KR20180020493A - Composition for anti-oxidation or anti-aging comprising 3-O-galloyl-3,3',5,5',7- pentahydroxyflavan - Google Patents

Abstract

The present invention relates to a composition for anti-oxidation comprising after-fermentation tea ketone fractions as active ingredients and, more specifically, to a composition for anti-oxidation or anti-aging comprising 3-O-galloyl-3,3′,5,5′,7-pentahydroxyflavan isolated from after-fermentation tea ketone fractions, isomers thereof, pharmaceutically allowable salts thereof, pro-drug thereof, hydrates thereof or solvates thereof as active ingredients. Accordingly, the composition of the present invention can be used in various fields associated with anti-oxidation and anti-aging.

Description

3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan. 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan}

This specification includes 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan To an antioxidant or anti-aging composition.

The human skin undergoes changes due to various internal and external factors as it gets older. Internally, the secretion of various hormones that regulate metabolism is decreased, the function of immune cells and the activity of cells are lowered, and the biosynthesis of immune proteins and biocompatible proteins necessary for the living body is reduced. As a result, As the amount of ultraviolet rays reaching the surface of the light beam increases and environmental pollution becomes more intense, free radicals and active harmful oxygen are increased, thereby reducing the thickness of the skin, increasing the wrinkles and reducing the elasticity, It becomes worse, skin troubles frequently occur, and there are various changes such as spots, freckles and black blotches.

As the aging progresses, the content and arrangement of collagen, elastin, hyaluronic acid, and glycoprotein, which constitute the skin, are changed or decreased, and oxidative stress due to free radicals and active noxious oxygen is experienced. Cox-2 (cyclooxygenase), an enzyme that produces proinflammatory cytokines, known to cause inflammation in most of the cells that make up the skin by aging or ultraviolet light The biosynthesis of matrix metalloproteinase (MMP), which is an enzyme that decomposes the skin tissue by these inflammatory factors, is increased, and nitric oxide (NO) production by iNOS (inducible nitric oxide synthase) . In other words, decrease of cellular activity due to natural endogenous aging and decrease of biosynthesis of substrate material by microinflammation, increase of stress due to various harmful environment, increase of active oxygen species by sunlight, As the degradation and denaturation are accelerated, the skin substrate is destroyed and thinned, and various symptoms of skin aging appear. Therefore, it is a reality that many researches have been conducted on active ingredients that can prevent and improve such aging phenomena.

On the other hand, active oxygen generated by various physical, chemical, and environmental factors such as in vivo enzymatic system, reductive metabolism, chemical agents, pollutants and photochemical reaction, etc. is non-selective, irreversible It is known to cause various diseases including cancer or aging cells by destructive action. In addition, lipid peroxides and various peroxides such as lipid peroxides produced as a result of lipid peroxidation by these active oxygen also cause oxidative destruction to cells and cause various dysfunctions and cause various diseases. Therefore, antioxidants such as free radical scavengers or peroxide formation inhibiting substances are expected to be used as agents for inhibiting or treating various diseases caused by these oxides. In particular, Antioxidant and anti-aging studies are continuing.

Korean Patent Registration No. 10-0975199

In one aspect, the present invention relates to an antioxidant or an anti-aging substance derived from a natural product, and to provide a composition for antioxidant or anti-aging comprising a post-fermentation tea fraction.

In another aspect, the present invention provides a pharmaceutical composition comprising 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyplasmin (3-O-galloyl- 5,5 ', 7-pentahydroxyflavan) as an active ingredient.

In one aspect, the present invention provides a pharmaceutical composition comprising 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyplasuban (3-O-galloyl-3,3 ' , 5 ', 7-pentahydroxyflavan), an isomer thereof, an acceptable salt thereof, a prodrug thereof, a hydrate thereof, a solvate thereof, or a post fermented tea ketone fraction containing the same as an active ingredient do.

In one aspect, the present invention provides a composition comprising a post fermented tea ketone fraction and a composition comprising a specific compound, and can be widely used in antioxidative and anti-aging related fields.

FIG. 1 shows the result of analysis of the post-fermentation tea extract by high performance liquid chromatography, with or without the acetone fraction.
Figure 2 shows a 10: 1 (v / v) fraction of caffeine-free chloroform: methanol divided by 10 sub-fractions using high-performance countercurrent chromatography (HPCCC).
Figure 3 shows fractions 1 to 5 of the 10 lower fractions fractioned using HPCCC.
Figure 4 shows fractions 6 to 10 of the 10 lower fractions fractioned using HPCCC.
FIG. 5 shows the oxidation inhibitory effect of trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane on DPPH (1,1-diphenyl-2-picryl hydrazyl) .
6 is a nuclear magnetic resonance (NMR) graph of trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane.
FIG. 7 is a graph showing that trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflasane shows an inhibitory effect on MMP-1 expression in HaCat cells.

Hereinafter, the present invention will be described in detail.

As used herein, the term " post-fermentation tea " includes fermentation by a separate microorganism or substance other than an enzyme present in tea leaves.

As used herein, the term " ketone fraction " includes fractions obtained by fractionation or fractionation of a specific substance or extract using a ketone solvent, and re-extraction thereof with a specific solvent. Any kind of ketone may be used, but acetone is preferable, and the fractionation method and the extraction method may be any method known to those skilled in the art.

As used herein, the term "isomers" refers in particular to optical isomers (for example, essentially pure enantiomers, essentially pure diastereomers or mixtures thereof) as well as morphological isomers (i. e., isomers differing only in the angle of one or more chemical bonds), position isomers (especially tautomers) or geometric isomers (e.g., cis-trans isomers) do.

As used herein, "essentially pure ", when used in reference to an enantiomer or partial isomer, means that at least about 90%, preferably at least about 95%, of a specific compound, such as an enantiomer or partial isomer, , More preferably at least about 97% or at least about 98%, even more preferably at least about 99%, even more preferably at least about 99.5% (w / w).

As used herein, " acceptable "is intended to refer to a substance that is approved by a government or equivalent regulatory agency that is capable of being used by an animal, more specifically a human, by avoiding significant toxic effects when used in conventional or medical dosage forms, Food codes, health functional foods, or those listed in general pharmacopoeias or as described in other general literature.

As used herein, " acceptable salt " means a salt according to one aspect of the present invention which is conventional or pharmaceutically acceptable and has the desired activity of the parent compound. The salt may be (1) formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; (3-hydroxybenzoyl) benzoic acid, or an acetic acid, propionic acid, hexanoic acid, cyclopentenepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, Benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4- chlorobenzenesulfonic acid, 2- 4-methylbicyclo [2,2,2] -oct-2-en-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert Acid addition salts formed with organic acids such as butylacetic acid, laurylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid; Or (2) salts formed when the acidic proton present in the parent compound is substituted.

As used herein, the term " prodrug " refers to a drug whose physical and chemical properties have been controlled by chemically altering a certain substance. Although it does not exhibit physiological activity itself, it does not exhibit chemical, physiological, The drug can be replaced with the original drug, and the drug efficacy can be demonstrated.

As used herein, " hydrate " means a compound to which water is bound, and is a broad concept that includes an inclusion compound having no chemical bonding force between water and the compound.

As used herein, " solvate " means a higher order compound formed between a molecule or ion of a solute and a molecule or ion of a solvent.

In one aspect, the present invention provides a pharmaceutical composition comprising 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan (3-O-galloyl-3,3 ' 5 ', 7-pentahydroxyflavan), an isomer thereof, an acceptable salt thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, a solvate thereof, or a post fermented tea ketone fraction containing the same Antioxidant or anti-aging composition.

According to one embodiment, the post-fermentation tea ketone fraction may be a ketone fraction for a water-insoluble extract of a post-fermentation tea.

According to another embodiment, the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane is a trans-3-O- , 5,5 ', 7-pentahydroxyflavan (Trans-3-O-galloyl-3,3', 5,5 ', 7-pentahydroxyflavan). The term " flavan " or "Flavan" in the present specification refers to the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan or trans- , 5,5 ', 7-pentahydroxyflasane.

According to another embodiment, the ketone fraction may be an alcohol extract of the ketone fraction.

According to another embodiment, the ketone is selected from the group consisting of acetone, carvone, pulegone, isolongifolanone, 2-heptanone, 2-pentanone, 3-hexanone, 3- , 2-octanone, 3-octanone, 2-nonanone, 3-nonanone, 2-undecanone, 2-tridecanone, methyl isopropyl ketone, ethyl isoamyl ketone, Aminocyclopentene, 2,3-heptanedione, 2,3-heptanedione, 2,3-heptanedione, 2,3-heptanedione, 2,3-heptanedione, 2-n-heptylcyclopentanone, cis-jasmone, dihydrojasmone, methylcorylon, 2-tert-butylcyclohexylcyclohexanone, Cyclohexanone, p-tert-butylcyclohexanone, 2-sec-butylcyclohexanone, celery ketone, krypton, p-tert- pentylcyclohexanone, methylcyclocitronone, 2-pentanone, oxide ketone, imicosporon, Methylacetophenone, p-methylacetophenone, propiophenone, acetophenone,? -Dyne, p-methoxyphenylacetone, p-methoxyphenylacetone, p-methoxyacetophenone, Dynascone, litorone, ionone, pseudoionone, methylionone, methylylitone, 2,4-di-tert-butylcyclohexanone, allylionone, 2 -Acetyl-3,3-dimethylnorbornane, verbenone, fenchone, cyclopentadecanone, cyclohexadecenone, and the like, and ketones as solvents which can be generally used in the art and their And mixtures thereof, preferably acetone.

According to another embodiment, the alcohol may be an alcohol alone or a mixture which can be generally used in the art, preferably ethanol.

According to another embodiment, the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone or trans-3-O- 5 ', 7-pentahydroxyplasane, isomers thereof, acceptable salts thereof, pharmaceutically acceptable salts thereof, prodrugs thereof, hydrates thereof or solvates thereof may be used in an amount of 0.000001% by weight or more, 0.000005 At least 0.001 wt.%, At least 0.00001 wt.%, At least 0.001 wt.%, At least 0.0001 wt.%, At least 0.0002 wt.%, At least 0.0005 wt.%, At least 0.001 wt.%, At least 0.005 wt.%, At least 0.01 wt. At least 1 wt.%, At least 1.5 wt.%, At least 1.8 wt.%, At least 2 wt.%, At least 2.2 wt.%, At least 2.5 wt.%, At least 2.8 wt.%, At least 3.2 wt.%, At least 3.5 wt.%, At least 3.8 wt.%, At least 4 wt.%, At least 4.2 wt.%, At least 4.5 wt.%, Or at least 4.8 wt.%, At most 5 wt. , 4.5 wt% Not more than 4.2 wt%, not more than 3.8 wt%, not more than 3.5 wt%, not more than 3.2 wt%, not more than 3 wt%, not more than 2.8 wt%, not more than 2.5 wt%, not more than 2.2 wt% , Not more than 1.5 wt%, not more than 1.2 wt%, not more than 1 wt%, not more than 0.5 wt%, not more than 0.3 wt%, not more than 0.1 wt%, not more than 0.05 wt%, not more than 0.01 wt%, not more than 0.005 wt% , 0.0005 wt% or less, 0.0002 wt% or less, 0.0001 wt% or less, 0.00005 wt% or less, 0.00001 wt% or less, or 0.000005 wt% or less. And preferably 0.000001 to 5% by weight.

According to another embodiment, the post-fermentation tea ketone fraction in the composition may be present in the composition in an amount of at least 0.01 wt%, at least 0.1 wt%, at least 1 wt%, at least 5 wt%, at least 10 wt%, at least 15 wt% , At least 25 wt%, at least 30 wt%, at least 35 wt%, at least 40 wt%, at least 45 wt%, at least 50 wt%, at least 55 wt%, at least 60 wt% At least 95 wt%, at least 75 wt%, at least 80 wt%, at least 85 wt%, at least 90 wt%, or at least 95 wt% Not more than 80 wt%, not more than 75 wt%, not more than 70 wt%, not more than 65 wt%, not more than 60 wt%, not more than 55 wt%, not more than 50 wt%, not more than 45 wt%, not more than 40 wt% Up to 25 wt.%, Up to 20 wt.%, Up to 15 wt.%, Up to 10 wt.%, Up to 5 wt.%, Up to 1 wt.%, Up to 0.1 wt.%, Or up to 0.05 wt.% have. And preferably 0.01% by weight to 100% by weight.

According to another embodiment, the fraction is selected from the group consisting of 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan or trans-3-O- 5 ', 7-pentahydroxyflasane, an isomer thereof, an acceptable salt thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof or a solvate thereof in an amount of not less than 0.0001% At least 0.0005% by weight, at least 0.001% by weight, at least 0.005% by weight, at least 0.01% by weight, at least 0.05% by weight, at least 0.1% by weight, at least 0.5% by weight, at least 1% by weight, at least 1.5% At least 4 wt.%, At least 4.5 wt.%, At least 5 wt.%, At least 4.5 wt.%, At most 4 wt.%, At most 3.5 wt.%, Not more than 3 wt%, not more than 2.5 wt%, not more than 2 wt%, not more than 1.5 wt%, not more than 1 wt%, not more than 0.5 wt%, not more than 0.1 wt%, not more than 0.05 wt% 0.001 wt% or less, 0.0005 wt% or less, and Can be 0.0002% by weight or less. Preferably from 0.0001 wt% to 5 wt%.

According to another embodiment, the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan or trans-3-O- , The isomer thereof, an acceptable salt thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof, is not less than 0.0001 g / kg / day, 0.001 g / day kg / day, at least 0.01 g / kg / day, at least 0.1 g / kg / day, at least 1 g / kg / day, at least 2 g / kg / day, at least 3 g / Day or more, 6 g / kg / day, 6 g / kg / day, 7 g / kg / day, 8 g / kg / day, 9 g / kg / day, 10 g / day or more, 13 g / kg / day or more, or 14 g / kg / day or more, 15 g / kg / day or less, 14 g / kg / day, 13 g / kg / day, 12 g / No more than 10 g / kg / day, no more than 9 g / kg / day, no more than 8 g / kg / day, no more than 7 g / kg / day, no more than 6 g / kg / day or less, 3 g / kg / day or less, 2 g / kg / day or less, 1 g / kg / day or less, 0.1 g / 0.01 g / kg / day or less, 0.001 g / kg / day or less, or 0.0005 g / kg / day or less. Preferably 0.0001 g / kg / day to 15 mg / kg / day.

According to another embodiment, the antioxidant may be due to oxidation inhibition of the radical or reduction of active oxygen.

According to another embodiment, the antioxidant may be at least one selected from the group consisting of anti-aging, skin elasticity improvement, and skin wrinkle improvement.

According to another embodiment, the anti-aging may be skin aging, and may be at least one selected from the group consisting of skin elasticity improvement and skin wrinkle improvement.

According to another embodiment, at least one selected from the group consisting of anti-aging, skin elasticity improvement, and skin wrinkle improvement may be one which inhibits the expression of collagenase. According to one embodiment, the collagenase may be MMP-1.

According to one embodiment of the present invention, the organic radical DPPH (1,1-diphenyl-2- (4-fluorophenyl) picryl hydrazyl). As a result, it was found that the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane of the present invention is a synthetic antioxidant or anti- As shown in Table 1, the antioxidant of the present invention can be used for antioxidant, anti-aging, skin elasticity improvement, and skin wrinkle improvement because of its remarkable antioxidant activity than Trolox (see Table 1). The main cause of aging is active oxygen or free radicals, so antioxidants can inhibit aging. Particularly, since aging of skin is directly related to skin elasticity and wrinkles, antioxidative and anti-aging actions can be realized when the above-mentioned substances of the present invention are applied to the human body or skin, and thus skin elasticity and skin wrinkles can be improved can do.

In addition, the inhibition of collagenase (MMP-1) expression using the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasubane according to an embodiment of the present invention (Fig. 5 and Fig. 7). As shown in Fig. 5 and Fig. As a result, it can be seen that the above materials can be used for anti-aging, skin elasticity improvement, and skin wrinkle improvement. As aging progresses, collagen and the like decrease, and accordingly, various symptoms such as decrease in elasticity and increase in wrinkles are accompanied. Therefore, anti-aging, skin elasticity improvement and skin wrinkle improvement can be promoted by inhibiting the expression of collagenase .

The composition according to one aspect of the present invention may be a pharmaceutical composition. The pharmaceutical composition may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, and the like. Formulations for oral administration may be in the form of tablets, pills, soft and hard capsules, granules, powders, granules, solutions, emulsions or pellets, It is not. Formulations for parenteral administration may be, but are not limited to, solutions, suspensions, emulsions, gels, injections, drops, suppositories, patches, ointments or spraying agents. The formulations may be readily prepared according to conventional methods in the art and may be prepared by conventional means including, but not limited to, surfactants, excipients, wetting agents, emulsifying accelerators, suspending agents, salts or buffers for controlling osmotic pressure, colorants, spices, stabilizers, preservatives, Other adjuvants may also be included.

The application amount or dosage of the composition according to one aspect of the present invention will vary depending on the age, sex, weight, pathological condition and the severity of the subject to be administered, the route of administration, or the judgment of the prescriber. Determination of the amount of application based on these factors is within the level of those skilled in the art and the daily dose of the active ingredient may be, for example, from 0.001 μg / kg / day to 5000 mg / kg / day, more specifically from 0.0001 mg / kg / / kg / day. < / RTI >

According to another embodiment, the composition may be a food composition or a health food composition.

The formulation of the food composition or the health food composition may be formulated into, for example, a tablet, a granule, a pill, a powder, a capsule, a liquid such as a drink, a caramel, a gel, a bar, . The food composition or health food composition of each formulation may be formulated by those skilled in the art without difficulty depending on the purpose of formulation or use, in addition to the active ingredient, and the synergistic effect Can happen.

The composition may be administered by various methods such as simple ingestion, drinking, injection administration, spray administration or squeeze administration.

In a food composition or a health food composition according to one aspect of the present invention, the dosage determination of the active ingredient is within the level of those skilled in the art, and its daily dose is, for example, from 0.0001 mg / kg / day to 15 mg / / Day, but it is not limited thereto, and may be varied depending on various factors such as the age, health condition, and complications of the subject to be administered.

The food composition or the health food composition according to one aspect of the present invention may be used in various foods such as chewing gum, caramel product, candy, ice cream, confectionery, beverage such as soft drink, mineral water, alcoholic beverage, And the like.

In addition to the above, the food composition or the health food composition according to one aspect of the present invention may contain flavors, coloring agents and enhancers (cheese, chocolate, etc.) such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the food compositions according to one aspect of the present invention may include natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally comprised in the range of 0 to about 50 parts by weight per 100 parts by weight of the composition according to one aspect of the present invention.

According to another embodiment, the composition may be an external preparation for skin. The composition for external application for skin may be a composition such as a cosmetic, an oral, a detergent, a medicine and a pharmaceutical, but is not limited thereto. The external preparation for skin is not particularly limited in its formulation. For example, the external preparation for skin is not particularly limited, and examples thereof include softening lotion, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, Packs, powders, body lotions, body creams, body oils, and body essences.

In addition, the composition for external application for skin of the present invention may further comprise at least one selected from a salt for a functional purpose and a pH adjusting agent for pH adjustment. At this time, the salt may be selected from inorganic salts, organic salts and / or organic-inorganic salts for ion shielding, moisturizing, ultraviolet shielding and the like. For example, the salt may be selected from sodium chloride (NaCl), sodium phosphate (Na3PO4) and calcium chloride (CaCl2). The pH adjusting agent is selected from an acid or base and may be selected from the group consisting of hydrochloric acid, sulfuric acid, tartaric acid, citric acid, phosphoric acid, acetic acid, lactic acid solution, sodium lactate, sodium hydroxide, potassium hydroxide, Oleamine, ammonia, and the like.

The external preparation for skin of the present invention may be a cosmetic, pharmaceutical or quasi-drug composition. The cosmetic, pharmaceutical or quasi-drug composition may further contain adjuvants such as preservatives, stabilizers, wettable or emulsifying accelerators, salts for controlling osmotic pressure and / or buffers, and other therapeutically useful substances. The composition may be formulated into lotions, creams, ointments or gels. The skin external composition composition is preferably transdermally administered.

The dosage of the active ingredient of the pharmaceutical or quasi-drug composition will vary depending on the age, sex, and weight of the subject to be treated, the particular disease or condition to be treated, the severity of the disease or pathological condition, the route of administration and the judgment of the prescriber. Dosage determinations based on these factors are within the level of those skilled in the art. In general, the dosage of the active ingredient may range from 0.0001 mg / kg / day to 15 mg / kg / day, but is not limited thereto.

In one aspect of the present specification, the composition is not particularly limited in formulation, but may be a cosmetic composition. The cosmetic composition may be, for example, a composition for skin and / or hair, for example, a softening agent, a convergent lotion, a nutritional lotion, a nutritional cream, a massage cream, an eye cream, an eye essence, a cleansing cream, a cleansing lotion, It may have formulations such as cleansing water, packs, powders, body lotions, body creams, body essences, body cleansers, hair dye preparations, shampoos, rinses, conditioners, hair tonics, ointments, gels, creams, patches, But is not limited thereto.

 In addition, in the respective formulations, the ingredients other than the above-mentioned essential ingredients can be suitably selected and blended by those skilled in the art depending on the kind of the external preparation, the purpose of use, and the like.

The cosmetic compositions according to the present disclosure may be provided in all formulations suitable for topical application. For example, it may be provided as a formulation of a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an oil phase in water, a suspension, a solid, a gel, a powder, a paste, a micro needle, a foam or an aerosol composition have. Compositions of such formulations may be prepared according to conventional methods in the art.

The cosmetic composition according to the present invention may further contain the components included in the functional additive and the general cosmetic composition in addition to the compounds or fractions of the present invention. The functional additives may include water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts. The cosmetic composition according to the present disclosure may contain other ingredients, which may preferably have a synergistic effect on the main effect, without impairing the main effect. In addition, the cosmetic composition according to the present invention may further include a moisturizing agent, an emollient agent, a surfactant, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a perfume, a cold agent or a limiting agent. The compounding amount of the above components can be easily selected by those skilled in the art within a range not to impair the objects and effects of the present invention, and the amount thereof may be 0.001 to 10% by weight, specifically 0.01 to 3% by weight, have.

Hereinafter, the configuration and effect of the present invention will be described in more detail with reference to examples, experimental examples, and formulation examples. However, these examples are provided for illustrative purposes only in order to facilitate understanding of the present specification, and the scope and range of the present specification are not limited by the following examples.

Example 1 Preparation of Post-Fermented Tea Samples

Water was added to green tea made from green tea ( Camellia sinensis var. Yabukita ) leaf to adjust the moisture content to 40% by weight. Bacillus subtillis (5 × 10 ⁶ cfu / g) was inoculated therein, fermented at 50 ° C. for 3 days, and fermented at 80 ° C. for 4 days. The fermented tea was dried to a moisture content of 4% by weight to 6% by weight with hot air at 70 캜 to 80 캜, and aged at 10 캜 for 100 days.

The aged tea samples were pulverized for 15 seconds, and then sieved with a stainless steel sieve having a mesh size of 1 mm. Then, 50 mg of the pulverized product was added to a 1.5 ml Eppendorf tube, and 1 ml of deionized water was added thereto. The mixture was stirred at a constant rate for 30 minutes in a constant temperature water bath at 60 ° C, and centrifuged at 25,000C for 13 minutes at 13,000 rpm. Only the non - soluble portion of the dried fermented green tea extract was isolated.

≪ Example 2 > Obtained fractions and compound separation

150 g of the post-fermentation tea sample was fractionated with acetone to remove catechin derivatives and caffeine as much as possible, and a soluble matter with flavonoids was obtained. 40 g of the acetone solubles were eluted first with silica gel column chromatography using a mixture of chloroform and methanol as a solvent in the order of 10: 1, 5: 1, 1: 1 (v / v) .

As a result of analysis by high performance liquid chromatography (HPLC, Waters Alliance 2695 system, Waters, USA) of 12 g of a chloroform: methanol 10: 1 (v / v) fraction, , Caffeine was still present as the main compound, and it was confirmed that caffeine was removed after fractionation using acetone as described above (FIG. 1).

8.9 g of the caffeine-free chloroform: methanol 10: 1 (v / v) fraction was fractionated using high-performance countercurrent chromatography (HPCCC, Dynamic Extractions Ltd, UK). The solvent used was n-hexane-TBME (Methyl tert-butyl ether) -BuOH-MeCN-Water (0.25: 3: 1: 1: 5, v / v) and the flow rate was 25 ml / min. A total of 10 sub-fractions were divided using the above conditions (FIG. 2), and analyzed by HPLC to confirm that most of the compounds contained in each fraction were flavonoid series (FIGS. 3 and 4).

From this fraction, trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone (Trans-3-0- were separated pentahydroxyflavan, C ₂₂ H ₁₈ O _10, molecular weight of 442.0900), the structure of the isolated compound are as follows.

(2)

Methanol-d3 was used as a solvent in the case of ¹ H nuclear magnetic resonance (NMR). The instrument was a Bruker Advance DPX-500 (BRUKER, USA) The NMR data of the -3,3 ', 5,5', 7-pentahydroxyflasane are as follows, and a specific graph is shown in FIG.

^{1 H NMR δ (500Hz) 7.01} (s, 2H), 6.89 (s, 1H), 6.76 (s, 2H), 6.04 (s, 1H), 5.96 (s, 1H), 5.36 (m, 1H), 5.13 (m, 1 H), 2.83 (m, 1 H), 2.73 (m, 1 H)

Experimental Example 1: Antioxidant effect test (DPPH test)

The antioxidant activity of the trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflasane was examined.

Specifically, a method of evaluating the antioxidant ability through the change of the absorbance generated by the reduction of the organic radical, DPPH (1,1-diphenyl-2-picryl hydrazyl) (the antioxidant is oxidized) was used. DPPH was inhibited and the absorbance was reduced compared with the control, and the concentration showing an absorbance of 50% or less of the absorbance of the control group was evaluated as the effective antioxidant concentration.

After adding 190 μL of 100 μM DPPH solution to ethanol, 10 μL of trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflurane and a control sample were added, After incubation at 37 ° C for 30 minutes, absorbance was measured at 540 nm. As a control sample, a synthetic antioxidant Trolox, which is widely used, was used. The results are shown in the following table.

DPPH analysis results (% inhibition) (IC ₅₀ : sample concentration when absorbance was reduced by 50% by the added sample) sample IC ₅₀ ([mu] M) Trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane 6.01 Trolox 8.66

As can be seen from the above Table 1, the trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasuban according to the present invention has more antioxidant ability than trolox I could. Thus, the present invention provides the use of trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane of the present invention for antioxidative use, antioxidative use, maintenance and enhancement of skin elasticity through antioxidation, It is possible to use it for the purpose of removing and wrinkling the skin through the skin.

<Experimental Example 2> Inhibitory effect of MMP-1 (matrix metalloproteinase-1, collagenase) on human skin fibroblasts

Human fibroblasts (KF-4109, obtained from Klebosha) were cultured on a 60 mm dish with 10% FBS-DMEM (Dulbecco's Modified Eagle's Media) and starvated with 0% FBS-DMEM for 24 hours. After washing twice with PBS, ultraviolet light was irradiated. The trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflagane was treated with 2 g / ml of the medium while changing the medium with 0% FBS-DMEM, (Control group treated with DMSO). MMP-1 was identified by Western blotting by collecting the medium, loading the same volume of medium, and extracting proteins from the cells to equally load the cell lysate and Western blotting to confirm the expression of alpha-tubulin.

As a result, it was confirmed that trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane (Flavan in FIG. The expression of MMP-1 was not significantly different according to the treatment. However, when irradiated with ultraviolet light (UV), trans- Treatment of 3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflagane reduced the expression of MMP-1 (Figure 5). On the other hand, in the case of alpha-tubulin, there was almost no difference in expression according to trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasuban treatment. That is, it was confirmed that trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflasane inhibits the expression of MMP-1 (collagenase). Thus, the trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan of the present invention can be used for the prevention of collagen decomposition, antioxidation, prevention and alleviation of skin aging, It can be used for the purpose of improving the appearance, alleviating the wrinkles of the skin and removing it.

<Experimental Example 3> Inhibitory effect of MMP-1 (collagenase) on HaCaT cells

HaCaT cells (provided by Prof. Dr. NE Fusenig, DKFZ Heidelberg, Germany), a human keratinocyte cell line, were cultured on a 60 mm dish with 10% FBS-DMEM (Dulbecco's Modified Eagle's Media) Starvation with FBS-DMEM. After washing twice with PBS, ultraviolet light was irradiated. The medium (flavan, i.e., trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflurane) was added to the medium at a concentration of 2 ug / ml After treatment, the cells were cultured for 48 hours (control group treated with DMSO). MMP-1 was identified by Western blotting by collecting the medium, loading the same volume of medium, extracting protein from the cells, loading the cell lysate equally, and confirming the expression of alpha-tubulin by Western blotting. In Fig. 7, "No UV" means that UV (ultraviolet) treatment was not performed.

As a result, it was found that trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavan inhibited the expression of MMP-1 (collagenase) (Fig. 7). Thus, the trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan of the present invention can be used for the prevention of collagen decomposition, antioxidation, prevention and alleviation of skin aging, It can be used for the purpose of improving the appearance, alleviating the wrinkles of the skin and removing it.

&Lt; Formulation Example 1 >

A soft capsule filling liquid was prepared by mixing 150 mg of the post-fermented tea acetone fraction according to Example 2 of the present invention, 440 mg of lactose, 430 mg of corn starch and 2 mg of magnesium stearate. Separately from the above, a soft capsule sheet was prepared in a ratio of 66 parts by weight of gelatin, 24 parts by weight of glycerin and 10 parts by weight of sorbitol solution, and filled with the filling liquid to prepare a soft capsule.

In addition, 3 mg of trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyplasmin, 440 mg of lactose, 450 mg of corn starch and 2 mg of magnesium stearate according to the present invention were mixed, Capsule filling liquid is prepared. Then, the soft capsule sheet was prepared as described above and filled with the filling liquid to prepare a soft capsule.

&Lt; Formulation Example 2 >

The mixture of 150 mg of the post-fermented tea acetone fraction according to Example 2 of the present invention, 15 mg of vitamin E, 15 mg of vitamin C, 250 mg of galactooligosaccharide, 60 mg of lactose and 140 mg of maltose was granulated and granulated using a fluidized bed drier, sugar ester) was added. The composition was tableted by a conventional method to prepare tablets.

Further, 3 mg of trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyplasone of Example, 15 mg of vitamin E, 15 mg of vitamin C, 259 mg of galactooligosaccharide, 140 mg of maltose were mixed and granulated using a fluidized bed drier and 8 mg of sugar ester was added. The composition was tableted by a conventional method to prepare tablets.

&Lt; Formulation Example 3 >

Fermented tea acetone fractions according to Example 2 of the present invention, 9 mg of vitamin E, 9 mg of vitamin C, 10 g of glucose, 0.6 g of citric acid and 25 g of liquid oligosaccharide were mixed and then filled with 400 ml of purified water. And then sterilized at 30 DEG C for 4 to 5 seconds to prepare a drink.

In addition, 0.1 mg of trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyplazone, 9 mg of vitamin E, 9 mg of vitamin C, 10 g of glucose, 0.6 g of citric acid, 25 g of oligosaccharide were mixed, and 400 ml of purified water was added to fill each bottle with 200 ml each. And then sterilized at 30 DEG C for 4 to 5 seconds to prepare a drink.

&Lt; Formulation Example 4 >

150 mg of the post-fermented tea acetone fraction according to Example 2 of the present invention, 9 mg of vitamin E, 9 mg of vitamin C, 250 mg of anhydrous crystalline glucose and 550 mg of starch were mixed and molded into granules using a fluidized bed granulator, .

Further, 3 mg of trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyplasone of Example, 9 mg of vitamin E, 9 mg of vitamin C, 250 mg of anhydrous crystalline glucose, The mixture was granulated into granules using a fluidized bed granulator, and filled into a bag to prepare granules.

&Lt; Formulation Example 5 >

 Injection was prepared according to a conventional method according to the composition shown in Table 2 below.

Compounding ingredient content The post-fermented tea acetone fraction according to Example 2  100 mg Sterile sterilized water for injection Suitable amount pH adjusting agent Suitable amount

In addition, an injectable preparation was prepared by using 3 mg of the trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflasane of the example instead of the post-fermented tea acetone fraction.

 &Lt; Formulation Example 6 >

Healthy foods were prepared in a conventional manner according to the compositions shown in Table 3 below.

ingredient content The post-fermented tea acetone fraction according to Example 2  150 mg Vitamin mixture Vitamin A acetate 70 [mu] g Vitamin E 1.0 mg Vitamin B1 0.13 mg Vitamin B2 0.15 mg Vitamin B6 0.5 mg Vitamin B12 0.2 [mu] g Vitamin C 10 mg Biotin 10 [mu] g Nicotinic acid amide 1.7 mg Folic acid 50 [mu] g Calcium pantothenate 0.5 mg Mineral mixture Ferrous sulfate 1.75 mg Zinc oxide 0.82 mg Magnesium carbonate 25.3 mg Potassium monophosphate 15 mg Dicalcium phosphate 55 mg Potassium citrate 90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg

Further, a health food was prepared by using 3 mg of the trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflasane of the example instead of the post-fermented tea acetone fraction.

Although the composition ratio of the vitamin and the mineral mixture is comparatively comparatively mixed with the ingredient suitable for the health food, the compounding ratio may be arbitrarily changed. The ingredients are mixed according to a conventional method for producing a healthy food, Can be used for the production of health food compositions.

&Lt; Formulation Example 7 >

ingredient content The post-fermented tea acetone fraction according to Example 2  50 mg Citric acid 1000 mg oligosaccharide 100 g Plum concentrate 2 g Taurine 1 g Purified water Balance Total volume 900 ml

As shown in Table 4, the remaining amount of purified water was added to a total volume of 900 ml, and the above components were mixed according to a conventional health drink manufacturing method. After stirring for about 1 hour at 85 캜, the solution was filtered Sterilized 2 liter container, sealed sterilized, and stored in a refrigerator to prepare a health drink.

Further, a health drink was prepared by using 0.1 mg of the trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflasane of the example instead of the post-fermented tea acetone fraction.

&Lt; Formulation Example 8 > Flexible longevity (skin lotion)

  3% by weight of the post-fermented tea acetone fraction according to Example 2, 1.00% by weight of L-ascorbic acid-2-phosphate magnesium salt, 1.00% by weight of water soluble collagen (1% aqueous solution), 0.10% by weight of sodium citrate, , 0.20% by weight of licorice extract, 3.00% by weight of 1,3-butylene glycol, and the remaining amount of purified water was used to prepare a softening lotion (skin lotion).

In addition, a softening longevity was prepared by using 0.1 wt% of trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane of the example instead of the post-fermented tea acetone fraction.

&Lt; Formulation Example 9 >

3.00% by weight of post-fermented tea acetone fractions according to Example 2, 2.00% by weight of polyethylene glycol monostearate, 5.00% by weight of self emulsifying monostearate glycerin, 4.00% by weight of propylene glycol, 6.00% by weight of squalane, 6.00% by weight of glyceryl, 1.00% by weight of sphingoglycolipids, 7.00% by weight of 1,3-butylene glycol, 5.00% by weight of beeswax, and purified water were used.

In addition, a cream-type preparation was prepared by using 0.1 wt% of trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflasane of the Example instead of the post-fermented tea acetone fraction.

&Lt; Formulation Example 10 >

3.00% by weight of the post-fermented tea acetone fraction according to Example 2, 13.00% by weight of polyvinyl alcohol, 1.00% by weight of L-ascorbic acid-2-phosphate magnesium salt, 1.00% by weight of lauroyl hydroxyproline, % Aqueous solution), 3.00% by weight of 1,3-butylene glycol, 5.00% by weight of ethanol, and purified water.

Further, a pack was prepared by using 0.1 wt% of trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflasane of the Example instead of the post-fermented tea acetone fraction.

Having described specific portions of the disclosure in detail, those skilled in the art will appreciate that these specific embodiments are merely exemplary and are not intended to limit the scope of the disclosure. Accordingly, the actual scope of the disclosure will be defined by the appended claims and their equivalents.

Claims (15)

3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan (3-O-galloyl-3,3 ' , An isomer thereof, an acceptable salt thereof, a prodrug thereof, a hydrate thereof, a solvate thereof, or a post-fermentation tea ketone fraction containing the same as an active ingredient
[Chemical Formula 1]

. 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan (3-O-galloyl-3,3 ' , An isomer thereof, an acceptable salt thereof, a prodrug thereof, a hydrate thereof, a solvate thereof, or a post-fermentation tea ketone fraction containing the same as an active ingredient
[Chemical Formula 1]

. 3. The composition of claim 1 or 2, wherein the post-fermentation tea ketone fraction is a ketone fraction for a water-insoluble extract of a post-fermentation tea. 3. The composition of claim 1 or 2, wherein the ketone is acetone. The method according to claim 1 or 2, wherein the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane is trans-3-O- 3,3 ', 5,5', 7-pentahydroxyflavan (Trans-3-O-galloyl-3,3 ', 5,5'
(2)

. The composition of claim 1 or 2, wherein the amount of 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane, an isomer thereof, an acceptable salt thereof, The hydrate thereof, or a solvate thereof, is 0.000001 wt% to 5 wt%, based on the total weight of the composition. The composition of claim 1 or 2, wherein the content of post fermented tea ketone fractions in the composition is from 0.01% to 100% by weight relative to the total weight of the composition. 3. The composition of claim 1 or 2, wherein said fraction is selected from the group consisting of the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflasane, the isomer thereof, , Hydrate thereof, or solvate thereof, is contained in an amount of 0.0001 wt% to 5 wt% based on the total weight of the fraction. The method according to claim 1 or 2, wherein the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyplazane (3-O-galloyl-3,3 ' Wherein the dosage is from 0.0001 mg / kg / day to 15 mg / kg / day, or an isomer thereof, an acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof. The composition of claim 1, wherein the antioxidant is by inhibiting oxidation of the radical. The composition according to claim 2, wherein the anti-aging is at least one selected from the group consisting of skin elasticity improvement and skin wrinkle improvement. 12. The composition according to claim 11, wherein the skin elasticity improvement and skin wrinkle improvement are achieved by inhibiting the expression of collagenase. 3. The composition of claim 1 or 2, wherein the composition is a food composition. 3. The composition according to claim 1 or 2, wherein the composition is a pharmaceutical composition. The composition of claim 1 or 2, wherein the composition is a cosmetic composition.

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