KR20240087883A - Compositions for anti-inflammation of Skin Using an Extract of Hydrilla verticillata - Google Patents

Abstract

The present invention has the activity of suppressing the production of inflammatory response-inducing cytokines and chemokines such as IL-6 and MCP-1 when treated with HaCaT cells, a human keratinocyte cell line in which an inflammatory response is induced by TNF-α/IFN-γ. Disclosed is a composition for inhibiting skin inflammatory response using black horse extract.

Description

Compositions for anti-inflammation of Skin Using an Extract of Hydrilla verticillata}

The present invention relates to a composition for inhibiting skin inflammatory response using black horse extract.

Inflammation is a protective response of immune cells such as macrophages against infection. Inflammation itself is a normal defense function and an essential defense system that maintains life. It promotes tissue regeneration or heals infections, but if excessive, it can also cause tissue damage.

Skin exists on the outermost layer 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 the epidermis, dermis, subcutaneous fat, and muscle. The epidermis, which constitutes the outermost layer of the skin, performs the primary barrier function to protect the skin from external stimuli. The epidermis is composed of 95% keratinocytes. These keratinocytes produce skin barrier factors through the differentiation process to perform the skin barrier function, and play an important role in inflammation and immune responses when exposed to stimulation. Dermatology in Clinical Practice Springer 2010: 1-15). When keratinocytes are exposed to TNF-α and IFN-γ, abnormal expression of cytokines and chemokines such as IL-1β, TNF-α, IL-6, IL-8, and MCP-1 (monocyte chemoattractant protein 1) is induced. Inflammatory cells such as white blood cells infiltrate into the inflammatory lesion site (J Allergy Clin Immunol. 2006; 118(1): 178-89). TNF-α and IFN-γ have been reported to induce the production of inflammatory cytokines not only in primary human keratinocytes but also in HaCaT cells, a human keratinocyte cell line ((J Dermatol Sci. 43(2): 75-84, 2006 ; J Drugs Dermatol 7:1038-1043, 2008; J Dermatol Sci 67: 101-110, 2012), excessive production of these inflammatory cytokines reduces the damage and cell proliferation rate of keratinocytes, leading to skin aging and atopic dermatitis. It causes various inflammatory skin diseases such as dermatitis, psoriasis, etc. ((Int J Dermatol. 2008; 47(3): 219-24; Clin Exp Rheumatol 24: S1-6, 2006). For this reason, TNF-α/IFN- HaCaT cells stimulated with γ are mainly used in research on inflammatory skin diseases such as atopic dermatitis (Biochem Biophys Res Commun. 2012; 427(2): 236-41).

The present invention discloses the anti-inflammatory activity of black horse extract in HaCaT cells stimulated with NF-α/IFN-γ.

The purpose of the present invention is to disclose a composition for inhibiting skin inflammatory response using black horse extract.

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

As confirmed in the Examples and Experimental Examples below, the present invention relates to the treatment of IL-6, MCP- It is completed by suppressing the production of inflammatory response-inducing cytokines and chemokines.

Considering the above, the present invention can be viewed as a composition for inhibiting skin inflammatory response containing black horse extract as an active ingredient.

In this specification, “black horseradish extract” refers to the stem, leaf, fruit, flower, root, underground part, above-ground part, plant or a mixture thereof of black horseradish, which is the subject of extraction, with water and a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.) , methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butylacetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or these. refers to an extract obtained by leaching using a mixed solvent, an extract obtained using a supercritical extraction solvent such as carbon dioxide, pentane, or a fraction obtained by fractionating the extract. The extraction method is based on the polarity, degree of extraction, and degree of preservation of the active substance. Considering this, any method such as cold immersion, reflux, heating, ultrasonic radiation, or supercritical extraction can be applied. In the case of a fractionated extract, the extract is suspended in a specific solvent and then mixed with a solvent of different polarity to form a fraction obtained by adsorbing the crude extract onto a column filled with silica gel and then mixed with a hydrophobic solvent, a hydrophilic solvent, or a mixture of these. It is meant to include fractions obtained using the mobile phase. In addition, the meaning of the extract includes concentrated liquid extract or solid extract from which the extraction solvent has been removed by methods such as freeze-drying, vacuum drying, hot air drying, and spray drying. Preferably, an extract obtained using water, ethanol, or a mixed solvent thereof as an extraction solvent (particularly a mixed solvent extract of water and ethanol), and a solid extract obtained by removing the extraction solvent from the extract are suspended in water and mixed with hexane. , refers to the fractions obtained when fractionated with dichloromethane or ethyl acetate, or each fraction obtained when sequentially fractionated with these fractionation solvents. Here, 'sequentially fractionated' means that the remaining water layer after fractionation is continuously used to fractionate with the fractionation solvent in the order listed above.

In addition, in this specification, “active ingredient” refers to an ingredient that exhibits the desired activity alone or can exhibit activity in combination with a carrier that is not active on its own.

In addition, as used herein, "inhibition of skin inflammatory response" is meant to include improvement (alleviation of symptoms), treatment, and prevention (inhibition or delay of onset) of skin diseases accompanied by inflammatory responses, as defined below.

In addition, in this specification, "skin disease accompanied by an inflammatory reaction" refers to bacterial skin infection, bacterial skin infection (e.g., acne due to skin infection by the acne bacteria Propionibacterium acnes ), fungal skin infection, conjunctivitis, iritis, scleritis, uveitis, This includes dermatitis (including atopic dermatitis), skin eczema, and psoriasis.

The composition for inhibiting skin inflammatory response of the present invention may contain the active ingredient in any amount (effective amount) as long as it can indicate the skin inflammatory disease for which it is intended to be treated, depending on the use, formulation, purpose of formulation, etc. Silver will be determined within the range of 0.001% by weight to 15% by weight based on the total weight of the composition. Here, the "effective amount" means that when the composition of the present invention is administered to mammals, preferably humans, to which it is applied during the administration period recommended by medical experts, etc., the intended medical and pharmacological effects, such as the effect of improving skin inflammatory diseases, are achieved. It refers to the amount of active ingredient contained in the composition of the present invention that can be expressed. Such effective amounts can be determined experimentally within the scope of the ordinary ability of those skilled in the art.

In addition to the active ingredients, the composition of the present invention has already been verified for safety in the art in order to increase and reinforce skin inflammatory response activity and to improve convenience of taking, ingesting, and using by adding similar activities such as anti-allergic activity. It may further include any compounds or natural extracts known to have the corresponding activity.

These compounds or extracts include compounds, extracts, and pharmaceuticals listed in compendial documents such as the Pharmacopoeia of each country (in Korea, the “Korean Pharmacopoeia”) and the Code of Health Functional Foods of each country (in Korea, it is the “Standards and Specifications for Health Functional Foods” notified by the Ministry of Food and Drug Safety). The laws of each country that govern the manufacture and sale of compounds or extracts and health functional foods that have received product approval in accordance with the laws of each country (in Korea, this is the Pharmaceutical Affairs Act) (in Korea, it is the “Health Functional Foods Act”) 」), compounds or extracts whose functionality has been individually recognized are included. For example, MSM (dimethylsulfonylmethane) with 'arthritis improvement' functionality according to the Korean Health Functional Foods Convention, N-acetylglucosamine with 'arthritis improvement' functionality and 'skin moisturizing' functionality, and 'Act on Health Functional Foods' in Korea. Enterococcus faecalis heat-treated dried powder, which has been individually recognized for its functionality in alleviating hypersensitive immune responses, complexes such as guava leaf extract, complexes such as Actinidia extract, lobule extract, and picaopreto powder, and PLAG (1-palmitoyl-2-linoleoyl- 3-acetyl-rac-glycerol), L. sakei Probio 65, which has been individually recognized for its functionality for 'improving sensitive skin conditions', oil containing gamma-linolenic acid, L. plantarum CJLP133, a lactic acid bacteria derived from fruit and vegetables, and probiotic ATP μ are these compounds or It would correspond to an extract.

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 viewed as a food composition.

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

In addition, the food composition of the present invention can have any product classification in terms of legal and functional classification as long as it complies with the enforcement laws and regulations at the time of manufacture and distribution. For example, it is a health functional food according to the Korean 「Act on Health Functional Foods」, or confectionery, beans, tea, and beverages according to each food type according to the food code of the Korean 「Food Sanitation Act」 (Ministry of Food and Drug Safety Notification 「Food Standards and Specifications」) , special purpose food, etc.

The food composition of the present invention may contain food additives in addition to the active ingredients. Food additives can generally be understood as substances that are added to, mixed with, or infiltrated into food when manufacturing, processing, or preserving food. Since they are consumed daily and for a long period of time with food, their safety must be guaranteed. In the food additive code of each country that regulates the manufacturing and distribution of food (in Korea, it is the Food Sanitation Act), food additives with guaranteed safety are limited in terms of ingredients or functions. In the Korea Food Additive Code (Ministry of Food and Drug Safety Notification “Food Additive Standards and Specifications”), food additives are classified into chemical synthetics, natural additives, and mixed preparations in terms of composition. These food additives are classified into sweeteners and flavors in terms of function. It is classified into preservatives, emulsifiers, acidulants, thickeners, etc.

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

Flavoring agents can be used to improve taste or aroma, and both natural and synthetic ones can be used. Preferably, natural products are used. When using natural products, they can serve the purpose of enhancing nutrition in addition to flavor. Natural flavoring agents may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or may be obtained from green tea leaves, coriander leaves, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, etc. You can also use things obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo nuts. Natural flavoring agents may be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents may be used, such as esters, alcohols, aldehydes, terpenes, etc.

Preservatives include calcium sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc., and emulsifiers include acacia gum, carboxymethyl cellulose, xanthan gum, Examples include pectin, and acidulants include acidic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and phosphoric acid. In addition to improving taste, acidulants may be added to ensure that the food composition has an appropriate acidity for the purpose of suppressing the growth of microorganisms.

As a thickening agent, a suspending agent, settling agent, gel forming agent, bulking agent, etc. may be used.

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

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

The food composition of the present invention may contain the above-described food additives in an appropriate amount to achieve the purpose of addition depending on the product type.

Regarding other food additives that can be included in the food composition of the present invention, each country's food code or food additive code can be referred to.

The composition of the present invention may be considered a pharmaceutical composition in other specific embodiments.

The pharmaceutical composition of the present invention contains a pharmaceutically acceptable carrier in addition to the active ingredient and can be prepared into an oral formulation or a parenteral formulation depending on the route of administration by a conventional method known in the art. Here, the route of administration may be any suitable 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 when two or more types of drugs according to the administration route are combined, for example, when one drug is administered firstly through an intravenous route and the other drug is administered secondarily through a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or dosage form, and for specifics, reference can 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, it can be prepared as powder, granules, tablets, pills, sugar-coated tablets, capsules, solutions, gels, syrups, suspensions, and wafers along with a suitable carrier according to methods known in the art. It can be manufactured in a dosage form 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, Cellulose such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, glycerol. etc. can be mentioned. When formulating, appropriate binders, lubricants, disintegrants, colorants, diluents, etc. can be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, and wax. As a lubricant, sodium oleate is used. , sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium and calcium salts, polydethylene glycol, etc. Disintegrants include starch, methyl cellulose, Examples include agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt. Additionally, diluents include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc.

When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it can be formulated in the form of injections, transdermal administration, nasal inhalation, and suppositories along with a suitable carrier according to methods known in the art. When formulated as an injection, an aqueous isotonic solution or suspension can be used as a suitable carrier. Specifically, an isotonic solution such as PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or 5% dextrose can be used. . When formulated for transdermal administration, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, paste preparations, linear preparations, and aerol preparations. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide. When formulated as a suppository, the carrier is Wethepsol ( witepsol), Tween 61, polyethylene glycols, cocoa fat, laurel paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, etc. can be used.

Regarding the specific formulation of pharmaceutical compositions, it is known in the art, and references can be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). The above documents are considered a 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, gender, age, patient's severity, and administration route. It may be in the /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 invention in any respect.

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

Even if the composition of the present invention is recognized as a cosmetic composition, the cosmetic composition may be classified as an arbitrary product in terms of its use and by law, and is specifically classified into functional cosmetics with uses such as improving skin troubles and atopic dermatitis, and non-functional cosmetics. It may be general cosmetics, etc. The product form can take any product form, specifically solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax. It can take the form of products such as foundation or spray. Specific product forms may include softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder formulation.

In addition to the active ingredient, the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions, such as stabilizers, solubilizers, surfactants, vitamins, conventional auxiliaries such as colorants and fragrances, and carriers.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier ingredient. You can.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, when the formulation is a spray, chlorofluorohydrocarbon and propane may be used as carrier ingredients. /May contain propellants 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 ester, polyethylene glycol, fatty acid ester of sorbitan, etc. can be used.

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

When the formulation of the present invention is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, and fatty acid amide. Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester can be used.

The cosmetic composition of the present invention can be prepared according to a cosmetic composition manufacturing method commonly used in the art, except that it contains an active ingredient that exhibits skin inflammatory reaction inhibition activity.

As described above, according to the present invention, a composition for inhibiting skin inflammatory response using black horse extract can be provided.

The composition of the present invention can be commercialized into foods, cosmetics, drugs, etc. for the purpose of suppressing skin inflammatory reactions.

Figure 1 shows the results of evaluating the cytotoxicity of the black horse extract in HaCaT cells in which an inflammatory response was induced by TNF-α/IFN-γ.
Figure 2 shows the results of evaluating the effect of black horse extract on the level of production of the inflammatory cytokine IL-6 in HaCaT cells in which an inflammatory response was induced by TNF-α/IFN-γ.
Figure 3 shows the results of evaluating the effect of black horse extract on the level of production of the chemokine MCP-1 in HaCaT cells in which an inflammatory response was induced by TNF-α/IFN-γ.

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

<Example> Inhibitory activity of skin inflammatory response of black horse extract

1. Preparation of black horse extract

10 times the weight of 70% ethanol was added to the dried powder of black horse (extraction part: whole plant), leached at room temperature for 24 hours, filtered with Whatman filter paper No. 2, and the filtrate was concentrated under reduced pressure and frozen. A solid black horse extract was prepared by drying. The extract was suspended in distilled water and then sequentially solvent-fractionated with hexane, dichloromethane, ethyl acetate, and butanol to prepare a total of five fractions, including the fraction of the remaining water layer.

2. Toxicity evaluation in inflammation-induced HaCaT cells

HaCaT cells were plated in a 96-well plate at 2×10 ⁴ cells/well and the cells were stabilized for 24 hours. Afterwards, all culture supernatants were removed and 200 μL of SFM was added to maintain serum deprivation.

After 24 hours, all culture supernatants were removed, 180 μL of new SFM (Serum Free Media) and 20 μL of sample diluted to an appropriate concentration were added and reacted for 1 hour. SFM with 0.1% DMSO and 20 μg/mL dexamethasone, an anti-inflammatory agent, were used as negative control (NC) and positive control (PC), respectively, to compare the activity of the samples.

To induce inflammation, 20 μL of a mixed solution (T+I; 10 ng/mL each) of tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) was added and re-cultured.

After 24 hours, to measure cell viability, all culture medium was removed, 100 μL of 0.5 mg/mL MTT solution was added, and the cells were cultured in an incubator for 30 minutes.

After removing all of the MTT solution, 100 μL of DMSO was added to dissolve the pigment generated within the cells, stirred for 1 minute on a plate shaker, and absorbance was measured at 550 nm. Cytotoxicity was measured by measuring the cells in the inflammation-induced group (T+I). Survival rate (cell viability, %) was calculated.

The results are shown in Figure 1. In Figure 1, number 1 is the hexane fraction, number 2 is the dichloromethane fraction, number 3 is the ethyl acetate fraction, number 4 is the butanol fraction, number 5 is the fraction of the residual water layer, and number 6 is the 70% ethanol extract. Referring to Figure 1, the hexane fraction and dichloromethane fraction did not show cytotoxicity at the treatment concentrations of 12 and 25 μg/mL, but showed cytotoxicity at 50 μg/mL. The 70% ethanol extract and other fractions generally did not show any special cytotoxicity even at 50 μg/mL, and the positive control group did not show any special cytotoxicity at a treatment concentration of 20 μg/mL.

Based on these cytotoxicity results, subsequent experiments were conducted with 25 μg/mL as the highest treatment concentration.

3. Evaluation of inhibition of inflammatory response in HaCaT cells - Evaluation of inflammatory cytokine content using ELISA method

HaCaT cells were plated in a 96-well plate at 2×10 ⁴ cells/well and the cells were stabilized for 24 hours. Afterwards, all culture supernatants were removed and 200 μL of SFM was added to maintain serum deprivation.

After 24 hours, all culture supernatants were removed, 180 μL of new SFM and 20 μL of sample diluted to the appropriate concentration were added, and reacted for 1 hour. SFM with 0.1% DMSO and 20 μg/mL dexamethasone were used as negative control (NC) and positive control (PC), respectively, and compared with the activity of the sample.

To induce inflammation, 20 μL of a mixed solution (T+I; 10 ng/mL each) of tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) was added and re-cultured.

The content of IL-6 (interleukin-6) and MCP-1 (monocyte chemoattractant protein-1) released in the cell culture supernatant was measured using each ELISA set according to the method provided by the manufacturer, and the results were prepared using the provided standard materials. The content (pg/mL) was calculated using a standard curve.

The results are shown in Figures 2 and 3. Referring to Figures 2 and 3, referring to Figure 1, the No. 1 hexane fraction, No. 2 dichloromethane fraction, and No. 3 ethyl acetate fraction significantly produced IL-6 and MCP-1 in a concentration-dependent manner. , and the 70% ethanol extract also inhibited the production of IL-6 and MCP-1 in a concentration-dependent manner, but there was no significant difference in the inhibition of IL-6 production.

<Please note that IL-8 data has been deleted due to inconsistent results>

Claims (6)

A composition for inhibiting skin inflammatory response comprising black horse extract as an active ingredient.
According to paragraph 1,
A composition wherein the black horse extract is any one of (a) to (c):
(a) Water, ethanol, or mixed solvent extract of black horse;
(b) each fraction obtained when the water and ethanol mixed solvent extract of black horse was fractionated with hexane, dichloromethane or ethyl acetate; and
(c) Each fraction obtained when the water and ethanol mixed solvent extract of black horse was sequentially fractionated with hexane, dichloromethane, or ethyl acetate.
According to paragraph 1,
The inhibition of the skin inflammatory response is improvement (alleviation of symptoms), treatment, and prevention of skin diseases accompanied by an inflammatory response,
A composition characterized in that the skin disease accompanying the inflammatory reaction is acne, atopic dermatitis, or skin eczema caused by skin infection with the acne bacteria Propionibacterium acnes .
According to any one of claims 1 to 3,
The composition is characterized in that it is a food composition.
According to any one of claims 1 to 3,
The composition is characterized in that it is a cosmetic composition.
According to any one of claims 1 to 3,
The composition is characterized in that it is a pharmaceutical composition.