Skin improvement cosmetic composition comprising extracts of polygala tenuifolia, angelica dahurica and elsholtzia splendens

Abstract

The present invention relates to a skin improvement cosmetic composition comprising extracts of Polygala tenuifolia, Angelica dahurica, and Elsholtzia splendens. The cosmetic composition exhibits excellent effects of alleviating skin inflammation and providing antioxidants, skin antibacterial action and skin moisturization, and thus can be used as a cosmetic for improving the skin and, further, as a pharmaceutical composition for treating inflammatory skin diseases.

Description

Cosmetic composition for skin improvement comprising extracts of ginseng, white paper and flower oil

[0001] This invention was made under the support of the Ministry of SMEs and Startups under the task identification number 1425174040 and task number S3289410, and the research management specialized organization of the said task is the Korea Technology Information Promotion Agency for SMEs, the research project name is “2022 Small and Medium Business Technology Innovation Development Project `Market Response Type (General Project)` Second Half Implementation Plan Announcement”, the research project name is “Development of a cosmetic composition including flower oil, white paper, and fermented extract of wild paper that helps improve atopy and itching by utilizing anti-inflammatory and skin moisturizing effects”, the main organization is Suikei Co., Ltd., and the research period is 2022.07.25-2023.12.31.

[0002] The present invention relates to a cosmetic composition for improving skin comprising extracts of Polygala tenuifolia, Angelica dahurica and Elsholtzia splendens, and more particularly, to a cosmetic composition having effects of improving skin inflammation, antioxidation, antibacterial and moisturizing the skin.

[0003] Due to concerns about the side effects and chronic use of chemical compounds, environmentally friendly plant-derived natural substances have recently been gaining attention as cosmetic ingredients. Research is actively underway on the diverse bioactive components of these natural products. Natural products contain a variety of bioactive components, including vitamins, minerals, and polyphenols, and are known for their excellent efficacy and stability. Medicinal plants used for cosmetic purposes are diverse, and with increasing interest in their efficacy and effectiveness, active research is underway to develop these materials. Accordingly, in response to the desire to promote health and maintain beautiful skin, natural products with efficacy such as alleviating itching, promoting hair growth, improving wrinkles, whitening, moisturizing, exfoliating, and treating acne, blemishes, and improving skin texture are being applied in cosmetics.

[0004] However, plant extracts used in existing cosmetic compositions often have limitations, such as limited efficacy, stability issues, and potential skin irritation. For example, some extracts can cause allergic reactions when applied directly to the skin, while others may lose efficacy with long-term storage. Furthermore, natural product-based cosmetic compositions face challenges in ensuring consistent quality due to inconsistent manufacturing methods and difficulties in standardizing raw materials. These issues highlight the need for new approaches to ensure the effective and safe use of natural product-based cosmetics.

[0005] Accordingly, there is a growing need for new natural raw materials and cosmetic compositions using them that can improve the problems of existing natural raw materials-based cosmetics.

[0006] The present inventors prepared a composition containing extracts of Polygala tenuifolia, Angelica dahurica, and Elsholtzia splendens as active ingredients, and confirmed that the composition according to the present invention has excellent effects in improving skin inflammation, antioxidant effects, skin antibacterial effects, and skin moisturizing effects.

[0007] Accordingly, the purpose of the present invention is to provide a cosmetic composition for improving skin comprising extracts of ginseng, white paper and flower oil.

[0008] Another object of the present invention is to provide a composition for treating inflammatory skin diseases comprising extracts of rhizome, white paper and flower oil.

[0009] Another object of the present invention is to provide a food composition for improving inflammatory skin diseases, which comprises extracts of raw materials, white paper, and flower oil.

[0010] The present invention relates to a cosmetic composition for improving skin comprising extracts of Polygala tenuifolia, Angelica dahurica and Elsholtzia splendens, and the composition according to the present invention has excellent effects of improving skin inflammation, antioxidant, antibacterial and moisturizing the skin.

[0011] Hereinafter, the present invention will be described in more detail.

[0012]

[0013] One aspect of the present invention is a cosmetic composition for improving skin, comprising Polygala tenuifolia extract, Angelica dahurica extract, and Elsholtzia splendens extract as active ingredients.

[0014] The term "Polygala tenuifolia" used herein refers to a perennial herb belonging to the Polygalaceae family. The core of the root is removed and dried for use. Polygala tenuifolia has been reported to have potential for development as a neuroprotective, antidepressant, analgesic, antifungal, and anticancer agent.

[0015] The term "Angelica dahurica" ​​used herein refers to the dried root of the Umbeliferae family, which is effective for sedation, fever reduction, and analgesic properties. It contains a large amount of sugars, minerals, essential oils, and over 20 types of coumarin. Research on Angelica dahurica has reported its ability to inhibit drug-metabolizing enzymes, inhibit metabolism, and have antithrombotic effects.

[0016] The term “Elsholtzia splendens” in this specification refers to an annual herbaceous plant belonging to the genus Elsholtzia in the family Labiatae, a Korean aromatic edible plant with a unique fragrance emitted from the whole plant. It is known to be effective for coughing, sweating, pain, inflammation, fever, etc., and has been used as an ingredient in folk remedies.

[0017] The term "cosmetic composition" of the present invention may be a formulation selected from the group consisting of, but is not limited to, a solution, an external ointment, a cream, a foam, a nourishing toner, an emollient toner, a perfume, a pack, an emollient, a milky lotion, a makeup base, an essence, a soap, a liquid cleanser, a bath agent, a sunscreen cream, a sun oil, a suspension, an emulsion, a paste, a gel, a lotion, a powder, a soap, a surfactant-containing cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation, a patch, and a spray.

[0018] The term "skin improvement" in this specification may be any one or more selected from the group consisting of skin moisturizing, skin soothing, skin regeneration, skin inflammation improvement, wrinkle improvement, elasticity improvement, skin whitening, antioxidant, anti-aging, skin protection from external stimuli, sensitive skin soothing, sensitive skin improvement, erythema relief, itching relief, skin barrier function strengthening, skin irritation relief, skin antibacterial, and skin pain relief, but is not limited thereto.

[0019] The term “skin inflammation” as used herein refers to diseases that include various inflammatory reactions that occur on the skin, and these diseases show symptoms such as skin redness, itchiness, rash, and swelling, and are sometimes accompanied by pain. The causes of skin inflammation are diverse, including allergic reactions, infections, autoimmune reactions, and environmental factors, and may be caused by an individual’s lifestyle habits or genetic factors. In the present invention, skin inflammation may be, but is not limited to, dermatitis, allergic dermatitis, irritant dermatitis, seborrheic dermatitis, atopic dermatitis, sensitive skin disease, pruritus, eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, rash, and acne.

[0020] In the present invention, the raw material extract may be extracted by selecting at least one selected from the group consisting of leaves, roots, stems, and fruits, but is not limited thereto.

[0021] In the present invention, the white paper extract may be extracted by selecting at least one selected from the group consisting of leaves, roots, stems, and fruits, but is not limited thereto.

[0022] In the present invention, the flower oil extract may be extracted by selecting at least one selected from the group consisting of leaves, roots, stems, and fruits, but is not limited thereto.

[0023] In the present invention, the raw material extract according to the present invention may include a solvent crude extract, a specific solvent-soluble extract (solvent fraction), and a solvent fraction of the solvent crude extract, and may be a crude extract obtained by extracting at least one selected from the group consisting of leaves, roots, stems, and fruits of raw materials with at least one solvent selected from the group consisting of water and straight-chain or branched alcohols having 1 to 4 carbon atoms.

[0024] In the present invention, the white paper extract according to the present invention may include a solvent crude extract, a specific solvent-soluble extract (solvent fraction), and a solvent fraction of the solvent crude extract, and may be a crude extract obtained by extracting at least one selected from the group consisting of leaves, roots, stems, and fruits of white paper with at least one solvent selected from the group consisting of water and straight-chain or branched alcohols having 1 to 4 carbon atoms.

[0025] In the present invention, the flower oil extract according to the present invention may include a solvent crude extract, a specific solvent-soluble extract (solvent fraction), and a solvent fraction of a solvent crude extract, and may be a crude extract obtained by extracting at least one selected from the group consisting of leaves, roots, stems, and fruits of flower oil with at least one solvent selected from the group consisting of water and straight-chain or branched alcohols having 1 to 4 carbon atoms.

[0026] In the present invention, when a mixture of water and alcohol is used to prepare a crude extract, it may be a straight-chain or branched alcohol aqueous solution having 1 to 4 carbon atoms in an amount of 70% or more and less than 100%.

[0027] Additionally, the alcohol aqueous solution may be at least one selected from the group consisting of a methanol aqueous solution, an ethanol aqueous solution, a propanol aqueous solution, and a butanol aqueous solution.

[0028] The extract of the original paper, white paper or flower oil according to the present invention may be a solvent fraction obtained by fractionating a solvent crude extract with an additional solvent, and for example, may be a solvent fraction obtained by using at least one solvent selected from the group consisting of chloroform, ethyl acetate, and hexane in the solvent crude extract.

[0029] In the present invention, the cosmetic composition may be a mixture of a root extract, a white paper extract, and a flower oil extract in a specific ratio.

[0030] In one embodiment of the present invention, the mixing ratio of the root extract, the white paper extract, and the flower oil extract may be mixed in a weight ratio of 0.05-3:0.05-3:0.05-3 based on the weight ratio (w / w), and for example, the mixing ratio of the root extract, the white paper extract, and the flower oil extract may be mixed in a weight ratio of 0.5-1.5:1.5-2.5:0.05-0.15 based on the weight ratio (w / w).

[0031] In a non-limiting embodiment of the present invention, the mixing ratio of the root extract, the white paper extract, and the flower oil extract may be mixed in a weight ratio of 0.5:1.5:1, 1.5:0.5:1, 0.1:1.95:0.95, 0.5:0.5:2, 0.95:1.95:0.1, or 1.45:0.1:1.45 based on the weight ratio (w / w).

[0032] The cosmetic composition of the present invention may additionally include one or more cosmetically acceptable carriers that are incorporated into general skin cosmetics, and may appropriately incorporate conventional ingredients such as oil, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc., but is not limited thereto.

[0033] When the cosmetic formulation of the present invention is an ointment, paste, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. may be used as a carrier component, but are not limited thereto. These may be used alone or in combination of two or more.

[0034] When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizer or an emulsifier may be used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, and the like. In particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol fatty acid esters, polyethylene glycol or fatty acid esters of sorbitan may be used, but are not limited thereto. These may be used alone or in combination of two or more.

[0035] When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth may be used as carrier components, but are not limited thereto. These may be used alone or in combination of two or more.

[0036] When the formulation of the present invention is soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. may be used as carrier components, but are not limited thereto. These may be used alone or in combination of two or more.

[0037] When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or a mixture thereof may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether may be additionally included.

[0038] The cosmetic composition according to the present invention has excellent anti-inflammatory effects as well as skin antioxidant effects by inhibiting DPPH (1-1-diphenyl-2-picryl-hydrazyl) and ABTS (2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) activities and NO (nitric oxide: NO) production. Specifically, it was confirmed that the composition according to the present invention has excellent antioxidant and anti-inflammatory effects at all mixing ratios, and in particular, when mixed at a specific ratio, its effects were confirmed to be particularly excellent (see FIGS. 1, 2 and 4 of the present invention).

[0039] It was confirmed that the cosmetic composition according to the present invention has excellent moisturizing effects by promoting hyaluronic acid production in keratinocytes as well as antioxidant and anti-inflammatory effects, and excellent antibacterial activity against specific strains (see Fig. 6 and Table 2). In particular, it was confirmed that the cosmetic composition according to the present invention has excellent antibacterial activity against strains of a specific genus (Staphylococcus).

[0040] Another aspect of the present invention is a pharmaceutical composition for treating, preventing, alleviating or suppressing skin inflammatory diseases, comprising Polygala tenuifolia extract, Angelica dahurica extract and Elsholtzia splendens extract as active ingredients.

[0041] In the present invention, the pharmaceutical composition comprises extracts of the original paper, white paper, and flower oil, similar to the cosmetic composition according to the present invention described above, and common contents between the two inventions are omitted to avoid excessive complexity of the present specification.

[0042] The term "comprising as an active ingredient" as used herein means comprising an amount sufficient to achieve the efficacy or activity of the extract. The quantitative upper limit of the extract included in the composition of the present invention can be selected and implemented within an appropriate range by those skilled in the art. According to one embodiment of the present invention, the composition of the present invention can be implemented as a pharmaceutical composition, cosmetic composition, or food composition comprising extracts of the rootstock, white paper, and flower oil as active ingredients.

[0043] The content of the extract as an effective ingredient in the composition according to the present invention can be appropriately adjusted depending on the form and purpose of use, the patient's condition, the type and severity of symptoms, etc., and may be 0.001 to 99.9 wt%, or 0.1 to 99.9 wt%, for example, 1 to 99 wt%, based on the solid weight, but is not limited thereto.

[0044] The daily dosage of the composition according to the present invention can be appropriately adjusted depending on the form and purpose of use, the patient's condition, the type and severity of symptoms, etc., and can be 1 to 1000 μg / ml based on the content of the active ingredient, for example, 0.001 to 10000 mg / kg, but is not limited thereto.

[0045] The composition according to the present invention can be administered to mammals, including humans, via various routes. Any commonly used route of administration may be used, including oral, transdermal, intravenous, intramuscular, and subcutaneous administration. For example, oral administration may be used.

[0046] The composition according to the present invention can be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, ointments, suspensions, emulsions, syrups, aerosols, etc., or in the form of parenteral formulations such as transdermal agents, suppositories, and sterile injectable solutions, respectively, according to conventional methods.

[0047] In one embodiment of the present invention, the composition may further contain, in addition to the mixed extract, pharmaceutically suitable and physiologically acceptable carriers, excipients, and diluents, and other auxiliary agents.

[0048] Carriers, excipients and diluents that may be included in the composition of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

[0049] When formulating, diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrating agents, and surfactants commonly used in the art can be used. Solid preparations for oral administration may include tablets, pills, powders, granules, capsules, etc., and such solid preparations may be prepared by mixing the extract with at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used, but are not limited thereto.

[0050] Oral preparations include suspensions, solutions, emulsions, syrups, and ointments. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included, but are not limited to these.

[0051] Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, suppositories, and transdermal preparations. Non-aqueous solutions and suspensions include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

[0052] Suppository formulations may include, but are not limited to, witepsol, macrogol, tween 61, cocoa butter, laurin butter, and glycerogelatin.

[0053] Another aspect of the present invention is a food composition for alleviating, suppressing or improving skin inflammatory diseases, comprising Polygala tenuifolia extract, Angelica dahurica extract and Elsholtzia splendens extract as active ingredients.

[0054] In one embodiment of the present invention, the food composition may be, but is not limited to, various foods, beverages, food additives, etc.

[0055] The food composition according to the present invention may include ingredients commonly added during food manufacturing, and may include, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavorings, but is not limited thereto.

[0056] Carbohydrates that may be included in the food composition according to the present invention include, but are not limited to, monosaccharides such as glucose and fructose, disaccharides such as maltose, sucrose, and oligosaccharides, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.

[0057] Flavoring agents that may be included in the food composition according to the present invention may include, but are not limited to, natural flavoring agents such as thaumatin and stevia extract, and synthetic flavoring agents such as saccharin and aspartame.

[0058] Another aspect of the present invention is a method for improving skin, comprising administering to a subject a composition comprising an extract of Polygala tenuifolia, an extract of Angelica dahurica, and an extract of Elsholtzia splendens.

[0059] Another aspect of the present invention is a method for treating a skin inflammatory disease, comprising administering to a subject a composition comprising an extract of Polygala tenuifolia, an extract of Angelica dahurica, and an extract of Elsholtzia splendens.

[0060] As used herein, the term "administration" refers to providing a given substance to a subject by any suitable method. The composition of the present invention for alleviating, suppressing, treating, or preventing allergic diseases may be administered orally or parenterally via any conventional route, as long as it can reach the target tissue. Furthermore, the composition of the present invention may be administered using any device capable of delivering the active ingredient to target cells.

[0061] The term "subject" as used herein means a mammal, specifically a human, including, but not limited to, a human, monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit or guinea pig.

[0062] In one embodiment of the present invention, the extract composition of the present invention may be administered alone, but may generally be administered mixed with a pharmaceutical carrier selected in consideration of the administration method and standard pharmaceutical practice.

[0063] For example, the composition containing the herbal extract of the present invention may be administered orally, intrabuccally or sublingually in the form of tablets containing starch or lactose, or in the form of capsules alone or containing excipients, or in the form of elixirs or suspensions containing flavoring or coloring chemicals. Such liquid preparations may be formulated with pharmaceutically acceptable excipients such as suspending agents (e.g., methylcellulose, semi-synthetic glycerides such as witepsol, or glyceride mixtures such as mixtures of apricot kernel oil and PEG-6 esters or mixtures of PEG-8 and caprylic / capric glycerides).

[0064] The dosage of the composition containing the herbal extract of the present invention may vary depending on the patient's age, body weight, sex, dosage form, health condition, and disease severity, and may be administered once or several times a day at regular intervals at the discretion of a doctor or pharmacist.

[0065] For example, the daily dosage may be 1 to 1000 μg / ml based on the active ingredient content, but this is an average case and the dosage may be higher or lower depending on individual differences.

[0066] The present invention relates to a cosmetic composition for improving skin, comprising extracts of Polygala tenuifolia, Angelica dahurica and Elsholtzia splendens, which has excellent effects of improving skin inflammation, antioxidant, antibacterial and moisturizing the skin, and can be used as a cosmetic for improving skin, and further, as a pharmaceutical composition for treating inflammatory skin diseases.

[0067] Figure 1 is a graph showing the results of measuring the DPPH radical scavenging activity of a mixed extract sample of raw paper, white paper, and flower oil according to one embodiment of the present invention.

[0068] FIG. 2 is a graph showing the results of measuring the ABTS radical scavenging activity of a mixed extract sample of raw paper, white paper, and flower oil according to one embodiment of the present invention.

[0069] FIG. 3 is a graph showing the results of measuring the survival rate of cells after treating Raw 264.7 cells with a mixed extract sample of raw paper, white paper, and flower oil according to one embodiment of the present invention.

[0070] FIG. 4 is a graph showing the results of measuring the amount of NO produced by Raw 264.7 cells after treating the cells with a mixed extract sample of raw paper, white paper, and flower oil according to one embodiment of the present invention.

[0071] FIG. 5 is a graph showing the results of measuring the cell viability of human keratinocyte cell line (HaCaT) cells after treating the cells with a mixed extract sample of raw paper, white paper, and flower oil according to one embodiment of the present invention.

[0072] FIG. 6 is a graph showing the results of measuring the amount of hyaluronic acid produced by human keratinocyte cell line (HaCaT) cells after treating the cells with a mixed extract sample of raw paper, white paper, and flower oil according to one embodiment of the present invention.

[0073] A cosmetic composition for improving skin inflammation, anti-oxidation, anti-bacterial or moisturizing the skin, comprising Polygala tenuifolia extract, Angelica dahurica extract and Elsholtzia splendens extract as active ingredients.

[0074] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only intended to illustrate the present invention, and the scope of the present invention is not limited by these examples.

[0075]

[0076] Example 1: Materials and Methods

[0077] 1-1. Reagents and equipment

[0078] Reagents included ascorbic acid (AA, Sigma-Aldrich, USA), 1,1-diphenyl-2-picryl-hydrazyl (DPPH, Sigma, USA), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid; ABTS, Sigma, USA), Dulbecco's Modified Eagles Medium (DMEM, Welgene, Korea), fetal bovine serum (FBS, Gibco, USA), FBS (Samchun, Korea), penicillin / streptomycin (Lonza, Switzerland), water-soluble tetrazolium salt (WST)-8 (Biomax, Korea). Lipopolysaccharide (LPS; SigmaAldrich, USA), Griess reagent system (Promega, USA), and hyaluronic acid (HA) enzyme-linked immunosarbent assay (ELISA) kit (Minneapolis, USA) were used.

[0079] The experimental equipment used included a grinder (Hanil, Korea), a rotary vacuum concentrator (BUCHI, Germany), an incubator (Thermo, USA), a microplate reader (BioTek, USA), and a pipette (Eppendorf, Germany).

[0080]

[0081] 1-2. Materials

[0082] The raw materials, white paper, and flower oil used for extraction were purchased from Jecheon Oriental Medicine (Jecheon, Korea) after being washed and dried, and then ground before use.

[0083]

[0084] 1-3. Sample extraction and mixing

[0085] Extraction was performed by adding 400 mL of 70% ethanol to 100 g each of crushed raw paper, white paper, and flower oil at room temperature for 48 hours. The extract was filtered through a filter paper and concentrated at 45°C using a rotary vacuum evaporator. The concentrated powder was stored at -50°C and used in the experiment.

[0086] The mixing ratio of the raw paper, white paper, and flower oil was set to seven mixing ratio conditions (Table 1) according to the content ratio of the raw paper extract, white paper extract, and flower oil extract, and the mixture was prepared and used.

[0087] Polygala teuifolia, Angelicae dahurica, Elsholtzia splendens, M1111, M20.51.51, M31.50.51, M40.11.95, M50.50.52, M60.951.95, M71.4 ...

[0088]

[0089] 1-4. Antioxidant efficacy evaluation

[0090] DPPH radical scavenging assay

[0091] Antioxidant activity was measured using the DPPH radical scavenging activity assay based on Blois's method. This assay is one of the generation methods, and DPPH is purple in its radical state, and changes color to yellow DPPH-H when reduced by accepting electrons. 100 μL of 0.2 mM DPPH solution and 80 μL of samples at various concentrations were mixed in a 96-well plate, reacted for 30 minutes at room temperature in the dark, and the absorbance was measured at 517 nm. The radical scavenging activity was calculated according to the following mathematical equation 1. Ascorbic acid, an antioxidant, was used as a positive control, and a group in which no sample was treated with the DPPH solution was used as a negative control.

[0092]

[0093] [Mathematical Formula 1]

[0094] DPPH scavenging activity (DPPH radical scavenging assay, %)

[0095] =(1-absorbance of sample added group / absorbance of sample not added group) × 100

[0096]

[0097] ABTS radical scavenging assay

[0098] The antioxidant activity using 2,2-azino-biazoline-6-sulphonic acid (ABTS) radical was measured with reference to the Van den Berg method. This method utilizes the principle that the blue-green ABTS+ radical becomes transparent when an antioxidant substance is present. 7 mM ABTS and 2.4 mM potassium persulfate were mixed and left at room temperature in the dark for 24 hours to form ABTS. The absorbance was measured at 734 nm, and the resultant was adjusted to 0.7 ± 0.1 before use in the experiment.

[0099] 100 μL of ABTS working solution and 100 μL of samples at various concentrations were mixed and reacted in a dark room for 10 minutes, and the absorbance was measured at 732 nm to calculate the radical scavenging activity as shown in Equation 2 below. Ascorbic acid, an antioxidant, was used as a positive control group, and a group in which no sample was treated with ABTS solution was used as a negative control group.

[0100]

[0101] [Equation 2]

[0102] ABTS radical scavenging activity (ABTS radical scavenging assay, %)

[0103] =(1-absorbance of sample added group / absorbance of sample not added group) × 100

[0104]

[0105] 1-5. Cell culture

[0106] Raw 264.7 cells were obtained from the Korea Cell Line Bank (KCLB; Seoul, Korea), and human keratinocyte cell line (HaCaT) cells were provided by Amore Pacific (Korea). Cells were cultured in DMEM containing 10% FBS and 1% penicillin / streptomycin, and cultured in an incubator maintained at 37°C and 5% carbon dioxide.

[0107]

[0108] 1-6. Cell viability test

[0109] Cells were seeded at 1 × 10⁴ cells / well in 96-well plates and cultured in an incubator for 24 hours to ensure complete cell attachment. Samples were treated for 48 hours. WST-8 was added at a concentration of 10%, reacted for 2 hours, and absorbance was measured at 450 nm using a microplate reader.

[0110]

[0111] 1-7. Measurement of nitric oxide (NO) inhibitory activity

[0112] Raw 264.7 cells were seeded at 1 × 10 in a 96-well plate. 4 After dispensing cells per well, the cells were cultured in an incubator for 24 hours to ensure complete cell attachment, and the samples were treated for 48 hours. LPS was used as a stimulus at a concentration of 10 ng / mL. The culture supernatant was collected, and the amount of NO production was quantified using the Griess reagent system according to the manufacturer's manual.

[0113]

[0114] 1-8. Hyaluronic acid (HA) measurement

[0115] HaCaT cells were seeded at 1 × 10 in 96-well plates. 4 After dispensing cells per well, the cells were cultured in an incubator for 24 hours to ensure complete cell attachment, and the samples were treated for 48 hours. Retinoic acid was used as a positive control. After collecting the supernatant from the cultured cells, an ELISA assay was performed according to the manufacturer's manual to measure the amount of hyaluronic acid produced.

[0116]

[0117] 1-9. Antibacterial activity

[0118] The antibacterial effect was measured against Staphylococcus aureus (S. aureus). The paper disc method was used to measure the antibacterial activity of the samples. One platinum piece of each strain cultured on a solid plate medium was inoculated into 4 mL of liquid medium and cultured for 20 hours to activate it. Then, 500 μL of the strain was inoculated into 3 mL of liquid medium and cultured for 4 hours. The number of bacteria was 1 × 10 6 - 5 × 10 6After inoculating the strain onto a solid plate medium to reach / mL, it was evenly spread using a spreader. Each sample was prepared at different concentrations, 50 μL was absorbed onto a paper disc (Toyo, Japan) with a diameter of 8 mm, and placed on the solid plate medium on which the strain was spread. After culturing at 37°C for 20 hours, the diameter of the clear zone (mm) around the disc was measured.

[0119]

[0120] 1-10. Statistical processing

[0121] All experiments were repeated three times and presented as the mean ± standard deviation. Using SPSS 18.0, independent-samples t-test and one-way ANOVA were performed. A p value less than 0.05 was considered statistically significant.

[0122]

[0123] Example 2: DPPH and ABTS radical scavenging activity

[0124] Radicals are unstable and readily react with cellular components, causing non-selective and irreversible damage. The DPPH radical, which exhibits a purple color, is a relatively stable compound and is used to test radical activity through its reaction with antioxidants. The DPPH and ABTS radical scavenging activities of the extracts are shown in Figures 1 and 2.

[0125] DPPH radical scavenging activity was highest in M6, which had a high mixing ratio of the original plant and flower oil, at 9.05±2.78-75.37±3.79% at concentrations of 50-1,000 μg / mL. Ascorbic acid, used as a positive control (PC), showed antioxidant activity of 87.21±0.56% at a concentration of 50 μg / mL. Other mixed extracts showed high DPPH radical scavenging activity in the order of M7 2.10±4.91-67.83±3.44%, M1 2.96±1.71-58.21±2.81%, M5 1.34±4.48-57.74±1.87%, M3 2.48±3.58-56.21±1.48%, M4 0.47±4.12-52.95±3.29%, and M2 1.73±1.33-51.35±0.36% at concentrations of 50-1,000 μg / mL, and increased in a concentration-dependent manner in all samples.

[0126] ABTS radical scavenging activity increased with increasing concentration in all mixed extracts, and in particular, M6 showed the highest activity of 22.32±2.62-99.19±0.20% at concentrations of 50-1,000 μg / mL, showing a similar trend to DPPH radical scavenging activity. ABTS radical scavenging activity increased in a concentration-dependent manner in all mixed extracts, and ascorbic acid, used as a positive control (PC), showed antioxidant activity of 99.68±0.21% at a concentration of 50 μg / mL. At a concentration of 500 μg / mL, M6 showed significantly high activity of 94.61±0.13%, followed by M1 92.30±1.04%, M3 87.87±1.32%, M2 80.42±0.11%, M4 80.31±1.22%, M5 76.27±1.44%, and M7 70.59±2.06% in order of ABTS radical scavenging activity.

[0127]

[0128] Example 3: Raw 264.7 cell viability

[0129] To determine the effect of the mixed extracts of the rootstock, white paper, and flower oil on the viability of Raw 264.7 cells, a WST-8 assay was performed, and the results are presented in Fig. 3. The concentrations of all mixed extracts were set to 50, 100, and 200 μg / mL, and since no toxicity was observed at any concentration, the maximum concentration was set to 200 μg / mL, and the amount of NO production was measured.

[0130]

[0131] Example 4: NO production in Raw 264.7 cells

[0132] To compare the effects of mixed extracts at various ratios on NO production, Raw 264.7 cells induced with an inflammatory response by LPS were treated with mixed extracts at concentrations of 50, 100, and 200 μg / mL and the Griess assay was performed. As shown in Fig. 4, the NO production inhibition ability was high in a concentration-dependent manner in M4 and M6, and in particular, M6 showed a statistically significant production inhibition ability of approximately 50% compared to the LPS-treated group at a concentration of 200 μg / mL.

[0133]

[0134] Example 5: HaCaT cell viability

[0135] To determine the effects of the mixed extracts of the rootstock, white paper, and flower oil on the viability of HaCaT cells, a WST-8 assay was performed. The concentrations were set at 12.5, 25.0, and 50.0 μg / mL, and as shown in Figure 5, all mixed extracts showed no toxicity within the concentration range of 12.5-50.0 μg / mL, so the maximum concentration was set at 50.0 μg / mL, and the amount of hyaluronic acid produced was measured.

[0136]

[0137] Example 6: Hyaluronic acid production in HaCaT cells

[0138] In order to investigate the effect of mixed extracts at various ratios on the moisturizing effect, HaCaT cells were treated with each mixed extract at 12.5, 25.0, and 50.0 μg / mL, and the culture fluid was harvested after 48 hours. The amount of hyaluronic acid (HA) produced was measured using an ELISA kit, which is shown in Fig. 6. The content of HA contained in skin tissue reaches 50% of the total HA in the human body, and when HA in the skin decreases due to various factors, it is directly related to skin aging, such as loss of skin moisture. This HA is widely used as a general indicator for the search for skin moisturizing materials in various in vitro models. When the mixed extracts were treated with HaCaT in a concentration range that did not show cytotoxicity and the amount of HA production was evaluated, all mixed extracts showed concentration-dependent HA production promotion activity in the concentration range of 12.5-50.0 μg / mL, and at a concentration of 50 μg / mL, M3 and M6 showed statistically significantly increased HA production ability (1.47-fold and 1.49-fold) compared to the control group.

[0139]

[0140] Example 7: Antibacterial test results

[0141] The disc diffusion test is a method in which a disc containing a sample is placed on an agar plate medium on which bacteria have been spread, and then cultured. The diameter of the inhibition zone (clear zone) of the sample that has diffused around the disc to the extent of inhibiting the growth of bacteria is measured.

[0142] The experimental results for the Staphylococcus aureus strain are shown in Table 2. Methylparaben, used as a positive control, was 8.5 mm at 4 mg / mL, and M4 and M6 were 8.75±0.25 and 9.75±0.25 mm at 8 mg / mL, respectively, demonstrating the effect of inhibiting the growth of the Staphylococcus aureus strain.

[0143] Sample concentration (mg / mL) Clear zone (mm) Methyl Paraben 48.5 M18-M28-M38-M48 8.75±0.25 M58-M68 9.75±0.25 M78-

[0144] The present invention relates to a cosmetic composition for improving skin comprising extracts of Polygala tenuifolia, Angelica dahurica and Elsholtzia splendens, and more particularly, to a cosmetic composition having effects of improving skin inflammation, antioxidation, antibacterial and moisturizing the skin.

Claims

1. A cosmetic composition for improving skin inflammation, anti-oxidation, anti-bacterial or moisturizing the skin, comprising Polygala tenuifolia extract, Angelica dahurica extract and Elsholtzia splendens extract as active ingredients.

2. A cosmetic composition for improving skin inflammation, antioxidation, skin antibacterial or skin moisturizing, wherein in paragraph 1, the original paper extract, the white paper extract and the flower oil extract are extracted with at least one solvent selected from water, alcohol having 1 to 4 carbon atoms and mixed solvents thereof.

3. In the first paragraph, the cosmetic composition is a cosmetic composition for improving skin inflammation, antioxidation, skin antibacterial or skin moisturizing, which inhibits DPPH (1-1-diphenyl-2-picryl-hydrazyl) and ABTS (2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) activity.

4. In the first paragraph, the cosmetic composition is a cosmetic composition for improving skin inflammation, antioxidation, skin antibacterial or skin moisturizing, which suppresses the production of NO (nitric oxide: NO).

5. A cosmetic composition for improving skin inflammation, antioxidation, skin antibacterial or skin moisturizing, wherein in the first paragraph, the mixing ratio of the original paper extract, the white paper extract and the flower oil extract is mixed in a weight ratio of 0.05-3:0.05-3:0.05-3 based on the weight ratio (w / w).

6. In the first paragraph, the cosmetic composition has antibacterial activity against Staphylococcus genus strains, is a cosmetic composition for improving skin inflammation, antioxidation, skin antibacterial or skin moisturizing.

7. In the first paragraph, the cosmetic composition is at least one formulation selected from the group consisting of serum, toner, essence, paste, mask pack, patch, gel, cream, lotion, nutritional lotion, nutritional cream, moisturizing cream, massage cream, powder, soap, cleanser, oil, foundation, makeup base, wax, and spray, a cosmetic composition for improving skin inflammation, antioxidation, skin antibacterial, or skin moisturizing.

8. A skin antibacterial or skin moisturizing cosmetic composition according to claim 1, wherein the skin inflammation is at least one selected from the group consisting of dermatitis, allergic dermatitis, irritant dermatitis, seborrheic dermatitis, atopic dermatitis, sensitive skin disease, pruritus, eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, rash, and acne.

9. A pharmaceutical composition for treating, preventing, alleviating or suppressing skin inflammatory diseases, comprising Polygala tenuifolia extract, Angelica dahurica extract and Elsholtzia splendens extract as active ingredients.

10. A pharmaceutical composition for treating, preventing, alleviating or suppressing a skin inflammatory disease, wherein the skin inflammatory disease in claim 9 is at least one selected from the group consisting of dermatitis, allergic dermatitis, irritant dermatitis, seborrheic dermatitis, atopic dermatitis, sensitive skin disease, pruritus, eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, drug rash and acne.

11. A pharmaceutical composition for treating, preventing, alleviating or suppressing a skin inflammatory disease, wherein in paragraph 9, the mixing ratio of the original paper extract, the white paper extract and the flower oil extract is mixed in a weight ratio of 0.05-3:0.05-3:0.05-3 based on the weight ratio (w / w).

12. A food composition for alleviating, suppressing or improving skin inflammatory diseases, comprising Polygala tenuifolia extract, Angelica dahurica extract and Elsholtzia splendens extract as active ingredients.

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