KR20190009164A - A composition for skin smoothing comprising an extract of fermented ponciri fructus - Google Patents

Abstract

The present invention relates to a skin soothing composition comprising an extract of fermented Ponciri Fructus as an active ingredient and a method to produce the extract. According to one aspect of the skin soothing composition, the extract of fermented Ponciri Fructus fermented by a strain in the genus Aspergillus is capable of soothing the skin as the extract shows anti-inflammatory functions and has effects of alleviating atopic dermatitis, moisturizing skin, or decreasing red spots. According to the other aspect of the method to produce the extract of fermented Ponciri Fructus, since the extract of fermented Ponciri Fructus having a skin soothing effect can be produced, the extract can be used in diverse fields such as cosmetics, foods or pharmacological compositions.

Description

[0001] The present invention relates to a skin-soothing composition comprising a fermented persimmon extract,

≪ / RTI >

In the pharmacology book, which is made by a Chinese Ming herbalist, Ishijin, describes the efficacy of Zisil in the herbaceous gangbok, "The cold affects the skin and causes papular papules (papules), which are very itchy." . At this time, Ponciri Fructus is a deciduous shrub ( Poncirus trifoliata ) of dried fruit.

Studies on various physiological activities of Zypsil have been carried out. For example, there is a previous study in which trifoliate orange extract inhibits proinflammatory cytokines, such as IL-1β, IL-6 and TNF-α, Lee, Korean J. Plant Res. 29 (3): 305-312, 2016). Also, Korean Patent Registration No. 1662719 discloses a composition for preventing or treating prostate-related diseases, which comprises a root extract. However, the research on fermentation of persimmon fermented products is insufficient.

Korean Patent No. 1662719.

 Eun Lee, Korean J. Plant Res. 29 (3): 305-312, 2016.

An aspect of the present invention provides a composition for dermatological soothing which comprises a zygos fermented product as an active ingredient.

Another aspect is a method for producing fermented soybean milk, And contacting the fermentation fiber compartment with a C1-C6 alcohol, water, or a mixture thereof.

An aspect of the present invention provides a composition for dermatological soothing which comprises a zygos fermented product as an active ingredient.

Another aspect provides a composition for skin soothing which comprises an extract of a fermentation fiber obtained by fermenting a flaxseed with a strain of Aspergillus genus as an active ingredient.

The skin soothing may be anti-inflammatory, atopic dermatitis, skin moisturizing, or erythema reduction. Since the extract of the fermented persimmon extract has an anti-inflammatory effect, atopic dermatitis improvement effect, skin moisturizing effect, or erythema reduction effect compared with the extract of the non-fermented persimmon leaf, the composition comprising the fermented persimmon extract has anti-inflammation, atopic dermatitis improvement , Skin moisturizing, or erythema reduction.

The term " anti-inflammatory " can be used interchangeably with " inflammation inhibition or improvement ", and may mean all actions in which the immune response is alleviated and NO production is suppressed.

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

The term " erythema reduction " may refer to any action in which the skin is reddosed by various extrinsic and / or intrinsic stimuli, or in which the redness is reduced in whole or in part by erythema caused by the expansion of blood vessels.

The above " Ponciri Fructus " refers to Poncirus < RTI ID = 0.0 > trifoliata ) of dried fruit. The fuselage may be one obtained by a commonly available method, for example, a commercially available one. The support may be a sterilized support, and the sterilization method may be performed in a manner customary in the art.

In one embodiment, the strains are Aspergillus germanium (Aspergillus niger), Aspergillus duck claim (Aspergillus oryzae), Aspergillus fumigatus (Aspergillus fumigatus), Aspergillus Playa booth (Aspergillus flavus), Aspergillus Terre Uz (Aspergillus terreus , and the like, but are not limited thereto.

In one embodiment, the strain may be an Aspergillus niger strain. The term " Aspergillus niger " is a type of black fungus, which forms black spores and is also called black germ. The Aspergillus niger can be separated from food, soil, sea and the like.

In 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. Solid fermentation can compensate for some of the disadvantages of conventional liquid fermentations such as (Silvia Martins et al., Biotechnology Advances, 29, 365-373, 2011). First, conventional liquid fermentation has a risk of corruption and contamination by E. coli and anaerobic bacteria harmful to the human body due to fermentation conditions of high water content. Secondly, due to the fermentation conditions having a high water content, stirring and sterilization are required during the fermentation process, which increases the process cost. Third, a large amount of chemicals harmful to the environment are required for the immersion fermentation process. In contrast, the composition according to one embodiment of the present invention is advantageous in that the solid fermentation method can be applied to the fuselage to overcome the problems of the conventional fermentation technology, thereby avoiding contamination of germs, reducing the processing cost, and recovering the product easily Can be. In one embodiment, it was confirmed that the content of ponciretin, which is an active ingredient in the solid fermentation product of the fingernail, increased more than twice as much as before fermentation. This is probably due to the increase in the expression of glucosease, which converts the glycoside poncirin into the non-parasitic poncho retin. Microorganisms exhibit a higher level of liquid culture relative to liquid culture when expressing the extracellular glucosease (Subramaniyam, R. and Vimala, R., Society for Science and Nature, 3 (3), 480-486, 2012). On the other hand, lactic acid bacteria such as Saccharomyces cerevisiae or lactobacilli such as Lactobacillus plantarum have no activity of the polysaccharide degrading enzyme and can not effectively decompose the cell walls of the medicinal material during solid fermentation. Therefore, The fermentation is carried out on the extract of the present invention. In contrast, Aspergillus niger can effectively decompose the cell wall of the zygosity by the extracellular enzyme and convert the pysyline in the zygosyl to the non-glycosylated vyschretin component. Therefore, the zygosity itself, which is not the zygomycin extract, Lt; / RTI >

In one embodiment, the fermentation chamber may be solid fermented by inoculating Aspergillus niger, in particular Aspergillus niger spore suspension, into the follicle.

In one embodiment, the pseudokinesin can be converted to ponceetin by the extracellular enzyme of Aspergillus niger through the fermentation. Examples of the extracellular enzyme include, but are not limited to, beta-glucosidase, alpha-Rhamnosidase, and the like. 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 niger ( A. niger ) for a long time. Therefore, in order to maximize the expression phase of the extracellular enzyme of Aspergillus niger ( A. niger ) and to increase the non-glycosidic component of the fibril, the fermentation temperature, time, and humidity are appropriately selected . The term " non-glycosides " may refer to a substance in which glycosides are removed through fermentation or hydrolysis. Thus, the unglycosylated component of the fibril can be pornhistetin.

The fermentation fiber compartment may be fermented after calibrating the initial moisture content of fermentation to 40 to 50%. If the initial moisture content is less than 40%, the fermentation time is terminated early in less than 48 hours. If the initial moisture content is above 50%, the moisture content is high and the pore between the fungi is decreased, and the growth of the absolute aerobic strain Aspergillus niger is slowed There is a problem that the probability of contamination increases.

The fermentation fibrils may be fermented at 25 to 35 ° C, 28 to 35 ° C, 30 to 35 ° C, 31 to 34 ° C, and 32 to 34 ° C. 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 level can be relatively lowered. On the contrary, when the temperature is lower than 25 ° C, the enzyme reaction rate is lowered, Can be lowered.

The fermentation chamber may be fermented for 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 fermentation chamber has a humidity of 10 to 99.9%, a humidity of 20 to 99.9%, a humidity of 30 to 99.9%, a humidity of 40 to 99.9%, a humidity of 50 to 99.9%, a humidity of 50 to 90%, a humidity of 60 to 90% Humidity, 75-85% humidity, or 80-85% humidity conditions. If the humidity is less than 75%, the time for spore formation of the strain becomes faster so that the enzyme activity and expression amount can be lowered. On the contrary, when the humidity is higher than 85%, the moisture content is high, The growth of Aspergillus niger ( A. niger ) may be slowed and the rate of contamination by other microorganisms may be increased.

For example, the fermentation broth may be fermented at 25-35 ° C for 60-65 hours at 75-85% humidity.

The fermentation fiber compartment may be the entire fermentation fiber compartment, a part thereof, or a material derived therefrom. The fermentation fiber used for the extraction may be a whole, a part thereof, or a material derived therefrom, which may be pulverized or chopped or suitably dried. The fermentation fiber compartment can be dried to have a moisture content of less than 20%, 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, 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 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 used in an amount of 5 to 15 (vol / g) times, such as 5 to 13 (vol / g) times, 5 to 11 (vol / , 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 fescue, 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 milk powder may be subjected to ultrasonic extraction by immersing in an extraction solvent for 2 to 4 hours, or may be extracted 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 from 0.001% to 80%, such as from 0.01% to 60%, from 0.01% to 40%, from 0.01% to 30%, from 0.01% to 20% %, 0.01% to 10%, 0.01% to 5%, 0.05% to 60%, 0.05% to 40%, 0.05% to 30%, 0.05% to 20% From 0.05% to 10%, from 0.05% to 5%, from 0.1% to 60%, from 0.1% to 40%, from 0.1% to 30%, from 0.1% to 20% % To 10 wt%, or 0.1 wt% to 5 wt%.

In one embodiment, a fraction of the extract of the fermentation pouch can be contained as an active ingredient instead of the fermentation pouch extract. In one embodiment, it may further comprise a fraction of the extract of the fermentation compartment. The term " fraction " refers to a substance in which the extract of the fermentation fiber compartment is divided into components of a part thereof, that is, a fractionated substance. The fraction may be obtained by solvent fractionation. The solvent fractionation may be a step of mixing the fermented yeast 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 successively fractionating the fermented persimmon extract in water and then fractionating it with methylene chloride, ethyl acetate, butanol, and water.

Specifically, the methylene chloride fraction is prepared by mixing the fermented yeast 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 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, agitation, etc., may be as described for the extracts used to prepare the fermented soybean extract described above. 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 extract may be one containing ponciretin. In one embodiment, the extract of the fermentation sponge may comprise about 2 times or more of the concentration of pon smear than the flax seed extract.

The composition according to one embodiment may be one that inhibits the expression of IL-8, IL-6, or TNF- ?, specifically inhibits the expression of NO, IL-6, or TNF-? Lt; / RTI >

The composition according to one embodiment may be one that inhibits the expression of TSLP (Thymic stromal lymphopoietin), specifically, inhibits the expression of TSLP, thereby improving atopic dermatitis.

The composition according to one embodiment of the present invention may exhibit a skin moisturizing effect by enhancing the recovery rate of percutaneous water loss in the human skin application experiment.

The composition according to one embodiment may exhibit an effect of suppressing skin erythema in a human skin application test.

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 dermal release according to one embodiment may further comprise cosmetically, pharmaceutically, or pharmaceutically acceptable excipients or carriers. 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. It can also be applied in various forms such as liquid, cream, paste, and solid phase. 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 can be easily selected by those skilled in the art within a range not to impair the object and effect of the present invention, and the amount thereof is 0.001 to 5% by weight, specifically 0.01 to 5% % ≪ / 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 5 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 diseases may be skin inflammation diseases, diseases caused by skin barrier function damage, atopic dermatitis, skin aging, or skin eruption. The term " prevention " includes inhibiting the occurrence of a disease. The term " treatment " includes inhibiting, alleviating, or eliminating the development of the disease.

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 skin moisture retention, skin moisture loss prevention, skin barrier enhancement, and anti-inflammatory effects.

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 can prevent or treat damage to the skin barrier function by increasing the recovery rate of transdermal water loss.

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 aging of the skin due to the effects of strengthening the skin barrier, maintaining skin moisture, preventing the loss of skin moisture, and the like.

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

Specifically, the method is a method of improving or treating inflammation of an individual, a method of improving or treating atopic dermatitis of an individual, a method of effectively maintaining skin moisture or preventing moisture loss of an individual, .

The terms " administering ", " introducing " and " transplanting " are used interchangeably and refer to a method of delivering a composition according to one embodiment ≪ / RTI > may refer to the arrangement of the composition according to the invention. 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 requiring skin soothing, for example, anti-inflammatory, atopic dermatitis treatment, skin moisturizing, or erythema reduction effect.

The administration can be carried out 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, 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 1000 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 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.

In another aspect, there is provided a method for producing fermented soybeans, comprising fermenting a rice bran with a strain of Aspergillus sp. And contacting the fermentation fiber compartment with a C1-C6 alcohol, water, or a mixture thereof.

The extracts of the above fuselage, fermentation, fermentation fungus, and fermentation fungus are the same as those described above.

In one embodiment, the strain may be Aspergillus niger .

In one embodiment, the fermentation may be a solid fermentation.

In one embodiment, the fermentation can be performed at 25-35 ° C for 60-65 hours at 75-85% humidity conditions.

The fermented zipper extract prepared by the above method can exhibit an anti-inflammatory effect by inhibiting the expression of IL-8, IL-6, or TNF- ?, suppressing the expression of TSLP, and exhibiting an effect of improving atopic dermatitis , The rate of recovery of transdermal water loss can be increased to show a skin moisturizing effect and skin erythema can be reduced. The antiinflammatory, atopic dermatitis, skin moisturizing, and erythema reduction effects of the extract of the fermentation fungus are superior to those of the unfermented fungus extract. Accordingly, the fermented zipper extract prepared by the above method can be used for various purposes such as cosmetics, foods, pharmaceutical compositions, and the like.

According to one aspect of the composition for calming skin, the extract of the fermentation fungus fermented with Aspergillus sp. Can be used for anti-inflammation, atopic dermatitis, skin moisturization, or erythema reduction.

According to another aspect of the present invention, it is possible to produce an extract of a fermented persimmon having a skin soothing effect, so that the extract can be applied to various fields such as cosmetics, foods, and pharmaceutical compositions.

Fig. 1 shows the mechanism of the conversion of pseudo-lysine, which is a major component of the fibril, into β-glucosidase and α-lamb- nocodase enzymes by solid phase fermentation using Aspergillus niger strain.
FIG. 2 is a graph showing changes in the content of the pon smectin component contained in the product before and after the solid fermentation of the pongee according to the strain type: Comparative Example 1; Comparative Example 2: Extract of fermented zipper fermented with saccharomyces cerevise; Comparative Example 3, Extract of Fermented Fibers Fermented with Lactobacillus plantarum; And Example 1, an extract of a fermentation zipper fermented with Aspergillus niger.
FIG. 3 is a graph showing the relative level of mRNA of TNF-α in keratinocytes as a result of confirming the anti-inflammatory effect of the fermentation fungus. (-): negative control; (+): PolyI: C and IL-4 treated group; Dex: a positive control group dexamethasone treated group; Comparative Example 1: Taxol extract treatment group; And Example 1: Extract-treated group of fermented zipper.
FIG. 4 is a graph showing the relative level of mRNA of IL-6 in keratinocytes as a result of confirming the anti-inflammatory effect of the fermentation fungus. (-): negative control; (+): PolyI: C and IL-4 treated group; Dex: a positive control group dexamethasone treated group; Comparative Example 1: Taxol extract treatment group; And Example 1: Extract-treated group of fermented zipper.
FIG. 5 is a graph showing the relative level of mRNA of IL-8 in keratinocytes as a result of confirming the anti-inflammatory effect of the fermentation fungus. (-): negative control; (+): PolyI: C and IL-4 treated group; Dex: a positive control group dexamethasone treated group; Comparative Example 1: Taxol extract treatment group; And Example 1: Extract-treated group of fermented zipper.
FIG. 6 is a graph showing the relative level of mRNA of TSLP in keratinocytes as a result of confirming the effect of improving the atopic dermatitis of the fermented wipe. (-): negative control; (+): PolyI: C and IL-4 treated group; Dex: a positive control group dexamethasone treated group; Comparative Example 1: Taxol extract treatment group; And Example 1: Extract-treated group of fermented zipper.
FIG. 7 is a graph showing recovery rate (%) of transdermal water loss as a result of confirming the moisturizing effect and the skin barrier improvement effect of the essence including the fermentation fudge.
8 is a graph showing the improvement rate (%) of erythema of the skin as a result of confirming the erythema-improving effect of the essence containing the fermentation fudge.
FIG. 9 is a clinical picture after application of the product as a result of confirming the erythema-improving effect of the essence including the fermentation wil.

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: Preparation of Aspergillus niger Fermentation Fibrosis and Its Extracts

Solid fermentation was carried out with Aspergillus niger ( A. niger ) to produce a fermentation waste, and an extract of the fermentation waste was prepared.

First, we purchased and prepared Jisil from Kyungdong market (Seoul, Korea). Water (0.62 kg) was added to 1 kg of the wiping paper having a moisture content of 11% to make the water content 45%. After sterilization at 121 ° C for 30 minutes, A. niger was prepared from spore suspension in a potato agar medium and incubated at 33 ° C for 60 to 65 hours at 75 ° C to 85% Lt; RTI ID = 0.0 >%< / RTI > The solid fermented fibrils were dried and pulverized at 60 캜 until the moisture content was less than 12%.

100 g of the pulverized fermentation feces was placed in an extraction container and immersed in 1 L of a 70% by weight aqueous ethanol solution as an extraction solvent and allowed to stand 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 5 to 7 g of concentrated extract. The obtained concentrated extract was used for in vitro skin test, added to the essence formulation, and used for clinical use.

Comparative Example 1: Preparation of goat cheese extract

We purchased and prepared Jisil from Kyungdong market (Seoul, Korea). Water (0.62 kg) was added to 1 kg of the wiping paper having a moisture content of 11% to make the water content 45%. Sterilized at 121 占 폚 for 30 minutes, dried and pulverized at 60 占 폚 until the moisture content became less than 12%.

100 g of powdered groundwater was placed in an extraction vessel, immersed in 1 L of 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 4-6 g of concentrated extract. The obtained concentrated extract was used for in vitro skin test, added to the essence formulation, and used for clinical use.

Comparative Example 2: Preparation of Saccharomyces cerevisiae fermented zipper and its extract

Saka access to my serenity Wiese (Saccharomyces The fermented waste was prepared by solid fermentation of the fibrils with.

First, we purchased and prepared Jisil from Kyungdong market (Seoul, Korea). Water (0.62 kg) was added to 1 kg of the wiping paper having a moisture content of 11% to make the water content 45%. After sterilization at 121 DEG C for 30 minutes, Saccharomyces cerevisae grown in Potato Dextrose Broth (BD Difco, USA) was prepared as a seed solution and inoculated into a sterilized zipper to obtain 33 Lt; 0 > C for 60 to 65 hours and 75 to 85% relative humidity. The solid fermented fibrils were dried and pulverized at 60 캜 until the moisture content was less than 12%.

100 g of the pulverized fermentation feces was placed in an extraction container and immersed in 1 L of a 70% by weight aqueous ethanol solution as an extraction solvent and allowed to stand 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 4-6 g of concentrated extract. The obtained concentrated extract was used for comparison of the active ingredients

Comparative Example 3: Preparation of Lactobacillus plantarum fermented soybean and its extract

Lactobacillus plantarum ( Lactobacillus plantarum ) was used for solid fermentation to produce a fermentation chamber and an extract of fermentation chamber was prepared.

First, we purchased and prepared Jisil from Kyungdong market (Seoul, Korea). Water (0.62 kg) was added to 1 kg of the wiping paper having a moisture content of 11% to make the water content 45%. On the 121 ℃ sterilized 30 minutes, MRS broth to prepare a Lactobacillus Planta room (L. Plantarum) grown in (MRS Broth, BD Difco, USA) to the seed solution, and inoculated them in sterilized jisil 33 ℃, 60 To 65 hours, at a humidity of 75 to 85%. The solid fermented fibrils were dried and pulverized at 60 캜 until the moisture content was less than 12%.

100 g of the pulverized fermentation feces was placed in an extraction container and immersed in 1 L of a 70% by weight aqueous ethanol solution as an extraction solvent and allowed to stand 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 4-6 g of concentrated extract. The obtained concentrated extract was used for comparison of the active ingredients

Experimental Example 1: Comparison of effective components of the feedstock and the fermented feedstock

The fermented zipper of Example 1, the zipper extract of Comparative Example 1, and the fermentation zipper of Comparative Examples 2 and 3 were compared and analyzed for the change of active ingredient under the following high-performance liquid chromatography (HPLC) conditions. The active ingredient was analyzed for glycosides poncirin and non-glycoside ponciretin. Standard reagent Voncietin was purchased from Core Science (http://www.coresciences.co.kr/), dissolved in 80% methanol, filtered through a 0.45 μm filter, and used as a standard solution.

<HPLC analysis conditions>

- HPLC model: HPLC-UV-Vis detector (Walnut Creek, CA, USA)

- Column: Shiseido Capcell Pak C18 column (Chuoku, Japan)

- Solvent Gradient mode:

Time (min) Solvent A (%) Solvent B (%) 0 60 40 10 60 40 20 50 50 30 40 60 35 60 40

- solvent A; 0.05% phosphoric acid solution B; MeOH

- Injection volume: 10 μl

- Run time: 35 min

- Wavelength: 246 nm

- Flow rate: 1 ml / min

- Standard concentration for calibration curve: 0, 1, 2, 4, 8 ppm

Fig. 1 shows the mechanism of the conversion of pseudo-lysine, which is a major component of the fibril, into β-glucosidase and α-lamb- nocodase enzymes by solid phase fermentation using Aspergillus niger strain.

FIG. 2 is a graph showing changes in the content of the pon smectin component contained in the product before and after the solid fermentation of the pongee according to the strain type: Comparative Example 1; Comparative Example 2: Extract of fermented zipper fermented with saccharomyces cerevise; Comparative Example 3, Extract of Fermented Fibers Fermented with Lactobacillus plantarum; And Example 1, an extract of a fermentation zipper fermented with Aspergillus niger.

As shown in Fig. 2, there was a change in the pornhistein component in Comparative Example 1 and Example 1 before and after fermentation. However, in the case of the fermentation fibrils of Comparative Examples 2 and 3 using saccharomyces cerevisiae or lactobacillus plantarum, there was almost no component change. Unlike the fungal Aspergillus niger, Saccharomyces cerevisiae or Lactobacillus plantarum lacks the activity of the extracellular polysaccharide degradation enzyme and can not effectively decompose the cell walls of the fibrillar during fermentation . For this reason, when yeast or lactic acid bacteria are used for fermentation, an extract of a pharmacologically degraded medicinal cell wall is cultured. In contrast, an additional non-glycosyl picricin component was produced in the fermentation chamber of Example 1 fermented with Aspergillus niger. It was judged that the extracellular enzyme of Aspergillus niger effectively decomposes the cell wall of the zygosity and at the same time produces the non-glycosylated picrystine component from the pysyline in the zygos. Thus, Aspergillus niger has been identified as a strain suitable for solid fermentation of the waste yarn itself, not the extract of zygosum.

Experimental Example 2: Improvement of Inflammatory Factor of Fermentation Fibrosis

2-1. Human skin keratinocyte cell line HaCaT cell culture

Human keratinocytes were cultured in DMEM containing 10% fetal bovine serum (FBS) and antibiotics. The culture was carried out in a 75T-flask and a 6-well plate, and cultured in a 37 ° C incubator with 5% CO ₂ . The culture medium was changed every 3 to 4 days. When the cells were overproduced, they were subcultured. Cells were dispensed into 6-well plates and washed with phosphate-buffered saline solution (PBS) 24 hours after incubation. The fermented zygote extract of Example 1 and the zygomycetum extract of Comparative Example 1 were diluted to a final concentration of 10 ppm with PolyI: C and IL-4, respectively, in DMEM without FBS and added to each well. Positive controls were treated with 1 μM dexamethasone. After 4 hours, cDNA was synthesized and PCR was performed to evaluate gene expression level.

2-2. Identification of inflammatory cytokine abatement effect using real-time PCR

The cDNA synthesized in Experimental Example 2-1 was used as a template and subjected to real-time PCR using a SYBR green master mix (cyanine dye) TNF-α, IL-6, and IL-8 gene expression levels were evaluated. Expression of the gene was compared with the relative value by calibrated quantification of the internal control gene beta-actin by the Ct (threshold cycle) method. Table 2 shows primer information used in real-time PCR.

gene SEQ ID NO: Name of the primer primer  direction Primer sequence TNF-a One TNF-alpha primer-F Forward 5'-GCTATCTGGTGCCCAGGCTAT-3 ' 2 TNF-alpha primer-R Reverse 5'-CGACGCCACAATCCTTGTAAT-3 ' IL-6 3 IL-6 primer-F Forward 5'-TACCCCCAGGAGAAGATTCC-3 ' 4 IL-6 primer-R Reverse 5'-TTTTCTGCCAGTGCCTCTTT-3 ' IL-8 5 IL-8 primer-F Forward 5'-CCAACACAGAAATTATTGTAAAGC-3 ' 6 IL-8 primer-R Reverse 5'-CGACGCCACAATCCTTGTAAT-3 '

FIG. 3 is a graph showing the relative level of mRNA of TNF-α in keratinocytes as a result of confirming the anti-inflammatory effect of the fermentation fungus. (-): negative control; (+): PolyI: C and IL-4 treated group; Dex: a positive control group dexamethasone treated group; Comparative Example 1: Taxol extract treatment group; And Example 1: Extract-treated group of fermented zipper.

FIG. 4 is a graph showing the relative level of mRNA of IL-6 in keratinocytes as a result of confirming the anti-inflammatory effect of the fermentation fungus. (-): negative control; (+): PolyI: C and IL-4 treated group; Dex: a positive control group dexamethasone treated group; Comparative Example 1: Taxol extract treatment group; And Example 1: Extract-treated group of fermented zipper.

FIG. 5 is a graph showing the relative level of mRNA of IL-8 in keratinocytes as a result of confirming the anti-inflammatory effect of the fermentation fungus. (-): negative control; (+): PolyI: C and IL-4 treated group; Dex: a positive control group dexamethasone treated group; Comparative Example 1: Taxol extract treatment group; And Example 1: Extract-treated group of fermented zipper.

As shown in FIGS. 3 to 5, it was confirmed that TNF-α, IL-6 and IL-8 overexpressed by poly I: C and IL-4 were significantly inhibited by the fermented zucchini extract treatment ( - p <0.01 compared to the control group + **: p <0.01 compared to the control group). Therefore, it was confirmed that the extract of the fermentation fungus was excellent in the anti-inflammatory effect.

Experimental Example 3: TSLP Inhibition Test of Fermentation Waste

3-1. Human skin keratinocyte cell line HaCaT cell culture

TSLP (thymic stromal lymphopoietin) is classified as a top-level cytokine that initiates type 2 helper T-cell mediated immune deficits that are recognized as the cause of atopic dermatitis. The degree of gene expression was evaluated to confirm the effect of the extract of the fermentation fungus of Example 1 on the expression of TSLP produced in human keratinocyte HaCaT.

Specifically, keratinocyte HaCaT, a constitutive cell line of human skin, was cultured in a DMEM culture medium containing 10% fetal bovine serum (FBS) and an antibiotic. The culture was carried out in a 75T-flask and a 6-well plate, and cultured in a 37 ° C incubator supplied with 5% CO ₂ . The culture medium was changed every 3 to 4 days. When the cells were overproduced, they were subcultured. Cells were dispensed into 6-well plates and washed with phosphate-buffered saline solution (PBS) 24 hours after incubation. The fermented zygote extract of Example 1 and the zygomycetum extract of Comparative Example 1 were diluted to a final concentration of 10 ppm with PolyI: C and IL-4, respectively, in DMEM without FBS and added to each well. The positive control group was treated with 1 μM of dexamethasone. After 4 hours, cDNA was synthesized and PCR was performed to evaluate gene expression level.

3-2. Identification of TSLP abatement effect using real-time PCR

The cDNA synthesized in Experimental Example 3-1 was used as a template and real-time PCR was performed using a SYBR green master mix (cyanine dye) The degree of gene expression of TSLP was evaluated. Expression of the gene was compared with the relative value by calibrated quantification of the internal control gene beta-actin by the Ct (threshold cycle) method. Table 3 shows the primer information used in the real-time PCR.

gene SEQ ID NO: primer  designation primer  direction Primer sequence TSLP 7 TSLP primer-F Forward 5'-GCTATCTGGTGCCCAGGCTAT-3 ' 8 TSLP primer-R Reverse 5'-CGACGCCACAATCCTTGTAAT-3 '

FIG. 6 is a graph showing the relative level of mRNA of TSLP in keratinocytes as a result of confirming the effect of improving the atopic dermatitis of the fermented wipe. (-): negative control; (+): PolyI: C and IL-4 treated group; Dex: a positive control group dexamethasone treated group; Comparative Example 1: Taxol extract treatment group; And Example 1: Extract-treated group of fermented zipper.

As shown in FIG. 6, it was confirmed that the TSLP overexpressed by poly I: C and IL-4 was significantly inhibited by the fermented zucchini extract treatment (- **: p <0.01 compared to the control group).

Formulation Example 1 and Comparative Formulation Examples 1 to 3: Preparation of Essence

Essences were prepared by a conventional method according to the composition components and the composition ratios as shown in Table 4, and they were Formulation Example 1 and Comparative Formulation Examples 1, 2 and 3, respectively.

Specifically, the formulation to which the extract of the fermentation fiber compartment according to Example 1 was added had the formulation according to Formulation Example 1 and Comparative Example 1, the formulation with the extract of Comparative Example 1, Comparative Example 1, Was added to Comparative Formulation Example 2, and the negative control to which the root extract was not added was Comparative Formulation Example 3.

Clear (content:% by weight) Formulation Example 1 Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 Cetearyl alcohol 0.5 0.5 0.5 0.5 Glyceryl stearate / PEG-100 stearate 0.5 0.5 0.5 0.5 PEG-40 stearate 0.8 0.8 0.8 0.8 C14-22 alcohol / C12-20 alkyl glucoside 0.2 0.2 0.2 0.2 Caprylic / capric triglyceride 3 3 3 3 Hydrogenated polydecene One One One One Cyclopentasiloxane 3 3 3 3 Purified water 53.1 50.1 50.1 50.1 glycerin One One One One Dipropylene glycol 4 4 4 4 Carbomer 8.5 8.5 8.5 8.5 Xanthan gum 10 10 10 10 Acrylate / C10-30 alkyl acrylate crosspolymer 8.5 8.5 8.5 8.5 Disodium EDTA 0.4 0.4 0.4 0.4 Tromethamine 4.5 4.5 4.5 4.5 Fermented persimmon extract 3 0 0 0 Ginseng extract 0 3 0 0 Raw material extracts Cosmetic ingredients 0 0 3 0 1,2-hexanediol One One One One Sum 100 100 100 100

Experimental Example 4: Confirmation of skin soothing effect of fermentation wipes

Experiments were conducted to evaluate the applicability of human body to Formulation Example 1 and Comparative Formulation Examples 1 to 3.

Specifically, five healthy adults (mean age 35.8 years) were evaluated for efficacy as follows.

① Transepidermal Water Loss (TEWL) was measured before the damage of normal skin on the test site (inner part of hump).

② After loading 30 μl of 1% SLS solution into each IQ chamber, damage was induced by applying the test site for 24 hours.

③ After removing the patches, the test area was washed with running water and then allowed to stand in a constant temperature and humidity room for 10 minutes.

④ The transdermal water loss was measured immediately after removal of the patch (= after damage), 1 day of product use, 2 days of product use, 6 days of product use and 14 days of product use. The recovery rate of transdermal water loss was used in the same sense as the recovery rate of damaged skin barrier.

⑤ Skin erythema of 1 day, 2 days of product use, 6 days of product use and 14 days of product use were measured immediately after removal of the patch (= after damage).

The measuring instrument was a Tewameter (CK electronic, Germany) and a Mexameter (CK electronic, Germany). For the analysis of the results, the recovery rate (%) and the reduction rate (%) of transdermal water loss in each measurement test after the use of the product were calculated according to the following formula 1 or 2, and the efficacy was evaluated.

[Equation 1]

Percutaneous water loss recovery rate (%) = (TEWL immediately after injury TEWL) / (TEWL immediately after injury-TEWL before injury) * 100

[Equation 2]

Reduced rate of erythema (%) = {(erythema index after impairment - erythema index at measurement) / TEWL after damage} * 100

FIG. 7 is a graph showing recovery rate (%) of transdermal water loss as a result of confirming the moisturizing effect and the skin barrier improvement effect of the essence including the fermentation fudge.

8 is a graph showing the improvement rate (%) of erythema of the skin as a result of confirming the erythema-improving effect of the essence containing the fermentation fudge.

FIG. 9 is a clinical picture after application of the product as a result of confirming the erythema-improving effect of the essence including the fermentation wil.

As shown in Figs. 7 to 9, the recovery rate of transdermal water loss was the highest in the formulation containing the fermentation fibrils, and the efficacy was also excellent in the erythema reduction rate.

<110> Cosmax Co., Ltd. <120> A composition for skin smoothing an extract of          fermented beetle fructus <130> PN117388 <160> 8 <170> KoPatentin 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> TNF-alpha primer-F <400> 1 gctatctggt gcccaggcta t 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TNF-alpha primer-R <400> 2 cgacgccaca atccttgtaa t 21 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> IL-6 primer-F <400> 3 tacccccagg agaagattcc 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> IL-6 primer-R <400> 4 ttttctgcca gtgcctcttt 20 <210> 5 <211> 24 <212> DNA <213> Artificial Sequence <220> IL-8 primer-F <400> 5 ccaacacaga aattattgta aagc 24 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> IL-8 primer-R <400> 6 cgacgccaca atccttgtaa t 21 <210> 7 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP primer-F <400> 7 gctatctggt gcccaggcta t 21 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP primer-R <400> 8 cgacgccaca atccttgtaa t 21

Claims (13)

A cosmetic composition for skin soothing, comprising an extract of a fermented soybean polysaccharide fermented with a strain of Aspergillus as an active ingredient. The composition of claim 1, wherein the skin soothing is anti-inflammatory, atopic dermatitis improvement, skin moisturizing, or erythema reduction. The composition according to claim 1, wherein the strain is an Aspergillus niger strain. The composition according to claim 1, wherein the fermentation is a solid fermentation. The composition of claim 1, wherein the extract is an extract of C1-C6 alcohol, water, or a mixture thereof. The composition according to claim 1, wherein the extract is an ethanol extract. The composition of claim 1, wherein the extract comprises ponciretin. A composition for skin-soothing food comprising an extract of a fermented soybean flour fermented with a strain of Aspergillus genus as an active ingredient. A pharmaceutical composition for preventing or treating skin inflammation diseases, atopic dermatitis, or skin erythema comprising an extract of a fermentation fungus fermented with a strain of Aspergillus genus as an active ingredient. Fermenting the strain to a strain of the genus Aspergillus to produce a fermentation waste; And
And contacting said fermentation fiber compartment with a C1-C6 alcohol, water, or a mixture thereof. 11. The method of claim 10, wherein the strain is an Aspergillus niger strain. 11. The method of claim 10, wherein the fermentation is a solid fermentation. 11. The method of claim 10, wherein the fermentation is performed at 25 to 35 DEG C for 60 to 65 hours at 75 to 85% humidity.

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