Cosmetic composition comprising extract extracted using eco-friendly natural eutectic solvent

Abstract

The present disclosure relates to a cosmetic composition which has lower cytotoxicity than an organic solvent extract by extracting a natural product using a co-crystal solvent of betaine and citric acid, and has improved safety and stability by preventing discoloration of the extract, and which can be used as a cosmetic composition for preventing discoloration of the extract. Therefore, the composition can be usefully used as a cosmetic for improving the skin condition and an external preparation for the skin.

Description

Technical Field

[0001] The present disclosure relates to a cosmetic composition. Background Art

[0002] Generally, organic solvents are used in liquid phase extraction. Organic solvents have the advantages of being cheap and having good extraction efficiency, but they also have the disadvantages of being harmful to the health of users or possibly causing environmental pollution problems. Therefore, attention to sustainable development is gradually increasing, and research related to extraction solvents that can replace organic solvents is also currently being actively carried out.

[0003] Among them, eutectic solvents are suggested as alternatives. Eutectic solvents are a kind of eutectic mixtures, which have a melting point lower than the melting point of each component by mixing two components. In the early stage of the eutectic solvent development step, ionic liquids were used as alternatives to organic solvents. However, it has been reported that eutectic solvents are not suitable for use as environmentally friendly solvents due to their low biodegradability, toxicity, and the amount of pollutants discharged in the preparation process. After that, deep eutectic solvents (Deep eutectic solvents), natural eutectic solvents (Natural eutectic solvents), etc. that make up for the shortcomings of existing ionic liquids are studied. However, deep eutectic solvents and natural eutectic solvents include many ingredients that cannot be used in cosmetics, such as choline chloride (Choline chloride), and have higher viscosity, and the solvent changes color in the eutectic solvent preparation process, so there are restrictions on their application in cosmetics and skin topical preparations.

[0004] Therefore, in order to solve the above problems, there is a need to develop environmentally friendly natural eutectic solvents. Summary of the invention Technical issues

[0005] In one aspect, a composition for improving skin condition is provided, which comprises a natural product extract extracted using an environmentally friendly natural eutectic solvent as an effective ingredient.

[0006] Another aspect provides a method for preparing an environmentally friendly natural eutectic solvent.

[0007] On the other hand, provided is a use of a natural product extract extracted using a natural eutectic solvent in preparing a cosmetic composition for improving skin condition, wherein the natural eutectic solvent comprises an amino acid or a derivative thereof, and an organic acid.

[0008] On the other hand, a use of a natural product extract extracted using a natural eutectic solvent in preparing a skin topical composition for improving skin condition is provided, wherein the natural eutectic solvent comprises an amino acid or a derivative thereof, and an organic acid.

[0009] On the other hand, provided is a use of a natural extract extracted using a natural eutectic solvent in preparing a pharmaceutical composition for preventing or treating skin inflammatory diseases, wherein the natural eutectic solvent comprises an amino acid or a derivative thereof, and an organic acid.

[0010] On the other hand, provided is a use of a natural extract extracted using a natural eutectic solvent in the preparation of a pharmaceutical composition for preventing or treating hair loss, or promoting hair growth or development, wherein the natural eutectic solvent comprises an amino acid or a derivative thereof, and an organic acid.

[0011] Another aspect provides a method for preventing, improving, or treating a condition in an individual, the method comprising treating an individual in need of the composition with an effective amount of the composition or applying the composition to the individual. Technical Solution

[0012] In one aspect, a composition for improving skin condition is provided, which comprises a natural extract extracted using an environmentally friendly natural eutectic solvent as an effective ingredient.

[0013] The natural eutectic solvent may be any one or more selected from the group consisting of amino acids or derivatives thereof, and organic acids.

[0014] The amino acid or its derivative can be, for example, betaine, alanine, cysteine, aspartic acid, glutamine, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, arginine, serine, threonine, valine, tryptophan, tyrosine, etc.

[0015] Furthermore, the organic acid may be, for example, citric acid, lactic acid, malic acid, maleic acid, pyruvic acid, fumaric acid, succinic acid, acetic acid, aconitic acid, tartaric acid, ascorbic acid, malonic acid, oxalic acid, glucuronic acid, neuraminic acid, sialic acid, shikimic acid, phytic acid, galacturonic acid, iduronic acid, hyaluronic acid, hydroxycitric acid, lactone derivatives, and the like.

[0016] According to one embodiment, the natural eutectic solvent may be a mixture of betaine and citric acid. The betaine and citric acid may be mixed in a weight ratio of 1:0.001 to 10. The betaine and citric acid may be mixed, for example, in a weight ratio of 1:0.001 to 10, 1:0.01 to 8, 1:0.05 to 7, 1:0.05 to 5, 1:0.01 to 10, 1:0.01 to 6, 1:0.1 to 4, 1:0.1 to 3, 1:1 to 10, 1:1 to 4, or 1:1 to 2. At this time, if the mixing weight ratio of the betaine and citric acid is lower than the range or exceeds the range, there is a problem that the betaine and citric acid precipitate, or the eutectic solvent becomes turbid or discolored.

[0017] According to another embodiment, the pH of the natural eutectic solvent may be 2.5 to 8. Generally, when betaine and citric acid are mixed, an acidic solution having a pH lower than 2.5 is obtained. The extract extracted using an acidic solvent has a lower acidity, and therefore, when the extract is applied to the skin, there is a high possibility of causing irritation or skin damage. Therefore, a pH adjuster may be added to the mixture of betaine and citric acid to adjust the pH of the natural eutectic solvent to 2.5 to 8. The pH of the natural eutectic solvent may be, for example, pH 2.5 to pH 8, pH 2.5 to pH 7.5, pH 3 to pH 7, pH 3 to pH 6.5, pH 3.5 to pH 6, pH 4 to pH 6, or pH 4.5 to pH 5.5. At this time, when the pH of the natural eutectic solvent is lower than the range, there is a problem of skin irritation due to high acidity, and when the pH exceeds the range, the natural product extraction efficiency is poor, and therefore there is a problem of not being able to fully exert the skin condition improvement effect caused by the active ingredient.

[0018] The pH adjuster may be a base or a buffer, for example, sodium citrate, sodium hydroxide, potassium hydroxide, sodium bicarbonate, arginine, tromethamine, triethanolamine, aminomethyl propanol, dipotassium phosphate, potassium phosphate, sodium anisate, sodium lactate, sodium carbonate, urea, etc.

[0019] The natural product can be selected from the group consisting of fig, Selaginella, black rice, dragon's blood, black bean, trumpet creeper, snow lotus, schoenensis, pollen, azolla, fragrant wood, wellhead grass, dicentrata, parasitic wood, creeping grass, glandular tooth blueberry, mulberry tinctoria, tomato, houttuynia cordata, tangerine peel, peony, chestnut flower, kudzu, and walnut.

[0020] The natural product extract can be obtained by finely chopping the dried natural product and then adding a natural eutectic solvent of 2 to 20 times the weight of the natural product for extraction. The extraction can be performed at an extraction temperature of 30° C. to 100° C. The extraction can be performed for an extraction time of 1 hour to 10 hours. As the extraction method, cold soaking, ultrasonic extraction, or circulating cooling extraction method can be used. The extraction can be repeated 1 to 5 times.

[0021] According to one embodiment, it has been confirmed that the safety and stability of the natural extract extracted by the natural eutectic solvent prepared by mixing betaine and citric acid are improved compared with the natural extract extracted by organic solvent, and it has excellent elimination ability for active oxygen and free radicals, and also shows excellent defense effect on cell damage caused by ultraviolet rays. In addition, compared with the natural extract extracted by organic solvent, the natural extract extracted by the natural eutectic solvent significantly inhibits the expression of hyaluronidase, inflammatory cytokines and / or inflammatory mediators, not only hinders tyrosinase activity and inhibits melanin synthesis, but also increases the expression of skin barrier or moisturizing, elasticity-related factors, and also inhibits the expression of acne or sebum, or hair loss-related factors.

[0022] The skin condition improvement or skin beauty improvement may be the inhibition or improvement of skin aging, wrinkles, elasticity, skin barrier strengthening, skin moisturizing, skin acne or sebum inhibition or improvement. Also, it may be the prevention, improvement or treatment of hair loss, or the promotion of hair growth or hair development.

[0023] In this specification, the term "skin aging" refers to the general term for visible and invisible changes that occur in the skin with age, for example, it refers to the phenomenon of reduced epidermal thickness, the number of cells or blood vessels in the dermis, the ability to recover from DNA damage, the cell alternation cycle, wound recovery, skin barrier function, epidermal moisture maintenance, sweat secretion, sebum secretion, vitamin D production, physical damage defense, chemical substance removal ability, immune response, sensory function, and a decrease in body temperature regulation.

[0024] The composition can be used to improve skin aging caused by exogenous factors or endogenous factors. The exogenous factors refer to various external factors, such as ultraviolet rays (light), and the endogenous factors are also called chronological factors, which mainly refer to factors generated by the passage of time. That is, specifically, the skin aging includes early aging symptoms induced by external stimuli such as ultraviolet rays, pollution, cigarette smoke, chemicals, and the like, and natural aging phenomena that occur as skin cell proliferation decreases with age, and both include concepts such as wrinkles, reduced elasticity, sagging skin, and dryness. In addition, wrinkles include wrinkles caused by changes in internal and external factors that cause stimuli that change the components of skin tissue.

[0025] The aging may be aging due to active oxygen or photoaging.

[0026] The "active oxygen" is generated by normal metabolic processes, harmful toxins such as ultraviolet rays, pesticides, cigarette smoke, air pollution, etc. Active oxygen causes damage to the components of the organism by leading to damage to skin cells and tissues and destroying the antioxidant defense network, such as lipid peroxidation, protein oxidation, activation of protease that destroys intercellular matrix components, chain breakage and abnormal cross-linking of collagen and elastin as elastic fibers, chain breakage of hyaluronic acid, promotion of melanin production reaction, and DNA oxidation.

[0027] Photoaging is a phenomenon caused by external environmental factors, the most representative of which is ultraviolet rays. Ultraviolet rays damage the components of biological bodies, such as the activation of protease and the scission and abnormal cross-linking of matrix proteins. The repetition of this mechanism also leads to obvious skin aging in appearance.

[0028] In this specification, the term "wrinkles" refers to the state in which the skin loses elasticity and becomes loose, such as skin folds. The "prevention or improvement of skin wrinkles" refers to all actions of inhibiting the expression of wrinkle-related factors to prevent or improve wrinkles or increase the total amount of collagen.

[0029] The term "skin barrier strengthening" refers to any action that improves the function of the skin barrier located on the outermost side of the skin and that prevents loss of moisture and nutrients.

[0030] The term "skin moisturizing" may refer to any action that maintains skin moisture or prevents moisture loss.

[0031] The "skin disease" may be a disease caused by damage to the skin barrier function, skin aging, skin wounds, skin scars, or skin inflammation. The term "prevention" includes inhibiting the occurrence of the disease. The term "treatment" includes inhibiting, alleviating, or eliminating the progression of the disease.

[0032] The skin barrier function damage may include all changes that occur in the skin due to the reduction or damage of the skin barrier function. For example, it may include increased skin wrinkles, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne, etc.

[0033] The skin inflammatory disease may be any one selected from the group consisting of skin wounds, dermatitis, atopic dermatitis, pruritus, eczematous skin diseases, dry eczema, erythema, urticaria, psoriasis, drug eruption, and acne.

[0034] In the present specification, the term "Alopecia" refers to a phenomenon in which hair falls off the scalp or a state in which hair becomes thinner or thinner, and "prevention or improvement of hair loss" refers to not only promoting the generation of new hair but also promoting the healthy growth of existing hair.

[0035] The hair loss can be, for example, androgenic alopecia, Alopecia Areata, Androgenetic Alopecia, Telogen Effluvium, Traumatic Alopecia, Trichotillomania, Pressure Alopecia, Anagen Efficacy, Nasal Alopecia, Alopecia Syphlltiac, Alopecia Seborrhecia, symptomatic alopecia, cicatricial alopecia, and congenital alopecia, etc.

[0036] In this specification, the term "hair growth" refers to the production of hair on the scalp, and "hair growth" refers to the growth of hair (i.e., Hair growth), and has the same meaning as other terms "hair care" used in the art.

[0037] The composition is effective for improving hair loss caused by exogenous or endogenous factors. Exogenous factors refer to various external factors, such as ultraviolet rays, and endogenous factors refer to factors that occur over time, including hair papilla cell apoptosis caused by telomeres and oxidative stress caused by reduced cell function.

[0038] In the present specification, the term "promoting hair growth or hair cultivation" means promoting the production and growth of hair to ultimately increase the proportion of growing hair in the entire hair. In addition, the hair growth or hair cultivation promotion means inhibiting hair loss caused by a decrease in the proportion of growing hair.

[0040] Based on the total weight of the composition, the composition may include 0.001 wt% to 80 wt%, for example, 0.01 wt% to 60 wt%, 0.01 wt% to 40 wt%, 0.01 wt% to 30 wt%, 0.01 wt% to % to 20 wt%, 0.01 wt% to 10 wt%, 0.01 wt% to 5 wt%, 0.05 wt% to 60 wt%, 0.005 wt% to 40 wt%, 0.05 wt% to 30 wt%, 0.05 wt% to 20 wt%, 0.05 wt% to 10 wt%, 0.05 wt% to 5 wt%, 0.1 wt% to 60 wt%, 0.1 wt% to 40 wt%, 0.1 wt% to 30 wt%, 0.1 wt% to 20 wt%, 0.1 wt% to 10 wt%, or 0.1 wt% to 5 wt% of a natural eutectic solvent extract. At this time, if the content of the extract is lower than the above range, there is a possibility that the following skin condition improvement effects cannot be fully achieved: skin aging inhibition, wound improvement, skin barrier strengthening, skin wrinkle inhibition, or skin moisturizing effect.

[0041] In the present specification, the term "comprising as an effective ingredient" means that the natural product extract of the present specification is added to an extent that the above-mentioned effect can be exerted, and means that various ingredients are added as auxiliary ingredients for drug delivery and stabilization, etc., and formulation is performed in various forms.

[0043] According to another embodiment, the composition may be a cosmetic composition.

[0044] The cosmetic composition may be, for example, a cosmetic dosage form having a softening lotion, a nourishing lotion, a massage cream, a nourishing cream, an essence, a mask, a gel, an ampoule, or a skin-adhesive type.

[0045] The ingredients included in the cosmetic composition may include ingredients generally used as active ingredients in cosmetic compositions other than the composition, for example, conventional auxiliary agents and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0047] According to another embodiment, the composition may be a composition for external skin application.

[0048] In the present specification, the skin topical agent can be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal patch, a medicated bandage, an emulsion, or a combination thereof. In addition, the skin topical agent can be a scalp skin care lotion, a scalp lotion, a scalp cream, a scalp serum, a scalp essence, a scalp ampoule, a scalp care, a scalp conditioner, a scalp shampoo, a scalp mask, a hair growth factor, a hair lotion, a hair cream, a hair spray, a mousse, a hair spray, a conditioner, a shampoo, a conditioner, a hair mask, a conditioner, an eyebrow growth agent, an eyelash growth agent, an eyelash nutrient, a pet shampoo, a pet conditioner, a hand sanitizer, a washing soap, a perfumed soap, a disinfectant cleaner, a wet wipe, a mask, an ointment, a patch, or a filter filler, etc. The skin preparation can be appropriately combined with ingredients commonly used in skin preparations such as cosmetics or medicines as needed, such as aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, or combinations thereof. The skin preparation can also be appropriately combined with metal chelators such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, such as caffeine, tannin, verapamil, licorice extract, glabridin, hot water extract of Kalin fruit, various medicinal herbs, tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts, vitamin C, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, kojic acid, such as glucose, fructose, trehalose and other sugars.

[0050] According to another embodiment, the composition may be a pharmaceutical composition.

[0051] The pharmaceutical composition may further include a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, and the lubricant may include magnesium stearate, talc, or a combination thereof. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, anhydrous calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinyl pyrrolidone, low-substituted hydroxypropyl cellulose, hydroxypropyl cellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

[0052] The pharmaceutical composition can be prepared into an oral or non-oral dosage form. The oral dosage form can be granules, powders, liquids, tablets, capsules, dry syrups, or a combination thereof. The non-oral dosage form can be an injection.

[0054] According to another embodiment, the composition may be a health functional food composition.

[0055] In the health functional food composition, the strain or its culture solution can be used alone, or can be used together with other foods or food ingredients, or can be appropriately used according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health or treatment). Usually, when preparing food or beverage, based on the raw material, the composition of this specification can be added in an amount of 15 parts by weight or less. The type of the health functional food is not particularly limited. Among the types of health functional foods, the beverage composition can contain various flavoring agents or natural carbohydrates as additional ingredients like conventional beverages. The natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extracts, or synthetic sweeteners such as saccharin and aspartame can be used. The health functional food composition may also include nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectin acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerol, alcohol, carbonating agents used in carbonated beverages, or combinations thereof. The health functional food composition may also include natural fruit juice, fruit juice beverages, pulp for preparing vegetable beverages, or combinations thereof.

[0057] On the other hand, a use of a natural product extract extracted using a natural eutectic solvent in preparing a cosmetic composition for improving skin condition is provided, wherein the natural eutectic solvent comprises an amino acid or a derivative thereof, and an organic acid. Regarding the use, the specific contents of skin condition, improvement, cosmetic composition, natural eutectic solvent, extraction, natural product, amino acid, derivative, organic acid, etc. are as described above.

[0058] On the other hand, a use of a natural product extract extracted using a natural eutectic solvent in preparing a skin topical composition for improving skin condition is provided, wherein the natural eutectic solvent comprises an amino acid or a derivative thereof, and an organic acid. Regarding the use, the specific contents of skin condition, improvement, skin topical composition, natural eutectic solvent, extraction, natural product, amino acid, derivative, organic acid, etc. are as described above.

[0059] On the other hand, a use of a natural product extract extracted using a natural eutectic solvent in preparing a pharmaceutical composition for preventing or treating skin inflammation diseases is provided, wherein the natural eutectic solvent comprises an amino acid or a derivative thereof, and an organic acid. Regarding the use, the specific contents of skin inflammation, prevention, treatment, pharmaceutical composition, natural eutectic solvent, extraction, natural product, amino acid, derivative, organic acid, etc. are as described above.

[0060] On the other hand, a use of a natural product extract extracted using a natural eutectic solvent in the preparation of a pharmaceutical composition for preventing or treating hair loss, or promoting hair growth or hair development is provided, wherein the natural eutectic solvent comprises an amino acid or a derivative thereof, and an organic acid. Regarding the use, the specific contents of hair loss, prevention, treatment, hair growth, hair development, pharmaceutical composition, natural eutectic solvent, extraction, natural product, amino acid, derivative, organic acid, etc. are as described above.

[0062] And, another aspect provides a method for preventing, improving, or treating a condition of an individual, the method comprising treating an individual in need of the composition with an effective amount of the composition or administering the composition to the individual.

[0063] The individual condition may be a skin-related condition or an inflammation-related condition.

[0064] In the present specification, the terms "administering", "introducing" and "implanting" are used interchangeably and may refer to placing a composition according to a specific embodiment into an individual by a method or route that causes the composition according to a specific embodiment to be at least partially localized to a desired site.

[0065] Administration can be carried out by methods known in the art. Administration can be carried out to an individual by any means, for example by routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. The administration can be carried out systemically or locally.

[0066] The subject may be a mammal, for example, a human, a cow, a horse, a pig, a dog, a sheep, a goat or a cat. The subject may be a subject in need of the following skin cosmetic improvements: skin moisturizing, skin barrier strengthening, skin inflammation suppression, and skin wrinkle improvement effects.

[0067] The administration may refer to administering 0.1 mg to 1000 mg of the composition according to one embodiment to each individual per day, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 The dosage can be 100mg to 200mg, 100mg to 50mg or 100mg to 25mg. However, the dosage can be prescribed differently according to factors such as preparation method, administration method, patient's age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity, and those skilled in the art can consider these factors to appropriately adjust the dosage. The number of times of administration can be once a day, or can be applied twice or more times within the scope of clinically acceptable side effects, and for the application site, it can be applied to one or two or more places, and the application interval can be every day or 2 days to 5 days, and the total number of days of application for a treatment can be 1 day to 30 days. If necessary, the same treatment can be repeated after the appropriate period. For animals other than humans, the same dosage per kilogram as humans can also be applied, or, for example, the amount obtained by converting the dosage based on the volume ratio (e.g., mean value, etc.) of the target animal and the organ (heart, etc.) of the human can be applied.

[0068] Another aspect provides a method for preparing a natural eutectic solvent, comprising: mixing betaine and citric acid to prepare a mixture; and adjusting the pH by adding any one selected from the group consisting of the following items to the mixture: Sodium citrate, sodium hydroxide, potassium hydroxide, sodium bicarbonate, arginine, tromethamine, triethanolamine, aminomethyl propanol, dipotassium phosphate, potassium phosphate, sodium anisate, sodium lactate, sodium carbonate, and urea.

[0069] In addition, a method for preparing a natural product extract using a natural eutectic solvent is provided, the method comprising extracting the natural product using amino acids, organic acids or a mixture thereof to prepare the extract. The specific contents of the natural product, the natural eutectic solvent, etc. are as described above.

[0070] The natural product extract prepared by the method has an excellent elimination rate for active oxygen and free radicals, and increases the expression of SPT, aquaporin 3 (AQP3), and ribonuclease A7 (RNase7) by inhibiting melanin synthesis, so it can be useful for improving skin conditions such as skin barrier strengthening, skin moisturizing, elasticity, and skin aging improvement. In addition, by reducing the expression of inflammatory cytokines, it can be useful for the prevention, improvement, or treatment of skin inflammatory diseases. Beneficial Effects

[0071] The cosmetic composition according to one aspect extracts natural substances by using a eutectic solvent of betaine and citric acid, and thus has lower cytotoxicity than an organic solvent extract, and has improved safety and stability by preventing discoloration of the extract, and thus can be usefully used as a cosmetic and a skin external preparation for improving skin conditions. DETAILED DESCRIPTION

[0072] Hereinafter, preferred embodiments are provided to help understanding of the present invention. However, the following embodiments are only provided to make the present invention more understandable and clear, and the content of the present invention is not limited to the following embodiments.

[0074] [Preparation Example]

[0075] Preparation Example 1: Preparation of natural eutectic solvents containing amino acids and organic acids

[0076] A natural eutectic solvent including betaine as an amino acid and citric acid as an organic acid was prepared. Specifically, betaine and citric acid were mixed in the same amount and stirred at 150 rpm for 2 hours at 60 to 100° C. to prepare a natural eutectic solvent having a pH of 2.

[0077] Preparation Example 2-4: Preparation of pH-adjusted natural eutectic solvent

[0078] A pH-adjusted natural eutectic solvent was prepared. Specifically, 1 M sodium citrate The aqueous solution was added to the natural eutectic solvent of Example 1 at 1, 5 and 10 kg to prepare pH 3 (Preparation Example 2), pH 5 (Preparation Example 3) and pH 7 (Preparation Example 4) natural eutectic solvents.

[0080] [Example]

[0081] Example 1. Preparation of Fig Fruit Cocrystal Solvent Extract

[0082] A natural eutectic solvent extract of fig fruit (Ficus carica fruit) was prepared. Specifically, 1 kg of dried and crushed fig fruit was extracted for 6 hours using the natural eutectic solvent of Preparation Example 1. Then, the eutectic solvent extract of fig fruit was prepared by filtering using Whatman #5 filter paper.

[0083] Example 2: Preparation of Selaginella cocrystal solvent extract

[0084] A Selaginella tamariscine cocrystal solvent extract was prepared by the same method as described in Example 1, except that Selaginella tamariscine was used.

[0085] Example 3, Preparation of Germinated Black Rice Cocrystal Solvent Extract

[0086] A germinated black rice co-crystal solvent extract was prepared by the same method as in Example 1 except that germinated black rice (Oryza sativa) was used.

[0087] Example 4: Preparation of Dragon Blood Cocrystal Solvent Extract

[0088] A eutectic solvent extract of Daemonorops draco was prepared by the same method as in Example 1 except that Daemonorops draco was used.

[0089] Example 5. Preparation of a co-crystal solvent extract of a mixture of fig fruit, Selaginella, germinated black rice and Sanguisorba officinalis

[0090] A eutectic solvent extract of a mixture of Ficus carica fruit, Selaginella, germinated black rice and Sanguisorba officinalis was prepared by the same method as described in Example 1, except that Ficus carica fruit, Selaginella, germinated black rice and Sanguisorba officinalis were used in the same amounts.

[0091] Examples 6 to 8: Preparation of pH-adjusted natural eutectic solvent extracts from fig fruit

[0092] The pH-adjusted natural eutectic solvent extract of fig fruit was prepared by the same method as in Example 1, except that the natural eutectic solvents of Preparation Examples 2 to 4 were used.

[0093] Examples 9 to 11: Preparation of pH-adjusted Selaginella natural eutectic solvent extract

[0094] The pH-adjusted Selaginella natural eutectic solvent extract was prepared by the same method as in Example 1, except that the natural eutectic solvents of Preparation Examples 2 to 4 were used.

[0095] Examples 12 to 14: Preparation of pH-adjusted germinated black rice eutectic solvent extract

[0096] The pH-adjusted germinated black rice eutectic solvent extract was prepared by the same method as in Example 1, except that the natural eutectic solvents of Preparation Examples 2 to 4 were used.

[0097] Preparation Examples 15 to 17: Preparation of pH-adjusted Dragon's Blood Natural Cocrystal Solvent Extract

[0098] The pH-adjusted Dragon's Blood natural eutectic solvent extract was prepared by the same method as in Example 1, except that the natural eutectic solvents of Preparation Examples 2 to 4 were used.

[0099] Preparation Examples 18 to 20, pH-adjusted eutectic solvent of a mixture of fig fruit, Selaginella, germinated black rice and dragon's blood Preparation of extracts

[0100] The eutectic solvent extract of the mixture of Ficus carica fruit, Selaginella, germinated black rice and Dragon's Blood was prepared by the same method as in Example 1 except that the natural eutectic solvent of Preparation Examples 2 to 4 was used.

[0102] [Comparative Example]

[0103] Comparative Example 1: Preparation of Fig Fruit Ethanol Extract

[0104] A fig fruit ethanol extract was prepared by the same method as in Example 1, except that 70% (v / v) ethanol was used.

[0105] Comparative Example 2: Preparation of Selaginella ethanol extract

[0106] The Selaginella ethanol extract was prepared by the same method as in Example 1, except that 70% (v / v) ethanol was used.

[0107] Comparative Example 3: Preparation of Germinated Black Rice Ethanol Extract

[0108] The germinated black rice ethanol extract was prepared by the same method as in Example 1 except that 70% (v / v) ethanol was used.

[0109] Comparative Example 4: Preparation of Dragon's Blood Ethanol Extract

[0110] The dragon's blood ethanol extract was prepared by the same method as in Example 1 except that 70% (v / v) ethanol was used.

[0112] [Experimental example]

[0113] Experimental Example 1: Confirmation of active ingredient content

[0114] The contents of phenol and flavonoids in the natural eutectic solvent extract according to one embodiment were confirmed. Specifically, 1 ml of each of Examples 1 to 20 and Comparative Examples 1 to 4 was added to 10 ml of distilled water, and 2 ml of Folin-Ciocalteu phenol reagent (Sigma) was added to mix. Then, the reaction was carried out at room temperature for 5 minutes, and 2 ml of 20% sodium carbonate was added to further react at room temperature for 1 hour. The total phenol content was determined by measuring the absorbance at 680 nm. Gallic acid was used as an indicator substance.

[0115] Then, 1.5 ml of each of Examples 1 to 20 and Comparative Examples 1 to 4 was mixed with 2% AlCl3H2O dissolved in the same amount of methanol, and reacted at room temperature for 10 minutes. Then, the total flavonoid content was determined by measuring the absorbance at 367 nm. Catechin was used as an indicator substance.

[0116] [Table 1]

[0117] As a result, it was confirmed that the total polyphenol content and the total flavonoid content in Examples 6 to 20 in which the pH was adjusted were significantly higher than those in Examples 1 to 5 in which the pH was not adjusted, as shown in Table 1. Specifically, in Examples 7, 10, 16, and 19 in which the pH was adjusted to 5, the total polyphenol content and the total flavonoid content were higher than those in the natural eutectic solvent in which the pH was adjusted to 3 or 7.

[0118] That is, the natural eutectic solvent according to one embodiment can maximize the active ingredient to be extracted at a specific pH.

[0119] Experimental Example 2: Confirmation of Cytotoxicity

[0120] The cytotoxicity of the natural eutectic solvent extract according to one embodiment to skin cells was confirmed. Specifically, Fibroblasts were diluted in Dulbecco's modified cell medium (DMEM) supplemented with 10% FBS at a rate of 1×10 5Cells / mL were injected into a 96-well plate and allowed to adhere for 24 hours. Then, Examples 1 to 20 and Comparative Examples 1 to 4 were diluted to appropriate concentrations and treated in each well and cultured for 24 hours. After 24 hours, the culture medium was removed and a medium containing MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was added. 200 μl of cell culture medium containing 2,5-diphenyl tetrazolium bromide solution (2.5 mg / ml) was added to each well and cultured in a 37°C, CO2 incubator for 2 hours. The culture medium was removed and 100 μl of dimethyl sulfoxide (DMSO) was added. After shaking for 5 minutes to dissolve the cells, the absorbance at 565 nm was measured using an ELISA reader. The cell survival rate (%) was determined according to the following mathematical formula 1, and the reagent concentration that did not affect cell survival was determined and the results are shown in the following Table 2.

[0121] [Mathematical formula 1]

[0122] Bo: Absorbance at 565 nm of the wells that developed color reaction with cell culture medium only

[0123] Bt: Absorbance at 565 nm of the wells that were not treated with reagents but were subjected to color development reaction

[0124] S: Absorbance at 565 nm of the wells treated with reagents and undergoing color development

[0125] [Table 2]

[0126] As a result, it was confirmed that the 100% cell survival concentration in Examples 1 to 20 was 5 to 40 times higher than that in Comparative Examples 1 to 4 as shown in Table 2. In particular, it was confirmed that the cell survival concentration of the natural eutectic solvent at pH 3 to 7 was significantly higher than that at pH 2.5. Specifically, for Examples 6 to 20 whose pH was adjusted, there was a trend that the cytotoxicity decreased as the pH increased, and in particular, it was confirmed that the 100% cell survival concentration in the natural eutectic solvent extract whose pH was adjusted to pH 5 was the highest.

[0127] Therefore, the natural eutectic solvent according to one embodiment has a low possibility of causing skin irritation and a wide range of useful concentrations, and thus can be used as a safe and effective solvent.

[0128] Experimental Example 3: Confirmation of stability at room temperature

[0129] The room temperature stability of the natural eutectic solvent extract according to one embodiment was confirmed. Specifically, the reagents were prepared by mixing the Examples 1 to 20 and Comparative Examples 1 to 4 with 60% by weight of dipropylene glycol. Then, the reagents were respectively put into glass containers and stored in a constant temperature bath kept at 45° C. for 4 weeks to measure the degree of discoloration. For the color change of the reagent, the color change (ΔE) was measured using a spectrophotometer (Minolta CR300), and the stability was evaluated and the results are shown in the following Table 3.

[0130] [Table 3]

[0131] As a result, it was confirmed that, as shown in Table 3, the color change at room temperature was less in Examples 6 to 20 than in the organic solvent extracts of Comparative Examples 1 to 4. Specifically, the natural product extracts extracted with the natural eutectic solvent adjusted to pH 5 (Examples 7, 10, 14, 16, and 19) showed less color change, thereby confirming that the highest stability was exhibited.

[0132] According to Examples 1 to 3, the extraction efficiency of Comparative Examples 1 to 4 was better than that of Examples 1 to 20, but the cytotoxicity was higher and the room temperature stability was poor. However, considering that the cytotoxicity and room temperature stability problems that may occur in organic solvent extraction are improved, according to one embodiment, it is usefully used as a solvent for preparing cosmetics and skin external preparations.

[0133] Experimental Example 4: Antioxidant Activity Confirmation (1)

[0134] Based on the results of Experimental Example 3, an activity experiment of a natural product extract extracted using a natural eutectic solvent at pH 5 was performed in the following Experimental Example.

[0135] The antioxidant activity of the natural eutectic solvent extract according to one embodiment was confirmed using the Nitro Blue Tetrazolium method. Specifically, 2.4 ml of 0.05 M Na2CO3, 0.1 mL of 3 mM xanthine solution, 0.1 ml of 3 mM ethylenediaminetetraacetic acid (EDTA), 0.1 ml of bovine serum albumin (BSA) solution, and 0.1 ml of 0.72 mM NBT solution were put into a vial, and the Examples 7, 10, 13, 16 and 19 and Comparative Examples 1 to 4 were added at 0.1% by weight and left to stand at 25 ° C for 10 minutes. Then, 0.1 ml of xanthine oxidase was added and stirred at high speed, incubated at 25 ° C for 20 minutes, and 0.1 ml of 6 mL of CuCl2 solution was added to stop the reaction, and then the absorbance (St) at 560 nm was measured. Distilled water Bt was used to replace the extract of the embodiment as a negative control group. As a blank control group (Blank) of the reagent solution, distilled water (Bo) was used instead of xanthine oxidase as a reaction enzyme, and the measurement results were calculated according to the following mathematical formula 2. Butylated hydroxytoluene was used as a positive control group.

[0136] [Mathematical formula 2]

[0137] St: absorbance at 560nm after the reagent enzyme reaction

[0138] Bt: absorbance at 560nm after enzyme reaction in negative control group

[0139] So: Absorbance of the reagent without enzyme at 560nm before enzyme reaction

[0140] Bo: absorbance at 560 nm of the negative control group without enzyme addition before enzyme reaction

[0141] [Table 4] Active oxygen elimination rate (antioxidant effect, %) Positive control group 86.15 Example 7 74.57 Example 10 91.15 Example 13 83.32 Example 16 90.47 Embodiment 19 90.07 Comparative Example 1 70.16 Comparative Example 2 88.64 Comparative Example 3 79.57 Comparative Example 4 82.45

[0142] As a result, as shown in Table 4, it was confirmed that the active oxygen elimination rates in Examples 7, 10, 13, 16 and 19 were significantly higher than those in Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0143] Therefore, the natural co-crystal solvent extract according to one embodiment has excellent antioxidant activity and thus can be usefully applied to prevent or improve aging.

[0144] Experimental Example 5: Antioxidant Activity Confirmation (2)

[0145] Using the DPPH method [2,2-di(4-tert-octylphenyl)-1-trinitrophenylhydrazine radical (2,2-Di(4-tert- octylpheny1-1-pycrylhydrazyl free radical))] was used to confirm the antioxidant activity of the natural eutectic solvent extract according to one embodiment. Specifically, Examples 7, 10, 13, 16, and 19 and Comparative Examples 1 to 4 were diluted to a concentration of 0.1% and placed in a 96-well plate. Then, DPPH prepared using 100 μl of methanol solution was added to adjust the total volume of the solution to 200 μl, and after standing at 37°C for 30 minutes, the absorbance was measured at 560 nm. The free radical elimination activity (%) was calculated according to the following mathematical formula 3, and the results are shown in the following Table 5. Butylated hydroxytoluene was used as a positive control group.

[0146] [Mathematical formula 3] Free radical elimination activity (%) = 100-(BA) × 100

[0147] A: Absorbance of untreated control wells

[0148] B: Absorbance of the experimental group wells

[0149] [Table 5] Free radical elimination rate (antioxidant effect, %) Positive control group 81.33 Example 7 70.51 Example 10 89.67 Example 13 81.35 Example 16 74.51 Embodiment 19 88.87 Comparative Example 1 61.34 Comparative Example 2 78.24 Comparative Example 3 69.47 Comparative Example 4 66.50

[0150] As a result, as shown in Table 5, it was confirmed that the radical elimination rates in Examples 7, 10, 13, 16 and 19 were significantly higher than those in Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0151] Therefore, the natural co-crystal solvent extract according to one embodiment has excellent antioxidant activity and thus can be usefully used for preventing or improving aging.

[0152] Experimental Example 6: Confirmation of the effect of protecting skin cells from ultraviolet rays

[0153] The mitigation effect of the natural eutectic solvent according to one embodiment on the cytotoxicity caused by ultraviolet rays was confirmed. 5 Cells / ml of fibroblasts were placed in a 96-well plate and allowed to attach for 24 hours. After 24 hours, the cells 7, 10, 13, 16, and 19 and Comparative Examples 1 to 4 were further cultured for 24 hours under the same culture conditions. Then, the culture medium was removed and each well was washed once with PBS, and 100 μl of PBS was placed in each well. Then, an ultraviolet B (UVB) lamp (model: F15T8, UV B 15W, Shanko Dennki, Japan) irradiation with ultraviolet light 10mJ / cm 2 After that, PBS was removed and 200 μl of DMEM supplemented with 10% FBS was added. Here, each reagent was used at a concentration that did not show cytotoxicity and then cultured for 24 hours. After 24 hours, the culture medium was removed, and 200 μl of MTT solution (2.5 mg / ml) containing cell culture medium was added to each well, and then cultured in a 37°C, CO2 incubator for 2 hours. The culture medium was removed and 100 μl of dimethyl sulfoxide (DMSO) was added to each well. After that, the cells were lysed by shaking for 2 minutes and the absorbance at 565 nm was measured in an ELISA instrument. The cell survival rate (%) and the mitigation rate of cytotoxicity caused by ultraviolet rays were calculated according to the following mathematical formula 4 and mathematical formula 5, respectively, and the results are shown in the following Table 6.

[0154] [Formula 4]

[0155] Bo: Absorbance at 565 nm of the wells that developed color reaction with cell culture medium only

[0156] Bt: Absorbance at 565 nm of the wells that were not treated with reagents but were subjected to color development reaction

[0157] St: Absorbance at 565 nm of the wells treated with the reagent and subjected to color development reaction

[0158] [Mathematical formula 5]

[0159] Bo: Cell viability in wells without UV and reagent treatment

[0160] Bt: Cell viability in wells irradiated with UV light but not treated with reagents

[0161] St: Cell viability in wells irradiated with UV light and treated with reagents

[0162] [Table 6] Mitigation rate of cytotoxicity (%) Example 7 25.9 Example 10 28.5 Example 13 32.7 Example 16 30.1 Embodiment 19 35.4 Comparative Example 1 12.8 Comparative Example 2 11.5 Comparative Example 3 14.6 Comparative Example 4 11.1

[0163] As a result, as shown in Table 6, it was confirmed that Examples 7, 10, 13, 16 and 19 had significantly higher cytotoxicity mitigation rates than Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0164] Therefore, the natural eutectic solvent extract according to one embodiment has excellent protective activity against cell damage caused by ultraviolet rays and can be usefully used to prevent or improve aging.

[0165] Experimental Example 7: Confirmation of the inhibitory effect on the activity of the enzymes involved in the induction of

[0166] The inflammation prevention, improvement or treatment effect of the natural eutectic solvent extract according to one embodiment was confirmed. Specifically, 100 μl of hyaluronidase solution (400U / ml) was put into each well and reacted at 37°C for 20 minutes, and then 100 μl of enzyme activation solution (compound 48 / 80CaCl22H2O, Sigma, 0.1 mg / ml) was added to 100 μl of Examples 7, 10, 13, 16, and 19 and Comparative Examples 1 to 4. After further reacting for 20 minutes at 37°C. Then, 250 μl of hyaluronic acid solution (0.4 mg / ml) was put into the reaction at 37°C for 40 minutes, and 100 μl of 0.4N NaOH was put into the reaction to terminate the reaction. After adding 100 μl of potassium borate solution and reacting at 95°C for 3 minutes and cooling, 3 mg of p-dimethylaminobenzaldehyde solution was put into the reaction and further reacted at 37°C for 20 minutes to develop color. Then, the absorbance was measured at 585 nm and the inhibition rate of hyaluronidase activity was measured. The oil-soluble licorice extract was used as a positive control group. The inhibition rate of hyaluronidase activity was calculated according to the following mathematical formula 6, and the results are shown in the following Table 7.

[0167] [Mathematical formula 6]

[0168] A: Enzyme activity of wells without reagent added

[0169] B: Enzyme activity in wells with added reagents

[0170] [Table 7] Inhibition rate of hyaluronidase activity (%) Positive control group 78.6 Example 7 65.2 Example 10 74.1 Example 13 59.2 Example 16 62.7 Embodiment 19 66.8 Comparative Example 1 53.7 Comparative Example 2 43.4 Comparative Example 3 46.7 Comparative Example 4 41.3

[0171] As a result, as shown in Table 7, it was confirmed that the inhibition rates of hyaluronidase activity in Examples 7, 10, 13, 16 and 19 were significantly higher than those in Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0172] Therefore, the natural co-crystal solvent extract according to one embodiment can be usefully used to prevent, improve or treat inflammatory diseases.

[0173] Experimental Example 8: Confirmation of the inhibitory effect on inflammatory cytokines caused by ultraviolet rays

[0174] The natural eutectic solvent extract according to one embodiment was confirmed to have a preventive, ameliorative or therapeutic effect on inflammation caused by ultraviolet rays. Specifically, the eutectic solvent extract was diluted to 3×10 5 3 ml of each keratinocyte cell line was divided into a culture medium supplemented with 10% fetal bovine serum (FBS) and cultured for 24 hours. Then, irradiation with 15 mJ / cm 2 UVB was applied and the Examples 7, 10, 13, 16 and 19 and Comparative Examples 1 to 4 were diluted in a medium containing 2% FBS to a concentration that did not show cytotoxicity, and further cultured for 24 hours. After culture, total RNA was extracted using RNeasy minikit (Qiagen, Hilden), and 1 μl of total RNA was heated at 37°C for 60 minutes using oligo(dT)15 primer, dNTP (0.5 μM), 1 unit of RNase inhibitor and 4 units of Omniscript reverse transcriptase (Qiagen, Hilden, Germany), and then heated at 93°C for 5 minutes to terminate the reaction to synthesize cDNA. PCR was performed to amplify interleukin-1α (IL-1α), tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), and glyceraldehyde phosphate dehydrogenase (GAPDH) from the cDNA. The products generated by PCR were electrophoresed in 2% agarose gel and confirmed by using an image analyzer. The density of each band was analyzed using the Image J program (NIH Image software, Maryland, USA). Madecassoside was used as a positive control group. The inhibition rate of the expression of inflammatory cytokines and inflammatory mediators was calculated according to the following mathematical formula 7, and the results are shown in the following Table 8.

[0175] [Mathematical formula 7]

[0176] Bo: Production of inflammatory cytokines or inflammatory mediators in wells without UV irradiation and reagent treatment

[0177] Bt: The amount of inflammatory cytokines or inflammatory mediators produced in the wells that were irradiated with UV light but not treated with reagents

[0178] St: The amount of inflammatory cytokines or inflammatory mediators produced in the wells irradiated with UV light and treated with reagents

[0179] [Table 8]

[0180] As a result, as shown in Table 8, it was confirmed that Examples 7, 10, 13, 16 and 19 had significantly higher expression inhibition rates of inflammatory cytokines and inflammatory mediators than Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0181] Therefore, the natural co-crystal solvent extract according to one embodiment inhibits the expression of inflammatory cytokines and / or inflammatory mediators, and thus can be usefully used to prevent, improve or treat inflammatory diseases caused by ultraviolet rays.

[0182] Experimental Example 9: Confirmation of the Tyrosinase Activity Inhibitory Effect

[0183] The whitening improvement activity of the natural eutectic solvent extract according to one embodiment has been confirmed. Specifically, L-tyrosine (Sigma) was dissolved in 0.05M sodium phosphate buffer solution (pH6.8) and made into a 0.1mg / ml solution, and 0.5ml of the prepared solution was put into a test tube. To this, 0.5ml of each of the embodiments 7, 10, 13, 16 and 19 and comparative examples 1 to 4, which were adjusted to an appropriate concentration and dissolved in 0.05M sodium phosphate buffer solution (pH6.8), was added and allowed to stand in a 37°C incubator for 10 minutes. Then, 0.5ml of 200 units / ml tyrosinase was added and reacted at 37°C for 10 minutes, and then the reaction was terminated by rapid cooling on ice. The absorbance at a wavelength of 475nm was measured using a spectrophotometer. 0.5ml of buffer was used as a negative control group, and an arbutin solution adjusted to an appropriate concentration and dissolved in 0.05M sodium phosphate buffer solution (pH6.8) was used as a positive control group. The inhibition rate (%) of tyrosinase was calculated according to the following mathematical formula 8, and the results are shown in the following Table 9.

[0184] [Mathematical formula 8]

[0185] A: Absorbance before reaction of the well to which the reagent was added

[0186] B: Post-reaction absorbance of the wells to which the reagents were added

[0187] C: Absorbance before reaction in wells without reagent added

[0188] D: Post-reaction absorbance of wells without reagent added

[0189] [Table 9] Inhibition rate of tyrosinase activity (%) Positive control group 92.4 Example 7 64.1 Example 10 56.8 Example 13 73.1 Example 16 68.9 Embodiment 19 63.3 Comparative Example 1 49.7 Comparative Example 2 51.2 Comparative Example 3 62.7 Comparative Example 4 52.4

[0190] As a result, as shown in Table 9, it was confirmed that Examples 7, 10, 13, 16 and 19 had significantly higher inhibition rates on tyrosinase activity than Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0191] Therefore, the natural eutectic solvent extract according to one embodiment can be usefully used to improve skin whitening.

[0192] Experimental Example 10: Confirmation of the inhibitory effect on melanin synthesis

[0193] The whitening improvement activity of the natural eutectic solvent extract according to one embodiment was confirmed. Specifically, melanoma cells B16F10 (Accession No.: ATCC6323) were cultured at 2×10 6 After the cells were separated and attached to a 6-well plate at a concentration of 10 cells / ml, the Examples 7, 10, 13, 16, and 19 and Comparative Examples 1 to 4 were treated at a concentration that did not cause toxicity and cultured for 72 hours. Then, the cells were separated using trypsin-EDTA, and the number of cells was determined and the cells were recovered by centrifugation. Slightly modified Lotan [Canser Res., 40, 3345-3350 (1980)] was used to quantify intracellular melanin. After washing the cell mass once with phosphate buffered saline (PBS), 1 ml of homogenization buffer (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM phenyl methane sulphonylfluoride (PMSF)) was added and vortexed for 5 minutes to lyse the cells. 1N NaOH 10% dimethyl sulfoxide (DMSO) was added to the cell filtrate obtained by centrifugation (3,000 rpm, 10 minutes) to dissolve the extracted melanin. Then, the absorbance of melanin at 405 nm was measured using an ELISA instrument, and the melanin was quantified to determine the inhibitory rate (%) of the reagent on melanin production. The inhibition rate of melanin production was calculated according to the following mathematical formula 9, and the results are shown in the following Table 10. Arbutin was used as a positive control group.

[0194] [Mathematical formula 9]

[0195] A: Melanin amount in the well without reagent added

[0196] B: Amount of melanin in the wells where the reagent was added

[0197] [Table 10]

[0198] As a result, as shown in Table 10, it was confirmed that Examples 7, 10, 13, 16, and 19 had significantly higher inhibition rates on melanin production than Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0199] Therefore, the natural eutectic solvent extract according to one embodiment can be usefully used to improve skin whitening.

[0200] Experimental Example 11: Confirmation of Skin Barrier Strengthening Effect

[0201] The skin barrier strengthening effect of the natural eutectic solvent extract according to one embodiment was confirmed. Specifically, human keratinocyte HaCaT cells were cultured in DMEM medium (Dulbecco's modified Eagles'Medium, Gibco 11965-092) including 10% fetal bovine serum, and all cultures were performed in a 37°C, 5% CO2 incubator. The cultured cell lines were treated with Examples 7, 10, 13, 16 and 19 and Comparative Examples 1 to 4 and further cultured for 24 hours. Then, total RNA was isolated from the cells of each reagent using RNeasy mini kit (Qiagen, Germany) and RNA was quantified at 260 nm using nanodrop, and cDNA was synthesized in an amplifier using 2 μg of RNA respectively (C1000 Thermal Cycler, Bio-Rad, USA). The synthesized cDNA was used as a template to add the target genes, i.e., SPT and AQP3, factors related to skin barrier strengthening and moisturizing, and primers and cDNA of antimicrobial peptide RNase7 to perform real-time polymerase chain reaction in a real-time PCR machine (Mic qPCR, biomlecular system, Australia). The gene expression level was finally analyzed by modifying the GAPDH gene, and the results are shown in the following Table 11.

[0202] [Table 11]

[0203] As a result, it was confirmed that, as shown in Table 11, the expressions of ceramide synthase SPT, AQP related to skin moisturizing and barrier strengthening, and RNase7 of skin antimicrobial peptide were significantly higher in Examples 7, 10, 13, 16 and 19 than in Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0204] Therefore, the natural eutectic solvent extract according to one embodiment can be usefully used for skin barrier strengthening.

[0205] Experimental Example 12: Confirmation of the Effects on Skin Wrinkles and Moisturizing

[0206] The natural eutectic solvent extract according to one embodiment was confirmed to have skin wrinkle and moisturizing improving activities. Specifically, 10% fetal bovine serum (FBS) was added to DMEM / F-12 (3:1) medium and 1×10 4 Cells / cm 2 Concentration cultured human dermal fibroblasts (Modern Tissue Technology, South Korea). After growing 70 to 80%, the strains were divided and subcultured at a ratio of 1:3, and the cells subcultured 3 to 4 times were used in the experiment. After the fibroblasts were cultured in a 48-well plate for at least 90%, the Examples 7, 10, 13, 16 and 19 and Comparative Examples 1 to 4 were applied at a concentration that did not show cytotoxicity, and after 24 hours, the amount of free procollagen in the culture medium was determined using the procollagen type-1C-peptide EIA kit (MK101, Takara, Japan). Ascorbic acid was used as a positive control group.

[0207] [Table 12] Type I procollagen production rate (%) Positive control group 65.4 Example 7 43.5 Example 10 35.3 Example 13 63.1 Example 16 41.8 Embodiment 19 56.8 Comparative Example 1 25.4 Comparative Example 2 21.5 Comparative Example 3 43.6 Comparative Example 4 24.7

[0208] As a result, as shown in Table 12, it was confirmed that the procollagen production rates of Examples 7, 10, 13, 16 and 19 were significantly higher than those of Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0209] Therefore, the natural eutectic solvent extract according to one embodiment can be usefully used for the prevention or improvement of skin wrinkles and moisturizing.

[0210] Example 13: Confirmation of the effect of improving acne and sebum or preventing or improving hair loss

[0211] The natural eutectic solvent extract according to one embodiment was confirmed to have a preventive or ameliorative effect on acne and sebum, or hair loss. Specifically, 3H (tritiume) was added to each microplate hole. 0.1 μCi of radiolabeled testosterone and the above-mentioned Examples 7, 10, 13, 16 and 19 and Comparative Examples 1 to 4, 10,000 fibroblasts were then inoculated and cultured. After 24 hours of culture, the supernatant was obtained and ethyl acetate-cyclohexane was used to (1:1) 1 ml of extraction solvent was used to obtain steroids. The steroids were placed on a thin film chromatography plate and developed using a chloroform / methanol mixture (98 / 2 (v / v)). The radioactivity of the points corresponding to testosterone and dihydrotestosterone (DHT) was measured using a densitometer to calculate the conversion rate, and the results were compared with the negative control group. The 5α -Reductase inhibition rate.

[0212] [Formula 10]

[0213] A: Conversion rate from testosterone to dihydrotestosterone without adding reagents

[0214] B: Conversion rate from testosterone to dihydrotestosterone when adding reagent

[0215] [Table 13] 5α-reductase inhibition rate (%) Example 7 57.1 Example 10 61.3 Example 13 30.2 Example 16 51.4 Embodiment 19 55.3 Comparative Example 1 38.5 Comparative Example 2 46.7 Comparative Example 3 28.6 Comparative Example 4 34.8

[0216] As a result, as shown in Table 13, it was confirmed that Examples 7, 10, 13, 16 and 19 had significantly higher 5α-reductase inhibition rates than Comparative Examples 1 to 4 in which the same extraction object was extracted using an organic solvent.

[0217] Therefore, the natural eutectic solvent extract according to one embodiment inhibits the conversion of dihydrotestosterone caused by 5α-reductase, and can be used not only to prevent or improve acne or sebum, but also to prevent, improve or treat hair loss, or promote hair growth or development.

[0218] Example 14: Confirmation of skin elasticity improvement effect

[0219] The skin elasticity improving effect of the cosmetic composition including the natural eutectic solvent extract according to one embodiment was confirmed. First, a cosmetic composition in a cream dosage form including the ingredients and contents of Table 14 below was prepared. Specifically, Phase B of Table 14 below was heated and stored at 70°C, and Phase A was added for pre-emulsification. Then, after emulsification by homogenization with a high-speed stirrer, the cosmetic composition was slowly cooled to prepare. In this specification, unless otherwise specified, the content is weight %. Twenty women aged 20 to 35 years old used Formulations 1 to 6 and Comparative Formulations 1 to 4 twice a day for two months.

[0220] Then, the skin elasticity was measured using a cutometer (SEM575, C+K Electronic Co., Germany). The skin elasticity before and after use of the formulation examples 1 and 5 and the comparative formulation 1 was measured. The measured values ​​were recorded as R7 values ​​of Cutometer SEM575, and the R7 value represents the viscoelasticity of the skin.

[0221] [Table 14]

[0222] [Table 15] Skin viscoelasticity Dosage Form Example 1 0.33 Dosage Form Example 5 0.31 Comparative dosage form example 1 0.11

[0223] As a result, it was confirmed that the skin viscoelasticity in Formulation Examples 1 and 5 was significantly increased compared to Comparative Formulation Example 1 as shown in Table 15.

[0224] Therefore, the natural product extract of the natural eutectic solvent according to one embodiment can be usefully used for improving skin elasticity.

[0225] Experimental Example 15: Confirmation of skin wrinkle improvement effect

[0226] The skin wrinkle improvement effect of the cosmetic composition including the natural eutectic solvent extract according to one embodiment was confirmed. Specifically, the same subjects were made to use the formulation examples 1 and 5 of Example 14 and the comparative formulation example 1 in the same way. Then, after making a replica made of silicon, an image analyzer visiometer (SV60, C+ Electronic Co., registered patent 10-1905517-13-Germany) was used. The skin wrinkle status after using the formulation examples 1 and 5 and the comparative formulation example 1 was measured. The measured value is expressed as the average of the parameter value before use subtracted from the parameter value after two months of use. If the measured value is a negative number, it means that the wrinkle improvement effect is high.

[0227] [Table 16]

[0228] R1: The value of the difference between the highest and lowest values ​​of the wrinkle contour

[0229] R2: The average of the R1 values ​​after the wrinkle contour is randomly divided into 5 grids

[0230] R3: The highest value among the R1 values ​​divided into 5 grids

[0231] R4: The average value obtained by subtracting the peak and valley values ​​from the baseline of the wrinkle contour

[0232] R5: The average value of the value obtained by subtracting each wrinkle contour from the baseline of the wrinkle contour

[0233] As a result, it was confirmed that, as shown in Table 16, the parameter values ​​showing the skin wrinkle state in Formulation Examples 1 and 5 were significantly lower than that in Comparative Formulation Example 1.

[0234] Therefore, the natural product extract of the natural eutectic solvent according to one embodiment can be usefully used for improving skin wrinkles.

[0235] Experimental Example 16: Confirmation of Skin Whitening Improvement Effect

[0236] The skin whitening improvement effect of the cosmetic composition including the natural eutectic solvent extract according to one embodiment was confirmed. Specifically, the same subjects were made to use the formulation examples 1 and 5 of Example 14 and the comparative formulation example 1 in the same way. Then, an image analyzer and a spectrophotometer (Minolta CR300) were used. The skin color brightness change (L) of the applied part was measured. In addition, objective naked eye observation was performed by a plurality of skilled personnel and subjective naked eye observation was performed by the test subjects, and the degree of whitening efficacy was evaluated according to the following scale.

[0237] -3: Extremely worse -2: Worsened -1: Slightly worsened 0: No change 1: Slightly improved 2: Improved 3: Extremely improved

[0238] [Table 17]

[0239] As a result, it was confirmed that the skin color lightness change in Formulations 1 and 5 significantly increased compared to Comparative Formulation 1 as shown in Table 17. Specifically, it was confirmed that the skin color lightness change in Formulation 5 in which the acidity of the natural eutectic solvent was adjusted to pH 5 significantly increased.

[0240] Therefore, the natural product extract of the natural eutectic solvent according to one embodiment can be usefully used for skin whitening improvement.

[0242] The descriptions recorded in the present invention are intended to provide examples, and those skilled in the art should be able to understand that the present invention can be modified into other specific forms without changing the technical ideas or essential features of the present invention. Therefore, the embodiments described above should be understood as exemplary rather than restrictive.

Claims

1. A cosmetic composition for improving skin condition, comprising a natural extract extracted using a natural eutectic solvent as an effective ingredient, wherein: The natural eutectic solvent includes amino acids or their derivatives, and organic acids.

2. The composition according to claim 1, wherein The natural products are selected from the group consisting of fig, Selaginella, black rice, dragon's blood, black bean, trumpet creeper, snow lotus, schoenensis, pollen, azolla, fragrant wood, wellhead grass, dicentrata, parasitic wood, creeping grass, glandular tooth blueberry, mulberry tinctoria, tomato, houttuynia cordata, tangerine peel, peony, chestnut flower, kudzu vine, and walnut.

3. The composition according to claim 1, wherein The pH of the natural eutectic solvent is pH 2.5 to pH 8.

4. The composition according to claim 1, wherein The amino acid or its derivative is selected from the group consisting of betaine, alanine, cysteine, aspartic acid, glutamine, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, arginine, serine, threonine, valine, tryptophan, and tyrosine.

5. The composition according to claim 1, wherein The organic acid is selected from the group consisting of citric acid, lactic acid, malic acid, maleic acid, pyruvic acid, fumaric acid, succinic acid, acetic acid, aconitic acid, tartaric acid, ascorbic acid, malonic acid, oxalic acid, glucuronic acid, neuraminic acid, sialic acid, shikimic acid, phytic acid, galacturonic acid, iduronic acid, hyaluronic acid, hydroxycitric acid, and lactone derivatives.

6. The composition according to claim 1, wherein The amino acid or its derivative and the organic acid are mixed in a weight ratio of 1:0.001 to 10.

7. The composition according to claim 1, wherein The skin condition is skin aging, moisturizing, wrinkles, elasticity, skin barrier strengthening, acne or sebum secretion.

8. The composition according to claim 7, wherein The skin aging is caused by ultraviolet rays or active oxygen.

9. A skin external preparation composition for improving skin condition, comprising a natural extract extracted using a natural eutectic solvent as an active ingredient, wherein: The natural eutectic solvent is amino acid or its derivatives, and organic acid.

10. A pharmaceutical composition for preventing or treating skin inflammatory diseases, comprising a natural extract extracted using a natural eutectic solvent as an effective ingredient, wherein: The natural eutectic solvent is an amino acid or its derivatives, and an organic acid.

11. The composition according to claim 8, wherein The inflammatory disease is selected from the group consisting of skin wounds, dermatitis, atopic dermatitis, pruritus, eczematous skin diseases, dry eczema, erythema, urticaria, psoriasis, drug eruption, and acne.

12. A pharmaceutical composition for preventing or treating hair loss, or promoting hair growth or development, comprising a natural extract extracted using a natural eutectic solvent as an active ingredient, wherein: The natural eutectic solvent is amino acid or its derivatives, and organic acid.

13. A method for preparing a natural eutectic solvent, comprising: preparing a mixture by mixing an amino acid or a derivative thereof, and an organic acid; A pH adjuster selected from the group consisting of sodium citrate, sodium hydroxide, potassium hydroxide, sodium bicarbonate, arginine, tromethamine, triethanolamine, aminomethyl propanol, dipotassium phosphate, potassium phosphate, sodium anisate, sodium lactate, sodium carbonate, and urea is added to the mixture.

14. Use of a natural product extract using a natural eutectic solvent extract in preparing a cosmetic composition for improving skin condition, wherein: The natural eutectic solvent includes amino acids or their derivatives, and organic acids.

15. Use of a natural product extract extracted using a natural eutectic solvent in the preparation of a skin external preparation composition for improving skin condition, wherein: The natural eutectic solvent includes amino acids or their derivatives, and organic acids.

16. Use of a natural extract extracted using a natural eutectic solvent in the preparation of a pharmaceutical composition for preventing or treating skin inflammatory diseases, wherein: The natural eutectic solvent includes amino acids or their derivatives, and organic acids.

17. Use of a natural extract extracted using a natural eutectic solvent in the preparation of a pharmaceutical composition for preventing or treating hair loss, or promoting hair growth or development, wherein: The natural eutectic solvent includes amino acids or their derivatives, and organic acids.

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