KR20200088674A - Composition for preventing or improving skin wrinkles comprising green tea extract which has modified amounts of ingredients - Google Patents

Abstract

The present invention discloses a composition for preventing or alleviating skin wrinkles containing a green tea extract with a varied ingredient content. Specifically, an extract and a composition according to an aspect of the present invention have less skin irritation, and are safe by being derived from environmentally friendly natural plant, thereby being able to effectively inhibit the expression of MMP-1 and exhibit excellent skin wrinkle prevention or alleviation efficacy.

Description

COMPOSITION FOR PREVENTING OR IMPROVING SKIN WRINKLES COMPRISING GREEN TEA EXTRACT WHICH HAS MODIFIED AMOUNTS OF INGREDIENTS}

The present invention relates to a composition for preventing or improving skin wrinkles comprising green tea extract having a changed component content.

The skin is a primary protective layer of the human body and protects organs in the body from changes in temperature and humidity, irritation of external environment such as ultraviolet rays and pollutants, and plays an important role in maintaining bio homeostasis such as body temperature control. This important skin, like other organs in the human body, gradually progresses with age. There are two main causes of skin aging, which can be classified into internal factors such as genetic variation of cells and tissue changes and external factors such as ultraviolet rays and humidity (Masamitsu Ichihashi, Fragrance Journal, Vol. 32, No. 5, 24~30). ). In the skin damaged by internal and external factors, skin resilience occurs, changes in the environment around the cell, and weakening of the intercellular network occur, resulting in scattered skin structures, resulting in loss of elasticity of the skin, keratinization, wrinkle formation, and skin transparency. Symptoms such as deterioration and skin atrophy appear.

The cause of the biggest change in aging skin is a decrease in the growth factor that was active in young skin. Growth factor acts as the first switch in the process of regeneration in the skin, so a decrease in growth factor in the skin causes a variety of deterioration in the skin, resulting in loss of components in the skin and disruption of the skin structure. In fact, the skin of a young baby whose growth factor is most active in the skin recovers quickly and easily even if it is wounded and has an amazing regenerating ability that does not leave any scars. In order to restore the wound when the wound occurs, the driving force of the skin and blood vessel cells to gather around the wound and create new cells to create a new skin structure with fibers such as collagen and elastin can be considered as growth factors. The types of growth factors participating in this series of processes include TGF-α (Transforming Growth Factor-α) and EGF (Epidermal Growth Factor), which participate in the entire regeneration process, and FGF (Fibroblast Growth), which plays a role in cell regeneration and activation. Factors include vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF) and insulin-like growth factor (IGF).

In addition, the aging phenomenon caused by ultraviolet light is called photoaging, and this photoaging phenomenon generates active oxygen radicals (ROS: Reactive Oxygen Radical) inside the cell by ultraviolet light, and this active oxygen is a dermal layer through a signaling system that causes an inflammatory reaction. Promote the synthesis of proteolytic enzymes (MMP-1, MMP-3 and MMP-9, etc.) such as collagen and elastin, which are elastic fibers of (Fisher GJ et al., Nature, Vol. 379, 335-339) of the dermal layer It is known that skin wrinkles are caused by reducing elasticity.

Accordingly, research activities have been continuously conducted to develop a physiologically active substance having an effect of improving wrinkles in order to prevent this skin aging phenomenon and maintain a healthier and more elastic skin. Typically, tretinoin (trans-retinoic acid) was approved by the U.S. FDA as a treatment for improving photo-aged skin in 1995, and retinol, which has fewer side effects, began to be used in cosmetic ingredients in the late 1990s and began in earnest. Wrinkle improvement cosmetics have begun to be released. Since then, female hormone-like substances and antioxidants extracted from various plants have been introduced into cosmetics as wrinkle-improving raw materials.

Although consumers' interest in and demand for cosmetics related to skin aging is rapidly increasing, the side effects of functional cosmetic products made of synthetic materials are continuously being raised, and research on the development of cosmetics using natural products that are effective and non-toxic is being actively conducted.

Republic of Korea Registered Patent Publication No. 10-1863053

An object of the present invention is to provide a composition derived from a natural plant that has no toxicity and is excellent in preventing or improving skin wrinkles and a method for manufacturing the same.

In order to achieve the above object, the present invention, in one aspect, based on the total weight of the extract, 4 to 15% by weight of (-)-gallocatechin gallate ((-)-gallocatechin gallate, GCG) and 4 to 15 It provides a composition for preventing or improving skin wrinkles, comprising green tea extract as an active ingredient, containing (%)-epigallocatechin gallate (EGCG) by weight (-)-epigalocatechin gallate.

The extracts and compositions according to one aspect of the present invention are safe from environmentally friendly natural plants with little irritation to the skin, thereby effectively suppressing the expression of MMP-1 and showing excellent skin wrinkle prevention or improvement efficacy.

FIG. 1 shows a chromatogram for the noticed green tea extract of Example 1 (Sample 1).
Figure 2 shows a chromatogram for the high-temperature treated green tea extract (sample 2) according to an aspect of the present invention.

In the present specification, "green tea extract", regardless of the extraction method, the extraction solvent, or the form of the extracted component or extract, is an extract of Bacillus subtilis or Bacillus subtilis , which is an evergreen tree belonging to the family Caraceae ( Camellia sinensis ). spp.), and extracts from fermented tea leaves, etc., and fractions of the extracted ones with a specific solvent. The tea includes one or more selected from the group consisting of tea tree leaves, flowers, stems, fruits, roots, stems, and cores of the roots, and preferably may be leaves. In addition, the form of the extract may be preferably a powder form. The extraction or fractionation can be performed using water, an organic solvent, or a mixed solvent thereof. The organic solvent may use alcohol, isopropanol, acetone, hexane, ethyl acetate, carbon dioxide, or a mixed solvent of two or more of them, but is not limited thereto, and the active ingredient of green tea is not destroyed or extracted by heating at room temperature or in a minimized condition Or fractionation. The alcohol may be a lower alcohol of C ₁ ~ C ₅ . The number or method of extraction or fractionation is not particularly limited, and for example, methods such as cold immersion extraction, ultrasonic extraction, reflux cooling extraction, and hot water extraction may be used. Preferably, the active ingredient is extracted or fractionated by cold immersion or warming. After filtration, the filtrate is concentrated under reduced pressure to obtain a green tea extract of the present invention.

As used herein, "epi catechin" refers to epigallocatechin (EGC), (-) epicatechin ((-)epicattechin, EC), (-)-epigalocatechin gallate ((-)-epigallocatechin gallate, EGCG), and epicatechin 3-O-gallate (ECG).

As used herein, the term "epi catechin epimer" refers to gallocatechin (GC), catechin (C), (-)-gallocatechin gallate (GCG) and catechin gallate (catechin) gallate, CG).

In one aspect, the present invention, based on the total weight of the extract, 4 to 15% by weight of (-)-gallocatechin gallate ((-)-gallocatechin gallate, GCG) and 4 to 15% by weight of (-)-epi It may be related to a composition for preventing or improving skin wrinkles comprising green tea extract containing gallocatechin gallate ((-)-epigallocatechin gallate, EGCG) as an active ingredient.

In one aspect, the GCG is 4% by weight or more, 5% by weight or more, 5.3% by weight or more, 5.59% by weight or more, 5.7% by weight or more, 6% by weight or more, 7% by weight or more, 8% by weight based on the total weight of the extract % Or more, 9% or more, 10% or more, 11% or more, 12% or more, 13% or more, or 14% or more. In another aspect, the GCG is 15 wt% or less, 14 wt% or less, 13 wt% or less, 12 wt% or less, 11 wt% or less, 10 wt% or less, 9 wt% or less, 8 wt% based on the total weight of the extract %, 7% or less, 6% or less, 5.7% or less, 5.59% or less, 5.3% or less, or 5% or less. When the GCG is included in the content range, it may exhibit excellent MMP-1 inhibitory activity.

In one aspect, the EGCG is based on the total weight of the extract 4 wt% or more, 5 wt% or more, 5.2 wt% or more, 5.27 wt% or more, 5.5 wt% or more, 6 wt% or more, 7 wt% or more, 8 It may be at least 10% by weight, at least 9% by weight, at least 10% by weight, at least 11% by weight, at least 12% by weight, at least 13% by weight, or at least 14% by weight. In another aspect, the EGCG is 15 wt% or less, 14 wt% or less, 13 wt% or less, 12 wt% or less, 11 wt% or less, 10 wt% or less, 9 wt% or less, 8 wt% based on the total weight of the extract %, 7% or less, 6% or less, 5.5% or less, 5.27% or less, 5.2% or less, or 5% or less.

As another embodiment, the total content of GCG and EGCG in the extract may be 30% by weight or less based on the total weight of the extract. In one aspect, the total content of the GCG and EGCG is 30% by weight or less, 25% by weight or less, 20% by weight or less, 18% by weight or less, 16% by weight or less, 15% by weight or less, 14% by weight based on the total weight of the extract % Or less, 12% or less, 10.5% or less, 10% or less, or 9% or less. In another aspect, the total content of the GCG and EGCG is 8% by weight, 9% by weight, 10% by weight, 10.5% by weight, 12% by weight, 13% by weight, 14% by weight based on the total weight of the extract % Or more, 16% or more, 18% or more, 20% or more, or 25% or more.

As another embodiment, the epicatechin content in the extract may be 20% by weight or less based on the total weight of the extract. In one aspect, the epicatechin content is 20 wt% or less, 18 wt% or less, 16 wt% or less, 15 wt% or less, 14 wt% or less, 12 wt% or less, 11 wt% or less based on the total weight of the extract, or It may be 10% by weight or less. In another aspect, the epicatechin content is at least 9% by weight, at least 10% by weight, at least 11% by weight, at least 12% by weight, at least 13% by weight, at least 14% by weight, at least 16% by weight based on the total weight of the extract, Or 18% or more.

As another embodiment, the total content of 8 catechins in the extract, i.e., EGCG, (-)-epigallocatechin (EGC), (-) epicatechin ((-)epicattechin, in the extract) EC), the total content of epicatechin 3-O-gallate (ECG), GCG, gallocatechin (GC), catechin (C) and catechin gallate (CG) Silver may be 19 to 30% by weight based on the total weight of the extract. In one aspect, the total content of the eight catechins is 19% by weight or more, 21% by weight or more, 23% by weight or more, 24% by weight or more, 24.5% by weight or more, 25% by weight or more, 26% by weight based on the total weight of the extract % Or more, 27% or more, 28% or more, or 29% or more. In another aspect, the total content of the eight catechins is 30% by weight or less, 29% by weight or less, 28% by weight or less, 27% by weight or less, 26% by weight or less, 25% by weight or less, 24.5% by weight based on the total weight of the extract % Or less, 24% or less, 23% or less, or 21% or less.

As another embodiment, the weight ratio of GCG:EGCG in the extract may be 1:0.5 to 2. In one aspect, the weight ratio of GCG:EGCG in the extract may be 1:0.5, 1:0.8, 1:1, 1:1.2, 1:1.5, 1:1.8 or 1:2.

As another embodiment, the extract may be an extract extracted one or more times by any one or more of water and C1 to C4 alcohol. In one aspect, the alcohol may be ethanol. In another aspect, the alcohol may be 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, or 70% or more ethanol. In another aspect, the alcohol may be 70% or less, 60% or less, 50% or less, 40% or less, or 30% or less ethanol.

As another embodiment, the content of the green tea extract in the composition for preventing or improving skin wrinkles may be 1% to 100% by weight based on the total weight of the composition. In one aspect, the content of the extract in the composition is 1% by weight, 10% by weight, 20% by weight, 30% by weight, 40% by weight, 50% by weight, 60% by weight, 70% by weight Or more, 80% by weight or more, or 90% by weight or more. In another aspect, the content of the extract in the composition is 100% or less, 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 40% or less, 30% or less It may be less than or equal to 20% by weight.

As another embodiment, the dosage of the active ingredient may be 5mg/kg/day to 1000mg/kg/day. In one aspect, the dosage is 5 mg/kg/day or more, 100 mg/kg/day or more, 200 mg/kg/day or more, 300 mg/kg/day or more, 400 mg/kg/day or more, 500 mg/kg/day or more, It may be 600 mg/kg/day or more, 700 mg/kg/day or more, 800 mg/kg/day or more, or 900 mg/kg/day or more. In another aspect, the dosage is 1000 mg/kg/day or less, 900 mg/kg/day or less, 800 mg/kg/day or less, 700 mg/kg/day or less, 600 mg/kg/day or less, 500 mg/kg/day or less, It may be 400 mg/kg/day or less, 300 mg/kg/day or less, 200 mg/kg/day or less, 100 mg/kg/day or less, 50 mg/kg/day or less, or 10 mg/kg/day or less.

As an embodiment, the extract may suppress the expression of matrix metalloproteinase-1 (MMP-1). That is, the extract or composition may exhibit a skin wrinkle prevention or improvement effect by suppressing the expression of MMP-1.

As an example embodiment, the composition may be a food, pharmaceutical or cosmetic composition.

The formulation of the food composition is not particularly limited, for example, tablets, granules, pills, powders, liquids such as drinks, caramels, gels, bars, tea bags, etc. may be formulated. The food composition of each formulation can be formulated by appropriately selecting the ingredients commonly used in the field other than the active ingredients according to the formulation or purpose of use without difficulty, and synergistic effect when applied simultaneously with other ingredients. Can happen. In addition, the food may be a health functional food.

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

In the food composition according to one aspect of the present invention, the dosage of the active ingredient is within the level of those skilled in the art, and may vary depending on various factors such as age, health condition, and complications of the subject to be administered. .

Food composition according to an aspect of the present invention, for example, chewing gum, caramel products, candy, ice cream, confectionery, various foods, soft drinks, mineral water, alcoholic beverages, beverage products, vitamins, minerals, etc. It may be a functional food product.

In addition to the above, the food composition according to one aspect of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, colorants and enhancers (cheese, chocolate, etc.), pectic acid and the like. Salts, alginic acids and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid used in carbonated beverages, and the like. In addition, food compositions according to an aspect of the present invention may include natural fruit juice and fruit juice for the production of fruit juice drinks and vegetable drinks. These ingredients can be used independently or in combination. The proportion of these additives is not so important, but is generally included in the range of 0 to about 60 parts by weight per 100 parts by weight of the composition according to one aspect of the present invention.

The pharmaceutical composition according to an aspect of the present invention may be administered by oral, parenteral, rectal, topical, transdermal, intravenous, intramuscular, intraperitoneal, subcutaneous, and the like. Formulations for oral administration may be tablets, pills, soft and hard capsules, granules, powders, granules, liquids, emulsions or pellets, but are not limited thereto. It is not. Formulations for parenteral administration may be solutions, suspensions, emulsions, gels, injections, drops, suppositories, patches or sprays, but are not limited thereto. The formulations can be readily prepared according to conventional methods in the art, surfactants, excipients, hydrating agents, emulsifying accelerators, suspending agents, salts or buffers for controlling osmotic pressure, colorants, spices, stabilizers, preservatives, preservatives or Other commercial adjuvants may be further included.

The composition according to an aspect of the present invention may include a pharmaceutically acceptable salt, which salt is formed of an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; Or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl) Benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2,2,2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert Acid addition salts formed from organic acids such as butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid; Or (2) a salt formed when the acidic proton present in the parent compound is substituted.

The applied amount or dosage of the pharmaceutical composition according to an aspect of the present invention will vary depending on the age, gender, weight, pathology and severity of the subject to be administered, the route of administration or the judgment of the prescriber. The determination of the active ingredient dosage based on these factors is within the level of those skilled in the art.

The cosmetic composition according to an aspect of the present invention may contain a cosmetic or dermatologically acceptable medium or base. These are all formulations suitable for topical application, e.g. solutions, gels, solids, dough anhydrous products, emulsions, suspensions, microemulsions, microcapsules, microgranules, or ionic (liposomes) obtained by dispersing an oil phase in an aqueous phase, and It may be provided in the form of a non-ionic vesicle dispersant, or in the form of a cream, skin, lotion, powder, ointment, spray or conceal stick. These compositions can be prepared according to conventional methods in the art. The cosmetic composition may also be used in the form of an aerosol composition further containing a propellant compressed in the form of a foam.

The cosmetic composition is not particularly limited in its formulation, and may be appropriately selected according to the purpose. For example, skin lotion, skin softener, skin toner, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing foam, Cleansing lotions, cleansing creams, cleansing water, powders, body lotions, body creams, body oils, body cleansers, and body essences can be formulated into formulations.

When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. may be used as a carrier component. Can.

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 as a carrier component, and in particular, in the case of a spray, additionally chlorofluorohydrocarbon, propane /Propellant 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 a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of ,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used.

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

In addition to the green tea extract, the cosmetic composition may further include functional additives and components included in the general cosmetic composition. The functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingo lipids, and seaweed extract.

In addition to the above-mentioned functional additives, the components contained in the general cosmetic composition may be blended with the composition. Other ingredients included include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, fungicides, antioxidants, plant extracts, pH regulators, alcohols, pigments, flavors, blood circulation Accelerators, cooling agents, limiting agents, purified water, and the like.

Hereinafter, examples, experimental examples, and formulation examples, the composition and effects of the present specification will be described in more detail. However, these examples are provided for purposes of illustration only to aid understanding of the present specification, and the scope and scope of the present specification are not limited by the following examples.

[Example 1] Preparation of general notice green tea extract and high temperature treated green tea extract

To 100 g of green tea (Camellia sinensis, Jeju Osulloc Farm), 1000 ml of 50% ethanol was added and stirred under reflux at 60° C. for 1 hour. The solution obtained by lowering the temperature of the sample to room temperature and filtering was distilled under reduced pressure to obtain 23 g of noticeable green tea extract (GT-LE-35CAT, sample 1) as a dark brown powder (yield 23%).

Meanwhile, 1000 ml of 50% ethanol was added to 100 g of green tea (Camellia sinensis, Jeju Osulloc Farm) to prepare a high-temperature-treated green tea extract, followed by stirring at reflux at 60° C. for 1 hour. After concentration, it was stirred for 1 to 7 hours at 1.5 kgf/cm ² of steam. Thereafter, the temperature was lowered to room temperature, the insoluble material was filtered, and concentrated under reduced pressure to obtain 10 g of a hot-treated green tea extract. At this time, the change in the content of 8 catechins was measured using the apparatus shown in Table 1 below for the high-temperature treated green tea extract obtained for each stirring time period (1-7 hours), and heat conversion of EGCG to GCG by heat at 5 hours. It was confirmed that the most conversion, the amount of 8 kinds of catechins did not decrease any more. At this time, the obtained high temperature processed green tea extract 10g (High temperature processed green tea extract, HTP-GTE) was used as Sample 2.

The conditions and results of analyzing the composition of the obtained two types of extracts are shown in Table 1 (composition analysis conditions of Samples 1 and 2), Table 2 (composition analysis results of Sample 1), and Table 3 (composition analysis results of Sample 2), respectively. ). In addition, the chromatograms for the two types of extracts are shown in Fig. 1 (Sample 1) and Fig. 2 (Sample 2). That is, it was confirmed that the composition of Sample 2 is different from the existing green tea extract. Specifically, Sample 2 contains significantly lower EGCG (5.27% by weight), EGC (3.53% by weight), and total catechin (24.41% by weight) than Sample 1, whereas Sample 1 has four components that are not found. It was confirmed that additional epicatechin epimers of.

column Thermofisher C18 5um, 4.6 X 250mm Detector UV 280nm Dilution Water in Gradient A-0.1% Trifluoro acetic acid (TFA) B-Acetonitrile Gradient profile 0 min A(90): B(10)
30min A(85): B(15)
42min A(80): B(20)
44min A(5): B(95)
49min A(90): B(10) Flow rate 1ml/min Injection volumn 20μl

Sample 1 GC EGC Caffeine C EC GCG EGCG CG ECG Total amount of epicatechin Epicatechin Epimer Total 8 kinds of catechins 0 9.16 3.21 0 3.63 0 20.93 0 2.62 36.34 0 36.34

Sample 2 GC EGC Caffeine C EC GCG EGCG CG ECG Total amount of epicatechin Epicatechin Epimer Total 8 kinds of catechins 4.9 3.53 3.9 1.57 0.95 5.59 5.27 1.3 1.3 11.05 13.36 24.41

(In Tables 2 and 3, GC is gallocatechin, EGC is epigallocatechin, C is catechin, EC is (-) epicatechin, GCG is gallocatechin gallate, EGCG is epigallocatechin gallate, CG is Catechin gallate, ECG is epicatechin 3-O-gallate)

(All of the units in Tables 2 and 3 above are green tea extracts (samples 1 and 2) by weight of the substance in relation to the total weight)

[Test Example 1] MMP-1 (Matrix metalloproteinase-1) inhibitory activity evaluation test

Effects of the green tea extracts of Sample 1 and Sample 2 of Example 1 on the inhibition of the production of MMP-1, a skin tissue degrading enzyme, in aging human skin fibroblasts, the weight ratio of EGCG and GCG, and MMP-1 according to the content of GCG Evaluation of MMP-1 inhibitory activity was carried out to confirm production inhibitory efficacy. Specifically, it proceeded through the following steps.

1-1. Cell lines and cell culture

Human dermal fibroblasts, normal human dermal fibroblast (NHDF; Lonza, Switzerland), were cultured in DMEM medium (Dulbecco's modified Eagle's Medium, Gibco 1210-0038) containing 10% fetal bovin serum. , 5% CO ₂ incubator. Cells with 6 to 9 times of subculture were used as young cells, and 37 passages were cultured by counting the number of cells for each passage and reculturing about 1/5 of the total number of cells again during passage culture. Thus, aging dermal fibroblasts were obtained. The EGCG and GCG were treated according to the ratio of EGCG:GCG and the concentration of GCG to aged dermal fibroblasts, or Sample 1 or Sample 2 was treated.

1-2. MMP-1 production measurement

An enzyme-linked immunosorbent assay (ELISA) method was used to confirm the amount of MMP-1 produced (Amersham, USA). The 1-1. After collecting the culture of the cells treated with the samples in step, centrifuged at 1000xg to separate the supernatant, the culture solution was put into a 96-well plate coated with MMP-1 antibody and reacted, followed by a secondary antibody (horse reddish peroxidase, HRP) was reacted at room temperature for 1 hour and washed with 100 μL PBS. After the reaction using TMB substrate solution (Pierce, USA), the amount of MMP-1 was confirmed by measuring the absorbance at 490 nm wavelength to confirm the color development, and the MMP-1 inhibitory activity (%) was calculated through the following formula. .

MMP-1 inhibitory activity (%) = (O.D. at 490 nm in the sample-added group/ O.D. at490 nm in the sample-free group) x 100

The results are shown in Tables 4 and 5 below.

Throughput (μg/ml) MMP-1 inhibitory activity (%) Immediate processing After 24 hours Sample 1 200 66±10.2 10±1.0 Sample 2 200 83±12.1 78±6.9

Throughput (μg/ml) EGCG:GCG MMP-1 inhibitory activity (%) EGCG GCG Immediate processing After 24 hours 3 0 - 56±9.7 6±1.0 2.4 0.6 4:1 53±5.4 9.4±0.9 2 One 2:1 67±6.1 22±1.1 1.8 1.2 1.5:1 76±5.2 54 ± 3.1 1.5 1.5 1:1 74±4.2 57±0.8 1.2 1.8 1:1.5 73 ±3.7 60 ± 6.3 0 3 - 55±9.2 10±1.0

From the results in Table 4, the GCG according to the present invention contains 5.59% by weight, EGCG at 5.27% by weight, 11.05% by weight of epicatechin, 13.36% by weight of epicatechin epimer, and 24.41% by weight of 8 catechins. In the case of the high-temperature-treated green tea extract containing (Sample 2), it was confirmed that the inhibitory activity of MMP-1 was superior to that of the normal green tea extract (Sample 1). Particularly, when the aged fibroblasts are treated after being left at room temperature for 24 hours, MMP-1 inhibition of the high-temperature treated green tea extract according to the present invention is inhibited, despite the total content of 8 catechins is less than that of the general green tea extract. It was confirmed that the activity was remarkably excellent.

In addition, from the results of Table 5, when the weight ratio of EGCG:GCG is 2:1, 1.5:1, 1:1 or 1:1.5, only EGCG is processed, only GCG is processed, or the weight ratio of EGCG:GCG is 4 It showed excellent MMP-1 inhibitory activity compared to :1, and among them, it was found that it exhibited particularly excellent MMP-1 inhibitory activity when the weight ratio of EGCG:GCG was 1.5:1, 1:1 or 1:1.5. there was.

As a result, according to the present invention, in the case of a high-temperature treated green tea extract containing 4 to 15% by weight of GCG and 4 to 15% by weight of EGCG, it can be effectively prevented or improved skin wrinkles through excellent MMP-1 inhibitory activity. In addition, it was also found that when the GCG and EGCG are included in a specific weight ratio according to the present invention, the effect of preventing or improving skin wrinkles according to MMP-1 inhibitory activity can be further improved.

[Formulation Example 1] Soft capsule

Sample 2 according to Example 1 was prepared as 150 mg, and a soft capsule filling solution was prepared by mixing lactose 440 mg, corn starch 430 mg, and magnesium stearate 2 mg. Then, separately from the above, a soft capsule sheet was prepared at a ratio of 66 parts by weight of gelatin, 24 parts by weight of glycerin, and 10 parts by weight of sorbitol solution, and the filling solution was filled to prepare a soft capsule.

[Formulation Example 2] Tablet

Sample 2 according to Example 1 was prepared as 150 mg, mixed with vitamin E 15 mg, vitamin C 15 mg, galactooligosaccharide 250 mg, lactose 60 mg, and maltose 140 mg, granulated using a fluid bed dryer, and then sugar ester (sugar ester) 8mg was added. Tablets were prepared by tableting the composition in a conventional manner.

[Formulation Example 3] Drink

Sample 2 according to Example 1 was prepared in 80mg, vitamin E 9mg, vitamin C 9mg, glucose 10g, citric acid 0.6g, and liquid oligosaccharide 25g was mixed, and 400ml of purified water was added to fill it. After filling the bottle, sterilized at 30°C for 4 to 5 seconds to prepare a drink.

[Formulation Example 4] Granules

150 mg of Sample 2 according to Example 1 was prepared, and mixed with vitamin E 9 mg, vitamin C 9 mg, anhydrous crystalline glucose 250 mg, and starch 550 mg, molded into granules using a fluidized bed granulator, and then filled into a bag to form granules. Was prepared.

[Formulation Example 5] Health Food

Prepare 150 mg of Sample 2 according to Example 1 above, vitamin mixture (70 μg of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.5 mg of vitamin B6, 0.2 mg of vitamin B12, vitamin C 10 mg, biotin 10 μg, nicotinic acid amide 1.7 mg, folic acid 50 μg) and mineral mixture (ferric sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium phosphate 15 mg, dicalcium phosphate 55 mg , 90 mg of potassium citrate, 100 mg of calcium carbonate, and 24.8 mg of magnesium chloride) were combined to prepare a health food.

[Formulation Example 6] Healthy Drink

50 mg of Sample 2 according to Example 1 was prepared, and 1000 mL of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, 1 g of taurine, and residual amount of purified water were added to prepare 900 mL of healthy beverage.

Since specific parts of the present specification have been described in detail above, it is obvious that for those skilled in the art, this specific technology is only a preferred embodiment, and the scope of the present specification is not limited thereto. Accordingly, the substantial scope of the present specification will be defined by the appended claims and their equivalents.

Claims (8)

Based on the total weight of the extract, 4-15% by weight of (-)-gallocatechin gallate ((-)-gallocatechin gallate, GCG) and 4-15% by weight of (-)-epigalocatechin gallate (( -)-epigallocatechin gallate, EGCG) containing green tea extract as an active ingredient, skin wrinkle prevention or improvement composition. According to claim 1, EGCG in the extract, epigallocatechin (epigallocatechin, EGC), (-) epicatechin ((-)epicattechin, EC), epicatechin 3-O-gallate (epicatechin 3-O-gallate, ECG ), the total content of GCG, gallocatechin (GC), catechin (C) and catechin gallate (CG) is 19 to 30% by weight based on the total weight of the extract. The composition of claim 1, wherein the weight ratio of GCG:EGCG in the extract is 1:0.5-2. The composition of claim 1, wherein the extract is an extract extracted one or more times with any one or more of water and C1 to C4 alcohols. The composition of claim 1, wherein the content of the extract in the composition is 1% to 100% by weight based on the total weight of the composition. According to claim 1, The dosage of the active ingredient is 5mg / kg / day to 1000mg / kg / day, the composition. The composition of claim 1, wherein the extract inhibits the expression of matrix metalloproteinase-1 (MMP-1). The composition according to any one of claims 1 to 7, wherein the composition is a food, pharmaceutical or cosmetic composition.

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