KR102560220B1 - Compositions for Skin Regeneration Using an Extract of Silene seoulensis Nakai and Compounds Isolated from the Same - Google Patents

Abstract

The present invention uses a siliendine A, B, and D compound of the following chemical formulas isolated from an extract of scutellum japonica, which was shown to have skin regeneration activity in a wound healing assay using keratinocytes. Disclosed is a composition for skin regeneration.

Description

Compositions for Skin Regeneration Using an Extract of Silene seoulensis Nakai and Compounds Isolated from the Same}

The present invention relates to a composition for skin regeneration using Silene seoulensis Nakai) extract and a compound isolated therefrom.

Skin exists in the outermost part of the human body and is in direct contact with the external environment, protecting the human body from external damage and preventing excessive evaporation of moisture and heat in the body. The skin is composed of three layers: epidermis, dermis, and subcutaneous tissue, and most of the outermost epidermis is composed of keratinocytes. Keratinocytes of the epidermis protect the skin by forming keratin through proliferation, migration, and differentiation from the basal layer, and are involved in inflammatory and immune responses by producing various cytokines. In addition, proliferation and migration of keratinocytes not only provide wound healing and skin regeneration, but also induce collagen synthesis to enhance skin elasticity and improve basement membrane formation so that skin cells can adhere well to connective tissue. It provides an environment, induces proliferation of skin cells, improves barrier function to selectively penetrate harmful substances, and provides interaction between epidermis and dermis. Due to this, the migration and proliferation of skin epidermal keratinocytes not only regenerates skin and heals wounds, but also promotes skin elasticity and helps maintain healthy skin.

Silene seoulensis Nakai belongs to the Caryophyllaceae family and is an annual plant special to Korea that grows in humid and shady places in the mountains throughout the country except Jeju Island. The stem is 30-60cm high, the bottom creeps sideways, and the upper part grows straight and has downward hairs. The leaves are opposite, egg-shaped, and the tip is pointed. Flowers form cyme at the end of stems from June to August, white or reddish white, about 1.2cm in diameter. The peduncle is slender and long. There are 5 petals, and the tip is split into two. Fruits are capsules, egg-shaped. It is also distributed in Japan and China. In addition, compared to plants of the genus Silene in Korea, the leaves are ovate and have petioles, and the lower part of the stem grows lying down. There are insufficient reports on active substances and physiological activities for slender jaundice.

The present invention discloses the skin regenerative efficacy of the extract of Possible Jang japonica extract.

An object of the present invention is to provide a composition for skin regeneration using an extract of gourd japonica.

Other objects or specific objects of the present invention will be presented below.

As confirmed in the following Examples and Experimental Examples, the present invention is a wound healing using dermal keratinocytes, in which the siliendine A, B, and D compounds of the formula below, isolated from the extract and its fraction, are It was completed by confirming that it exhibits distinct skin regeneration activity in the wound healing assay.

<Formula 1>

Siliendyne A

<Formula 2>

Siliendyne B

<Formula 3>

Shilliendyne D

Considering the foregoing, in one aspect, the present invention can be identified as a composition for skin regeneration containing an active ingredient, and in another aspect, a composition for healing skin wounds containing an active ingredient. In another aspect, it can be identified as a composition for improving skin wrinkles containing an active ingredient.

In the present specification, the term "extract of japonica japonica" refers to extracting the leaves, branches, aerial parts, rhizomes, roots, underground parts, or mixtures thereof of water, lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), methylene chloride , ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof It refers to an extract obtained by leaching using an extract obtained by leaching, an extract obtained by using a supercritical extraction solvent such as carbon dioxide or pentane, or a fraction obtained by fractionating the extract. , reflux, heating, ultrasonic radiation, supercritical extraction, etc. may be applied. In the case of the fractionated extract, after suspending the extract in a specific solvent, the fraction obtained by mixing and standing with a solvent of different polarity, and the crude extract are adsorbed on a column filled with silica gel, etc., and then a hydrophobic solvent, a hydrophilic solvent, or a mixed solvent thereof It is meant to include fractions obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent is removed by lyophilization, vacuum drying, hot air drying, spray drying, or the like. Preferably, an extract obtained by using water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof as an extraction solvent and a fraction of the extract, preferably the extract, is mixed with (i) water and (ii) hexane, dichloromethane, ethyl A fraction of any one solvent layer obtained by fractionation with any one solvent selected from acetate and butanol, more preferably water and ethanol mixed solvent extract, is suspended in water, and then sequentially using hexane, dichloromethane, ethyl acetate and butanol It means the fraction obtained when fractionated as Here, the sequential fractionation means fractionation with the solvents in the order listed above by continuously using the water fraction after fractionation.

In addition, in the present specification, "active ingredient" means a component that exhibits the desired activity alone or can exhibit activity in combination with a carrier having no activity itself.

The composition of the present invention may contain the active ingredient in an arbitrary amount (effective amount) as long as it can exhibit a skin regeneration effect, etc. according to the formulation, blending purpose, etc., and a typical effective amount is 0.001 weight based on the total weight of the composition. % to 15% by weight. Here, "effective amount" may indicate intended medical and cosmetic effects, such as skin regeneration, when the composition of the present invention is administered to a mammal, preferably a human, to which it is applied, during the administration period according to the advice of medical experts. It refers to the amount of the active ingredient included in the composition of the present invention. Such an effective amount can be determined empirically within the ordinary skill of the skilled artisan.

In addition to active ingredients, the composition of the present invention is used for skin regeneration activity, skin wrinkle activity, skin whitening activity, skin hypersensitivity reaction inhibition activity, skin protection activity (suppression of skin damage by ultraviolet rays, skin moisturizing, etc.), etc. In order to enhance the convenience of taking, ingesting, and using through the addition of similar activities, any compounds or natural extracts known to have safety and corresponding activities may be additionally included in the art.

These compounds or extracts include each country's Pharmacopoeia ("Korean Pharmacopoeia" in Korea), each country's Health Functional Food Codex (in Korea, it is "Health Functional Food Standards and Specifications", a notification by the Ministry of Food and Drug Safety), and each country's Functional Cosmetics Codex (in Korea, a notification by the Ministry of Food and Drug Safety Compounds or extracts listed in compendial documents such as "Functional Cosmetics Standards and Test Methods"), compounds or extracts approved as items under the laws of each country governing the manufacture and sale of pharmaceuticals ("Pharmaceutical Affairs Act" in Korea), health In accordance with the laws of each country regulating the manufacture and sale of functional foods (in Korea, it is the 「Health Functional Food Act」), the laws of each country regulating the manufacture and sale of compounds or extracts with recognized functionality, and the manufacture and sale of functional cosmetics (in Korea, the 「Cosmetics Act」 ”), compounds or extracts whose functionality has been recognized are included.

These ingredients include, for example, retinol, retinyl palmitate, adenosine, polyethoxylated amide, etc., which are recognized as skin moisturizing ingredients in the Cosmetics Code according to the Korean Cosmetics Act (Korean Ministry of Food and Drug Safety notice, 「Standards and Test Methods for Functional Cosmetics」). , Drometrizol, drometrizol trisiloxane, digaloyltrioleate, dimethicodyethylbenzalmalonate, diethylhexylbutamidotriazone, etc. recognized as UV protection ingredients in the same Korean Cosmetic Code, and the same Arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol, Oil Soluble Licorice (Glycyrrhiza) Extract, etc., recognized as skin whitening ingredients in the Cosmetics Code, and UV protection ingredients according to the Korean Health Functional Food Act Complexes such as pine bark extract, phosphatidylserine, finger root extract powder, complex extracts such as red ginseng and cornus officinalis, etc. recognized as L. sakei Probio 65, gamma linolenic acid-containing fat, fruit and vegetable-derived lactic acid bacteria L. plantarum CJLP133, probiotics ATP, and the like. One or more of these compounds or natural extracts may be included in the composition of the present invention together with the active ingredient.

In a specific aspect, the composition of the present invention can be regarded as a food composition.

The food composition of the present invention can be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, and ionic beverages, processed oils such as milk and yogurt, chewing gum, rice cakes, traditional Korean snacks, bread, snacks, noodles, etc. It can be manufactured into health functional food preparations such as foods, tablets, capsules, pills, granules, liquid, powder, flakes, pastes, syrups, gels, jellies, and bars.

In addition, the food composition of the present invention may take any product classification as long as it conforms to the enforcement regulations at the time of manufacture and distribution in legal and functional classification. For example, health functional food according to the Korean 「Health Functional Food Act」, or snacks, legumes, teas, beverages according to each food type according to the Food Code (MFDS notification 「Food Standards and Specifications」) of the Korea 「Food Sanitation Act」 , special purpose foods, etc.

The food composition of the present invention may include food additives in addition to the active ingredient. Food additives may generally be understood as substances that are added to, mixed with, or infiltrated into food in manufacturing, processing, or preserving food. Since food is consumed daily and for a long period of time, its safety must be guaranteed. According to the Food Additives Code under the laws of each country governing the manufacture and distribution of food (in Korea, it is the 「Food Sanitation Act」), safety-guaranteed food additives are limitedly regulated in terms of ingredients or functions. In the Korean Food Additives Codex (MFDS notification 「Standards and Specifications for Food Additives」), food additives are classified into chemical synthetic products, natural additives and mixed preparations in terms of ingredients and are regulated. It is divided into additives, preservatives, emulsifiers, acidulants, and thickeners.

Sweeteners are used to impart appropriate sweetness to foods, and both natural and synthetic sweeteners can be used in the composition of the present invention. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavors may be used to improve taste or aroma, and both natural and synthetic flavors may be used. Preferably, it is the case of using a natural one. In case of using natural ones, in addition to flavor, the purpose of enhancing nutrition can also be combined. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or obtained from green tea leaves, roundworms, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo, etc. can be used. Natural flavors can be liquid concentrates or solid extracts. In some cases, synthetic flavors may be used, and synthetic flavors may include esters, alcohols, aldehydes, terpenes, and the like.

As preservatives, sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. may be used, and as emulsifiers, acacia gum, carboxymethylcellulose, xanthan gum, pectin and the like, and examples of the acidulant include acid salt, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like. Acidulants may be added to the food composition to have an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to improving taste.

As the thickening agent, a suspending agent, a sedimentation agent, a gel forming agent, a swelling agent, and the like may be used.

In addition to the food additives described above, the food composition of the present invention may include physiologically active substances or minerals known in the art and whose stability is guaranteed as food additives for the purpose of supplementing or reinforcing functionality and nutrition.

Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, and the like. Examples of minerals include calcium preparations such as calcium citrate and magnesium stearate. magnesium preparations such as the like, iron preparations such as iron citrate, chromium chloride, potassium iodine, selenium, germanium, vanadium, zinc, and the like.

The food composition of the present invention may include the above-mentioned food additives in an appropriate amount to achieve the purpose of addition according to the type of product.

Regarding other food additives that may be included in the food composition of the present invention, reference may be made to national food codes or food additive codes.

The composition of the present invention can be understood as a pharmaceutical composition in another specific aspect.

The pharmaceutical composition of the present invention may be formulated into an oral formulation or a parenteral formulation according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. The route of administration herein may be any appropriate route including topical route, oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of a combination of two or more routes is a case in which two or more formulations of drugs are combined according to the route of administration, for example, one drug is firstly administered intravenously and the other drug is secondly administered through a topical route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or dosage form, and specifically, reference may be made to the pharmacopoeia of each country including the "Korean Pharmacopoeia".

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powder, granule, tablet, pill, dragee, capsule, liquid, gel, syrup, suspension, wafer according to a method known in the art together with a suitable carrier. It can be prepared in formulations such as Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, and wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Celluloses such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, Serol etc. are mentioned. In the case of formulation, appropriate binders, lubricants, disintegrants, coloring agents, diluents, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferrite, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, and the like, and oleic acid as a lubricant Sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polyethylene glycol, etc. may be mentioned. As disintegrants, starch, methyl cellulose , agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt, and the like. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and glycine.

When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it may be formulated in the form of an injection, transdermal administration, nasal inhalation, and suppository along with a suitable carrier according to a method known in the art. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, an isotonic solution such as phosphate buffered saline (PBS) containing triethanolamine, sterile water for injection, or 5% dextrose may be used. . When formulated as a transdermal formulation, it may be formulated in the form of ointments, creams, lotions, gels, external solutions, pastas, liniments, air rolls, and the like. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and when formulated as a suppository, the carrier is Witepsol ( witepsol), tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, and the like can be used.

Regarding the specific formulation of the pharmaceutical composition, it is known in the art, and for example, reference may be made to Remington's Pharmaceutical Sciences (19th ed., 1995) and the like. These documents are considered as part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, severity of the patient, and route of administration. /kg range. Administration can be done once a day or divided into several times. These dosages should not be construed as limiting the scope of the present invention in any respect.

In another specific aspect, the composition of the present invention can be identified as a cosmetic composition.

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified as an arbitrary product in terms of its use and law, and specifically, functional cosmetics and non-functional cosmetics with uses such as improving skin troubles and atopic dermatitis. It may be a general cosmetic or the like. In product form, it may take any product form, specifically, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax It may take the form of a product such as foundation or spray. In a specific product form, it may be a softening lotion, nutrient lotion, nutrient cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder formulation.

The cosmetic composition of the present invention may include, in addition to the active ingredient, ingredients commonly used in cosmetic compositions, for example, stabilizers, solubilizers, surfactants, vitamins, conventional adjuvants such as pigments and fragrances, and carriers.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide 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, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, specifically water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol, fatty acid esters of sorbitan, and the like may be used.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester, microcrystals Star cellulose, aluminum metahydroxide, bentonite, agar, and the like can be used.

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

The cosmetic composition of the present invention may be prepared according to a method for preparing a cosmetic composition conventionally performed in the art, except for including an active ingredient exhibiting skin regeneration activity.

As described above, according to the present invention, it is possible to provide a composition for skin rejuvenation using an extract of japonica japonica.

These compositions of the present invention can be commercialized as foods, cosmetics, pharmaceuticals, and the like.

1 is NMR data of siliendine A, B, and D, which are compounds isolated from an extract of A.
2 to 5 are experimental results obtained by evaluating the skin regeneration efficacy of human dermal keratinocytes of the extracts and fractions of the extracts and fractions by a wound healing assay.
6 to 8 are experimental results obtained by evaluating the skin regeneration efficacy of siliendine A, B, and D, which are compounds isolated from aphids, on human dermal keratinocytes by a wound healing assay.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these Examples and Experimental Examples.

<Example> Preparation of extract of goji japonica extract and separation of compounds

<Example 1> Preparation of extract of gourd japonica

To 450 g of the dried above-ground part of the pulverized intestinal lice, a 50% ethanol solution of 10 times the weight was added, left at room temperature for 24 hours, extracted twice, filtered, and the filtrate was concentrated under reduced pressure and lyophilized to obtain a solid extract of 66.5 g was obtained.

<Example 2> Separation of the compound from the extract of gourd japonica

After suspending the 50% ethanol extract in distilled water, sequential solvent fractionation was performed using hexane, dichloromethane, ethyl acetate and butanol to obtain each fraction. Among the fractions, the dichloromethane fraction (1.5 g) was separated into 8 fractions by medium pressure liquid chromatography (MPLC) using a column filled with reverse phase silica gel. After loading fraction 3 on Kromasil C18 (21.2 × 250 mm, 5 μm) column high-performance liquid preparative chromatography (prep-HPLC), acetonitrile (CH CN) aqueous solution containing 0.05% trifluoroacetic acid ( HPLC was performed with acetonitrile:water = 10-40:90-60) set at 10 mL/min, and siliendine B was separated at 3.2 mg at 9.01 minutes and siliendien A at 11.35 minutes. did

In addition, after loading fraction 2 of the fraction into a high-performance liquid preparative chromatography (prep-HPLC) equipped with a YMC pack ODS-A C18 (30 × 250 mm) column, 0.05% trifluoroacetic acid (trifluoroacetic acid) containing HPLC was performed by setting an acetonitrile (CH3CN) aqueous solution (acetonitrile: water = 20 to 40: 80 to 60) at 10 mL/min, and 1.5 mg of siliendine D was separated at 17.92 minutes and the separated compound The structure of was identified using 700 MHz NMR. NMR data are shown in FIG. 1 .

<Experimental Example> Evaluation of skin regeneration activity of the compound isolated from the extract of Astragalus japonica (Wound healing assay)

HaCaT cells, human keratinocytes, were purchased from Korea Cell Line Bank (KCLB, Seoul, Korea), DMEM (Dulbecco's) containing 100 units/ml P/C (penicilin/streptomycin) and 10% FBS (fetal bovine serum). Modified Eagle Medium) (WLGENE, Korea) was used and cultured in an incubator at 37°C and 5% CO ₂ . Hacat cells were dispensed at 5x10 ⁵ cells/ml in 500 µl each in a 24-well plate and cultured for 24 hours. Thereafter, the medium was replaced with a medium that did not contain FBS, and each well was scratched using a 200 μl tip, and the samples were treated by concentration. As a positive control, epidermal growth factor (EGF, 50 ng/ml) was used. Samples were treated by concentration and incubated in a CO ₂ incubator for 48 hours. After sample treatment, cell pictures were taken using a microscope at 0 and 48 hours, and the wound healing area was calculated using the Image J program at 0 and 48 hours to determine the closure rate (%). did

The results of measuring the sealing ratio of siliendine A, B, and D, which are compounds isolated from the extract and fractions thereof, are shown in FIGS. 2 to 8, respectively.

As shown in FIGS. 2 to 5, after making wound gaps in HaCaT cells, as a result of treating the extracts and fractions for 48 hours by concentration at which toxicity did not appear, the 50% ethanol extract of 50% ethanol extract of japonica japonica was used as a control group ( NOR), and in particular, at a concentration of 10 μg / mL, the closure rate was 50.65 ± 2.51%, which was better than the positive control, EGF (Epidermal growth factor). Hexane, dichloromethane, and ethyl acetate fractions also showed better activity than the control group (NOR).

In addition, as shown in FIGS. 6 to 8, as a result of evaluating the skin regeneration efficiencies of siliendine A, B and D, which are compounds isolated from the spleen, all three compounds have better skin than the control group (NOR). showed regenerative efficacy. In particular, siliendyne D showed higher activity than the positive control at 1, 5, and 10 μM.

Claims (9)

Skin rejuvenation containing as an active ingredient the water and ethanol mixed solvent extract of slender spinosa, the fraction of (a) or (b) below, siliendine A, siliendine B or siliendine D Composition for:
(a) a fraction of hexane, dichloromethane, or ethyl acetate obtained by fractionating the water and ethanol mixture solvent extract of S. chinensis with (i) water and (ii) a solvent selected from hexane, dichloromethane and ethyl acetate; and
(b) Fractions of hexane, dichloromethane or ethyl acetate when extracts of water and ethanol mixed solvent of St. John's wort were suspended in water and sequentially fractionated using hexane, dichloromethane, ethyl acetate and butanol.
Skin wounds containing as active ingredients the water and ethanol mixed solvent extract of slender sage, the fraction of (a) or (b) below, siliendine A, siliendine B or siliendine D Therapeutic pharmaceutical composition:
(a) a fraction of hexane, dichloromethane, or ethyl acetate obtained by fractionating the water and ethanol mixture solvent extract of S. chinensis with (i) water and (ii) a solvent selected from hexane, dichloromethane and ethyl acetate; and
(b) Fractions of hexane, dichloromethane or ethyl acetate when extracts of water and ethanol mixed solvent of St. John's wort were suspended in water and sequentially fractionated using hexane, dichloromethane, ethyl acetate and butanol.
According to claim 1,
The extract is a composition in which the extraction site is an aerial part.
delete delete According to claim 1,
The composition is a composition, characterized in that the cosmetic composition.
According to claim 1,
The composition is a composition, characterized in that the food composition.
According to claim 1,
The composition, characterized in that the composition is a pharmaceutical composition.



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