KR101991936B1 - Composition for Preventing or Improving Skin Wrinkle Comprising Enzyme Treated Extract of Gastrodia as Active Ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing or improving skin wrinkles comprising the hydrolyzed enzyme hydrolyzed cheonma extract as an active ingredient.
The compositions of the present invention exhibit the efficacy of inhibiting the expression of MMP-1 protein in human dermal fibroblasts and increasing the production of type I procollagen, thereby preventing the occurrence of skin wrinkles and improving the wrinkles of damaged skin. Overall, the compositions of the present invention can prevent and greatly improve skin wrinkles.

Description

Composition for Preventing or Improving Skin Wrinkle Comprising Enzyme Treated Extract of Gastrodia as Active Ingredient}

The present invention relates to a composition for improving skin wrinkles, and more particularly to a composition for preventing or improving skin wrinkles containing an enzyme-treated cheonma extract as an active ingredient.

Skin wrinkles are known to be affected by various factors such as skin moisture content, collagen content, and immunity to the external environment. The most important factors affecting the formation of double wrinkles are collagen production and collagenase, a collagen degrading enzyme. Naze expression and activity.

Collagen is present in the dermal layer of the skin and is known as a major ingredient that gives skin elasticity with elastin, which accounts for about 70-80% of the total dry weight of the skin. Collagen is reduced by internal factors such as decreased cellular activity due to natural aging, and biosynthesis is reduced or degradation is accelerated by external factors such as increased stress in various harmful environments or increased free radical species by sunlight. have.

On the other hand, overproduction of reactive oxygen species (ROS) has been reported in UVB exposure environments, and signaling systems associated with UVB exposure include MAPKs (mitogen activated protein kinases), extracellular signal-regulated kinase (ERK), It is associated with proteins such as p38 kinase, and c-Jun amino-terminal kinase (JNK), which induces increased expression of NF-ĸB and AP-1 transcription factors. In addition, AP-1 protein is associated with the activity of matrix metalloproteinase (MMP) gene, NF-ĸB protein affects the expression of proinflammatory cytokines such as interleukin.

The breakdown of collagen and elastin in aged skin is mainly caused by increased expression of its degrading enzymes, such as MMP proteins. In addition, pro-inflammatory cytokines interfere with the synthesis of collagen and promote the breakdown of collagen. Type I collagen is the most abundant protein in the extracellular matrix of connective tissue. Extracellular matrix includes other types of proteins such as type III, V, and VII collagen, elastin, proteoglycans, fibronectin.

Controlling the expression level of type I collagen protein is the most important factor in preventing skin photoaging. Irradiation of UVB reduces expression of collagen precursor procollagen in skin fibroblasts through inhibition of the TGF-β / Smad signaling system. In addition, the TGF-β1 signaling system is one of the important factors regulating the synthesis of extracellular matrix in human skin fibroblasts.

Thus, the generation of reactive oxygen species (ROS) regulated by AP-1 and MAPKs factors, and the expression of MMPs, ILs, TGF-β, and type I procollagen, serve as useful markers in skin photoaging studies.

As described above, various attempts have been made to search for natural substances that are effective in improving skin wrinkles by inhibiting collagen reduction. In order to take advantage of collagen's anti-wrinkle effect, products containing collagen were added to external skin composition such as cosmetics or ointment.However, these products are applied to the skin surface by applying collagen itself. There was no improvement. In order to solve these problems, interest in collagen synthesis promoters has been increased, and conventionally known collagen synthesis promoters include vitamin C, retinoic acid, transforming growth factor (TGF), and protein derived from placenta (JP8). -231370), betulinic acid (betulinic acid, JP8-208424), chlorella extract (JP9-40523, JP10-36283, fibroblast proliferation). However, the substance has a problem in that the amount of use is limited due to safety problems such as irritation and redness when applying the skin, or the effect thereof is insignificant, so that the anti-wrinkle effect cannot be expected. Therefore, there is an urgent need for the development of a new antiwrinkle composition which is safer in vivo than the conventional antiwrinkle composition and has a higher antiwrinkle effect.

Meanwhile, Gastrodia elata Blume, Gastrodia) is a perennial genus belonging to the Orchidaceae family, lacking chlorophyll, which is impossible for photosynthesis of nutritional substances using carbon assimilation, and does not have roots. Lee, Y. M (1990) In Oriental medicine dictionary, Sammundang, Seoul.p. 814, Sin, K. G (1980) In Sinssi Herbology, Summunsa, Kor. Pp.288-290) . In addition, Chunma is also known as red roots, ear poison, oaks, shoots, jeongpungcho, etc., and mainly refers to symptoms such as stress and fatigue, as well as severe adult diseases such as hypertension, headache, paralysis, neurological diseases and diabetes. It is known to be effective. In particular, studies on the anti-wrinkle effect of gastrodigenin (4-hydroxybenzyl alcohol) in the active ingredient of cheonma have been little.

Prior art related to the wrinkle improvement activity of Chunma Patent Registration No. 10-0893912 "Functional and functional cosmetics containing peony and cheonma extract as an active ingredient" is a hot water extract of the mixed powder of peony and cheonma and concentrated under reduced pressure , It is known that the extract produced through the ethanol extraction process is effective in improving wrinkles, and Patent Publication No. 10-2011-0116389 "Skin external preparation composition containing aloe and cheonma fermented filtrate" is aloe and cheonma It is disclosed that the fermentation filtrate prepared by extracting and culturing the fermentation broth obtained by adding lactic acid bacteria and filtering the fermentation broth has the effect of repairing damaged skin barrier, promoting skin cell growth, improving wrinkles, and anti-inflammatory.

However, there is no known technique that shows sufficient effect for improving skin wrinkles using cheonma as a single material.

Therefore, the inventors of the present invention intensively researched to develop a substance effective for improving skin wrinkles and preventing skin wrinkles while using a single agent of chemistry, and as a result, enzyme-treated cheonma extract was induced by UVB irradiation. In response to photoaging of dermal fibroblasts (NHDFs), the present invention has been found to have an excellent effect on preventing and improving skin wrinkles.

Therefore, the main object of the present invention is to provide a composition for preventing or improving skin wrinkles, which contains an enzyme-treated cheonma extract as an active ingredient having an excellent effect on preventing or improving skin wrinkles.

Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.

According to an aspect of the present invention, the present invention provides a composition for preventing or improving skin wrinkles, which comprises a glycolytic enzyme-treated cheonma extract as an active ingredient.

The present inventors have diligently researched to further improve the skin wrinkle effect of cheonma and increase the utility of the material. When cheonma extract is treated with glycolytic enzyme, 4-hydroxy benzyl alcohol and glucose are α-glyco. A significant increase in the component of alpha glucopyranosiloxy benzyl alcohol [4-alpha-D-glucopyranosyloxy benzyl alcohol] was observed (FIG. 1), it was found that the anti-wrinkle prevention and improvement effect of the enzyme-treated cheonma extract with increased alpha glucopyranosiloxy benzyl alcohol content was also significantly increased, and there was no problem in the safety when applied to the skin. .

The composition of the present invention is not cytotoxic, has an excellent antioxidant effect (Fig. 2), matrix metallo-proteinase-1 (hereinafter referred to as 'MMP') (Fig. 3 and Fig. 4) and type I There is a feature that can effectively suppress the improvement of skin wrinkles through increased expression of procollagen protein (FIGS. 5 and 6).

Gastrodia of the present invention elata Blume (Gastrodia) is a perennial herb belonging to the Orchidaceae family, also called red roots, poisonous cattle, oaks, shoots, and jeongpungcho.

As used herein, the term "extract" refers to an extract obtained by extracting a plant, a diluent or concentrate of the extract, a dried product obtained by drying the extract, a crude or purified product of the extract, a mixture thereof, or the like, It includes the extract itself and extracts of all formulations that can be formed using the extract. The extract of the present invention may be preferably prepared and used in the form of a dry powder after extraction.

The cheonma extract of the present invention can be extracted from natural, hybrid or mutant plants of cheonma, and can also be extracted from plant tissue culture.

The term "glycolytic enzyme" of the present invention means an enzyme that catalyzes the reaction. In the present invention, the term "glycolytic enzyme" means an enzyme that decomposes oligosaccharides such as cellulose, amylose, tannin, and pectin in a broad sense.

In the present invention, examples of the glycolytic enzyme include glucosyltransferase (CGTase, cyclodextrin glycosyltransferase), glucoamylase, cellulase, pectinase, glucosidase, and The enzyme selected from the group consisting of tanase enzymes or complex enzymes thereof may be used, but is not limited thereto. If the enzyme has an effect of enhancing the wrinkle improvement effect of the cheonma extract by enzyme treatment, Included in glycolytic enzymes. The enzymes may be used alone or in combination of these enzymes. Glycolytic enzyme-treated cheonma extract presented in the present invention, unless otherwise specified, includes all of the cheonma extract treated with glycolytic enzyme in addition to the enzymes exemplified above, it can be used as a wrinkle improving material.

As used herein, the term "skin wrinkle" refers to a streak caused by skin attenuation, which may be caused by a gene, a decrease in collagen present in the dermis of the skin, or an external environment.

In the present invention, "prevention or improvement of skin wrinkles" refers to inhibiting or inhibiting generation of wrinkles on the skin or alleviating wrinkles already produced.

The main functions of skin fibroblasts include the regeneration of damaged skin tissue through extracellular matrix synthesis and proliferation.Exogenous aging factors such as photoaging or accumulation of damage caused by free oxygen and cellular aging due to telomeres breakdown They lower the physiological activity of dermal fibroblasts in the dermis. Type I collagen, which accounts for more than 95% of the extracellular matrix of the skin, plays a very important part in the physiological activity of fibroblasts through interactions and signal exchanges with cell adhesion molecules and cytoskeleton. When the synthesis of is decreased, the amount of collagen in the skin tissue is reduced, thereby reducing the surface tension to reduce the skin elasticity and wrinkles of the skin, as well as reducing the bioactive function, including the proliferation and regeneration of the cells Cause a vicious cycle.

Therefore, collagen of dermal fibroblasts can be seen to be directly related to the repair or regeneration of tissue during skin regeneration, skin elasticity, skin wrinkle formation and skin damage.

That is, when the synthesis of collagen or procollagen of skin fibroblasts or the synthesis of matrix metallo-proteinase (hereinafter referred to as 'MMP') that breaks down collagen is inhibited, skin elasticity improvement, skin regeneration, The effects of improving skin wrinkles, healing wounds, repairing and regenerating damaged skin tissue, and preventing skin aging can be obtained.

The composition comprising the glycolytic enzyme-treated cheonma extract of the present invention as an active ingredient is characterized by increasing the expression of type I procollagen protein and inhibiting the expression of MMP-1 protein.

Specifically, the composition comprising the enzyme-treated cheonma extract of the present invention is a matrix protease-1 (matrix metallo-) to reduce cytotoxicity and promote collagen degradation in normal human dermal fibroblasts (NHDF) irradiated with ultraviolet light inhibits the expression of proteinase and increases the production of type I procollagen, which has effects such as increased skin elasticity, skin regeneration, skin wrinkle improvement, wound healing, repair and regeneration of damaged skin tissue, and prevention of skin aging. Can be. Specifically, it may have an effect of preventing or improving skin wrinkles (see FIGS. 3 to 6).

Enzyme-treated cheonma extract contained in the cosmetic composition for preventing or improving skin wrinkles of the present invention is effective in removing skin wrinkles by itself, and mixed with materials known to be effective in preventing and improving skin wrinkles. Even when used, it can continuously maintain the effect of preventing and improving skin wrinkles.

The material that is effective in improving skin wrinkles is not limited thereto, but chamomile flower extract, rosemary leaf extract, pasqueflower leaf extract, Ersnior extract, ginseng extract, Yukaritus leaf extract, geranium flower extract, ginseng extract, thyme extract , Nettle extract, yarrow extract, clove extract, fennel extract, papaya leaf extract, sunflower extract, and angelica extract.

In addition, the cosmetic composition of the present invention may further include a water-soluble vitamin, a fat-soluble vitamin, a polymer peptide, a polymer polysaccharide, a sphingolipid and a seaweed extract.

The water-soluble vitamins may be any one that can be formulated in 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 salts thereof (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 present invention. Included in water-soluble vitamins. The water-soluble vitamin can be obtained by a conventional method such as microbial conversion, microbial culture purification, enzyme method or chemical synthesis.

The fat-soluble vitamins may be any one that can be formulated in cosmetics, but preferably vitamin A, carotene, vitamin D2, vitamin D3, vitamin E, and the like, and derivatives thereof (ascorbic palmitate, ascorbic acid stearate). Bin, dipalmitinate, ascorbate, dl-alpha tocopherol acetate, dl-alpha tocopherol vitamin E, DL-pantothenyl alcohol, D-pantotenyl alcohol, pantothenyl ether, etc.) do.

The polymer peptide may be any compound that can be formulated in cosmetics, and preferably collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin and the like. The polymer peptide can be purified and obtained by a conventional method such as purification method of microbial culture solution, enzyme method or chemical synthesis method, or can be purified and used from natural products such as dermis such as pig or cow and silk fiber of silkworm.

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

The sphingolipide may be any compound as long as it can be blended into cosmetics, and preferably, ceramide, phytosphingosine, sphingose sugar lipid, and the like. The 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 one that can be formulated in cosmetics, but may preferably include brown algae extract, red algae extract, green algae extract, and the like, and calginane, arginic acid, and arginic acid purified from these seaweed extracts. Sodium, 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 the above essential components, if necessary, with other ingredients normally blended into the cosmetic.

Examples of the blending component that may be added include oil or fat components, humectants, emollients, ultraviolet absorbers, pH adjusters, flavoring agents, blood circulation accelerators, cooling agents, limiting agents, purified water and the like.

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

Examples of the ester fats and oils include glyceryl tri2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl mystinate, isopropyl palmitate, ethyl stearate, octyl palmitate, and isocetyl isostearate, Butyl stearate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocyanate isocetyl, isostearyl mysterylate, isostearyl palmitate, octylate acid octyldodecyl, isocetyl isostearate, diethyl sebacate, Diisopropyl adipropyl, isoalkyl neopentane, tri (capryl, capric acid) glyceryl, trimethyl ethyl trimethylolpropane, trimethyl sterol trimethylolpropane, tetra 2-ethylhexanopentane Slitol, Cetyl Caprylic Acid, Decyl Laurin, Hexyl Lauric Acid, Mycetic Decyl, Myristin Mitrisyl, Myristin Cetyl, Stearyl Stearate, Decyl Oleate, Ricinooleic Acid Cetyl, isostearyl laurate, isotridecyl myristin, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyl dodecyl oleate, octyl dodecyl linoleate, isopropyl isopropyl acid, 2-ethylhexanoic acid cetostearyl, 2-ethylhexanoic acid stearyl, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicapric acid, propylene glycol dicaprylic acid, neopentyl glycol dicapric acid Neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentane, isostearyl octanoate, octyl isonanoate , Hexyldecyl neodecanoate, octyldodecyl neodecanoate, isetyl isostearate, isstearyl isostearate, octyldecyl isostearate, polygly Linoleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactic acid, cetyl lactate, octyl lactate, triethyl citrate, acetyl triethyl citrate, Acetyl tributyl citrate, trioctyl citrate, diisostearyl malic acid, 2-ethylhexyl hydroxystearate, diethyl 2-ethylhexyl, diisobutyl adipicate, diisopropyl sebacinate, dioctyl sebacate, stearic acid Cholesteryl, isostearic acid cholesteryl, hydroxystearic acid cholesterol, oleic acid cholesteryl, oleic acid dihydrocholesteryl, isostearic acid pitsteryl, oleic acid pitsteryl, 12-steloylhydroxy Isocetyl Stearate, 12-Steloylhydroxystearate Stearyl, 12-Steloylhydroxystearate Isostea And the like ester, such as.

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

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

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

The animal or plant oils include avocado oil, almond oil, olive oil, sesame oil, rice bran 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, moon sour colostrum, marker demia nut oil, egg yolk oil, tallow, horse oil, mink oil, orange rape oil, jojoba oil, canderry wax, carnauba wax, liquid ranol And animal or plant fats and oils such as 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.

As the water-soluble low molecular moisturizer, 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 mentioned.

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

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

Examples of the ultraviolet absorber include paraaminobenzoic acid, ethyl paraaminobenzoate, amyl paraaminobenzoic acid, octyl paraaminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylate, Cinnamic acid benzyl, paramethoxy cinnamic acid-2-ethoxyethyl, paramethoxy cinnamic acid octyl, diparamethoxy cinnamic acid mono-2-ethylhexane glyceryl, paramethoxy cinnamic acid isopropyl, diisopropyl diisopropyl cinnamic acid ester mixture , Urocanoic acid, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and salts thereof, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone disulfonate, dihydroxy Benzophenone, tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p- (carbo-2'-ethylhexyl-1'-oxy) -1,3,5-triazine, 2- (2-hi And the like can be mentioned hydroxy-5-methylphenyl) benzotriazole.

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

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

Cosmetics prepared by containing the composition of the present invention may take the form of solutions, emulsions, viscous mixtures and the like.

In addition, the components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the components as an active ingredient, for example, conventional auxiliaries such as stabilizers, pigments and natural flavors and It may further include a carrier.

The composition of the present invention can be used in various ways, such as cosmetics or face wash having a skin wrinkle prevention and improvement effect.

As a product to which the composition of the present invention can be added, for example, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream , Cosmetics such as foundation, essence, nutrition essence, pack, and soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

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 a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent 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.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or improving skin wrinkles comprising a glycolytic enzyme-treated cheonma extract as an active ingredient.

The term "glycolytic enzyme" of the present invention means an enzyme that catalyzes the reaction. In the present invention, the term "glycolytic enzyme" means an enzyme that decomposes oligosaccharides such as cellulose, amylose, tannin, and pectin in a broad sense.

In the present invention, the glycolytic enzyme is a glucosyltransferase (CGTase, cyclodextrin glycosyltransferase), glucoamylase (glucoamylase), cellulase (cellulase), pectinase (glucosidase), and tangana The enzyme selected from the group consisting of a tannase enzyme or a complex enzyme thereof may be included, but is not limited thereto. If the enzyme has an effect of enhancing the antiwrinkle effect of the cheonma extract by enzymatic treatment, the sugar-valent moisture of the present invention Harm is included in the enzyme.

In addition, in the pharmaceutical composition for preventing or improving skin wrinkles of the present invention, the hydrolase treated cheonma extract is characterized by increasing the expression of type I procollagen protein and inhibiting the expression of MMP-1 protein. .

The composition of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the compositions of the invention may also be used in the form of their pharmaceutically acceptable salts, or may be used alone or in combination with other pharmaceutically active compounds.

Carriers, excipients and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In formulating the composition of the present invention, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.

The composition of the present invention is administered directly to the skin in the form of an external preparation for application to the skin in the form of an external preparation for creams, gels, patches, sprays, ointments, warnings, lotions, linens, pastas or cataplasmas. It may be, but is not limited thereto.

Preferred dosages of the mixed compositions of the present invention vary depending 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 of administration, and may be appropriately selected by those skilled in the art, preferably 0.01 μg / ml to 25 It may be administered at a dose of μg / ml. Topical administration may be administered once a day or may be divided several times.

Since the cosmetic and pharmaceutical compositions of the present invention have an effect of preventing or improving skin wrinkles per se, they can be directly used as a composition for preventing or improving skin wrinkles, and other natural extracts are effective for preventing and improving skin wrinkles. It is also available in the form of mixing with the ingredients is expected to be able to enhance the existing skin wrinkle prevention or improvement more.

As described above, the composition comprising the glycolytic enzyme-treated cheonma extract of the present invention as an active ingredient inhibits the expression of MMP-1 in human dermal fibroblasts and increases the production of type I procollagen, It is effective in preventing the occurrence and improving wrinkles of damaged skin. Overall, the compositions of the present invention can prevent and greatly improve skin wrinkles.

In addition, the composition of the present invention is not only very safe for human body, but also excellent in stability.

1 is a chromatographic result showing the changes in the components of the cheonma extract according to the hydrolase treatment.
Figure 2 is a graph showing the DPPH radical inhibitory activity of glycolytic enzyme-treated cheonma extract.
Figure 3 is a graph showing the effect of glycolytic enzyme-treated cheonma extract on the expression of MMP-1 protein in human fibroblasts.
Figure 4 is a graph showing the effect of glucose hydrolase treated cheonma extract on the expression of MMP-1 protein in UVB-irradiated human fibroblasts.
5 is a graph showing the effect of glycolytic enzyme-treated cheonma extract on the expression of type I procollagen protein in human fibroblasts.
Figure 6 is a graph showing the effect of glycolytic enzyme-treated cheonma extract on the expression of type I procollagen protein in UVB-irradiated human fibroblasts.

Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Example  One: Sugar hydrolysis  Preparation of Enzyme Treated Chunma Extract

After crushing 15kg of cheonma outpost, 50% ethanol was added 10-fold and refluxed for 4 hours at 70 ° C. to obtain an extract of about 0.92 kg (yield 6.13%).

HPLC analysis of 1 g of the extract showed that the content of 4HBA (4-hydroxy benzyl alcohol) was 12 mg (1.2% of the extract).

As a sugar acceptor, 300 g (4 HBA standard: 3.6 g) of cheonma extract was used as a sugar receptor, and 4.6 g of soluble starch was used as a sugar donor. The prepared sugar receptor and donor were mixed in 0.91 mL of 0.1 CaCl ₂ solution, 2,250 U of CGTase (Cyclodextrin glycosyltransferase) was added, and 0.1M phosphate acetate buffer solution (pH 6.0) was added thereto to make 700 mL.

The above reaction solution was reacted for 48 hours while stirring in a dark at 50 ° C., and then heated in a boiling water bath for 10 minutes to inactivate CGTase enzyme.

Subsequently, the maltooligosylglucosides produced by the CGTase enzyme reaction were adjusted to pH 5.0 with 1.0 M acetic acid aqueous solution to hydrolyze monoglucoside, and added with glucoamylase (80 U) and stirred at 50 ° C. Hydrolysis for hours.

After completion of the hydrolysis reaction, the reaction solution was autoclaved for 10 minutes to inactivate glucoamylase, and the supernatant was lyophilized.

Extracted in the same manner as above, lyophilized untreated enzyme was used as a comparative example.

Example  2. Comparative analysis of the components of cheonma extract with and without enzyme treatment

In addition, the enzyme-treated cheonma extract and the enzyme-free cheonma extract components prepared in Example 1 were analyzed using chromatography (Dionex Chromelon Chromatography, Dionex, USA) equipment. The column (Sun) C18 (Waters, 4.6x250mm, 5micron) was used as a column, the column oven temperature was 30 ℃, the detector was analyzed with a injection volume of 10uL, flow rate 1.0mL / min.

As a result, the enzyme-treated cheonma extract and the enzyme-free cheonma extract were found to be significantly different in the main component content, the enzyme-treated cheonma extract in the 4-hydroxy benzyl alcohol (4HBA) and glucose α-glycoside Significant increase in alpha glucopyranosiloxy benzyl alcohol (alpha-gastrodine, 4-alpha-D-glucopyranosyloxy benzyl alcohol) in the form of (FIG. 1).

Example  3. Enzymatically treated  Antioxidant Effect of Chunma Extract

Sang S et al., J. Agric. Food Chem. 50 (8), p2459-2463, 2002, et al., Using a reducing power of DPPH (1,1-diphenyl-2-picrylhydrazyl, Sigma, USA). According to the described method, the antioxidant activity of the enzyme-treated cheonma extract according to the present invention was measured.

180 μl of 0.2 mM DPPH solution dissolved in methanol, 20 μl of distilled water, cheonma extract (negative control), glycolytic enzyme treated cheonma extract, and arbutin (positive control) (1, 10, 50, 100, and 250 μg / ml) After incubating for 30 minutes at 37 ° C in a 96-well microplate, the amount of reduced DPPH was measured using a spectrophotometer (Precision microplate reader; Molecular Devices, VersaMax) and absorbance at 520 nm. The activity was compared. The experiment was performed three times (see FIG. 2).

As a result, it was observed that the cheonma extract of the hydrolase treated example showed a very good antioxidant effect in all 1, 50, 100, and 250 µg / ml treatment groups, compared to the unenzymatic cheonma extract. The levels of these antioxidant effects were also superior to those of the positive control arbutin.

Example  4. MMP -1 protein synthesis inhibition

2 ml of DMEM was added to a 40 mm cell culture dish, and human fibroblasts were inoculated at a concentration of about 1.2 × 10 ⁵ , followed by incubation for 24 hours at 37 ° C. and 5% CO ₂ . Thereafter, UVB was irradiated under 144mJ / cm ² condition, and then cultured for 3 days by replacing with a medium containing the glycolytic enzyme-treated cheonma extract prepared in Example 1 (1, 10, and 100 µg / ml, respectively). . Afterwards, the culture was harvested and centrifuged at 4 ° C. and 7500 rpm for 5 minutes using an ELISA method, MMP-1 (Human Total MMP-1 kit, R & D Systems, Inc., Minneapolis, MN, USA). The change in protein expression amount was confirmed (see FIGS. 3 and 4).

First, in the human fibroblast experiment without UV irradiation, it can be seen that the glycolytic enzyme-treated cheonma extract treatment group inhibits the production of MMP-1 protein in a concentration-dependent manner. This effect was found to be superior compared to the group treated with the unenzymatic cheonma extract, in which the cheonma extract of the example hydrolyzed enzyme was treated in the 1, 10, and 100 μg / ml treated groups. It was observed to show the effect of inhibiting the production of a significantly improved MMP-1 protein compared to the cheonma extract did not (Fig. 3).

In addition, human fibroblasts treated with ultraviolet radiation at a dose of 144 mJ / cm ² increased the production of MMP-1 protein by more than two times as compared to the group without UV treatment. Thus, the increase in the production of MMP-1 protein due to the ultraviolet irradiation treatment showed a phenomenon that decreases by the treatment of cheonma extract. The increased MMP-1 protein expression decreased by UV irradiation of cheonma extract was superior to the enzyme treated cheonma extract, compared to the group treated with ethanol extract, 1, 10, and 100 ㎍. In the / ml treated group, it was observed that the cheonma extract of the hydrolyzed enzyme treated example showed a significantly improved inhibitory effect on the production of MMP-1 protein compared to the unenzymatic cheonma extract (FIG. 4).

Through the above results, it was found that the glycolytic enzyme-treated cheonma extract of the present invention can be effectively used to prevent and improve skin wrinkles by effectively inhibiting the production of MMP-1 protein that causes wrinkles. .

Example  5. Type 1 Procollagen  Promote the synthesis of protein

2 ml of DMEM was added to a 40 mm cell culture dish, and human fibroblasts were inoculated at a concentration of about 1.2 × 10 ⁵ , followed by incubation for 24 hours at 37 ° C. and 5% CO ₂ . Thereafter, UVB was irradiated under 144mJ / cm ² condition, and then cultured for 3 days by replacing with a medium containing the glycolytic enzyme-treated cheonma extract prepared in Example 1 (1, 10, and 100 µg / ml, respectively). . Afterwards, the culture was harvested and centrifuged at 4 ° C. and 7500 rpm for 5 minutes to change protein expression levels of type 1 procollagen (Procollagen Type IC Peptide EIA Kit, Takara, Shiga, Japan) using the ELISA method. Was confirmed (see FIGS. 5 and 6).

First, in the human fibroblast experiment without ultraviolet irradiation, it was found that the group treated with Glycolytic enzyme-treated cheonma extract increased the production of type 1 procollagen protein in a concentration-dependent manner. This effect indicates a significantly improved production of type 1 procollagen protein compared to the group treated with unenzymatic cheonma extract, resulting in an increased effect of type 1 procollagen protein upon treatment with enzyme-treated cheonma extract. Larger (Figure 5).

In addition, human fibroblasts treated with UV radiation at a dose of 144 mJ / cm ² reduced the production of type 1 procollagen protein by 2.3 times compared to the UV-free group. The decrease in production of type 1 procollagen protein due to UV irradiation showed a phenomenon of increasing again by the treatment of cheonma extract. At this time, the chemihydrolase treated Chunma extract treated group showed a significantly improved production of type 1 procollagen protein compared to the non-enzymatically treated group (FIG. 6).

Through the above results, the composition comprising the glycolytic enzyme-treated cheonma extract of the present invention acts on the TGF-β / Smad signaling system to promote the production of type 1 procollagen, MMP- collagen degrading enzyme Through the action of suppressing the expression of 1, it could be confirmed that there is an effect to effectively prevent the generation of wrinkles, and improve the generated wrinkles.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto.

Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (9)

Enzymatic reaction product produced by the second enzymatic treatment with glucoamylase enzyme is effective on the enzymatic reaction product produced by the first enzymatic treatment of water-soluble starch and cheonma extract with glucosyltransferase (CGTase, cyclodextrin glycosyltransferase) enzyme. Cosmetic composition for preventing or improving skin wrinkles contained as a component. delete The method of claim 1,
The enzyme-treated cheonma extract increases the expression of type I procollagen protein, cosmetic composition for preventing or improving skin wrinkles, characterized in that to suppress the expression of MMP-1 protein. delete delete Enzymatic reaction product produced by the second enzymatic treatment with glucoamylase enzyme is effective on the enzymatic reaction product produced by the first enzymatic treatment of water-soluble starch and cheonma extract with glucosyltransferase (CGTase, cyclodextrin glycosyltransferase) enzyme. Pharmaceutical composition for preventing or improving skin wrinkles comprising as an ingredient. delete The method of claim 6,
The enzyme-treated cheonma extract is a pharmaceutical composition for preventing or improving skin wrinkles, characterized in that to increase the expression of type I procollagen protein, inhibit the expression of MMP-1 protein. delete

Images