KR20080105526A - Composition comprising an extract of magnolia officinalis rehd. et wils. or 4-o-methylhonokiol isolated therefrom having anti-inflammatory, anti-allergy and anti-wrinkle activity - Google Patents

Abstract

A pharmaceutical composition for the skin preparation for external use, and a cosmetic composition are provided to improve antiinflammatory effect, anti-allergic effect and wrinkle prevention. A pharmaceutical composition for the skin preparation for external use contains an extract of Magnolia officinalis Rehd. et Wils., or the 4-O-methyl honokiol represented by the formula I separated from the extract as an effective ingredient. The extract is extracted from the stem bark, fruits, leaves or roots of Magnolia officinalis Rehd. et Wils.

Description

Anti-inflammatory, anti-allergic and anti-wrinkle composition comprising hydroxya extract or 4-O-methyl-honokiol isolated therefrom as an active ingredient. Composition comprising an extract of Magnolia officinalis Rehd. et Wils. or 4-O-methylhonokiol isolated therefrom having anti-inflammatory, anti-allergy and anti-wrinkle activity}

      The present invention relates to an anti-inflammatory, anti-allergic and anti-wrinkle composition containing a thick extract or a compound isolated therefrom as an active ingredient.

      Allergy is a type of hypersensitivity reaction due to imbalance, which is a biochemical phenomenon that shows a specific and modified response to foreign substances (antigens). Diseases include atopic dermatitis, allergic asthma, rhinitis and food allergies. 10-40% of the world's population suffers from various allergic diseases. In the normal human immune system, the Th1 response regulates cellular immunity and the Th2 response regulates humoral immunity. In allergic patients, an overly Th2 response can cause this balance to break, leading to a variety of adverse reactions, such as excessive increases in blood IgE (immunoglobulin E) or excessive secretion of histamine.

      Upon exposure to antigen, it is delivered to T-lymphocytes, releasing cytokines such as interleukin 4, 5, 6 and granulocyte-macrophage stimulator (GM-CSF). Cytokines react with B-lymphocytes in lymph nodes, and B-lymphocytes self-modify plasma cells to secrete IgE antibodies. IgE antibodies are attached to mast cells and basophils, resulting in histamine, serotonin, leukotriene B4, C4, D4, E4, platelet activator, prostaglandin D, interleukin 1, 3, 4, 5, 6, GM-CSF, tumor necrosis factor -a, bradykinin, etc. are released to mediate the inflammatory response.

      In order to treat or prevent allergies, excessive Th2 responses should be suppressed, Th1 responses should be efficiently induced to stabilize the T-immune response, and directly or indirectly act on B cells producing excessive IgE to block inflammatory responses mediated by IgE. shall.

      The inflammatory response induced by allergies is as follows.

     Inflammation is an immune response of the human body in response to wounds or diseases, and oxidative stress such as ultraviolet rays, free radicals, and free radicals activate inflammatory factors, causing various diseases and aging of the skin. One of the hallmarks of inflammation is the increased oxygenation of arachidonic acid, which is metabolized by the cyclooxygenase (COX) producing prostaglandins and the 5-lipoxygenase pathway producing leukotriene. Prostaglandins and leukotrienes are mediators of inflammation. Thus, therapies designed to inhibit cyclooxygenase and / or lipoxygenase activity are of great interest. Cyclooxygenase enzymes come in two forms: cyclooxygenase-1 and cyclooxygenase-2. The latter form, ie, cyclooxygenase-2, appears to play an important role in the progression of inflammation. Thus, inhibiting cyclooxygenase-2 enzyme may be an effective way to reduce inflammation without the side effects associated with irreversible cyclooxygenase-1 inhibition.

      Another powerful inflammatory mediator, nitric oxide (NO), is produced from L-arginine by NO synthetase (NOS) and is produced in many types of cells by UV-like stress, endotoxins, and cytokines. . These inflammatory stimuli increase the expression of inducible NOS in the cell, generating NO, activating the macrophages and causing an inflammatory response.

      Interleukin-8 (IL-8) is a chemotactic factor produced by several cell types, including monocytes, fibroblasts, endothelial cells and keratinocytes. Its production from endothelial cells is induced by interleukin-1, Tumor Necrosis Factor or lipopolysaccharide. IL-8 stimulates numerous functions in vitro. It is shown to have chemical attracting properties for neutrophils, T-lymphocytes and basophils. In addition, it not only induces histamine release from basophilic cells of both normal and atopic individuals, but also induces lysosomal enzyme release and respiratory explosion from neutrophils. IL-8 has also been shown to increase the surface expression of Mac-1 (CD11b / CD18) on neutrophils without protein neosynthesis, which may contribute to increased adhesion of neutrophils to vascular endothelial cells. Many diseases are characterized by massive neutrophil infiltration.

The outermost layer of skin, the epidermal layer, is produced from the dermal layer, which consists of collagen and elastin. After giving moisture to the skin, it gives elasticity inde two components of the dermis, the enzyme that breaks down the enzyme collagenase and elastin to break down collagen elastase I are active, and losing skin elasticity causing wrinkles (Funtional Cosmetology , society of cosmetic chemists of Japan , p163-180 ) . In addition, Matrix Metallo-Preoteinase (MMP) -1 is a metalloproteinase having zinc at its active center and is secreted in the form of latent protease (zymogen) in vivo. In order to have enzymatic activity, structural modifications occur, resulting in cleavage and activation of amino terminal sites. The activated MMPs are regulated by inhibitors such as 2-macroglobulin or TIMPs (tissue inhibitors of metalloproteinase) and the skin keratinocytes ( Many cells, including keratinocytes and fibroblasts, secrete MMP (Fisher, GJ et al., Photochem . Photobiol . 69 , 154 , 1999). Even in one UV irradiation, MMP activity in the skin is increased, and the collagen in the skin is significantly disrupted, so that MMPs affect collagen breakdown of the dermis and play a very important role in light aging.

       Existing therapeutic agents can be broadly divided into steroidal anti-inflammatory drugs or nonsteroidal anti-inflammatory drugs and antihistamines or antileukotrienes. Most of the former have a strong immunosuppressive action, so it may have a temporary sedative effect in a short time, but from mild side effects such as nausea to severe side effects such as growth inhibition or osteoporosis, the liver takes many problems. The latter may show a temporary relief, but there are side effects such as drowsiness and dizziness, which limits its use. Therefore, there is an urgent need for substances derived from natural products that can be applied more safely as well as efficacy and effect.

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 ( Machilus thunbergii, Machilus ), an evergreen tree belonging to the family Lauraceae. 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. Korean Patent Application No. 2003-0027093 has reported about a cosmetic composition for whitening containing crude extract or non-polar solvent soluble extract of Machilus thunbergii , which is a domestic thick pepper belonging to Camphoraceae. However, Magnolia still belongs to Magnolia officinalis ) and The anti-inflammatory, anti-allergic and anti-wrinkle effect of 4-O-methylhonokiol has not been reported.

 Therefore, the inventors of the present invention, the extract of the hoobak extract and compounds isolated therefrom inhibit the iNOS activity, iNOS, COX2 gene expression inhibitory effect, PGE2, IL-6, IL-8 production inhibitory effect, antihistamine activity enhancement, elastase inhibitory effect and MMP The present invention was completed by confirming that there is an excellent effect on anti-inflammatory, anti-allergic and wrinkle improvement through -1 expression inhibition effect.

It is an object of the present invention to provide an anti-inflammatory, anti-allergic and anti-wrinkle pharmaceutical composition and cosmetic composition containing the thick extract or a compound separated therefrom as an active ingredient.

      In order to achieve the above object, the present invention is an anti-inflammatory, anti-inflammatory containing 4-O-methyl honokiol (4-O-methyl honokiol) represented by the following formula (I) Provides an external skin pharmaceutical composition for allergy and wrinkle improvement.

      In addition, the present invention is an anti-inflammatory, anti-allergic and anti-wrinkle cosmetics containing 4-O-methyl honokiol represented by the following formula (I) isolated from the thick extract or an active ingredient as an active ingredient To provide a composition.

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

      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 hexane Contains extracts available for use.

       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 leaf or stem bark of the thick gourd of about 2 to 20 times, 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 2 to 20 times, preferably about 2 to 5 times the volume of water of the crude thin extract obtained above, and then 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 an anti-inflammatory, anti-allergic and anti-wrinkle pharmaceutical composition containing the thick extract obtained by the above method or a compound separated therefrom as an active ingredient.

       The anti-inflammatory, anti-allergic and anti-wrinkle composition 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. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are 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 are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspending agents, liquid solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The anti-inflammatory, anti-allergic and anti-wrinkle composition containing the thick extract of the present invention or a 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, It may be used as a pharmaceutical composition in the form of a lotion, linen, pasta or cataplasma external preparation, 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.

      The present invention also provides an anti-inflammatory, anti-allergic and anti-wrinkle cosmetic composition containing a thick extract obtained by the above manufacturing process or a compound separated therefrom as an active ingredient.

      The anti-inflammatory, anti-allergic and anti-wrinkle composition containing the thick extract of the present invention or a compound separated therefrom as an active ingredient comprises 0.1 to 50% by weight of the extract or compound, based on the total weight of the composition.

      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-irritant sample in skin irritation test You can use it with confidence.

      The composition containing the thick extract of the present invention or a compound separated therefrom can be used in various ways, such as cosmetics and shampoos for anti-inflammatory, anti-allergic and wrinkle improving 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 microorganism culture, enzyme or chemical synthesis.

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

      The 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 can be normally purified from a mammal or fish.

      The sphingolipid may be any compound as long as it can be blended into cosmetics. Preferably, ceramide, phytosphingosine, sphingosaccharide lipid, etc. may be mentioned. Sphingolipids 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.

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 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, triisooctyl citrate, lauryl lactate, myritic lactate, octyl lactate, octyl lactate Ethyl, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malic acid, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipicate, diisopropyl sebacinate, Dioctyl sebacate, cholesteryl stearate, cholesteryl isostearic acid, 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.

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 B (polymerization degree n = 2 or more), polypropylene Glycol (polymerization degree n = 2 or more), polyglycerol B (polymerization degree n = 2 or more), lactic acid, lactic acid salt, etc. are mentioned.

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

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

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

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

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

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

      Anionic surfactants include fatty acid soaps, alpha-acyl sulfonates, alkyl sulfonates, alkyl allyl sulfonates, alkyl naphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, and alkylamide phosphates. , Alkyloylalkyl taurine salt, N-acylamino acid salt, POE alkyl ether carboxylate, alkyl sulfosuccinate, sodium alkyl sulfo acetate, acyl hydrolyzed collagen peptide salt, perfluoroalkyl phosphate ester, etc. are mentioned. .

      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, oxide Inorganic pigments such as aluminum, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, coloramine 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-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.

      Moreover, the compounding component which may be added other than this is not limited to this, In addition, although all the 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 percentage with respect to gross weight. More preferably 0.01-3% by weight.

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

       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 anti-inflammatory, anti-allergic and anti-wrinkle cosmetic compositions of the present invention may be prepared in any formulation commonly prepared in the art, for example, emulsion, cream, lotion, pack, foundation, lotion, essence, hair cosmetic, etc. Can be mentioned.

      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 carriers, vegetable fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide, etc. may be used as carrier components. Can be.

      When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular, in the case of spray, additionally chlorofluorohydrocarbon, Propellant such as propane / butane or dimethyl ether.

      When the formulation of the present invention is a solution or emulsion, a solvent, solvating or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

       When the dosage form of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can 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 (hc-1)  Produce

3 kg of stem bark obtained from the dried and shredded bark of Magnolia officinalis Rehd. Et Wils. , Which was obtained from Kyungdong Market, was added with 9 L of 95% ethanol and extracted three times at room temperature, and the filtrate was filtered under reduced pressure. It was concentrated under reduced pressure (EYELA, N-1000, Japan) to obtain 450 g of dried crude extract, which was used in the following experimental example (hereinafter, referred to as hc-1).

Example 2. Preparation of Thick Non-Polar Solvent Soluble Extract (hc-2)

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

Example 3. Separation of 4-O-Methyl Honokiol from Thick Extract

      After dissolving 70 g of the hydrated hexane extract obtained in Example 2 in 300 ml of methanol and adsorbed on a 300 g C18 column, a fraction was obtained using a mixed solution of methanol: water = 4: 1 (v / v). The eluent 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  30 ℃ Flow rate  18 mL / min Detector  UV 254 nm 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  30 ℃ Flow rate  1.0 mL / min Detector  UV 254 nm Injection volume  20 μl Mobile phase  80% methanol

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

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

Experimental Example 1. Experiment on Cytotoxicity

      By measuring the cell viability of the human fibroblasts (ATCC 2076) using the hupak extract prepared in Examples 1 to 3 or a compound isolated therefrom as a sample, the experiment as follows to determine whether they are toxic Was performed.

Human fibroblasts (ATCC 2076) were inoculated into a 24-well culture plate of IMDM (Iscove's Modified Dulbecco's Medium, GIBCO) containing 10% bovine serum at a concentration of 5x10 ⁴ cells / ml, and serum-free 24 hours after inoculation. After replacement with medium, test samples were added at a concentration of 0.5, 1, 5 or 10 ug / ml, incubated in a 37 ° C., 5% CO ₂ incubator for one day, then the supernatant was removed and again 5% PBS (phosphate buffered saline). After washing by adding 200ul each, MTT solution was added 1.0ml per well, MTT was removed after 4 hours, and then incubated at 37 ℃ overnight with DMSO 1.0ml per well, and then measured for absorbance at 570nm Cell viability was calculated using 1, and the results are shown in Table 4 below.

Cell viability (%) = (B OD _{570 λ} value / A OD _{570 λ} value) × 100

A: control group (test group without sample)

B: sample (hc-1 of Example 1, hc-2 of Example 2 or 4-O-methylhonokiol of Example 3) having a concentration of 0.1, 0.5, 1, 5 or 10 ug / ml

sample Cell survival rate (%) Control 100  0.1 ug / ml of hc-1 of Example 1 110.3  0.5 ug / ml of hc-1 of Example 1 103.4 1 ug / ml of hc-1 of Example 1 96.4 5 g / ml of hc-1 of Example 1 97.2 Hc-1 10ug / ml of Example 1 100.6  0.1 ug / ml of hc-2 of Example 2 112.1  0.5ug / ml of hc-2 of Example 2 98.8 Hc-2 1ug / ml of Example 2 97.3 Hc-2 5ug / ml of Example 2 100.2  10 ug / ml of hc-2 of Example 2 107.4 0.1 ug / ml 4-O-methylhonokiol of Example 3 108.1  0.5 ug / ml 4-O-methylhonokiol of Example 3 99.8 1 ug / ml 4-O-methylhonokiol of Example 3 102.9 5 ug / ml 4-O-methylhonokiol of Example 3 105.3 10 ug / ml 4-O-methylhonokiol of Example 3 100.7

      As a result, as shown in Table 4, the fibroblasts treated with 4-O-methyl-honokiol were 99.8 to 108.1%, the highest cell viability, hc-1 of Example 1, hc of Example 2 All of the fibroblasts treated with -2 or 4-O-methylhonokiol of Example 3 had a cell viability of 95% or more. Therefore, it was found that the thick extract and the compound separated therefrom are safe substances with little toxicity.

Experimental Example 2. Anti-allergic experiment

      In order to measure the anti-allergic effect of the hupak extract prepared in Examples 1 to 3 and the compound separated therefrom, the experiment was performed as follows.

1 x 10 ⁵ RBL-2H3 mast cells were suspended in 15% FBS-MEM (Fetal Bovine Serum, Fetal Bovine Serum, Modified Eagle Medium, Gibco) and inoculated in a 24-well culture plate. , Overnight with 0.5 ug / ml of IgE, discarded medium and siraganian buffer I (pH 7.2, 119 mM sodium chloride, 5 mM potassium chloride, 0.4 mM magnesium chloride, 25 mM PIPES, 40 mM) After washing with 0.5 ml of sodium hydroxide, 160ul of Shiraganian Buffer II (addition of 5.6mM glucose, 1mM calcium chloride, 0.1% BSA) to Shilaganian Buffer I was reacted at 37 ° C for 10 minutes, and 20ul of sample was treated. After reacting at 37 ° C for 10 minutes, 20ul of 1ug / ml DNP-BSA was treated and reacted at 37 ° C for 30 minutes.The solution was transferred to an e-tube, centrifuged, and 20ul of supernatant was added. 20 μl of the substrate, 1 mM ρ-nitrophenyl-N-acetyl-β-D-glucosaminide, was added to a 96-well plate at 37 ° C. After 1 h, stop solution (0.1M sodium carbonate) After treating 200ul and measuring the absorbance at 405nm, the anti-allergic effect was obtained using Equation 2 below, and the results are shown in Table 5 below.

Anti-allergic effect (%) = {(D OD _{405 λ} value-B OD _{405 λ} value-C OD _{405 λ} value) / (A OD _{405 λ} value-B OD _{405 λ} value-C OD _{405 λ} value)} × 100

A: control group (test group without sample)

B: Blank (Experiment group with only sample and substrate added to ELISA plate)

C: Spontaneous (Experimental group without added DNP-IgE and sample)

D: Sample (hc-1 of Example 1, hc-2 of Example 2 or 4-O-methylhonokiol of Example 3 having a concentration of 1, 5 or 10 ug / ml)

sample Anti-allergic effect (%)  1 ug / ml of hc-1 of Example 1 25  5 g / ml of hc-1 of Example 1 33  Hc-1 10ug / ml of Example 1 51 Hc-2 1ug / ml of Example 2 29  Hc-2 5ug / ml of Example 2 48  10 ug / ml of hc-2 of Example 2 74  1 ug / ml 4-O-methylhonokiol of Example 3 37  5 ug / ml 4-O-methylhonokiol of Example 3 66  10 ug / ml 4-O-methylhonokiol of Example 3 102 DSCG (Disodium Chrymoglycate) 100uM 55

      As a result, as shown in Table 5, hc-1 of Example 1 and hc-2 of Example 2 is 10 ug / ml concentration, the 4-O-methyl honokiol of Example 3 is 5ug / ml At the concentration of

The concentration required to inhibit allergy by 50% was exceeded IC _50, and the 4-O-methyl-honochiol of Example 3 completely inhibited the degranulation of mast cells with an antiallergic effect of 102% at a concentration of 10 ug / ml. . Therefore, it was found that the thick extract and the compound separated therefrom have an excellent effect on inhibiting the allergic reaction.

Experimental Example 3. Inhibition of iNOS Activity

     In order to investigate the inhibitory effect of the extract and the compounds isolated therefrom, experiments to measure the activity of iNOS (Inducible Nitric Oxide Synthase) in Raw264.7 cells, which are macrophages of mice, It was performed together.

Suspend cells at a density of 8 x 10 ⁵ in 10% FBS-DMEM (Fetal Bovine Serum, Fetal Bovine Serum, Dulbecco's Modified Eagle Medium, Gibco) medium containing no phenol-red. Inoculate the plate by incubation, and after 24 hours, the samples were treated for 1, 5 or 10 ug / ml incubation for 18 hours, and then treated with 1 ug / ml of lipopolysaccharide (LPS, lipopolysaccharide, Sigma). After incubation for 24 hours, the supernatant was collected, 100ul each was placed in a 96-well plate, and the same amount of Gries' reagent (Griess reagent, Sigma Co., Ltd.) was added thereto, and the mixture was gently shaken at room temperature for 10 minutes to measure absorbance at 570 nm. A calibration curve was prepared using sodium nitrite as a standard, the control group was used as a control group only, and the production amount of nitric oxide (NO) in the lipopolysaccharide (LPS) treated group was 100%. The amount of nitric oxide (NO) produced was determined, and the results are shown in Table 6 below.

sample NO production rate (%) Control 8.64 Lipopolysaccharide (LPS) 100 LPS + ug-1 1ug / ml of Example 1 88.9 LPS + 5 ug / ml hc-1 of Example 1 61.3  LPS + 10 ug / ml hc-1 of Example 1 48.9 LPS + ug-2 1ug / ml of Example 2 79.8 LPS + 5ug / ml of hc-2 of Example 2 53.5  LPS + hc-2 10ug / ml of Example 2 20.7 LPS + 1 ug / ml 4-O-methylhonokiol of Example 3 70.1 LPS + 5 ug / ml 4-O-methylhonokiol from Example 3 44.5 LPS + 10 ug / ml 4-O-methylhonokiol from Example 3 9.7

      As a result, as shown in Table 6, the concentration of hc-1 of Example 1, hc-2 of Example 2, and 4-O-methylhonokiol of Example 3 treated with lipopolysaccharide (LPS) Dependently inhibited the activity of iNOS. In particular, the 4-O-methyl honokiol of Example 3 was confirmed that the NO production rate of 9.7% at a concentration of 10ug / ml almost no difference from the control, it was the most effective in inhibiting the production of nitric oxide. Therefore, it was found that the extract and the compound isolated therefrom have an excellent effect on inhibiting the inflammatory response by inhibiting NO production by inhibiting iNOS activity.

Experimental Example  4. iNOS  And COX  2 gene expression inhibition experiment

In order to investigate the inhibitory effect of the extracts and the compounds isolated therefrom, Lipopolysaccharide (LPS) was treated to Raw264.7 cells, which are macrophages of mice, to artificially increase the expression of the COX2 (Cyclooxygenase2) gene. Experiments to measure the activity of COX2 and iNOS (inducible Nitric Oxide Synthase) were carried out as follows using the RT-PCR method.

Cells were suspended in 10% FBS-DMEM (Fetal Bovine Serum, Fetal Bovine Serum, Dulbecco's Modified Eagle Medium, Gibcosa) at a density of 8 x 10 ⁵ , inoculated in 100 mm dishes, and inoculated in a 100 mm dish. hc-1, hc-2 of Example 2 or 4-O-methylhonokiol of Example 3 was incubated for 18 hours by treatment at a concentration of 1, 5 or 10 ug / ml, followed by 1 ug / ml of lipopolysac After incubation for 8 hours with the treatment of Lide (LPS, lipopolysaccharide, Sigma), the cell medium was removed and 1 ml of trizol (Trizol, Invitrogen) was added to the RNA isolation method of Invitrogen (Chromczynski, P. , and Sacchi, N. Anal . Biochem . 162 , p156, 1987), RNA was isolated and quantitated at 260 nm to perform reverse transcription polymerization (Ribber's transcription-polymerase chain reaction). Reverse transcription polymerization was used an all-in-one reverse transcription polymerization kit (All-in-one RT-PCR kit, Superbio), the primer conditions used in the experiments are described in Table 7 below, all-in-one reverse transcription polymerization The experiment was carried out according to the kit's manual (Gibbs, RA Anal. Chem., 62 (13) , p1202, Jul 1, 1990) to measure the activity effect (%) of COX2 and iNOS, and the results are shown in Table 8 below. Indicated.

 iNOS Primer Sense   5'-CAGTTCTGCGCCTTTGCTCAT-3 '(SEQ ID NO: 1) Antisense   5'-GGTGGTGCGGCTGGACTTT-3 '(SEQ ID NO: 2)  Reaction conditions   After reverse transcription at 50 ° C. for 30 minutes, the reverse transcriptase was inactivated at 96 ° C. for 3 minutes, followed by 32 polymerization reactions of 94 ° C. 30 seconds, 60 ° C. 30 seconds, and 72 ° C. for 1 minute.   COX 2 Primer  Sense   5'-CTGAAGCCCACCCCAAAC-3 '(SEQ ID NO: 3) Antisense   5'-AACCCAGGTCCTCGCTTATG-3 '(SEQ ID NO: 4) Reaction conditions   After reverse transcription at 50 ° C for 30 minutes, the reverse transcriptase was inactivated at 96 ° C for 3 minutes, followed by 32 polymerization reactions of 94 ° C 30 seconds, 55 ° C 30 seconds, and 72 ° C for 1 minute. Actin primer Sense 5'-GAGACCTTCAACACCCCAGCC-3 '(SEQ ID NO: 5) Antisense 5'-GGCCATCTCTTGCTCGAAGTC-3 '(SEQ ID NO: 6) Reaction conditions After reverse transcription at 50 ° C for 30 minutes, the reverse transcriptase was inactivated at 96 ° C for 3 minutes, followed by 22 polymerization reactions of 94 ° C for 1 minute, 62 ° C for 1 minute, and 72 ° C for 1 minute.

sample iNOS (%) COX2 (%) Control 100 100 Lipopolysaccharide (LPS) 141.3 133.6 LPS + ug-1 1ug / ml of Example 1 144.2 122.1 LPS + 5 ug / ml hc-1 of Example 1 132.6 123.6 LPS + 10 ug / ml hc-1 of Example 1 110.5 105.4 LPS + ug-2 1ug / ml of Example 2 137.8 123.2 LPS + 5ug / ml of hc-2 of Example 2 125.6 109.5 LPS + hc-2 10ug / ml of Example 2 103.3 97.4 LPS + 1 ug / ml 4-O-methylhonokiol of Example 3 120.9 110.9 LPS + 5 ug / ml 4-O-methylhonokiol from Example 3 100.7 105.1 LPS + 10 ug / ml 4-O-methylhonokiol from Example 3 86.3 77.4

      As a result of the experiment, as shown in Table 8, the concentration of hc-1 of Example 1, hc-2 of Example 2, and 4-O-methylhonokiol of Example 3 treated with lipopolysaccharide (LPS) Dependently inhibited the activity of iNOS and COX2. In particular, the 4-O-methyl honokiol of Example 3 showed iNOS and COX2 production rates of 86.3 and 77.4% at a concentration of 10 ug / ml, showing little difference from the control group, and had the largest iNOS and COX2 inhibitory effect. . Therefore, it was found that the extract and the compound isolated therefrom have an excellent effect of inhibiting the inflammatory response by inhibiting the activities of iNOS and CIX2.

Experimental Example 5. PGE ₂ , IL-6, IL-8 production inhibitory effect

Experiments were performed as follows to determine the inhibitory effect of the extract of gourd extract and the compound isolated therefrom by measuring the inhibitory effect of the production of inflammation mediators PGE ₂ , IL-6 and IL-8 in human fibroblasts.

Human fibroblasts were seeded in 6-well culture plates at a density of 1 × 10 ⁵ cells and incubated in a 37 ° C., 5% CO ₂ incubator for 24 hours, and then treated with 500 uM of hydrogen peroxide (H ₂ O ₂ ) in the wells for 24 hours. After stimulation, the hc-1 of Example 1, hc-2 of Example 2 or 4-O-methyl-honokinool sample of Example 3 were treated for 1, 5 or 10 ug / ml concentration and reacted for 48 hours. After the ELISA assay was performed by collecting the culture medium, PGE ₂ was tested using Asay Design's kit, and IL-6 and IL-8 were used by Endogen's kit. (P. Tijssen, in Practise and Theory of Enz. Immunoassays , Elsevier, Amsterdam, 1985), the following equation 3 was used to determine the effect of inhibiting the production of PGE ₂ , IL-6 and IL-8, the results It is shown in Table 9 below.

   Inhibitory effect = {1- (A-B) / (C-B)} X100

A: PGE ₂ , IL-6 or IL in the hc-1 of Example 1, hc-2 of Example 2, or 4-O-methylhonokiol of Example 3 treated with hydrogen peroxide (H ₂ O ₂ ) -8 yield (pg / ml)

B: Production amount of PGE ₂ , IL-6 or IL-8 in control treated only with medium (pg / ml)

C: Production amount of PGE ₂ , IL-6 or IL-8 in hydrogen peroxide (H ₂ O ₂ ) treated group (pg / ml)

sample PGE ₂ production amount (pg / ml) PGE ₂ , inhibitory effect IL-6 production amount (pg / ml) IL-6 inhibitory effect IL-8 production amount (pg / ml) IL-8 inhibitory effect Control 156.1 23.9 123.2 H ₂ O ₂ (500uM) 447.8 182.3 317.4 H ₂ O ₂ + hc-1 1 ug / ml of Example 1 401.2 15.97% 170.9 7.20% 298.0 9.99% H ₂ O ₂ + 5 ug / ml of hc-1 of Example 1 366.9 27.73% 160.8 13.57% 255.6 31.82% H ₂ O ₂ + hc-1 10 ug / ml of Example 1 310.4 47.1% 135.5 29.55% 209.3 55.66% H ₂ O ₂ + hc-2 1ug / ml of Example 2 302.5 49.81% 165.2 31.47% 280.4 19.05% H ₂ O ₂ + hc-2 5ug / ml of Example 2 290.8 53.82% 123.5 51.70% 233.9 43.00% H ₂ O ₂ + hc-2 10ug / ml of Example 2 220.4 77.96% 100.9 62.66% 191.0 65.09% H ₂ O ₂ + 4-ug-methylhonokiol of Example 3 1 ug / ml 300.1 50.63% 150.2 20.27% 259.4 29.87% H ₂ O ₂ + 4-ug-mlHonokiol of Example 3 5 ug / ml 205.4 83.10% 103.7 49.62% 199.2 60.87% H ₂ O ₂ + 10 ug / ml 4-O-methylhonokiol of Example 3 137.1 106.51% 5 5.9 79.80% 110.8 106.39%

As a result of the experiment, as shown in Table 9, hc-1 of Example 1, hc-2 of Example 2 and 4-O-methylhonokiol of Example 3 treated with hydrogen peroxide were PGE ₂ , IL in a concentration-dependent manner. Inhibited the production of -6 and IL-8. In particular, the 4-O-methylhonokiol of Example 3 treated with hydrogen peroxide had the greatest inhibitory effect of about 79.80 to 106.51% of PGE ₂ , IL-6 and IL-8 at a concentration of 10 ug / ml. Therefore, it was found that the extract and the compound isolated therefrom have an excellent effect of inhibiting the inflammatory response by inhibiting the activity of inflammatory mediators PGE ₂ , IL-6 and IL-8.

Experimental Example 6. Determination of antihistamine activity using animal experiment

In order to determine the anti-allergic effect of the extract and the compound isolated therefrom by measuring the histamine blocking effect through animal experiments, hc-1 of Example 1, hc-2 of Example 2 and 4-O- of Example 3 Experiments were performed as follows using a method for evaluating methylhonochiol for ileum isolated from guinea pigs (Akah PA et al, Jounal of Ethnopharmacology, 55 , pp87-92, 1997).

After feeding 400g of guinea pigs (Sam taco, 5 weeks old) without water for 48 hours, and killing the ileum on the day of the experiment, separated the ileum and took the 10cm ileum, and then divided the small ileum at 2.5cm intervals. In a water bath, connect the pressure transducer and record the trace using a multichannel polygraph. In the first step, starting from 0.5ug / 35ml with respect to histamine, up to 2.5ug / 35ml, After confirming that the tissue is actively reacting, the dissolved hc-1 of Example 1, hc-2 of Example 2, 4-O-methylhonokiol of Example 3 or an antiallergic composition DSCG (disodium Chrymoglycate) was added to 10ug, 100ug, 1mg, 2mg, 5mg or 10mg to determine the IC ₅₀ value to determine the histamine blocking effect, and the results are shown in Table 10 below.

sample IC ₅₀ (ug / ml) Hc-1 of Example 1 289 Hc-2 of Example 2 154 4-O-Methyl Honokiol of Example 3 43 DSCG (Disodium Chrymoglycate) 5

As a result, as shown in Table 10, the 4-O-methyl honokiol of Example 3 has an IC ₅₀ value of 43 ug / ml, even though it is higher than DSCG, an anti-allergic drug used directly in the clinic, anti-allergic as a cosmetic The effect was very good. Therefore, it was found that excellent anti-allergic effect was obtained by inhibiting hc-1 of Example 1, hc-2 of Example 2, and 4-O-methylhonokiol histamine of Example 3.

Experimental Example 7. Determination of anti-inflammatory effect using animal experiment

The experiment was performed as follows to determine the inflammation inhibitory effect of the extract of hoobak and the compound separated therefrom using an animal experiment.

Experimental method of Winter scan in (Winter CA, Risley EA, Nuss GW, Proc. Soc. Exp. Biol. Med., 111, p544, 1962), the carrageenan that causes acute inflammation mice (Sam tacos, 5-week-old) by using the After measuring the volume of the hind paws before and after carrageenan injection of the hc-1 of Example 1, hc-2 of Example 2, 4-O-methyl-honokiol of Example 3 or an indomethacin treatment group which is an anti-inflammatory substance, Inflammation inhibition was calculated using Equation 4, and the results are shown in Table 11 below.

  Inhibition of Inflammation = {1- (A-B) / (C-B)} X100

A: Volume of the hind paws in the hc-1 of carrageenan treated Example 1, hc-2 of example 2, 4-O-methylhonokiol or indomethacin treated group of Example 3

B: volume of hind paw in solvent treated control

C: volume of hind paw in carrageenan treatment group

sample % Inhibition 120 mg / kg of hc-1 of Example 1 68.7 120 mg / kg of hc-2 of Example 2 57.3 120 mg / kg of 4-O-methylhonokiol of Example 3 50.4 Indomethacin 120mg / kg 61.9

      As a result of the experiment, as shown in Table 11, hc-1 of Example 1, hc-2 of Example 2 and 4-O-methyl honokiol of Example 3 has a high inflammation inhibition rate of about 50.4 to 68.7% In particular, the hc-1 of Example 1 showed the highest inhibitory effect of hc-1 of 68.7%, showing a higher inhibition rate of inflammation than indomethacin, an anti-inflammatory substance. Therefore, it was found that hc-1 of Example 1, hc-2 of Example 2, and 4-O-methylhonokiol of Example 3 had an excellent effect on inhibiting the inflammatory response.

Experimental Example 8. Elastase inhibitory effect

In order to determine the effect of inhibiting the elastase inhibitory extract and the compound separated therefrom was effective to inhibit the generation of wrinkles was carried out as follows.

8-1. Preparation of the solution

     A buffer was made by adjusting the 0.267M Tris solution with 0.267M hydrochloric acid so that the pH was 8.0, and a succinyl-alanine-alanine-alanine-pi-nitroanilide standard was prepared. Elastase substrate solution was prepared at a concentration of 8.8 mM, and enzyme solution was prepared at a concentration of 10 µg / ml for Porcine Pancreatic Elastase.

8-2. Absorbance Measurement of Control

      Absorbance A using the blank obtained as a blank solution using 100 μl of purified water instead of the hc-1 of Example 1, hc-2 of Example 2, and 4-O-methyl-honochiol of Example 3 as a sample solution. The absorbance D was measured using 120 μl of purified water instead of the enzyme solution and the test solution as a color correction solution, and used in Equation 5 below.

8-3. Absorbance Determination of Thick Extract and Compound Isolated therefrom

      Into 60 μl of the buffer prepared in Experimental Example 8-1, 20 μl of substrate solution, hc-1 of Example 1, hc-2 of Example 2, or 100 μl of 4-O-methylhonokiol sample of Example 3 were mixed. After adding 20 µl of enzyme solution, shaking and reacting at 25 ° C. for 15 minutes, the amount of p-nitroaniline was measured by absorbance B at 410 nm, and 20 µl of substrate solution, 20 µl of purified water and 100 µl of sample were added to 60 µl of buffer. Absorbance C was measured using the solution obtained by the same method as the control solution, and the inhibition rate of elastase was calculated using Equation 5 below, and the results are shown in Table 12 below.

   % Elastase inhibition = [1-{(B-C) / (A-D)}] × 100

   A: absorbance A of Experimental Example 8-2

   B: Absorbance B of Experimental Example 8-3

   C: absorbance C of Experimental Example 8-3

   D: absorbance D of Experimental Example 8-2

sample Elastase Inhibition Rate (%)  Hc-1 10ug / ml of Example 1 15.9  1hc-1 50ug / ml of Example 33.5  1hc-1 100ug / ml of Example 48.8  10 ug / ml of hc-2 of Example 2 39.7  25ug / ml of hc-2 of Example 2 54,8  50 ug / ml of hc-2 of Example 2 59.0  10 ug / ml 4-O-methylhonokiol of Example 3 41.3  25 ug / ml 4-O-methylhonokiol of Example 3 60.6  50 ug / ml 4-O-methylhonokiol of Example 3 65.7

      As a result, as shown in Table 12, the hc-1 of Example 1, hc-2 of Example 2 and 4-O-methyl honokiol of Example 3 has an elastase inhibition rate of about 15.9 to 65.7% It showed a high elastase inhibitory effect, in particular, 4-O-methyl honokiol of Example 3 showed the highest elastase inhibitory effect of 65.7%. Therefore, it was found that hc-1 of Example 1, hc-2 of Example 2, and 4-O-methylhonokiol of Example 3 had an excellent effect of inhibiting wrinkle formation by inhibiting elastase.

Experimental Example 9. Inhibitory Effect of MMP-1 mRNA Expression

MMP-1 mRNA Expression Inhibitory Effect The experiment was carried out as follows to determine whether the thick extract and the compound isolated therefrom are effective in inhibiting wrinkle formation.

Human fibroblasts (ATCC 2076) were inoculated into 60 mm dishes at a concentration of 1 × 10 ⁶ cells / ml using medium using IMDM (Iscove's Modified Dulbecco's Medium, Gibco) containing 10% bovine serum, and then 24 hours later with PBS. Irradiated with UVA 6J and then replaced with a serum-free medium to replace 1, 5 or 10 ug / ml of hc-1 of Example 1, hc-2 of Example 2, or 4-O-methylhonokiol of Example 3 Incubated in a 5% CO ₂ incubator at 37 ° C. for one day, and then the medium of the cells was removed, and 1 ml of Trizol (Invitrogen) was added to the RNA separation method of Invitrogen (Chromczynski, P. , and Sacchi, N. Anal. Biochem. 162 , p156, 1987.), RNA was quantified at 260 nm using an ultraviolet detector, and then all-in-one reverse transcription polymerization kit (All-in-one RT). -PCR kit (Superbio) for reverse transcription-polymerase chain reaction And showed the primers and reaction conditions are the following Table 13, the test according to the manual of the all-in-one reverse transcription polymerase reaction kit (Gibbs, RA Anal. Chem. , 62 (13), p1202, Jul 1, 1990) is corrected for actin The degree of expression of MMP-1 was expressed as a value, and the results are shown in Table 14 below.

  MMP-1 Primer  Sense   5'-TGGGAGCAAACACATCTGA-3 '(SEQ ID NO: 7) Antisense  5'-ATCACTTCTCCCCGAATCGT-3 '(SEQ ID NO: 8) Reaction conditions   After reverse transcription at 50 ° C for 30 minutes, the reverse transcriptase was inactivated at 96 ° C for 3 minutes, followed by 29 polymerization reactions of 94 ° C for 1 minute, 48 ° C for 1 minute, and 72 ° C for 1 minute. Actin primer Sense 5'-GAGACCTTCAACACCCCAGCC-3 '(SEQ ID NO: 5) Antisense 5'-GGCCATCTCTTGCTCGAAGTC-3 '(SEQ ID NO: 6) Reaction conditions After reverse transcription at 50 ° C for 30 minutes, the reverse transcriptase was inactivated at 96 ° C for 3 minutes, followed by 22 polymerization reactions of 94 ° C for 1 minute, 62 ° C for 1 minute, and 72 ° C for 1 minute.

sample MMP-1 expression level (%, corrected for actin) Control 69.9 UVA 100 UVA + ug-1 1ug / ml of Example 1 101.7 UVA + 5ug / ml of hc-1 of Example 1 90.8 UVA + 10 ug / ml of hc-1 of Example 1 76.5 UVA + ug-2 1ug / ml of Example 2 90.2 UVA + 5ug / ml of hc-2 of Example 2 81.3 UVA + 10ug / ml of hc-2 of Example 2 71.1 UVA + 1 ug / ml 4-O-methylhonokiol of Example 3 88.4 UVA + 5 ug / ml 4-O-methylhonokiol of Example 3 70.9 UVA + 10 ug / ml 4-O-methylhonokiol of Example 3 66.1 UVA + EGCG 25uM 74.3

      As a result, as shown in Table 14, the concentration of hc-1 of Example 1, hc-2 of Example 2, and 4-O-methylhonokiol of Example 3 that induced the expression of MMP-1 by UVA Independently, MMP-1 activity was inhibited, and in particular, 4-O-methylhonokiol of Example 3 exhibited the highest MMP-1 activity by expressing 66.1% at 10 ug / ml. Therefore, it was found that hc-1 of Example 1, hc-2 of Example 2, and 4-O-methylhonokiol of Example 3 had an excellent effect of inhibiting the formation of wrinkles by inhibiting the activity of MMP-1. .

       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 Preparation of Flexible Lotion (Skin)

      Table 15 shows an example of preparation of a lotion (skin lotion) in a cosmetic containing a thick leaf extract or a compound separated therefrom.

number Raw material Formulation Example 1 Comparative Formulation Example 1 One   Hc-1 of Example 1 1.0 - 2 Butylene glycol 4.0 4.0 3 glycerin 8.0 8.0 4 Carboxy Vinyl Polymer 0.06 0.06 5 Disodium Ethylenediaminetetraacetic Acid 0.01 0.01 6 ethanol 7.0 7.0 7 Triethanolamine 0.2 0.2 8 Hydrogenated Castor Oil 0.4 0.4 9 Phenyltrimethicone 3.5 3.5 10 Paraoxybenzoic acid propyl 0.2 0.2 11 Methyl paraoxybenzoate 0.2 0.2 12 Purified water Remaining amount Remaining amount system 100.0 100.0

Formulation example  2. Preparation of nutrient lotion (lotion)

      Table 16 shows an example of the formulation of nutrient lotion in a cosmetic containing a thick extract or a compound isolated therefrom.

number Raw material Formulation Example 2 Comparative Formulation Example 2 One Hc-1 of Example 1 1.0 - 2 Stearic acid 1.0 1.0 3 Cetyl alcohol 1.0 1.0 4 Glyceryl Monostearate 1.0 1.0 5 Polyoxyethylene sorbitan monostearate 1.0 1.0 6 Sorbitan sesquioleate 1.0 1.0 7 Floating fly pin 4.0 4.0 8 Squalane 3.0 3.0 9 Caprylic / Capric Triglycerides 4.0 4.0 10 Carmoxy Vinyl Polymer 0.1 0.1 11 Butylene glycol 5.0 5.0 12 Triethanolamine 0.2 0.2 13 Paraoxybenzoic acid propyl 0.2 0.2 14 Methyl paraoxybenzoate 0.2 0.2 15 Purified water Remaining amount Remaining amount system 100.0 100.0

Formulation example  3. Preparation of nutrition cream

     Table 17 shows an example of the preparation of the nourishing cream in the cosmetic containing the thick extract or the compound isolated therefrom.

number Raw material Formulation Example 3 Comparative Formulation Example 3 One   Hc-1 of Example 1 1.0 - 2 Stearic acid 2.0 2.0 3 Cetyl alcohol 2.0 2.0 4 Glyceryl Monostearate 2.0 2.0 5 Polyoxyethylene sorbitan monostearate 0.5 0.5 6 Sorbitan sesquioleate 0.5 0.5 7 Glyceryl Monostearate / Glyceryl Tearate / Polyoxyethylene Stearate 1.0 1.0 8 Wax 1.0 1.0 9 Floating fly pin 4.0 4.0 10 Squalane 4.0 4.0 11 Caprylic / Capric Triglycerides 4.0 4.0 12 Carboxy Vinyl Polymer 0.3 0.3 13 Butylene glycol 5.0 5.0 14 glycerin 3.0 3.0 15 Triethanolamine 0.5 0.5 16 Paraoxybenzoic acid propyl 0.2 0.2 17 Methyl paraoxybenzoate 0.2 0.2 18 Purified water Remaining amount Remaining amount system 100.0 100.0

Formulation example  4. Massage  Manufacture of cream

     A formulation example of a massage cream in a cosmetic containing a thick leaf extract or a compound separated therefrom is shown in Table 18 below.

number Raw material Formulation Example 4 Comparative Formulation Example 4 One   Hc-1 of Example 1 1.0 - 2 Beeswax 10.0 10.0 3 Polysorbate 60 1.5 1.5 4 Sorbitan sesquioleate 0.8 0.8 5 Liquid paraffin 40.0 40.0 6 Squalane 5.0 5.0 7 Caprylic / Capric Triglycerides 4.0 4.0 8 Butylene glycol 3.0 3.0 9 Propylene glycol 3.0 3.0 10 Triethanolamine 0.2 0.2 11 antiseptic 0.4 0.4 12 Pigment 0.001 0.001 13 Spices 0.1 0.1 14 Purified water Remaining amount Remaining amount system 100.0 100.0

Formulation example  5. Preparation of Essence

      Formulations of nutritional essences in cosmetics containing thick extracts or compounds isolated therefrom are shown in Table 19.

number Raw material Formulation Example 5 Comparative Formulation Example 5 One   Hc-1 of Example 1 1.0 - 2 Sitosterol 1.70 1.70 3 Polyglyceryl 2-oleate 1.50 1.50 4 Ceteares-4 1.2 1.2 5 cholesterol 1.5 1.5 6 Dicetylphosphate 0.4 0.4 7 Concentrated glycerin 5.0 5.0 8 Carboxy Vinyl Polymer 0.2 0.2 9 Xanthan Gum 0.2 0.2 10 antiseptic 0.4 0.4 11 Spices 0.1 0.1 12 Purified water Remaining amount Remaining amount system 100.0 100.0

Formulation example  6. 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

clinical Experimental Example  1. Anti-inflammatory effect

      In order to determine the anti-inflammatory effect of the thick extract and the compound separated therefrom, the experiment was performed as follows.

      The left ear of the mouse is the control site, the right ear is washed with ethanol clean, and the sample (Formulation Examples 1 to 5 and Comparative Formulation Examples 1 to 5) 20 μl each day continuously applied for 4 days and last application 1 After 1 hour, ethanol was applied to the left ear and arachidonic acid was applied to the right ear at 2 mg / ear. After 1 hour, the ear edema was measured three times on both ears with a micrometer. acid) anti-inflammatory effect was measured using Equation 6 based on the treatment group, the results are shown in Table 21 below.

 Inhibition rate of inflammation (%) = (A-B) / A × 100

   A: Average thickness of control ears (thickness of arachidonic acid treated ears-thickness of ears before treatment)

   B: thickness of the sample-coating group ears (thickness of the sample-treated ears-thickness of the ears before the treatment)

sample density(%) menstruum Ear thickness (㎛) Inhibition rate of inflammation (%) Before sample processing After sample processing Arachidonic acid - ethanol 284 481 - Indomethacin One ethanol 276 342 66.50 One Formulation Example 1 of Formulation Example 1 100 ethanol 280 379 49.75 2 Comparative Formulation Example 1 of Formulation Example 1 100 ethanol 283 479 0.51 3 Formulation Example 2 of Formulation Example 2 100 ethanol 277 381 47.21 4 Comparative Formulation Example 2 of Formulation Example 2 100 ethanol 283 467 6.60 5 Formulation Example 3 of Formulation Example 3 100 ethanol 270 335 67.01 6 Comparative Formulation Example 3 of Formulation Example 3 100 ethanol 275 454 9.14 7 Formulation Example 4 of Formulation Example 4 100 ethanol 266 349 57.87 8 Comparative Formulation Example 4 of Formulation Example 4 100 ethanol 279 463 6.60 9 Formulation Example 5 of Formulation Example 5 100 ethanol 288 358 64.47 10 Comparative Formulation Example 5 of Formulation Example 5 100 ethanol 277 458 8.12

      As a result of the experiment, as shown in Table 21, cosmetics made from the raw material extract or the compound separated therefrom are 47.21 to 67.01%, which is excellent in inhibiting inflammation compared to 0.51 to 9.14% of products that are not. I could confirm the fact.

clinical Experimental Example  2. Skin irritation test

      The experiment was performed as follows to test the skin irritation of the thick extract and the compound separated therefrom.

      Formulation and 0.08% aqueous solution of sodium lauryl sulfate were applied to a healthy adult male and 50 females on a back site for a predetermined amount (0.2 g) of each sample (Formula Examples 1 to 5 and Comparative Formulation Examples 1 to 5) for 24 hours. After removing the pin chamber after 4 hours, the skin condition change was visually read, and the stimulus degree was calculated using Equation 7 below, and the results are shown in Table 22 below.

Stimulus degree = [{(+-) number × 1} + {(+) number × 2} + {(++) number × 3}] / number of subjects

sample Number of subjects Judgment result Stimulus ++ + +- - Sodium Lauryl Sulfate0.08% 50 One 5 12 32 0.50 Formulation Example 1 of Formulation Example 1 + Sodium Lauryl Sulfate 0.08% 50 0 2 5 43 0.18 Comparative Formulation Example 1 of Formulation Example 1 + Sodium Lauryl Sulfate 0.08% 50 One 4 10 35 0.42 Formulation Example 2 of Formulation Example 2 + Sodium Lauryl Sulfate 0.08% 50 0 2 6 42 0.2 Comparative Formulation Example 2 Formulation Example 2 + Sodium Lauryl Sulfate0.08% 50 One 2 13 34 0.40 Formulation Example 3 of Formulation Example 3 + Sodium Lauryl Sulfate0.08% 50 0 One 6 43 0.16 Comparative Formulation Example 3 of Formulation Example 3 + Sodium Lauryl Sulfate0.08% 50 One 3 5 41 0.28 Formulation Example 4 of Formulation Example 4 + Sodium Lauryl Sulfate 0.08% 50 0 One 8 41 0.2 Comparative Formulation Example 4 of Formulation Example 4 + Sodium Lauryl Sulfate 0.08% 50 0 6 10 34 0.44 Formulation Example 5 of Formulation Example 5 + Sodium Lauryl Sulfate 0.08% 50 0 2 3 45 0.14 Comparative Formulation Example 5 of Formulation Example 5 + Sodium Lauryl Sulfate 0.08% 50 One One 10 39 0.30  -; No erythema or unusual phenomena +-; Slightly redder than +; Markedly redder than ++; More reddening and swelling

      As a result of the experiment, as shown in Table 22, cosmetics made from the raw material extract or the compound separated therefrom are 0.14 to 0.2, and the skin irritation is much smaller than that of 0.51 to 9.14. I could confirm the fact.

clinical Experimental Example  3. Measurement of skin elasticity improvement effect by reducing wrinkle depth

      In order to determine the skin elasticity improvement effect of the thick extract and the compound separated therefrom, the experiment was performed as follows.

Formulation Example 3 of Formulation Example 3 was divided into two groups of 20 women, and applied to the left eye edge twice a day after washing the morning and evening. The control group applied the nourishing cream to the right eye edge 15 days and 30 days after application. The skin wrinkle depth of the surface was measured by a cutometer SEM 474, which is a measure of the depth of wrinkles, and the smaller the value, the better the elasticity. The average value of the 20 subjects is shown in Table 23 below.

sample Wrinkle Depth (%) 0 days 15th 30 days Formulation Example 3 of Formulation Example 3 0.51 0.43 0.29 Control 0.49 0.46 0.40

      As a result, as shown in Table 23, it can be seen that the cosmetic product made of the formulation example 3 of Formulation Example 3 has an excellent effect of improving skin elasticity by decreasing the wrinkle depth over time.

Clinical Trial 4. Confirmation of Skin Condition Improvement Effect

      In order to confirm the skin condition improvement effect on the essence made using Formulation Example 5 of Formulation Example 5, the experiment was carried out as follows.

       A healthy female group of 36 to 69 years old, divided into two groups of 30 people, the left side of the face contains the formulation example 5 essence of Formulation Example 5, and the right side of the face contains the essence of the control group every morning and evening twice a day for 4 weeks After application, the degree of improvement of skin condition was measured by the average score of 30 people using the grading method of Table 24, and the results are shown in Table 25 below.

(A) Rating of improvement effect on skin elasticity, sagging Was greatly improved  3 points Was improved  2 points Slightly improved  1 point No improvement  0 points Worsened -1 point (B) Rating of improvement effect on wrinkles and fine lines Was greatly improved  3 points Was improved  2 points Slightly improved  1 point No improvement  0 points Worsened -1 point

sample (A) skin elasticity, sagging (B) wrinkles, fine lines Formulation Example 5 (Formulation Example 5) 79 82 Control 40 41

As a result of the experiment, as shown in Table 25, the essence made using Formulation Example 5 of Formulation Example 5 has a much better skin improvement effect compared to the control group with a skin improvement effect of 79 to 82 points 40 to 41 points Could.

      Thickening extract of the present invention and compounds isolated therefrom are iNOS activity inhibitory effect, iNOS, COX2 gene expression inhibitory effect, PGE2, IL-6, IL-8 production inhibitory effect, antihistamine activity enhancement, elastase inhibitory effect and MMP- 1 It can be usefully used as an external skin pharmaceutical composition and cosmetic composition for anti-inflammatory, anti-allergic and wrinkle improvement because it shows the expression inhibition effect.

<110> bioland <120> composition comprising an extract of Magnolia officinalis Rehd.          et Wils. or 4-O-methylhonokiol isolated therefrom having          anti-inflammation, anti-allergy and anti-wrinkle activity <160> 8 <170> KopatentIn 1.71 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> forward primer for iNOS <400> 1 cagttctgcg cctttgctca t 21 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for iNOS <400> 2 ggtggtgcgg ctggacttt 19 <210> 3 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> forward primer for COX2 <400> 3 ctgaagccca ccccaaac 18 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for COX2 <400> 4 aacccaggtc ctcgcttatg 20 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> forward primer for actin <400> 5 gagaccttca acaccccagc c 21 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for actin <400> 6 ggccatctct tgctcgaagt c 21 <210> 7 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> forward primer for MMP-1 <400> 7 tgggagcaaa cacatctga 19 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for MMP-1 <400> 8 atcacttctc cccgaatcgt 20  

Claims (8)

Hug ( Magnolia officinalis Rehd . et Wils . A skin external pharmaceutical composition for improving anti-inflammatory, anti-allergic and antiwrinkle comprising 4-O-methyl honokiol represented by the following formula (I) as an active ingredient.

The skin external pharmaceutical composition according to claim 1, wherein the extract of the pear extract is extracted from a stem shell, seed, leaf or root of the pear extract. According to claim 1, wherein the extract is an external skin pharmaceutical composition, characterized in that the crude extract or non-polar solvent soluble extract. The skin external pharmaceutical composition according to claim 3, wherein the crude extract is an extract available in water including purified water, lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. According to claim 3, wherein the non-polar solvent soluble extract is an external skin pharmaceutical composition, characterized in that the extract soluble in a solvent selected from hexane, chloroform, methylene chloride, ethyl acetate, glycerin, propylene glycol, butylene glycol or ether. According to claim 1, wherein the extract or compound isolated therefrom The skin external pharmaceutical composition, characterized in that contained in 0.1 to 50% by weight relative to the total weight of the composition. Hug ( Magnolia officinalis Rehd . et Wils . An anti-inflammatory, anti-allergic and anti-wrinkle cosmetic composition containing 4-O-methyl honokiol represented by the formula (I) of claim 1 or an extract therefrom as an active ingredient.  According to claim 7, wherein the composition is a 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 Cosmetic composition, characterized in that the formulation selected from the group consisting of essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.