the magolol component, anti-anxiety inhibitor using the ovobatol component Patent (Hong Jin-tae et al., Korean Patent Application No. 2005-0037854) has been filed. In 1978, 4-O-methylhonokiol was first isolated from bakbak (El-Feraly, FS et al., Lloydia, 41 , p493, 1978). Known to have insecticidal effects on mosquito larvae and brine shrimp (Nitao JK et al., Phytochemistry , 30 , p2193, 1991), and have neuroprotective and neurotrophic effects. Also known is Tomoyuki E. et al., Bioorganic & Medicinal Chemistry Letters , 14 , p2621, 2004). Republic of Korea Patent Application No. 2003-0027093 is a domestic hobak ( Machilus) belonging to the camphor family It relates to a whitening cosmetic composition containing a crude extract of thunbergii ) or a non-polar solvent-soluble extract as an active ingredient. However, Magnolia still belongs to Magnolia officinalis ) and No whitening effect or tyrosinase inhibitory activity by 4-O-methyl honokiol has been reported.

Accordingly, the present inventors have completed the present invention by confirming that the extract of hoobac and the compound isolated therefrom have an excellent effect on skin whitening through tyrosinase inhibitory effect and melanin biosynthesis inhibitory effect in B16 melanoma cells.

An object of the present invention to provide a skin whitening pharmaceutical composition and cosmetic composition for skin whitening containing a thick extract or a compound isolated therefrom as an active ingredient.

      In order to achieve the above object, the present invention is for skin whitening containing 4-O-methyl honokiol (4-O-methyl honokiol) represented by Formula (I) Provides an external skin pharmaceutical composition.

      In addition, the present invention provides a cosmetic composition for skin whitening containing 4-O-methyl honokiol represented by the following formula (I), which is isolated from a thick leaf extract or an active ingredient.

      The grommets extract is characterized in that it is extracted from the stem bark, seeds, leaves or roots of the grommets, preferably stem bark or leaves, more preferably stem bark.

      Extracts as defined herein are characterized as being crude extracts or nonpolar solvent soluble extracts.

      The crude extract as defined herein includes extracts available in water, including purified water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof, preferably water and ethanol mixed solvents, more preferably 75 to 100% ethanol. do.

      The nonpolar solvent soluble extract as defined herein is a solvent selected from hexane, chloroform, methylene chloride, ethyl acetate, glycerin, propylene glycol, butylene glycol or ether, preferably hexane, chloroform or ethyl acetate, more preferably in hexane Contains available extracts.

       Hereinafter, a method of obtaining a thick extract of the present invention will be described in detail.

      The thick gourd extract of the present invention is dried from the leaves or stem bark of the gourd, about 1 to 20 times the weight of the water, preferably about 2 to 5 times the volume of water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, Preferably a mixed solvent of water and ethanol, more preferably about 75-100% ethanol at an extraction temperature of about 0-100 ° C., preferably about 10-30 ° C., about 1 hour to 15 days, preferably about 2 Extraction method such as hot water extraction, reflux cooling extraction, ultrasonic extraction for 7 to 7 days, preferably the extract extracted by the reflux cooling extraction method under reduced pressure filtration at about 20 to 100 ℃, preferably about 50 to 70 ℃ After concentration under reduced pressure, the crude thin extract of the present invention can be obtained.

       In addition, the thick thin non-polar solvent soluble extract of the present invention is dispersed in about 1 to 15 times, preferably about 1 to 10 times the volume of water of the obtained thick thin extract extract, hexane, chloroform, methylene chloride, ethyl acetate , Glycerin or propylene glycol, preferably hexane, chloroform or ethyl acetate, more preferably hexane is fractionated 1 to 5 times, preferably 2 to 4 times by adding a volume of about 0.1 to 0.5 times the amount of water. A nonpolar solvent soluble extract can be obtained.

        In addition, the compounds of the present invention can be separated as follows.

       The hexane solvent extract of the non-polar solvent extract obtained by the above obtaining method is dissolved in a lower alcohol having 1 to 4 carbon atoms, preferably methanol, and adsorbed onto a C18 column, and then lower alcohol having 1 to 4 carbon atoms, preferably methanol: water. 10: 1 to 4: 1 to obtain a fraction, adsorbed onto silica gel, and then subjected to silica gel column chromatography two to five times to separate the active fraction, and finally using high performance liquid chromatography. 4-O-methylhonokiol can be obtained.

       4-O-methylhonokiol of the present invention is commercially available or can be obtained by the above method.

       The present invention provides a skin whitening pharmaceutical composition for skin whitening containing a thick extract or a compound isolated therefrom as an active ingredient.

       The composition for skin whitening of the present invention comprises 0.1 to 50% by weight of the extract or a compound separated therefrom based on the total weight of the composition.

       The external skin pharmaceutical composition containing the thick extract of the present invention or the compound separated therefrom as an active ingredient may further include appropriate carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions.

      External skin pharmaceutical composition containing a thick extract according to the present invention or a compound isolated therefrom as an active ingredient, external preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc. It can be formulated and used in the form of sterile injectable solutions, and preferably provides an external skin pharmaceutical composition of creams, gels, patches, sprays, ointments, warnings, lotions, linens, pastas or cataplasmas .

      Carriers, excipients and diluents that may be included in the composition comprising the thick extract of the present invention or a compound separated therefrom as an active ingredient include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol , Starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate And mineral oils. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. Or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The skin whitening composition containing the thick extract of the present invention or the compound separated therefrom as an active ingredient is a skin external preparation formulation that can be applied to the skin as a cream, gel, patch, spray, ointment, warning agent, lotion, linen Preparation, or may be used as a pharmaceutical composition in the form of an external preparation for skin of pasta or cataplasma, but is not limited thereto.

       The preferred dosage of the composition containing the thick extract of the present invention or a compound separated therefrom as an active ingredient depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and is appropriately selected by those skilled in the art. Can be. However, for the preferred effect, the composition of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg per day. Topical administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

       In addition, the present invention provides a cosmetic composition for skin whitening containing a thick leaf extract obtained by the above manufacturing process or a compound separated therefrom as an active ingredient.

       In addition, as a medicinal herb that has been used for a long time as a herbal medicine and edible, as a medicinal herb extract of the present invention or a compound isolated therefrom also has no problems such as toxicity and side effects, and has been proved to be a non-irritating sample in the skin irritation test. You can use it with confidence.

      The composition containing the thick extract of the present invention or a compound isolated therefrom can be used in various ways, such as cosmetics and shampoos for skin whitening effect. Examples of products to which the present composition can be added include cosmetics such as various creams, lotions, skins, essences, shampoos, rinses, cleansing agents, face washes, soaps, treatments, packs, 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 vitamins may be any compound that can be incorporated into cosmetics, but preferably vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H, and the like. Their salts (thiamine hydrochloride, sodium ascorbate salt, etc.) and derivatives (ascorbic acid-2-sodium phosphate salt, ascorbic acid-2-magnesium phosphate salt, etc.) may also be used in the water-soluble vitamins that can be used in the present invention. Included. The water-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification from culture of microorganisms, enzymatic or chemical synthesis.

       Usable vitamins include vitamins such as vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol) Alpha-tocopherol vitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenyl ethyl laurate, and the derivatives thereof (palmitic acid ascorbin, stearic acid ascorbic acid, dipalmitic acid ascorbin, dl-alpha tocopherol acetate, Ether, etc.) are also included in the usable vitamins used in the present invention. Oil-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification of microorganism culture, enzyme or chemical synthesis.

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

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

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

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

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

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

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

      As ester fats and oils, glyceryl tri2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl mystinate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl acid isetyl, isostyl acid mystearyl, isostaryl palmitate, octylate acid octyldodecyl, isostearic acid isetyl, diethyl sebacinate, adidi Diisopropyl phosphate, isoalkyl neopentane, tri (capryl, capric acid) glyceryl, trimethyl ethyl trimethylolpropane, trimethyl stearate trimethylol propane, tetra 2-ethylhexanopentane Tol, cetyl caprylate, lauric acid decyl, lauric acid hexyl, myristic acid decyl, myristic acid myristyl, myristic acid cetyl, stearyl stearate, decyl oleate, lisinooleic acid Teyl, isostearyl laurate, isotridecyl myristin, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyl dodecyl oleate, octyl dodecyl linoleate, isopropyl isopropyl acid, 2-ethylhexanoate cetostearyl, 2-ethylhexanoate stearyl, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicapric acid, propylene glycol di (capryl, capric acid), Dicaprylic acid propylene glycol, dicapric acid neopentyl glycol, dioctanoate neopentyl glycol, tricaprylate glyceryl, triundecyl glyceryl, triisopalmitinate glyceryl, triisostearate glyceryl, octylate octadodode Sil, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isstearyl isostearate, Sothete octylate, polyglycerol oleate, polyglycerine isostearate, triisocetyl citrate, triisoalkyl citrate, triisoctyl citrate, lauryl lactate, myritic lactate, octyl lactate, octyl lactate, tricitric acid Ethyl, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di2-ethyl hexyl 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.

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

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

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

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

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

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

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

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

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

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

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

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

       Examples of the anionic surfactant include fatty acid soap, alpha-acylsulfonate, alkylsulfonate, alkylallylsulfonate, alkylnaphthalenesulfonate, alkylsulfate, POE alkylether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl phosphate, alkylamide phosphate , Alkyloyl taurine salts, N-acyl amino acid salts, POE alkyl ether carboxylates, alkylsulfosuccinates, sodium alkylsulfoacetates, acylated hydrolyzed collagen peptide salts, and perfluoroalkyl phosphate esters .

       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 carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amide sulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type and amideamine type. An amphoteric surfactant etc. are mentioned.

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

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

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

       As fungicides, hinokithiol, trichloric acid, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, ginxitlionone, benzalkonium chloride, photosensitive Sodium No. 301, sodium mononitroguicol, undecylenic acid, and the like.

       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, any of the above-mentioned components may be blended within the range not to impair the objects and effects of the present invention, but it is preferably 0.01 to 5% by weight based on the total weight, More preferably from 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 components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the thick extract or the compound separated therefrom as an active ingredient, for example, stabilizers, solubilizers, vitamins, pigments and Conventional adjuvants such as perfumes and carriers.

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

       Specifically, the cosmetic composition of the present invention skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisturizing cream, hand cream, foundation, essence, nutrition essence, Formulations of packs, soaps, cleansing foams, cleansing lotions, cleansing creams, hair lotions, hair tonics, hair essences, hair shampoos, hair rinses, hair treatments, body lotions and body cleansers.

       When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide are used as carrier components. Can be.

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

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

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

       When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

       However, the following Examples and Experimental Examples are only illustrative of the present invention, the contents of the present invention is not limited by the following Examples, Reference Examples and Experimental Examples.

Example  Thick Of crude extract (cm-1)  Produce

Buying groceries at Kyungdong Market ( Magnolia) officinalis Rehd . meat Wils . After drying and cutting the tree, 3L of stem bark was added with 9L of 95% ethanol and extracted three times at room temperature, and the filtrate was filtered and concentrated under reduced pressure with EYELA (N-1000, Japan). 450 g of the extract was obtained and used in the following experimental example (hereinafter referred to as cm-1).

Example  2. Thick non-polar solvent soluble extract ( cm -2) Preparation

      200 g of the crude thin extract obtained in Example 1 was suspended in 2 L distilled water, and then dissolved by adding 500 ml of hexane, and then fractionally extracted. This process was repeated twice to collect a portion dissolved in the hexane layer, followed by hexane. The layer was concentrated under reduced pressure to obtain 70 g of a thick hexane soluble extract, which was used in the following experimental example (hereinafter referred to as cm-2).

Example  3. 4-O- from Thick Extract Methylhonokiol  detach

      After dissolving 70 g of the hydrated hexane extract obtained in Example 2 in 300 ml of methanol and adsorbed on a 300 g of C18 column, a fraction was obtained using a mixed solution of methanol: water = 4: 1 (v / v), The eluate was concentrated under reduced pressure to give 40 g of a yellowish brown concentrate, which was dissolved in methylene chloride. Then, the mixture was adsorbed onto 1 kg of silica gel (Merck, 9385) packed in a column (4.5x40 cm) using a mixed solution of hexane: ethyl acetate = 9: 1, and the ratio of hexane and ethyl acetate was 9: 1. The silica gel column chromatography was carried out a total of two times while converting to 6: 4 in order to separate the fractions, and finally, 4-O-methylhonokiol having the following physical properties by using high performance liquid chromatography under the conditions shown in Table 1 below. 2 g was obtained and used in the following experimental example.

¹ H-NMR was analyzed by Varian (Garian, GEMINI, 400 MHz) and ¹³ C-NMR by Varian (Varian, GEMINI, 100 MHz), and analyzed by CDCl ₃ (Chloroform- d , Aldrich). ) Was used as the solvent, trimethylsilane was used as the internal standard, and the δ value (ppm) was used, and the NMR result was confirmed to be consistent with the published data (Shoji Y. et al., Chem) . . Pharm. Bull. 39, p2024 , 1991).

¹ H-NMR (400 MHz, CDCl ₃ ): δ ppm 3.45 (2H, d, J = 6.0 Hz, H-7 '), 3.45 (2H, d, J = 6.0 Hz, H-7), 3.90 (3H, s, OMe), 5.10 (4H, m, H-9 and H-9 '), 6.01 (2H, m, H-8 and H-8'), 6.95 (1H, dd, J = 8, 1.5 Hz, H-3 '), 6.97 (1H, dd, J = 8.0, 1.5 Hz, H-3), 7.05 (2H, m, H-2 and H-6'), 7.23 (1H, dd, J = 8.5, 1.6 Hz, H-6), 7.30 (1H, doublet of doublets, J = 8.5, 1.6 Hz, H-4 ′);

¹³ C-NMR (100 MHz, CDCl ₃ ): δ ppm 129.09 (C-1), 130.67 (C-2), 128.67 (C-3), 156.94 (C-4), 115.53 (C-5), 127.86 ( C-6), 34.22 (C-7), 136.48 (C-8), 115.47 (C-9), 127.83 (C-1 '), 150.79 (C-2'), 110.88 (C-3 '), 129.64 (C-4 '), 132.09 (C-5'), 130.46 (C-6 '), 39.36 (C-7'), 137.77 (C-8 '), 115.77 (C-9'), 55.47 ( OMe).

Column  Shim-Pak prep-ODS column, Shimachusa Column temperature
(column temperature) 30 ℃ Flow rate  18 mL / min Detector  UV 254 nm Injection volume
(injection volume)  400 μl Mobile phase  80% methanol

Reference Example  1. Purity Analysis

In order to determine the purity of the thick extract and 4-O-methyl honokiol obtained in Examples 1 to 3, purity analysis was performed using high performance liquid chromatography (HPLC) under the conditions described in Table 2 below. It is shown in Table 3 below.

Column Capsule Park C ₁₈ 4.6 X 250mm, Shiseido Column temperature
(column temperature)  30 ℃ Flow rate  1.0 mL / min Detector  UV 254 nm Injection volume
(injection volume)  20 μl Mobile phase  80% methanol

Experimental material Content of 4-O-Methyl Honokiol (%) Cm-1 of Example 1 12.50 Cm-2 of Example 2 30.28

     As a result, as shown in Table 3, the cm-2 of Example 2 was 30.28%, the content of 4-O-methyl honokiol was much higher than the cm-1 of Example 1 which is 12.50%.

Experimental Example  One. Mushroom Tyrosinase  Activity inhibitory effect

Mushroom tyrosinase activity by Vanni et al. (A. Vanni, Annali di Chimica , 80 , p35, 1990) to find out whether the thick extract or the compound isolated therefrom prepared in Examples 1 to 3 involved in the inhibition of melanin biosynthesis Inhibitory effect was measured.

1.0 ml of 0.1 M potassium phosphate buffer (pH 6.8), 1.0 ml of 0.3 mg / ml aqueous L-tyrosine solution and 0.1 ml of mushroom tyrosinase (Sigma, T-7755) at 1,250 U / ml concentration, followed by 0.2 ml of the sample solution prepared in Examples 1 to 3 was added at a concentration of 20, 30, 50, 100, 200 or 300 µg / ml, and the enzyme reaction was performed at 37 ° C for 10 minutes. (Sigma) was used. The absorbance of the reaction solution was measured at 480 nm, and the inhibition rate of tyrosinase activity was determined by Equation 1 below. The results are shown in Table 4 below. The result of inhibiting the tyrosinase activity by 50% was determined as an IC ₅₀ value. It is shown in Table 5 below.

      Mushroom tyrosinase inhibition rate (%) = [(A-B) / A] × 100

       A: Absorbance at 480 nm of reaction solution without sample

B: absorbance at 480 nm of reaction solution to which sample was added

Experimental material Concentration (µg / ml) Tyrosinase activity inhibitory effect (%) Cm-1 of Example 1 100 12.75 200 38.45 300 60.23 Cm-2 of Example 2 50 16.38 100 43.15 200 68.56 4-O-methylhonokiol of Example 3 20 25.07 30 49.01 50 67.12 Arbutin (control) 100 20.36 200 35.89 300 57.56

 As a result, as shown in Table 4, 4-O-methyl honokiol was found to be the most excellent inhibitory effect of tyrosinase activity compared to the concentration of 67.12% tyrosinase activity even at 50㎍ / ㎖.

Experimental material IC ₅₀ (μg / ml) Cm-1 of Example 1 256 Cm-2 of Example 2 163 4-O-methylhonokiol of Example 3 35 Arbutin (control) 278

As a result, as shown in Table 5, in the 4-O-methyl honokiol of Example 3, the IC ₅₀ value of 35 ㎍ / ㎖ showed the highest effect of inhibiting mushroom tyrosinase, cm-1, Both cm-2 of Example 2 showed a higher inhibitory effect on mushroom tyrosinase compared to the control. Therefore, by inhibiting the tyrosinase activity involved in the initial stage of melanogenesis, it was found that the thick extract of the present invention or 4-O-methylhonokiol isolated therefrom has an excellent effect in inhibiting melanin production and showing a whitening effect. .

Experimental Example  2. B16 Melanoma  Cytotoxicity Test Using Cells

By measuring the cell viability of B16 melanoma cells (ATCC CRL6323) using cm-1 of Example 1, cm-2 of Example 2 and 4-O-methylhonokiol of Example 3 as samples, Experiments were conducted as follows (Maeda and Fukuda, J. Soc. Cosmetic Chem ., 42 , p361, 1991).

B16 melanoma cells were inoculated in Dulbecco's modified Eagle's Medium (DMEM) medium with 10% bovine serum at a density of 1 × 10 ⁵ cells and cultured at 5% CO ₂ , 37 ° C. for one day. Thereafter, the cells were changed to fresh DMEM 10% medium, and the samples were treated with wells at a concentration of 20, 50, 100, 200, or 300 µg / ml, and then incubated for 3 days, and then the medium was removed and 0.33 mg / ml of MTT (( 1 ml of 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide) and Sigma M5655) were reacted at 37 ° C for 4 hours, and then MTT was removed and DMSO ( 1 ml of dimethyl sulfoxide) was added to measure absorbance at 575 nm to determine the color development. The results are shown in Table 6 below.

Experimental material Concentration (µg / ml) Cell survival rate (%) Cm-1 of Example 1 100 98.67 200 91.65 300 90.75 Cm-2 of Example 2 100 98.56 200 96.52 300 94.56 4-O-methylhonokiol of Example 3 20 99.62 50 97.56 100 95.12 Arbutin (control) 50 98.58 100 95.58 200 96.05

      As a result, as shown in Table 6, B16 melanoma cells cultured by treating cm-1 of Example 1 or cm-2 of Example 2 showed the best cell viability at 98.56 to 98.67% at 100 µg / ml. The B16 melanoma cells cultured by treatment of cm-1 of Example 1, cm-2 of Example 2, or 4-O-methylhonokiol of Example 3 had a cell viability of 90% or more. Therefore, it was found that the extract or the compound isolated therefrom is a safe substance with little toxicity.

Experimental Example 3. Inhibitory Effects of Melanin Biosynthesis Using B16 Melanoma Cells

The method of Maeda and Fucus in order to investigate the inhibitory effect of melanogenesis in the B16 melanoma cells of cm-1 of Example 1, cm-2 of Example 2 and 4-O-methylhonokiol of Example 3 Fukuda, J. Soc. Cosmetic Chem ., 42 , p361, 1991) was modified with the following experiment.

B16 melanoma cells (ATCC CRL6323) were seeded in Dulbecco's modified Eagle's Medium (DMEM) with 10% bovine serum at a density of 1 × 10 ⁵ cells and incubated at 5% CO ₂ , 37 ° C. for one day. Thereafter, the cells were changed to DMEM 10% medium treated with α-MSH (melanocyte stimulating hormone; melanocyte stimulating hormone, 0.34 μg / ml, Sigma), cm-1 of Example 1, cm-2 of Example 2, and The 4-O-methylhonokiol of Example 3 was treated with 1, 5, 10, 50 or 100 μg / ml in wells and incubated for 3 days. After 3 days, the medium was collected in a 15 ml flask tube and the cells were trypsinized. After treatment and removal, the medium was transferred to a collected tube, and then centrifuged at 3,000 rpm for 30 minutes to discard the supernatant, and treated with 250 ml of 1N sodium hydroxide for 10 minutes in boiling water, followed by light centrifugation. After sonication for 10 minutes to completely dissolve the melanin absorbance at 475 nm, the results are shown in Table 7 below.

Experimental material Concentration (µg / ml) Melanin production inhibitory effect (%) Cm-1 of Example 1 10 1.78 50 37.85 100 57.56 Cm-2 of Example 2 10 6.89 50 68.56 100 89.87 4-O-methylhonokiol of Example 3 One 5.05 5 48.07 10 78.86 Arbutin (control) 10 1.25 50 6.87 100 17.52

      As a result, as shown in Table 7, 4-O-methylhonokiol showed the most excellent melanogenesis inhibitory effect at 78.86% at 10 μg / ml. In addition, cm-1 of Example 1, cm-2 of Example 2 showed a higher melanogenesis inhibitory effect compared to the control group 1.78% to 1.78 to 6.89% at 10 μg / ㎖. Therefore, it was found that the extract of gourd and the compound separated therefrom have an excellent effect on skin whitening by inhibiting melanin production.

Experimental Example 4. Skin irritation test

In order to determine the skin irritation of the extracts of pear extracts or the compounds isolated therefrom, the New Zealand White male rabbits were used internally in the laboratory according to the Korea Food and Drug Administration Notice 1999-61, "Drug Toxicity Test Standard." A stimulation test was performed. Observation of skin irritation reactions such as erythema, scalp formation, edema formation to the patch site 24 hours after the patch.

As a result of applying cm-1 of Example 1, cm-2 of Example 2, and 4-O-methylhonokiol of Example 3 to the skin of rabbits, no general symptoms were observed. Was not observed. In addition, mild erythema was found as a result of erythema and skin formation, but as a result of stimulus evaluation, it was classified as a weak irritant and was evaluated as a substance with little irritation.

      Examples of the formulation of the composition are described below, but are not intended to limit the present invention but to explain in detail only.

Formulation example  1. flexible lotion ( skin )

       Butylene glycol, glycerin, polyoxyethylene (60) hardened castor oil, betaine, citric acid, sodium citrate, and preservatives were added to the purified water, followed by stirring and dissolving. The thick gourd extract obtained in Example 1 was added thereto, stirred well, and aged to prepare a flexible gourd extract containing a thick gourd extract. Table 8 shows the content of each component.

Raw material Formulation Example 1
Content (w / w%) Cm-1 of Example 1 50.0 Butylene Glycol 7.0 glycerin 5.0 Polyoxyethylene (60) hardening castor oil 0.2 ethanol 5.0 Betaine 2.0 Citric acid 0.02 Sodium citrate 0.06 antiseptic a very small amount Spices a very small amount Purified water Balance

Formulation example  2. Lotion ( emulsion )

 The thick gourd extract, butylene glycol, glycerin, carboxyvinyl polymer, arginine, preservative and purified water obtained in Example 2 were heated with stirring at 70 to 75 ℃. Squalane, butylene glycol dicaprylate / dicaprate, sorbitan stearate, polysorbate 60, glyceryl stearate and stearyl glycerate mixture, which were heated by stirring at 75 to 80 ° C. were added thereto, followed by emulsification. When the mixture was cooled to about 45 ° C. while stirring, the fragrance was added thereto, stirred, cooled to 30 ° C., and aged to prepare a thick extract extract-containing lotion. Table 9 shows the content of each component.

Raw material Formulation Example 2
Content (w / w%) Cm-2 of Example 2 40.0 Butylene glycol 8.0 glycerin 5.0 Squalane 10.0 Butylene Glycol Dicaprylate / Dicaprate 5.0 Sorbitan stearate 1.5 Polysorbate 60 1.0 Glyceryl stearate 0.5 Stearyl glycyrrhetinate 0.2 Carboxyvinyl polymer 0.1 Arginine 0.1 antiseptic a very small amount Spices a very small amount Purified water Balance

Formulation example  3. Essence

Cytocyterol, polyglyceryl 2-oleate, ceramide, ceteareth-4 and cholesterol were mixed by stirring, followed by emulsification by addition of a mixed solution of 4-O-methylhonokol, dicetylphosphate, concentrated glycerin and purified water. Then, when the mixture was cooled to 45 ° C. while stirring, the fragrance was added and stirred, and the mixture was cooled to 30 ° C. and aged. Carboxyvinyl polymer, xanthan gum and preservatives were added thereto, stabilized, and aged to prepare 4-O-methyl-honokinool-containing essence. Table 10 shows the content of each component.

Raw material Formulation Example 3
Content (w / w%) 4-O-methylhonokiol of Example 3 10.0 Cytostolol 1.70 Polyglyceryl 2-oleate 1.50 Ceramide 0.7 Setares-4 1.2 cholesterol 1.5 Dicetyl phosphate 0.4 Concentrated glycerin 5.0 Carboxyvinyl polymer 0.2 Xanthan gum 0.2 antiseptic a very small amount Spices a very small amount Purified water Balance

Formulation example  4. Cleanser (Cleansing Foam)

 Sodium N-acyl glutamate, glycerin, PEG-400, and propylene glycol were added to the purified water, followed by mixing. Then, a small amount of 4-O-methylhonokiol was added thereto, and EDTA-4Na was added thereto, followed by heating and stirring at 80 ° C. to dissolve. To this, POE (15) oleyl alcohol ether, laurin derivative and methyl paraben mixed solution heated at 80 ° C. were added and stirred, followed by the addition of fragrance, followed by gradual cooling to prepare 4-O-methylhonokiol-containing cleanser. Table 11 shows the content of each component.

Raw material Formulation Example 4
Content (w / w%) 4-O-methylhonokiol of Example 3 20.0 N-acyl glutamate 20.0 glycerin 10.0 PEG-400 15.0 Propylene glycol 10.0 POE (15) oleyl alcohol ether 3.0 Lauric derivatives 2.0 Methyl paraben 0.2 EDTA-4Na 0.03 Spices 0.2 Purified water Balance

Formulation example  5. Preparation of Hydrophilic Ointment

number Raw material Content (% by weight) Content (% by weight) One  4-O-methylhonokiol of Example 3 0.5 - 2   White vaseline 250 250 3   Stearyl alcohol 220 220 4   Ethyl (or methyl) p-oxybenzoate 0.25 0.25 5   Propylene glycol 120 120 6   Sodium lauryl sulfate 15 15 7   Propyl p-oxybenzoate 0.15 0.15

Thick leaf extract of the present invention and the compound isolated therefrom have tyrosinase inhibitory effect and melanin biosynthesis inhibitory effect in B16 melanoma cells, so it can be usefully used as a skin external pharmaceutical composition and cosmetic composition for skin whitening.

Claims (8)

delete delete delete delete delete delete Water and Ethanol Mixed Solvent Soluble Crude Extract of Magnolia officinalis Rehd. Et Wils. Stem Bark or Non-polar Solvent Soluble Extract Soluble in Solvents Selected from Hexane, Chloroform or Ethyl Acetate A cosmetic composition for skin whitening containing (4-O-methylhonokiol) as an active ingredient. delete