WO2019143055A1 - Composition, comprising extract of fermented beans obtained from fermentation by aspergillus fumigatus strain, for skin care improvement - Google Patents

Abstract

The present invention relates to a composition for skin care improvement. A composition for skin care improvement according to an embodiment shows excellent effects in skin moisturization, skin barrier enhancement, and skin regeneration, and causes no irritation to skin, by comprising: an extract of fermented beans obtained from fermentation by an Aspergillus fumigatus strain; a gum; and glyceryl glucoside, and thus the composition can be safely used.

Description

 A composition for improving skin beauty comprising an extract of fermented soybean fermented with Aspergillus cristatus strain

And a composition for improving skin beauty.

The epidermis, which is the outermost layer of the skin, is an organ that is always in contact with the external environment and serves as a protective barrier. The skin barrier, which exists in the stratum corneum of the epidermis, suppresses the loss of water and electrolytes, protects the human body from external physical damage and chemicals, and prevents bacteria and viruses from penetrating into the skin. The stratum corneum is composed of dead keratinocyte protein and lipid existing between keratinocyte. It is known that lipid bilayer between keratinocyte plays the most important role in maintaining skin barrier.

Aspergillus cristatus ) is a microorganism dominated by Fuzhuan brick tea, a post fermented tea in China, and it is called a golden flower fungus because the Chinese form spores like the gold flower. . A. cristatus has been used for a long time in the production of brick tea as a safe strain because it does not produce carcinogenic mycotoxins. In addition, Yongyi Ge et al. (2016) reported that the presence and expression of mycotoxin gene was not observed in A. cristatus isolated from the blastocyst .

Soybean is one of cosmetic raw materials, and it helps skin to be enriched and helps to improve blood circulation, and it gives the skin vitality. In one room, it has the effect of enhancing the immunity of the human body, and it is known that the blood pressure is lowered, blood is removed, blood is removed from the blood, and prevention and improvement of tumor are prevented. Therefore, beans have been used as raw materials for cosmetics.

However, there is no study on fermented soybeans fermented using A. cristatus .

One aspect is that the beans are produced by Aspergillus The present invention provides a composition for improving skin beauty, comprising an extract of fermented soybean fermented with a cristatus strain, gum, and glyceryl glucoside as an active ingredient.

Another aspect is that the beans are produced by Aspergillus the present invention provides a method for improving skin conditions comprising administering to an individual an effective amount of a composition comprising an extract of fermented soybean fermented with a cristatus strain, gum, and glyceryl glucoside.

Another aspect provides the use of a composition comprising an extract of a fermented soybean, a gum, and a glyceryl glucoside, wherein the soybean is fermented with a strain of Aspergillus cristatus , for the production of a skin condition improving agent.

One aspect is that the beans are produced by Aspergillus The present invention provides a composition for improving skin beauty, comprising an extract of fermented soybean fermented with a cristatus strain, gum, and glyceryl glucoside as an active ingredient.

The skin cosmetic improvement or skin condition improvement may be skin moisturizing, barrier strengthening, or skin regeneration. In one embodiment, a composition comprising fermented soybean extract, gum, and glyceryl glucoside, wherein the soybean has been fermented with the A. cristatus strain, has a skin moisturizing, barrier strengthening, or skin regenerating effect . Therefore, the composition containing the fermented soybean extract, gum, and glyceryl glucoside can be used as a composition for skin moisturizing, barrier strengthening, or skin regeneration.

The term "skin moisturizing" may mean any action that maintains skin moisture or prevents moisture loss.

The term "skin barrier enhancement" may refer to any action that is located at the outermost position of the skin, thereby enhancing the function of the skin barrier to prevent moisture and nutritional loss.

The term "skin regeneration" may mean any action that, when a portion of the skin is lost, replenishes that part or promotes the proliferation of skin cells.

The term "improvement" may mean any action that at least reduces the degree of symptomatology, such as a condition associated with relief or treatment of a condition.

The term " Aspergillus & cristatus "is a kind of yeast fungus that forms spores like the golden flower and is also called golden flower fungus. The above A. cristatus can be separated from food, soil, And may be, for example, isolated from Fuzhuan brick tea, a post fermented tea of China, for example.

In a composition according to one embodiment, the strain may be an Aspergillus cristatus Cosmax-GF strain (accession number: KCCM11820P). The Aspergillus cristatus Cosmax-GF strain is a novel strain isolated from a zygotic body manufactured in China, specifically, diluting a zygotic body with a NaCl solution; Culturing the diluted zygotes in potato agar medium supplemented with chloramphenicol; And a step of selecting strains which form gold colonies in the medium. The NaCl solution may contain from 0.50% w / w to 0.99% w / w, from 0.60% w / w to 0.95% w / w, from 0.70% w / w to 0.90% w / ), 0.80% (w / w) to 0.90 (w / w), or 0.84% (w / w) to 0.86% (w / w), for example 0.85% (w / w) NaCl solution. The culture may be performed at a temperature of 20 to 40 캜, 25 to 40 캜, 25 to 35 캜, 27 to 35 캜, or 28 to 32 캜. The culture may be performed for 10 to 100 hours, 20 to 90 hours, 30 to 80 hours, 40 to 70 hours, 48 to 65 hours, or 48 to 60 hours.

The term "soybean" is a plant of dicotyledonous roots and legumes, which is also called "soybeans ". The soybean can be used without limitation, and for example, various varieties such as black bean, black bean, brown bean, stain bean, and black bean can be used, but the present invention is not limited thereto. The soybeans may be those obtained by a conventionally available method, for example, those sold in the market. The soybean may be sterilized soybean, and the sterilization method may be carried out by a conventional method in the art.

In a composition according to one embodiment, the fermentation may be a solid fermentation. The term "solid fermentation" may mean fermenting microorganisms in a solid state raw material having low water content. The solid fermentation method may be advantageous in that it can be easily recovered without contamination of germs compared with the conventional liquid fermentation method or semi-solid fermentation method.

In the composition according to one embodiment, the fermented soybeans are soybean Aspergillus way may be one which loose Christa tooth (A. cristatus), specifically Aspergillus Christa tooth (A. cristatus) inoculated with the spore suspension on the solid fermentation .

In a composition according to one embodiment, the culture substrate can be converted to a functional material by extracellular enzyme of A. cristatus through the fermentation. Examples of the extracellular enzymes include, but are not limited to, alkaline phosphatase, esterase (C4), esterase lipase (C8), acid phosphatase, Beta-glucosaminidase, alpha-galactosidase, beta-galactosidase, beta-glucosidase, N-acetyl-beta-glucosaminidase ), Alpha-mannosidase, and the like, but are not limited thereto. Therefore, it is important to maintain the phase of conversion from mycelium to spore, which is the highest point of biosynthesis of the enzyme in the life cycle of Aspergillus cristatus . Therefore, in order to maximize the expression phase of extracellular enzyme of A. cristatus to maximize the non-glycosylated component of the soybean, the fermentation temperature, time, and humidity may be suitably selected .

The fermented soybeans may be fermented at 25 to 40 캜, 25 to 35 캜, 28 to 35 캜, 30 to 35 캜, 31 to 34 캜, 32 to 34 캜 or 33 캜. When the fermentation temperature is higher than 35 ° C, the time required for the strain to form spores is increased, so that the enzyme activity and expression amount can be lowered. On the contrary, when the temperature is lower than 30 ° C, the enzyme reaction rate is slowed, Can be lowered.

The fermented soybeans may be cultured for 10 to 100 hours, 20 to 100 hours, 20 to 90 hours, 30 to 90 hours, 30 to 80 hours, 40 to 80 hours, 40 to 70 hours, 48 to 65 hours, To 65 hours, or 60 to 65 hours. The fermentation time can be appropriately selected in accordance with the fermentation temperature so as to terminate until spores are formed in consideration of the growth of hyphae.

Wherein the fermented soybeans have a moisture content of 10 to 99.9%, a moisture content of 20 to 99.9%, a moisture content of 30 to 99.9%, a moisture content of 40 to 99.9%, a moisture content of 50 to 99.9%, a moisture content of 50 to 90%, a moisture content of 60 to 90% Humidity, 75-85% humidity, or 80-85% humidity conditions. When the humidity is less than 75%, the time required for the strain to form into the spore is increased, and the activity and expression amount of the enzyme can be lowered. On the contrary, when the humidity is higher than 85%, the moisture content is high, The growth of aerobic strain A. cristatus may be slowed and the incidence of contamination by other microorganisms may be increased.

For example, the fermented soybeans may be fermented at 30 to 35 DEG C for 48 to 65 hours at 75 to 85% humidity.

The fermented soybeans may be the entire fermented soybean, a part thereof, or a material derived therefrom. The fermented soybeans used for the extraction may be the whole, a part thereof, or a material derived therefrom, which may be pulverized or cut or suitably dried. The fermented soybeans may be dried to have a moisture content of less than 20%, or less than 15%, for example less than 12%.

The extract may be extracted with a hydrophilic solvent, for example, alcohol, water, or a combination thereof. The alcohol may be a compound having at least one-OH group of C1 to C10. The alcohol may be a C1 to C6 alcohol, or a C3 to C6 polyhydric alcohol. The alcohol may be methanol, ethanol, n-propanol, isopropanol, n-butanol, sec-butanol, isobutanol, tert-butanol, n-pentanol, n-hexanol or mixtures thereof. The solvent may be, for example, a mixture of water and an alcohol, that is, an aqueous solution of an alcohol. The alcohol concentration of the alcohol aqueous solution may be 1 to 100% (w / w), such as 1 to 99.5% (w / w), 10 to 100% (w / w), 20 to 100% (w / (W / w), 40 to 100% (w / w), 50 to 100% (w / w), 60 to 100% (W / w), 60 to 90% (w / w), 60 to 80% (w / w), 65 to 75% have. The aqueous alcohol solution may be methanol, ethanol, or an aqueous butanol solution. In a composition according to one embodiment, the extract may be an ethanol extract.

The extract may be extracted by a conventional method in the art such as warm extraction, pressure extraction, ultrasonic extraction, hot water extraction, reflux cooling extraction, subcritical extraction, or supercritical extraction.

The extraction solvent may be selected from the group consisting of 5 to 15 (volume / weight) times, such as 5 to 13 (volume / weight), 5 to 11 (volume / , 8 to 13 (volume / weight) times, 8 to 11 (volume / weight) times, or 9 to 11 (volume / weight) times. For example, 0.5 to 1.5 L of the extraction solvent may be added to 100 g of the whole fermented soybean, a part thereof, or a material derived therefrom.

The extraction may be carried out at a temperature of 4 ° C to 70 ° C, for example 4 ° C to 50 ° C, 4 ° C to 40 ° C, 4 ° C to 30 ° C, 10 ° C to 70 ° C, 15 ° C to 70 ° C, 10 C to 50 C, 10 C to 50 C, 4 to 40 C, 4 to 30 C, 10 to 40 C, 10 to 35 C, or 10 to 30 C or room temperature .

The extraction time can be appropriately selected according to the selected temperature and extraction method. For example, the extraction time may be from 1 hour to 3 days, from 1 hour to 2 days, from 1 hour to 1 day, from 5 hours to 3 days, from 5 hours to 2 days, from 5 hours to 1 day, from 10 hours to 3 days, from 15 Hour to 2 days, 15 hours to 36 hours, 18 hours to 30 hours, 1 day to 3 days, 1 day to 2 days, or 24 hours.

For example, the fermented soybean powder can be extracted by ultrasonic extraction by immersing the fermented soybean powder in the extraction solvent for 2 to 4 hours, or by immersing in a solvent for 24 to 72 hours at room temperature. The extraction may be one or more times, for example, one to five times, one to four times, one to three times, two to five times, or two to four times, each extraction being performed in the same way, It can be performed in other ways.

The above extraction can be performed by separating the plant residue and the extract by a known method such as filtration. The extraction can also include removing the solvent from the resulting extract by known methods such as concentration under reduced pressure. The extraction may also comprise preparing the dried extract by drying, such as lyophilization, of the resulting extract.

The extract may contain 1 ppm to 10000 ppm, 1 ppm to 1000 ppm, 1 ppm to 500 ppm, 10 ppm to 10000 ppm, 10 ppm to 1000 ppm, 10 ppm to 500 ppm, 50 ppm to 10000 ppm, 50 ppm to 1000 ppm , 50 ppm to 500 ppm, or 50 ppm to 150 ppm.

The extract may contain from 0.001% w / v to 80% w / v, such as 0.001% w / v to 50% w / v, 0.001% w / w / v), 0.001% (w / v) to 5% (w / v), 0.001% (w / v) to 1% (w / v) v), 0.01% (w / v) to 60% (w / v), 0.01% (w / v) to 40% (w / v), 0.01% , 0.01% (w / v) to 20% (w / v), 0.01% (w / v) to 10% (w / v), 0.01% May be included at a concentration of 0.01% (w / v) to 1% (w / v).

The composition according to one embodiment of the present invention may contain, as an active ingredient, a fraction of the fermented soybean extract instead of the fermented soybean extract. The composition according to one embodiment may further comprise a fraction of the extract of the fermented soybean. The term "fraction" refers to a substance in which the extract of the fermented soybean is divided into its components, that is, a fractionated substance. The fraction may be obtained by solvent fractionation. The solvent fractionation may be a step of mixing the fermented soybean extract with a solvent and separating the substance present in the solvent. The fraction may be a methylene chloride fraction, an ethyl acetate fraction, a butanol fraction, or a water fraction obtained by suspending the fermented soybean extract in water and sequentially fractionating the fraction with methylene chloride, ethyl acetate, butanol, and water.

Specifically, the methylene chloride fraction is prepared by mixing the fermented soybean extract with water, mixing the mixture with methylene chloride, allowing the mixture to stand for a predetermined time, separating the methylene chloride layer, separating the separated methylene chloride layer , ≪ / RTI > Separation of the fractions may include removing methylene chloride from the methylene chloride layer. Further, the ethyl acetate fraction was obtained by mixing the remaining water after the methylene chloride fraction was separated with ethyl acetate, leaving it for a predetermined time, separating the ethyl acetate layer, and separating the fraction from the separated ethyl acetate layer Lt; / RTI > Separation of the fractions can include removal of the ethyl acetate from the ethyl acetate layer. In addition, the butanol fraction may be obtained by mixing water remaining after the ethyl acetate fraction is separated again with butanol, leaving the butanol layer separated for a predetermined time, and separating the fraction from the separated butanol layer. Separation of the fractions may include removing butanol from the butanol layer. Conditions for such fractionation, such as temperature conditions, pressure conditions, time, amount or concentration of solvent used, stirring, etc., may be as described for the extracts used to prepare the above fermented soybean extract. The fractionation may be repeated one or more times, for example, 1 to 5 times.

Separation of the fractions may be accomplished by known methods such as filtration. The fractionation may also include removing the solvent from the fraction obtained by known methods such as concentration under reduced pressure. The fractionation may also include concentration and / or drying of the obtained fractions. The concentration may be reduced pressure concentrated. The drying may include vacuum drying, boiling drying, spray drying, room temperature drying or freeze drying.

The gum may be a polymer polysaccharide having elasticity, and the kind thereof is not limited. In one embodiment, it may be at least one selected from the group consisting of black locust bean gum, gum arabic, ghatti gum, karaya gum, guar gum, tragacanth gum, xanthan gum, carrageenan, mannan and glucomannan. In another embodiment, the black locust bean gum may be used.

1 ppm to 1000 ppm, 1 ppm to 1000 ppm, 10 ppm to 10000 ppm, 10 ppm to 5000 ppm, 10 ppm to 1000 ppm, 50 ppm to 10000 ppm, 50 ppm to 5000 ppm, 50 ppm to 1000 ppm, 100 ppm to 10000 ppm, 100 ppm to 5000 ppm, 100 ppm to 1000 ppm, 200 ppm to 5000 ppm, 200 ppm to 1000 ppm, 300 ppm to 5000 ppm, 300 ppm to 1000 ppm, 400 ppm To 5000 ppm, 400 ppm to 1000 ppm, or 400 ppm to 600 ppm.

(W / v) to 80% (w / v), such as 0.001% (w / v) to 50% (w / v), 0.001% / v), 0.001% (w / v) to 5% (w / v), 0.001% (w / v) to 1% (w / v), 0.01% ), 0.01% (w / v) to 60% (w / v), 0.01% (w / v) to 40% (w / v), 0.01% 0.01% (w / v) to 10% (w / v), 0.01% (w / v) to 5% (w / v), or 0.01% % (w / v) to 1% (w / v).

The above-mentioned "Glyceryl glucoside (2-GG)" may be used interchangeably with "Gluco-glycerol" and may mean a substance in which glycerol and glucose are combined.

Wherein the glyceryl glucoside is present in an amount ranging from 0.1 ppm to 1000 ppm, 0.1 ppm to 500 ppm, 0.1 ppm to 100 ppm, 0.1 ppm to 50 ppm, 1 ppm to 1000 ppm, 1 ppm to 500 ppm, 1 ppm to 100 ppm, ppm to 50 ppm, 5 ppm to 1000 ppm, 5 ppm to 500 ppm, 5 ppm to 100 ppm, 5 ppm to 50 ppm, or 5 ppm to 15 ppm.

The glyceryl glucoside may be present in an amount of from 0.0001% (w / v) to 10% (w / v), such as 0.0001% (w / v) to 5% (w / v) (w / v), 0.005% (w / v) to 10% (w / v), 0.005% (w / v) to 5% w / v), 0.001% (w / v) to 10% (w / v), 0.001% (w / v) v), 0.001% (w / v) to 0.1% (w / v), or 0.001% (w / v) to 0.01% (w / v).

The composition according to one embodiment may be one that increases the expression of HAS3, or AQP3, and more specifically, that increases the expression of HAS3, or AQP3, to exhibit skin moisturizing effects.

The composition according to one embodiment may be one which improves the skin barrier and enhances the skin barrier.

The composition according to one embodiment may be one which promotes proliferation of fibroblasts, specifically, one that exhibits a skin regeneration effect by promoting proliferation of fibroblasts.

The composition according to one embodiment of the present invention may exhibit skin moisturizing effect by maintaining skin moisture content for a long period of time.

The composition according to one embodiment may comprise the extract as an effective amount or as an active ingredient. The effective amount may be appropriately selected depending on the individual. The severity of the disease or condition, the age, weight, health, sex, sensitivity of the individual to the extract, time of administration, route and rate of excretion, duration of administration, factors including other compositions combined or co- May be determined according to factors well known in the art, physiology or medical field.

The composition for improving skin care according to one embodiment may further comprise a cosmetically, pharmaceutically or pharmaceutically acceptable excipient or carrier. 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 hydroxy cellulose, or a combination thereof. The disintegrant may be sodium starch glycolate, calcium monohydrogen phosphate anhydrous, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or combinations thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The composition may be formulated into a parenteral dosage form. The parenteral dosage form may be an injection or an external preparation for skin. The external skin preparation may be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal patch, a drug-containing bandage, a lotion, or a combination thereof.

The external preparation for skin is usually used as a component used in external skin preparations such as cosmetics or medicines such as an aqueous component, an oily component, a powder component, an alcohol, a moisturizer, a thickener, an ultraviolet absorber, a whitening agent, an antiseptic, , Coloring agents, various skin nutrients, and the like can be appropriately blended as needed.

The external preparation for skin may be a metal blocker such as sodium edetate, sodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate or gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridine, Vitamin C, ascorbic acid magnesium phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, fructose, fructose and other herbal medicines, various herbal medicines, tocopherol acetate, glycyrrhizic acid, Sugars such as trehalose and the like can also be appropriately compounded.

The composition according to one embodiment may be a cosmetic composition. In this case, the extract can be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutrition cream, a moisturizer cream, a hand cream, a foundation, , A soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion, a body cleanser, a suspension, a gel, a powder, a paste, a mask pack or a sheet or an aerosol composition. Compositions of such formulations may be prepared according to methods conventional in the art. The cosmetic composition may further contain preservatives, stabilizers, surfactants, solubilizers, moisturizers, emollients, ultraviolet absorbers, preservatives, bactericides, antioxidants, pH regulators, organic and inorganic pigments, fragrances, have. The amount of the additional component such as the above-mentioned moisturizing agent and the like can be easily selected by those skilled in the art within a range not to impair the purpose and effect of the present invention. The amount of the additional component is 0.001 to 5% by weight, % ≪ / RTI >

The composition according to one embodiment may be a food composition. In this case, it can be formulated into a conventional health functional food formulation known in the art. The food composition may be prepared by conventional formulations such as powders, granules, tablets, pills, capsules, suspensions, emulsions, syrups, And may be manufactured in the form of any health food such as confectionery, pizza, ramen, other noodles, gums, jellies, dairy products including ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamin complexes. A pharmaceutically acceptable carrier or excipient may be used for the formulation of the health food, and any carrier or additive known to be usable in the art for the preparation of the formulation to be produced may be used. As the above-mentioned additives, there can be used various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, Carbonating agents, and the like. In addition, it may contain natural fruit juice, fruit juice beverage and flesh for the production of vegetable beverages. These additive components may be used independently or in combination, and the proportion of the additive may be 0.001 to 5 wt%, specifically 0.01 to 3 wt%, based on the total weight of the composition.

The content of the extract in the food composition may be suitably determined according to the intended use (prevention or improvement). Generally, it may contain 0.01 to 15% by weight of the total food weight, and when it is prepared as a beverage, it may contain 0.02 to 10 g, specifically 0.3 to 1 g, based on 100 mL.

The beverage may further contain ingredients other than the above extract, and may further contain various flavors or natural carbohydrates commonly used in beverages. Examples of the natural carbohydrate include conventional sugars such as monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugars such as xylitol, sorbitol, erythritol, Of sugar alcohols may be contained. In addition, as a flavoring agent, a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.) may be contained. The ratio of the natural carbohydrate may be generally about 1 to 20 g, specifically about 5 to 12 g per 100 mL of beverage.

The composition according to one embodiment may be a pharmaceutical composition. Specifically, the pharmaceutical composition may be a pharmaceutical composition for preventing or treating a skin disease. The skin disease may be a disease caused by skin barrier function impairment, skin aging, skin wounds, skin scars, or skin inflammation. The term "prevention" includes inhibiting the occurrence of a disease. The term "treatment" includes inhibiting, alleviating, or eliminating the development of the disease.

Damage to the skin barrier function may mean all the changes that appear on the skin due to the degradation or damage of the skin barrier function. For example, it may include, but is not limited to, increased wrinkles, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne and the like. The pharmaceutical composition according to one embodiment of the present invention can prevent or treat damage to the skin barrier function by improving the skin barrier function by improving the skin layer of the skin.

The aging of the skin can mean all types and intangible changes that appear on the skin as the age goes on. For example, it may include, but is not limited to, increased skin wrinkles, drying, decreased wound healing ability, decreased skin immune function, and the like. The pharmaceutical composition according to one embodiment of the present invention can prevent or treat skin aging by the effects of strengthening skin barrier, maintaining skin moisture, preventing loss of skin moisture, and regenerating skin.

The skin wound or skin scar can mean any disease that can be improved or remedied by maintaining the continuity of skin cells regenerated, differentiated, proliferating, interfering with the synthesis of scarring collagen and promoting collagen degradation. But are not limited to, scars such as bruises, abrasions, lacerations, scars such as hypertrophic scars, keloid scars, and the like. The pharmaceutical composition according to one embodiment can prevent or treat skin wounds or skin scars due to the effects of skin moisture retention, prevention of skin moisture loss, and skin regeneration.

The skin inflammatory disease may mean all diseases caused by immune stimulation. But are not limited to, for example, atopic dermatitis, eczema, seborrheic dermatitis, psoriasis, acne, and the like. The pharmaceutical composition according to one embodiment of the present invention can prevent or treat skin inflammatory diseases by the effects of maintaining skin moisture, preventing loss of skin moisture, strengthening skin barrier, and regenerating skin.

Compositions according to one embodiment may be prepared according to conventional methods in the cosmetics, food, or pharmaceutical arts.

Another aspect provides a method of improving skin beauty of an individual comprising administering the composition to a subject. The composition is the same as described above.

Specifically, the method may be a method of efficiently maintaining skin moisture or preventing moisture loss of an individual, a method of enhancing a skin barrier function of an individual, or a method of improving or promoting skin regeneration of an individual.

The term " administering ", " introducing ", and "transplanting" are used interchangeably and refer to the administration of a composition according to one embodiment to a subject by at least partial localization of the composition to a desired site, May refer to the placement of the composition according to embodiments. May be administered by any suitable route of delivery of at least a portion of the extract or extract components of the composition according to one embodiment to the desired location in the living individual.

Administration can be by any method known in the art. Administration may be by any means directly administered to a subject, such as by intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration, . The administration can be systemically or locally administered.

The subject may be a mammal, such as a person, a cow, a horse, a pig, a dog, a sheep, a goat, or a cat. The subject may be an individual in need of skin aesthetic improvement, for example skin moisturizing, barrier enhancement, or skin regeneration effect.

The administration may comprise a combination of fermented soybean extract, gum, and glyceryl glucoside in an amount of 0.1 mg to 1,000 mg, such as 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg To 25 mg, 5 mg to 500 mg, 5 mg to 100 mg, or 1 mg to 5 mg, or 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 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 mg, 10 mg to 50 mg, or 10 mg to 25 mg. However, the dose may be variously prescribed depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate and responsiveness of the patient, These factors can be taken into account to appropriately adjust the dosage. The number of doses may be at least once per day or within the range of clinically acceptable side effects and may be administered at one or two or more doses at the site of administration and may be administered daily or every two to five days The number of days of administration can be administered from one day to 30 days at one time of treatment. If necessary, the same treatment may be repeated after the appropriate time. For an animal other than a human, the dosage may be the same as that of a human per kg, or the dosage may be converted into a dose in terms of a volume ratio (for example, an average value) One dose may be administered.

The composition according to one aspect of the present invention includes extracts of fermented soybean fermented with A. cristatus strain, gum, and glyceryl glucoside to improve skin moisturizing, barrier strengthening, and skin regeneration. It exhibits excellent effects and can be safely used because it does not irritate the skin.

FIG. 1 is a graph showing mRNA relative levels of the moisturizing factors HAS3 and AQP3. (-): untreated group; 100 ppm of hyaluronic acid: 100 ppm of hyaluronic acid treatment group; And Example 3: a group of treatments comprising fermented soybeans, locust bean gum and glyceryl glucoside.

FIG. 2 shows the results of immunohistochemical staining of artificial skin using AQP3 antibody to confirm the effect of increasing AQP3 expression in artificial skin. (-): untreated group; UVB +: UVB irradiated group; UVB + hyaluronic acid 100 ppm: hyaluronic acid 100 ppm treated group; UVB + Example 3: A group of treatments comprising fermented soybeans, locust bean gum and glyceryl glucoside.

FIG. 3 shows H & E staining results for confirming skin barrier improvement effect on artificial skin. (-): untreated group; 100 ppm of hyaluronic acid: 100 ppm of hyaluronic acid treatment group; Example 3: A group treated with a composition comprising fermented soybeans, locust bean gum and glyceryl glucoside.

FIG. 4 is a result of observing the degree of proliferation of cells after 24 hours after scraping the cells as a result of confirming the skin cell regeneration effect. (-): untreated group; Example 3: A group treated with a composition comprising fermented soybeans, locust bean gum and glyceryl glucoside.

5 is a graph showing the amount of cells per hour (hr) as a result of confirming the skin cell regeneration effect. (-): untreated group; Example 3: A group treated with a composition comprising fermented soybeans, locust bean gum and glyceryl glucoside.

FIG. 6 is a graph comparing skin moisture percentage (%) with time after applying Formulation Example 1 and Comparative Formulation Example 1. FIG.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  1: Aspergillus Christatus ( A. cristatus ) Isolation and Identification of Strain

Ten grams of a bicomponent manufactured in China was appropriately serially diluted with a 0.85% (w / w) NaCl solution and applied to a Potato Dextrose Agar (PDA, Himedia, India, M1942) supplemented with chloramphenicol And then cultured for 30 to 60 hours. Among the dominant strains, strains forming gold colonies were selected and identified. Identification was made by Charles River Laboratories Korea, Inc. and analyzed the Internal Transcribed Spacer (ITS2) part. As a result, 99.9% homology with Aspergillus cristatus was obtained and used in the present invention. The isolated strain was named A. cristatus Cosmax-GF, deposited at the Korean Microorganism Conservation Center on March 22, 2016, and assigned the deposit number KCCM11820P.

Example 2: Preparation of fermented soybean and extract

Soybean was fermented with A. cristatus isolated in Example 1 to prepare fermented soybeans, and fermented soybean extract was prepared.

Specifically, the beans used for the solid fermentation were prepared by washing after purchase in the market. The washed soybeans were sterilized at 121 DEG C for 30 minutes. Then, A. cristatus of Example 1, which had been grown on a potato agar medium, was prepared as a spore suspension, and the suspension was added to the sterilized soybeans and incubated at 33 ° C. for 60 to 65 hours at 75 to 85% Lt; / RTI > The solid fermented soybean was dried and pulverized at 60 ° C until the moisture content was less than 12%.

Powdered fermented soybeans (100 g) were placed in an extraction vessel, and immersed in 1 L of a 70 wt% ethanol aqueous solution as an extraction solvent and left at room temperature for 24 hours to obtain an extract. The supernatant, which was centrifuged at 3000 rpm for 10 minutes, was filtered through a 0.45 μm membrane to prepare a filtrate. The filtrate was concentrated under reduced pressure using a rotary evaporator to obtain 9 to 11 g of concentrated extract.

Example 3: Preparation of composition for improving skin care

(CAS No. 9000-40-2, Sigma-Aldrich), and glyceryl glucoside (CAS No. 22160-26-5, Jin Chem Co., Ltd.) prepared in Example 2, To prepare a composition for skin aesthetic improvement. Specifically, the extract of Example 2 was contained at a concentration of 100 ppm, i.e., 0.01% (w / v), and contained at a concentration of locust bean black 500 ppm, i.e., 0.05% (w / v) Glucoside was included at a concentration of 10 ppm, i.e. 0.001% (w / v).

number	ingredient	Concentration (ppm)
One	The extract of Example 2	100
2	Locust Blind Sword	500
3	Glyceryl glucoside	10

Experimental Example  One: Moisturizing factor  Confirmation of increased expression

Experiments were conducted to confirm the effect of increasing the moisturizing function of the composition prepared in Example 3 above.

First, human keratinocytes were inoculated in a 6-well cell culture dish at a density of 5 × 10 ⁵ cells and cultured in a 5% CO ₂ incubator at 37 ° C. for 24 hours. After the addition of the composition of Example 3, the cells were further cultured for 24 hours. The cells were recovered and RNA was isolated by adding 1 ml of trizol (RNA iso, DAKARA, Japan). RNA was quantified using Nanodrop 2000 (Thermo, USA), and cDNA was synthesized by reacting at 42 ° C for 55 minutes and at 70 ° C for 15 minutes (Reverse Transcriptase Mix, ELPIS biotech, Korea). The synthesized cDNA was synthesized using a template of hyaluronic acid synthase-3 (HAS3), aquaporin 3 (AQP3) and a cyanine dye (SYBR Green supermix, Applied Biosystems, USA) The expression of HAS3 and AQP3 genes was finally assessed by real-time PCR using a real-time PCR machine (Step One Plus, Applied Biosystems, USA) using real-time polymerase chain reaction. The primer sequences used in the real-time PCR were shown in Table 2 below. The real-time PCR reaction was carried out at 94 ° C for 5 minutes, after activation of the polymerase, at 40 ° C for 30 seconds at 95 ° C, 1 minute at 54 ° C, Respectively. The expression level of the gene was finally analyzed by correction for the β-actin gene. As a comparative example, purified water containing hyaluronic acid at a concentration of 100 ppm was used.

gene	SEQ ID NO:	primer  designation	primer  direction	primer  order
HAS3	One	HAS3-F	Forward	5'-CTTAAGGGTTGCTTGCTTGC-3 '
	2	HAS3-R	Reverse	5'-GTTCGTGGGAGATGAAGGAA-3 '
AQP3	3	AQP3-F	Forward	5'-AGACAGCCCCTTCAGGATTT-3 '
	4	AQP3-R	Reverse	5'-TCCCTTGCCCTGAATATCTG-3 '
β-Actin	5	Beta-Actin-F	Forward	5'-GGCCATCTCTTGCTCGAAGT-3 '
	6	Beta-Actin-R	Reverse	5'-GAGACCTTCAACACCCCAGC-3 '

FIG. 1 is a graph showing mRNA relative levels of the moisturizing factors HAS3 and AQP3. (-): untreated group; 100 ppm of hyaluronic acid: 100 ppm of hyaluronic acid treatment group; And Example 3: a group of treatments comprising fermented soybeans, locust bean gum and glyceryl glucoside.

As shown in Fig. 1, the expression of HAS3 and AQP3 was greatly increased in the composition-treated group of Example 3 as compared with the group treated with 100 ppm of hyaluronic acid. Therefore, it was found that the composition containing the fermented soybean, locust bean gum, and glyceryl glucoside exhibited excellent effect of increasing the moisturizing function.

Experimental Example 2: Improvement of moisture and skin barrier strengthening effect in artificial skin

Experiments were conducted to confirm the effect of improving the water content and skin barrier properties of the composition prepared in Example 3.

Experiments to confirm the effect of moisture improvement on artificial skin were performed as follows. First, UVB was applied to artificial skin to reduce AQP3. The composition of Example 3 and the comparative example, purified water containing 100 ppm of hyaluronic acid, were each treated for 3 days with artificial skin. Artificial skin was cut and fixed to paraffin. Immunohistochemical staining of skin tissue was performed using AQP3 antibody.

Experiments to confirm skin barrier enhancement effect on artificial skin were performed as follows. The composition of Example 3 and the comparative example, purified water containing 100 ppm of hyaluronic acid, were each treated for 3 days with artificial skin. Artificial skin was cut and fixed to paraffin. H & E staining was performed to confirm the change of skin layer structure of artificial skin.

FIG. 2 shows the results of immunohistochemical staining of artificial skin using AQP3 antibody to confirm the effect of increasing AQP3 expression in artificial skin. (-): untreated group; UVB +: UVB irradiated group; UVB + hyaluronic acid 100 ppm: hyaluronic acid 100 ppm treated group; UVB + Example 3: A group of treatments comprising fermented soybeans, locust bean gum and glyceryl glucoside.

As shown in FIG. 2, treatment of the composition of Example 3 showed an increase in the expression of AQP3 reduced by UVB, and the effect was found to be superior to that of 100 ppm of hyaluronic acid.

FIG. 3 shows H & E staining results for confirming skin barrier improvement effect on artificial skin. (-): untreated group; 100 ppm of hyaluronic acid: 100 ppm of hyaluronic acid treatment group; Example 3: A group treated with a composition comprising fermented soybeans, locust bean gum and glyceryl glucoside.

As shown in FIG. 3, the surface layer of the skin was significantly improved in the composition-treated group of Example 3 as compared with the non-treated group and the hyaluronic acid-treated group.

Experimental Example 3: Confirmation of skin cell regeneration effect

Experiments were conducted to confirm the skin cell regeneration effect of the composition prepared in Example 3 above.

First, human fibroblasts were inoculated in a 96-well cell culture dish at a density of 2 × 10 ⁴ cells and cultured in a 37, 5% CO ₂ incubator for 24 hours. After the center of the cultured cells was scraped off using a cell scraper, the composition of Example 3 was added and cultured for 24 hours. The extent of cell proliferation was measured using an IncuCyte ZOOM machine and the amount of cells per time was quantitated.

FIG. 4 is a result of observing the degree of proliferation of cells after 24 hours after scraping the cells as a result of confirming the skin cell regeneration effect. (-): untreated group; Example 3: A group treated with a composition comprising fermented soybeans, locust bean gum and glyceryl glucoside.

5 is a graph showing the amount of cells per hour (hr) as a result of confirming the skin cell regeneration effect. (-): untreated group; Example 3: A group treated with a composition comprising fermented soybeans, locust bean gum and glyceryl glucoside.

As shown in Figs. 4 and 5, it was found that the composition of Example 3 was excellent in skin cell proliferation effect.

Formulation Example 1 and Comparative Formulation Example 1: Preparation of cosmetic composition

The cosmetic composition of Formulation Example 1 containing the skin-beauty-improving composition of Example 3 was prepared. For comparison, the cosmetic composition of Comparative Formulation Example 1 which did not contain the composition for skin-care improvement of Example 3 was prepared. Table 3 below shows the specific components and contents of the cosmetic composition of Formulation Example 1 and Comparative Formulation Example 1.

number	ingredient	Formulation Example 1 (% by weight)	Comparative Formulation Example 1 (% by weight)
One	Example 3	5	-
2	Xanthan gum (KELTROL ® F)	0.03	0.03
3	Propanediol (ZEMEA ® SELECT PROPANDIOL)	5	5
4	1% EDTA-2Na	2	4
5	Betain (BETAFIN ® )	0.5	0.5
6	TEA (TEA) 10%	1.5	1.5
7	Polyacrylic acid polymer (CARBOPOL ® 940 POLYMER)	0.1	0.1
8	Purified water	Balance	Balance

Experimental Example 4: Skin moisturizing clinical evaluation

Skin moisturizing power of the cosmetic composition of Formulation Example 1 and Comparative Formulation Example 1 was evaluated clinically.

Specifically, five healthy adult women with an average age of 26.2 years were coated with Formulation Example 1 and Comparative Formulation Example 1, respectively, and the moisture resistance was measured using a Corneometer CM825 (CK electronic, Germany). The moisture content of the test area (22 cm ² ) was measured at the test site (20 cm ² ), and 20 μl of the product was applied to the test site and absorbed before the application, 2 hours, 4 hours, 8 hours The skin moisture was measured three times each. The results of the measurements were obtained by calculating the skin moisture content and the rate of change using the average value of the three measurements, and then compared with the non-attrition value.

FIG. 6 is a graph comparing skin moisture percentage (%) with time after applying Formulation Example 1 and Comparative Formulation Example 1. FIG.

As shown in Fig. 6, when the product of Formulation Example 1 including the composition of Example 3 was used, it was confirmed that the moisture of the skin was the most excellent and the moisturizing power was the highest.

Claims (15)

1. Aspergillus The present invention relates to a composition for improving skin beauty comprising an extract of fermented soybean fermented with a cristatus strain, gum, and glyceryl glucoside as an active ingredient.
2. The composition according to claim 1, wherein the improvement of the skin is moisturizing, barrier strengthening, or skin regeneration.
3. The composition of claim 1, wherein the strain is an Aspergillus cristatus Cosmax-GF strain (accession number: KCCM11820P).
4. The composition of claim 1, wherein the fermentation is a solid fermentation.
5. The composition for improving skin care according to claim 1, wherein the fermented soybean is fermented at 30 to 35 DEG C for 48 to 65 hours at 75 to 85% humidity.
6. The composition of claim 1, wherein the extract is an extract of C1-C6 alcohol, water, or a mixture thereof.
7. The composition of claim 1, wherein the extract is an ethanol extract.
8. The composition for improving skin care according to claim 1, wherein the extract is contained at a concentration of 0.001% (w / v) to 10% (w / v).
9. The composition for improving skin care according to claim 1, wherein the composition is at least one selected from the group consisting of black locust bean gum, gum arabic, ghatti gum, karaya gum, guar gum, tragacanth gum, xanthan gum, carrageenan, mannan and glucomannan .
10. The composition for improving skin care according to claim 1, wherein the black is contained at a concentration of 0.001% (w / v) to 10% (w / v).
11. The composition for improving skin care according to claim 1, wherein the glyceryl glucoside is contained at a concentration of 0.0001% (w / v) to 1% (w / v).
12. The composition according to claim 1, wherein the expression of HAS3 or AQP3 is increased.
13. The composition of claim 1, wherein the composition is a cosmetic, a food, or a pharmaceutical composition.
14. Aspergillus A method for improving skin condition comprising administering to an individual an effective amount of a composition comprising an extract of fermented soybean fermented with a cristatus strain, gum, and glyceryl glucoside.
15. Use of a composition comprising an extract of fermented soybeans obtained by fermenting soybeans with Aspergillus cristatus strain, gum, and glyceryl glucoside for the manufacture of a skin condition improving agent.

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