CN112438921A

CN112438921A - Composition for improving skin beauty comprising fermented notoginseng root extract obtained from aspergillus papulosus

Info

Publication number: CN112438921A
Application number: CN202010275128.4A
Authority: CN
Inventors: 庆瑞娟; 柳正镇; 吕炫周; 姜承贤; 朴明三
Original assignee: Cosmax Inc
Current assignee: Cosmax Inc
Priority date: 2019-08-27
Filing date: 2020-04-09
Publication date: 2021-03-05
Anticipated expiration: 2040-04-09
Also published as: KR102239415B1; CN112438921B; KR20210025316A

Abstract

The present invention relates to a composition for improving skin beauty comprising a fermented notoginseng root extract obtained by fermentation with a novel Aspergillus cristatus strain, and a method for preparing the same. According to the composition for improving skin beauty in one aspect of the present invention, the fermented notoginseng root extract fermented with the strain of aspergillus coronarius has the effects of improving skin wrinkles, enhancing skin elasticity and inhibiting skin aging, and thus is useful for improving skin beauty. According to the method for preparing the notoginseng root extract according to another aspect of the present invention, the fermented notoginseng root extract having the effect of improving skin beauty can be prepared, and the prepared extract can be applied to various fields such as cosmetics and foods.

Description

Composition for improving skin beauty comprising fermented notoginseng root extract obtained from aspergillus papulosus

Cross Reference to Related Applications

This application claims priority and benefit from korean patent application No. 10-2019-0105186, filed on 27.08.2019, which is incorporated herein by reference for all purposes as if fully set forth herein.

Technical Field

The present invention relates to a composition for improving skin beauty comprising a fermented notoginseng root extract.

Background

With age, human skin is affected by a number of intrinsic and extrinsic factors. That is, in an intrinsic aspect, since secretion of various hormones regulating metabolism is decreased and functions and cellular activities of immune cells are decreased, biosynthesis of immune proteins and organism constituent proteins required for a human body is decreased. Externally, the ultraviolet content reaching the earth surface from the sun increases due to the destruction of the ozone layer, and as environmental pollution progresses, radicals, active oxygen, and the like increase, causing various changes, for example, not only a decrease in the thickness of the skin, an increase in wrinkles, a decrease in elasticity, but also a darkening of the skin color, frequently causing problems in the skin, and an increase in moles, freckles, age spots, and the like.

The phenomena of wrinkles and decreased elasticity of the skin, drooping and dryness of the skin, etc., which generally occur in aged skin, are caused by the change or decrease in the content and arrangement of substances constituting the skin (e.g., collagen, elastin, hyaluronic acid and glycoprotein), and particularly, collagen has a function of controlling the tension and strength of the skin, so that the decrease of collagen has a very deep relationship with the aging of the skin and the formation of wrinkles. In addition, as the dermis functions in the skin to support the strength and morphology of the skin, changes in the dermis morphology play a critical role in wrinkle formation and skin sloughing as aging progresses. Therefore, a substance increasing dermal components such as collagen and elastin can be effectively applied to improve skin wrinkles, inhibit skin aging, or enhance skin elasticity.

On the other hand, Aspergillus cristatus (Aspergillus cristatus) is a microorganism that is dominant in fu zhu an brick tea (Fuzhuan brick tea) as post fermented tea (post fermented tea) in china, and is also called Golden flower Fungus (Golden flower Fungus) by chinese because it forms spores such as Golden flower. Eurotium cristatus (a. cristatus) is a safe strain that does not produce carcinogenic mycotoxins, and has been used in brick tea production for a long time. In addition, it was also reported that the presence and expression of mycotoxin genes were not observed in eurotium cristatum (a. cristatus) isolated from fu brick tea by Yongyi Ge et al (2016).

The Notoginseng radix root is root of Panax notoginseng (Burk) F.H.Chen) belonging to Araliaceae. Notoginseng radix has the effects of stopping bleeding and removing blood stasis, and can be used for treating various bleeding syndromes, such as hematemesis, epistaxis, hemoptysis, hematochezia, contusion, fracture, blood stasis, abdominal pain, abscess, pain, puerperal hemorrhage, etc. Pharmacological effects have been reported to include shortening of blood coagulation time, anti-fatigue effect, lowering of blood pressure, lowering of heart rate, increase of coronary blood flow, lipolysis, enhancement of liver function, activation of immune function, and the like.

Research is being conducted to find and develop food or cosmetic materials from natural products that have been used for a long period of time and have less side effects, but research on fermented notoginseng roots as a material is rare.

[ Prior Art document ]

[ patent document ]

(patent document 0001) korean registered patent No. 10-1419463.

[ non-patent literature ]

(non-patent document 0001) Youngyi Ge et al, "synthetic genetic and transfer microorganisms analytes of the fuzzy brick tea-transfer fungus Aspergillus cristatus", BMC genetics (2016) 17: 428.

disclosure of Invention

An aspect of the present invention provides a cosmetic composition for improving skin beauty, comprising a fermented notoginseng root extract obtained by fermenting notoginseng root using Aspergillus cristatus (Aspergillus cristatus) strain as an effective ingredient.

Another aspect of the present invention provides a food composition for improving skin beauty, comprising a fermented notoginseng root extract obtained by fermenting notoginseng root using Aspergillus cristatus (Aspergillus cristatus) strain as an effective ingredient.

Another aspect of the present invention also provides a method for preparing a fermented notoginseng root extract, which comprises: fermenting Notoginseng radix with Monascus coronarius strain to obtain fermented Notoginseng radix; and contacting the fermented notoginseng root with a C1-C6 alcohol, water or a mixture thereof.

The skin beautifying improving agent can be used for improving skin wrinkles, enhancing skin elasticity or inhibiting skin aging.

The term "improving skin wrinkles" or "enhancing skin elasticity" may refer to alleviating the degree of sagging or sagging of the skin, reducing or inhibiting the formation of skin wrinkles, or improving already formed wrinkles.

The term "improvement" may refer to the alleviation of the condition or at least all actions that reduce a treatment-related parameter, such as the degree of symptoms.

The term "inhibiting skin aging" may refer to inhibiting or delaying aging of the skin, regardless of the cause of aging.

The "Aspergillus cristatus" is an Aspergillus species which forms spores like the golden flower and is therefore also called golden flower fungus (golden flower fungus). The aspergillus guani (a. cristatus) may be, but not limited to, isolated from food, soil, sea, etc., and may be, for example, isolated from fu zhuan brick tea (Fuzhuan brick tea) which is post fermented tea (post fermented tea) in china.

In the composition according to an embodiment, the strain may be Aspergillus cristatus (GF 8 strain) (accession No.: KCCM 12432P). The aspergillus guani GF8 strain is a novel strain separated from Fuzhuan tea prepared from China, and particularly can be a strain separated by a method comprising the following steps: diluting Fuzhuan tea by using a sodium chloride solution; culturing the diluted Fuzhuan tea in potato dextrose agar medium supplemented with chloramphenicol (chloremphenicol); and screening the strains forming golden colonies in the medium. The concentration of the sodium chloride solution can be 0.50% (w/w) to 0.99% (w/w), 0.60% (w/w) to 0.95% (w/w), 0.70% (w/w) to 0.90% (w/w), 0.80% (w/w) to 0.90% (w/w), 0.84% (w/w) to 0.86% (w/w) (e.g., 0.85% (w/w)). The culturing may be carried out at a temperature of 20 ℃ to 40 ℃, 25 ℃ to 35 ℃, 27 ℃ to 35 ℃, 28 ℃ to 32 ℃. The culturing may be performed for 10 hours to 100 hours, 20 hours to 90 hours, 30 hours to 80 hours, 40 hours to 70 hours, 48 hours to 65 hours, 48 hours to 60 hours.

The Notoginseng radix root is root of Panax notoginseng (Burk) F.H.Chen) belonging to Araliaceae family. The notoginseng root can be obtained by a conventional method, for example, by purchasing a product sold in the market. The notoginseng root may be sterilized notoginseng root, and the sterilization method may be performed by a conventional method in the art.

In a composition according to a particular embodiment, the fermentation may be a solid fermentation. The term "solid fermentation" may refer to fermentation performed by inoculating a microorganism to a raw material having a solid state of low moisture. The solid fermentation method has advantages in that it is not contaminated with various bacteria and the product can be easily recovered, compared to the existing fermentation methods such as a liquid fermentation method or a semi-solid fermentation method.

In the composition according to a specific embodiment, the fermented notoginseng root may be subjected to solid fermentation by inoculating a aspergillus guani (a. cristatus), specifically, a suspension of spores of the aspergillus guani (a. cristatus), to the notoginseng root.

In the composition according to a specific embodiment, the culture substrate may be converted into a functional substance by the extracellular enzyme (extracellular enzyme) of aspergillus papulosus (a. cristatus) through the fermentation. The kind of the extracellular enzyme may include, for example, Alkaline phosphatase (alkali phosphatase), Esterase (C4)), Esterase Lipase (C8)), Acid phosphatase (Acid phosphatase), α -galactosidase (alpha-galactosidase), β -galactosidase (beta-galactosidase), β -glucosidase (beta-glucosidase), N-acetyl- β -glucosaminidase (N-acetyl-beta-glucosaminidase), α -mannosidase (alpha-mannosidase), and the like, but is not limited thereto. Therefore, it is important to maintain a phase (phase) of conversion from hyphae to spores, which is the highest point of the enzyme biosynthesis amount in the life cycle (life cycle) of aspergillus guani (a. cristatus), for a long time. Therefore, in order to maximally maintain the stage of high extracellular enzyme expression of aspergillus papulosus (a. cristatus) and increase useful components of notoginseng root, appropriate fermentation temperature, time, and the like may be selected.

The fermented notoginseng root can be fermented at a temperature of 25 ℃ to 40 ℃, 25 ℃ to 35 ℃, 25 ℃ to 32 ℃, 28 ℃ to 40 ℃, 28 ℃ to 35 ℃, 28 ℃ to 32 ℃ or 30 ℃. When the fermentation temperature exceeds 35 ℃, the activity and expression amount of the enzyme may be relatively decreased because the time required for the strain to sporulate becomes shorter, and on the contrary, when the fermentation temperature is lower than 25 ℃, the conversion rate to the functional ingredient may be decreased because the reaction rate of the enzyme is slowed down.

The fermented notoginseng root may be fermented for 0.5 to 20 days, 1 to 20 days, 1.5 to 20 days, 2.5 to 20 days, 3 to 20 days, 0.5 to 10 days, 1 to 10 days, 1.5 to 10 days, 2.5 to 10 days, 3 to 10 days, 0.5 to 9 days, 1 to 9 days, 1.5 to 9 days, 2.5 to 9 days, 3 to 9 days, 0.5 to 7.5 days, 1 to 7.5 days, 1.5 to 7.5 days, 2.5 to 7.5 days, 3 to 7.5 days, 0.5 to 7 days, 1 to 7 days, 1.5 to 7 days, 2.5 to 7 days, 3 to 7 days, or 4 to 6 days. The fermentation time may be appropriately selected in consideration of the growth of hyphae depending on the fermentation temperature so as to terminate before sporulation.

For example, the fermented notoginseng root may be fermented at 25 to 35 ℃ for 1 to 10 days.

The fermented notoginseng root may be the whole of, a part of, or a material derived from the fermented notoginseng root. The fermented notoginseng root used for extraction may be obtained by grinding or fine-cutting or appropriately drying the whole, a part thereof or a material derived therefrom. The fermented notoginseng root having a moisture content of 20% or less and 15% or less (for example, 12% or less) after drying can be used.

The extract can be extracted by using a hydrophilic solvent such as 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, a C3 to C6 polyol. The alcohol can 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 alcohol, i.e., an aqueous alcohol solution. The alcohol concentration of the alcohol aqueous solution may be 1% (w/w) to 100% (w/w), for example, 1% (w/w) to 99.5% (w/w), 10% (w/w) to 100% (w/w), 20% (w/w) to 100% (w/w), 30% (w/w) to 100% (w/w), 40% (w/w) to 100% (w/w), 50% (w/w) to 100% (w/w), 60% (w/w) to 100% (w/w), 70% (w/w) to 100% (w/w), 75% (w/w) to 100% (w/w), 60% (w/w) to 90% (w/w), 70% (w/w) to 90% (w/w), 75% (w/w) to 85% (w/w), or 80% (w/w). The alcohol aqueous solution may be a methanol aqueous solution, an ethanol aqueous solution, or a butanol aqueous solution. In a composition according to a particular embodiment, the extract may be an alcoholic extract.

The extract may be extracted by a conventional method in the art, such as heat extraction, pressure extraction, ultrasonic extraction, hot water extraction, reflux cooling extraction, subcritical extraction or supercritical extraction, etc. For example, the extraction solvent may be added to the whole, a part of, or a material derived from the fermented notoginseng root and subjected to sonication, followed by stirring in a rotary shaker.

The extraction solvent may be 0.5 to 10 (v/w), 0.8 to 10 (v/w), 0.5 to 5 (v/w), 0.8 to 5 (v/w), 0.5 to 3 (v/w), 0.8 to 3 (v/w), 0.5 to 2 (v/w), 0.8 to 2 (v/w), 0.5 to 1.5 (v/w) or 1 (v/w) times of the fermented notoginseng root. For example, it may include adding 50L to 50mL of the extraction solvent to 100g of the fermented notoginseng root as a whole, a part thereof, or a material derived therefrom.

The extraction time may be appropriately selected according to the selected temperature and extraction method. For example, the extraction time may be 1 hour to 3 days, 1 hour to 2 days, 1 hour to 1 day, 5 hours to 3 days, 5 hours to 2 days, 5 hours to 1 day, 10 hours to 3 days, 15 hours 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 extraction may be performed by stirring in a rotary shaker for 24 hours after adding an extraction solvent to the whole of the fermented notoginseng root, a part thereof, or a material derived therefrom and performing sonication for 10 minutes. The extraction may be more than one extraction, for example, 1 to 5, 1 to 4, 1 to 3, 2 to 5, or 2 to 4 extractions, each of which may be performed in the same method or in a different method.

The extraction may be by known methods such as filtration to separate the plant residue from the extraction liquid. The extraction may also include removing the solvent from the resulting extract by known methods such as concentration under reduced pressure. The extraction may also include preparing the resulting extract into a dried extract by drying, such as freeze drying.

The extract may be present in an amount of 0.001% to 80%, for example, 0.001% to 60%, 0.001% to 40%, 0.001% to 30%, 0.001% to 20%, 0.001% to 10%, 0.001% to 5%, 0.01% to 60%, 0.01% to 40%, 0.01% to 30%, 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%, 0.05% to 10%, 0.05% to 5%, 0.1% to 60%, 0.1% to 40%, 0.1% to 30%, 0.1% to 20%, 0.1% to 10%, or 0.1% to 5%, based on the total weight of the composition.

The composition according to an embodiment may include a fraction of the fermented notoginseng root extract instead of the fermented notoginseng root extract as an effective ingredient. The composition according to an embodiment may further include a fraction of the fermented notoginseng root extract. The term "fraction" refers to a substance of a component into which the fermented notoginseng root extract is divided, i.e., a substance that has been fractionated. The fraction may be obtained by solvent fractionation (fractionation). The solvent fractionation may be to mix the fermented notoginseng root extract with a solvent and separate substances present in the solvent. The fraction may be a dichloromethane fraction, an ethyl acetate fraction, a butanol fraction, or a water fraction obtained by suspending the fermented notoginseng root extract in water and then sequentially fractionating the suspended extract using dichloromethane, ethyl acetate, butanol, and water.

Specifically, the dichloromethane fraction may be obtained by mixing the fermented notoginseng root extract with water, mixing the mixture with dichloromethane again, standing for a certain time, then separating a dichloromethane layer, and separating a fraction from the separated dichloromethane layer. The separation of the fractions may include removing dichloromethane from the dichloromethane layer. Further, the ethyl acetate fraction may be one obtained by mixing water remaining after the separation of the dichloromethane fraction again with ethyl acetate, standing for a certain time, then separating an ethyl acetate layer, and separating a fraction from the separated ethyl acetate layer. The separation of the fractions may include removing ethyl acetate from the ethyl acetate layer. In addition, the butanol fraction may be obtained by mixing water remaining after separating the ethyl acetate fraction with butanol again, standing for a certain time, separating a butanol layer, and separating a fraction from the separated butanol layer. The separation of the fraction may include removing butanol from the butanol layer. The conditions of the fractionation, such as temperature conditions, pressure conditions, time, amount or concentration of solvent used, stirring, and the like, may be the same as described with respect to the extraction for preparing the fermented notoginseng root extract. The fractionation may be repeated more than once, for example 1 to 5 times.

The separation of the fractions may be accomplished by known methods such as filtration. The fractionation may also include removing the solvent from the resulting fraction by a known method such as concentration under reduced pressure. The fractionation may also include concentrating and/or drying the resulting fraction. The concentration may be concentration under reduced pressure. The drying may include drying under reduced pressure, boiling dry, spray drying, drying at room temperature, or freeze drying.

The composition according to an embodiment may increase the expression of at least one selected from the group consisting of type i collagen α 1(alpha-1type 1collagen, COL1a1), fibrin 1(fibrillin 1, FBN1) and elastin (elastin, ELN), and particularly, may exhibit effects of improving skin wrinkles, enhancing skin elasticity and inhibiting skin aging by increasing the expression of COL1a1, FBN1 and ELN.

A composition according to a particular embodiment may comprise an effective amount of the extract or comprise the extract as an active ingredient. The effective amount may be appropriately selected depending on the individual. The effective amount may be determined based on factors including the following, as well as other factors well known in the physiological to medical arts: severity of disease-to-condition (condition), age, weight, health, sex of the individual, sensitivity of the individual to the extract, time of administration, route and rate of excretion, period of administration, other compositions used in conjunction or concomitantly with the composition.

The composition for improving skin beauty according to an embodiment may further include a cosmetically, dietetically or pharmaceutically acceptable excipient or carrier. The carrier may be an excipient, disintegrant, binder, lubricant, or combination thereof. The excipient may be microcrystalline cellulose, lactose, low substituted hydroxycellulose, or a combination thereof. The disintegrant may be sodium starch glycolate, anhydrous dibasic calcium phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The formulation type of the composition may be a non-oral administration dosage form. The non-oral administration dosage form may be an injection or a skin external preparation. The skin external agent can be cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, medicated bandage, lotion or their combination.

The skin external preparation may be appropriately mixed with ingredients of skin external preparations generally used in cosmetics, pharmaceuticals, and the like, for example, aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, perfumes, colorants, various skin nutrients, and the like, as necessary.

The skin external preparation can be mixed with appropriate amount of disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, metal chelating agent such as gluconic acid, caffeine, tannin, verapamil, Glycyrrhrizae radix extract, glabridin, hot water extract of Cirsium japonicum fruit, various crude drugs, tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts, and saccharides such as vitamin C, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, kojic acid, glucose, fructose, and trehalose.

The composition according to a particular embodiment may be a cosmetic composition. At this time, the extract may be prepared into a dosage form including: lotions (lotions), emollients, lotions, astringents (astringents), lotions, milk lotions (milk lotions), moisturizing lotions, nutritional liquids, massage creams, nutritional creams, moisturizing creams, hand creams, foundations, essences, nutritional essences, films, soaps, facial foams, facial cleansers, body lotions, body cleansers, suspensions, gels, powders, pastes (pastes), masks or sheet masks, or spray compositions. Compositions of such dosage forms may be prepared according to methods conventional in the art. The cosmetic composition may further comprise preservatives, stabilizers, surfactants, solvents, moisturizers, emollients, ultraviolet absorbers, preservatives, bactericides, antioxidants, pH adjusters, organic and inorganic pigments, fragrances, cold sensates or antiperspirants. The mixing amount of the additional ingredients such as the humectant and the like can be easily selected by those skilled in the art within a range not impairing the object and effect of the present invention, and can be 0.001% to 5%, specifically, 0.01% to 3%, based on the total weight of the composition.

The composition according to a particular embodiment may be a food composition. At this time, a formulation of a conventional health food known in the art may be prepared. The food composition may be prepared in the general dosage forms such as powder, granule, tablet, pill, capsule, suspension, oil, syrup, maceration, liquid, extractant, etc., and in any health food forms such as meat, sausage, bread, chocolate, candy, snack, biscuit, pizza, stretched noodles, other noodles, chewing gum, jelly, dairy products including ice cream, various soups, beverages, tea, drinkable preparation, alcoholic beverage, and vitamin complex. In order to formulate the health food, a dietetically acceptable carrier or additive may be used, and any carrier or additive known in the art may be used to prepare a formulation desired to be prepared. The additives may include various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acids and salts thereof, alginic acids and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonating agents for carbonated beverages, and the like. In addition, the additives may include pulp for preparing natural fruit juice, fruit juice beverages, and vegetable beverages. These additive components may be used alone or in combination, and the proportion of the additive may be 0.001% to 5%, specifically 0.01% to 3%, based on the total weight of the composition.

The content of the extract in the food composition may be appropriately determined depending on the purpose of use (prevention or improvement). Generally, 0.01% to 15% by weight of the total weight of the food can be included, and when prepared as a beverage, a proportion of 0.02g to 10g, specifically, a proportion of 0.3g to 1g, can be contained on the basis of 100 mL.

The beverage may further include other ingredients than the extract, and may further include various flavors or natural carbohydrates, etc. generally used for beverages. The natural carbohydrate may include conventional sugars such as monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), polysaccharides (e.g., dextrin, cyclodextrin, etc.), and sugar alcohols such as xylitol, sorbitol, erythritol, etc. In addition, the flavors may include natural flavors (e.g., thaumatin, stevia extract, etc.) and synthetic flavors (e.g., saccharin, aspartame, etc.). The proportion of natural carbohydrates may typically be from about 1g to 20g, in particular from about 5g to 12g, per 100mL of beverage.

Another aspect of the present invention provides a method of improving the cosmetic appearance of skin of an individual comprising the step of applying the cosmetic, food composition to the individual. The composition is the same as described above.

In particular, the method may be a method of improving skin wrinkles, a method of enhancing skin elasticity, or a method of inhibiting skin aging in a subject.

The terms "administered," "introduced," and "transplanted" are used interchangeably and may refer to a method of achieving at least partial localization of a composition according to an embodiment to a desired site or to the placement of a composition according to an embodiment into an individual according to a route. The extract or at least a portion of the extract component of the composition according to a particular embodiment may be administered by any suitable route for delivery to a desired location in a living body.

Administration can be performed by any method known in the art. Administration can be carried out directly to the individual in any manner by routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal, or subcutaneous administration. The use may be performed systemically or locally.

The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat or cat. The subject may be a subject in need of an improvement in skin beauty such as improvement in skin wrinkles, enhancement in skin screen elasticity, or an effect of suppressing skin aging.

The administering can be daily administering the fermented notoginseng root extract to the individual in the following amounts: 0.1mg to 1000mg, e.g., 0.1mg to 500mg, 0.1mg to 100mg, 0.1mg to 50mg, 0.1mg to 25mg, 1mg to 1000mg, 1mg to 500mg, 1mg to 100mg, 1mg to 50mg, 1mg to 25mg, 5mg to 1000mg, 5mg to 500mg, 5mg to 100mg, 5mg to 50mg, 5mg to 25mg, 10mg to 1000mg, 10mg to 500mg, 10mg to 100mg, 10mg to 50mg, or 10mg to 25 mg. However, the administration amount may be variously prescribed depending on factors such as formulation method, administration method, age, body weight, sex, pathological condition, diet, administration time, administration route, excretion rate, and response sensitivity of the patient, and the like, and those skilled in the art will understand that the administration amount may be appropriately adjusted in consideration of these factors. The number of administrations may be 1 time per day or 2 or more times within a clinically acceptable side effect range, with respect to the administration site, one or two or more administrations may be given, and the total number of administration days per day or every 2 to 5 days as an interval may be 1 to 30 days at the time of one treatment. If desired, the same treatment can be repeated after an appropriate time. For animals other than humans, the administration may be performed in the same amount as humans per kg of the administration amount, or in an amount converted to the administration amount by a volume ratio (for example, average value) of organs (heart and the like) of the target animal and humans, or the like.

Another aspect of the present invention provides a method for preparing a fermented notoginseng root extract, which comprises: fermenting notoginseng root using Aspergillus cristatus (Aspergillus cristatus) strain to prepare fermented notoginseng root; and contacting the fermented notoginseng root with a C1-C6 alcohol, water or a mixture thereof.

The Notoginseng radix root, Aspergillus crustis strain, fermented Notoginseng radix root and fermented Notoginseng radix extract are the same as above.

According to a method of a specific example, the strain may be Aspergillus cristatus (GF 8 strain) (accession No.: KCCM 12432P).

According to a method of a particular embodiment, the fermentation may be a solid fermentation.

According to a method of a specific embodiment, the fermentation may be performed at 25 ℃ to 35 ℃ for 1 day to 10 days.

The fermented notoginseng root extract prepared by the method can exhibit effects of improving skin wrinkles, enhancing skin elasticity, and inhibiting skin aging by increasing the expression of at least one selected from COL1a1, FBN1, and ELN. The fermented notoginseng root extract has obviously better effects of improving skin wrinkles, enhancing skin elasticity and inhibiting skin aging than the unfermented notoginseng root extract. Therefore, the fermented notoginseng root extract prepared by the method can be used for various purposes, such as cosmetics, foods, and the like.

Drawings

FIG. 1 is a photograph of fermented Panax notoginseng roots solid-cultured with Aspergillus curvatus (A. cristatus) GF8 strain (accession No.: KCCM 12432P).

FIG. 2 shows the results of liquid chromatography-mass spectrometry (LC-MS) of Notoginseng radix in comparative examples 1 and 2 and example 1. Comparative example 1: notoginseng radix extract; comparative example 2: fermenting with Aspergillus curassis GF8 strain liquid for 4 days to obtain Notoginseng radix extract; example 1: solid fermenting with Aspergillus curassis GF8 strain for 4 days to obtain Notoginseng radix extract.

Fig. 3 is LC-MS results of notoginseng roots of comparative example 1 and examples 2 to 5. Comparative example 1: notoginseng radix extract; example 2 to example 5: a notoginseng root extract is solid-fermented with Aspergillus curassis GF8 strain for 2 days (example 2), 4 days (example 3), 6 days (example 4) or 8 days (example 5).

FIG. 4 is a graph showing the relative mRNA level (relative level) of dermal component gene COL1A1 in Normal Human Dermal Fibroblasts (NHDF) of skin origin at 20 and 65 years old. None (None): a negative control group; liquid Fermentation (LF)0 days and solid fermentation (SSF)0 days: the notoginseng root extract treatment group of comparative example 1; LF 4 days: the liquid fermented notoginseng root extract treatment group of comparative example 2; SSF 4 days: the solid fermented notoginseng root treatment group of example 1.

Fig. 5 is a graph of the relative mRNA levels of dermal component gene FBN1 in 20 and 65 year old skin-derived NHDF cells. None: a negative control group; LF 0 and SSF 0 days: the notoginseng root extract treatment group of comparative example 1; LF 4 days: the liquid fermented notoginseng root extract treatment group of comparative example 2; SSF 4 days: the solid fermented notoginseng root treatment group of example 1.

Fig. 6 is a graph of the relative mRNA levels of the dermal component gene COL1a1 in 20 and 65 year old skin-derived NHDF cells. None: a negative control group; SSF 0 day: the notoginseng root extract treatment group of comparative example 1; SSF 2 days, SSF 4 days, SSF 6 days, and SSF8 days: the solid-state fermentation notoginseng root treatment groups of example 2 to example 5, respectively.

Fig. 7 is a graph of the relative mRNA levels of dermal component gene FBN1 in 20 and 65 year old skin-derived NHDF cells. None: a negative control group; SSF 0 day: the notoginseng root extract treatment group of comparative example 1; SSF 2 days, SSF 4 days, SSF 6 days, and SSF8 days: the solid-state fermentation notoginseng root treatment groups of example 2 to example 5, respectively.

Fig. 8 is a graph of the relative mRNA levels of dermal component gene ELN in 20 and 65 year old skin-derived NHDF cells. None: a negative control group; SSF 0 day: the notoginseng root extract treatment group of comparative example 1; SSF 2 days, SSF 4 days, SSF 6 days, and SSF8 days: the solid-state fermentation notoginseng root treatment groups of example 2 to example 5, respectively.

Detailed Description

Hereinafter, the present disclosure will be described in more detail by examples. However, these embodiments are for the purpose of describing the present disclosure, and the scope of the present disclosure is not limited to these embodiments.

Comparative example 1: preparation of pseudo-ginseng root extract

The method comprises collecting 300g of Notoginseng radix (produced in Yunnan, China, available from Torrel Jingdong, Korea) and washing, adding water to make the water content of Notoginseng radix 45%, and soaking for 2 hr. Placing the Notoginseng radix with water into a metal sieve, and sterilizing at 121 deg.C for 60 min. Freeze-drying until the final water content is 12%, and pulverizing. 200mL of 80% ethanol was added to 200g of the pulverized sample, and the mixture was sonicated for 10 minutes, and then stirred in a rotary shaker at 200rpm for 24 hours and extracted. Then the extract is added intoCentrifuging at 4 deg.C and 2370G for 10 min, and filtering the supernatant with 0.22 μm filter (C)

Merck Millipore, usa). The filtered supernatant was dried in a vacuum concentrator, and then the dry weight was measured for use.

Comparative example 2: preparation of Liquid Fermentation (LF) Notoginseng radix extract

300g of the same notoginseng root as described in comparative example 1 was taken, washed and pulverized, and then 1.2L of distilled water was added and sterilized at 121 ℃ for 60 minutes. Then, an Aspergillus cristatus (Aspergillus cristatus) GF8 strain grown in a Potato Dextrose Agar (PDA) medium was made into a spore suspension, which was inoculated into a solution containing sterile notoginseng roots and liquid-fermented at 30 ℃ for 4 days. Freeze-drying until the final water content is 12%, and pulverizing. 200mL of 80% ethanol was added to 200g of the pulverized sample, and the mixture was sonicated for 10 minutes, and then stirred in a rotary shaker at 200rpm for 24 hours and extracted. The extract was centrifuged and filtered and the supernatant was dried in the same manner as described in comparative example 1, and then the dry weight was measured for use.

Example 1: preparation of Solid State Fermentation (SSF) Notoginseng radix extract

In the same manner as described in comparative example 1, 2X 10⁵A suspension of CFU (colony-forming unit)/mL of Aspergillus curdatus GF8 spores was inoculated into sterilized Notoginseng radix roots and cultured at 30 ℃ for 4 days to perform solid fermentation. Then freeze-dried until the final moisture content is 12% and pulverized. 200mL of 80% ethanol was added to 200g of the pulverized sample, and the mixture was sonicated for 10 minutes, and then stirred in a rotary shaker at 200rpm for 24 hours and extracted. The extract was centrifuged and filtered and the supernatant was dried in the same manner as described in comparative example 1, and then the dry weight was measured for use.

Example 2 to example 5: preparation of solid fermented notoginseng root extract

A solid fermented notoginseng root extract was prepared in the same manner as described in example 1, except that after the aspergillus papulosus GF8 spore suspension was inoculated to the sterilized notoginseng root, it was cultured at 30 ℃ for 2 days (example 2), 4 days (example 3), 6 days (example 4) or 8 days (example 5).

Preparation example 1: preparation of essence formulation added with notoginseng root or fermented notoginseng root extract

The essence formulation was prepared according to the composition and content of table 1 below by a conventional method.

[ TABLE 1 ]

Experimental example 1: liquid chromatography-mass spectrometry analysis of solid fermented pseudo-ginseng root metabolites

The change pattern of the pseudo-ginseng root metabolites after solid fermentation is researched by liquid chromatography-mass spectrometry (LC-MS).

Specifically, LC-MS analysis was performed using a UHPLC (Ultra-high-performance LC) mass spectrometer (UHPLC-Orbitrap-IT-MS/MS, Thermo Fisher Scientific, USA). The samples of comparative example 1, comparative example 2 and examples 1 to 5 were injected into a chromatography column (Hypersil Gold C18 column) at a rate of 0.3mL/min by 5. mu.L each, and the mobile phase consisted of solution A (0.1% (v/v) aqueous formic acid solution) and solution B (0.1% (v/v) acetonitrile formate solution), and the temperature of the Probe heater (Probe heater) was adjusted to 300 ℃ for analysis. The LC-MS patterns obtained from the analysis results are shown in fig. 2 and 3.

As a result, as shown in fig. 2, it can be seen that the notoginseng root metabolites vary depending on the fermentation method, i.e., liquid fermentation or solid fermentation. In addition, as shown in fig. 3, it can be seen that the notoginseng root metabolites vary according to the fermentation time in the solid fermentation.

Experimental example 2: confirming the Gene expression increasing action of dermal Components in solid fermented Panax notoginseng root

To confirm the gene expression increasing effect of the solid-fermented notoginseng root dermal components of the skin, real-time Polymerase Chain Reaction (PCR) was performed.

Specifically, Normal Human Dermal Fibroblasts (NHDF) collected from human skin of 20 and 65 years old were treated at 3.5X 10⁵The density of individual cells/well was plated in 6-well plates and incubated at 37 ℃ in an incubator with 5% carbon dioxide for 24 hours. Next, after removing the medium and washing with Dulbecco's Phosphate-Buffered Saline (DPBS) and changing to serum-free (serum) -medium, 10. mu.g/mL of each of the samples of comparative example 1, comparative example 2 and examples 1 to 5 was treated for further culture for 24 hours.

Then, the cells were recovered with Accutase (Merck, USA), and the cell recovery kit (RNAprep, USA) was used

Total RNAprep Kit, NEB, USA) to isolate RNA. After quantification of the isolated RNA at 260nm using NanoDrop, 2. mu.g of RNA was synthesized into complementary DNA (cDNA) using an amplification instrument (C1000Thermal Cycler, Bio-Rad, USA). Each of the synthesized cDNAs was mixed with the primers and SYBR-Green mixture (SYBR-Green supermix, Applied Biosystems, USA) of Table 2 below to perform real-time PCR. The real-time PCR reaction was performed for 40 cycles under the following conditions: after 5 minutes of polymerase activation at 94 ℃, polymerase activation was carried out for 30 seconds at 95 ℃, 30 seconds at 55 ℃ and 30 seconds at 72 ℃.

[ TABLE 2 ]

Relative mRNA levels of each of collagen type i α 1(alpha-1type 1collagen, COL1a1), fibrin 1(fibrillin 1, FBN1) and elastin (elastin, ELN) genes as human dermal components were corrected and analyzed based on mRNA expression amounts of β -actin (β -actin) genes, as shown in fig. 4 to 8.

As a result, as shown in fig. 4 and 5, it was confirmed that the relative mRNA expression amounts of COL1a1 and FBN1 in solid fermented notoginseng root (example 1) were significantly higher than those of notoginseng root (comparative example 1) and liquid fermented notoginseng root (comparative example 2) in both cells, and that the mRNA expression of COL1a1 and FBN1 was increased to a level similar to that of the control group of 25-year-old-derived NHDF cells in the naturally-aged 65-year-old-derived NHDF cell group treated with solid fermented notoginseng root.

In addition, as shown in fig. 6 and 7, the mRNA expression of COL1a1 and FBN1 was increased in notoginseng roots (examples 2 to 5) that were solid-fermented for 2 days, 4 days, or 6 days in 20-year-old-derived NHDF cells and in notoginseng roots (examples 2 to 5) that were solid-fermented for 2 days, 4 days, 6 days, or 8 days in 65-year-old-derived NHDF cells, as compared to notoginseng roots (comparative example 1) before fermentation.

As shown in fig. 8, mRNA expression of ELN was increased in the notoginseng roots that were solid-fermented for 2 days, 4 days, 6 days, or 8 days in 20-year-old-derived NHDF cells (examples 2 to 5) and in the notoginseng roots that were solid-fermented for 4 days, 6 days, or 8 days in 65-year-old-derived NHDF cells (examples 3 to 5), as compared to the notoginseng roots before fermentation (comparative example 1).

That is, as can be seen from the results shown in fig. 6 to 8, the expression increasing patterns of COL1a1, FBN1 and ELN are different depending on the solid fermentation time of notoginseng root, and the effect is excellent particularly in notoginseng root which is solid-fermented for 4 days or 6 days.

According to a composition for improving skin beauty in one aspect of the present invention, the use of fermented notoginseng root extract fermented with aspergillus guani (a. cristatus) strain can improve skin wrinkles, enhance skin elasticity, inhibit skin aging.

According to the method for preparing the fermented notoginseng root extract according to another aspect of the present invention, the fermented notoginseng root extract having the effect of improving skin beauty can be prepared, and the prepared extract can be applied to various fields such as cosmetics and foods.

Claims

1. A cosmetic composition for improving skin beauty comprises a fermented Panax notoginseng root extract obtained by fermenting Panax notoginseng root with a strain of Aspergillus coronarius as an effective ingredient.

2. The composition according to claim 1, wherein the improvement in skin beauty is improvement in skin wrinkles, enhancement in skin elasticity or inhibition of skin aging.

3. The composition according to claim 1, wherein the strain is Aspergillus papulosus GF8 strain (accession No: KCCM 12432P).

4. The composition of claim 1, wherein the fermentation is a solid fermentation.

5. The composition of claim 1, wherein fermenting the notoginseng root is carried out for 1 to 10 days.

6. The composition of claim 1, wherein the fermenting notoginseng root is performed at 25 ℃ to 35 ℃.

7. The composition of claim 1, wherein the extract is an extract of C1-C6 alcohol, water, or a mixture thereof.

8. The composition of claim 1, wherein the extract is an ethanol extract.

9. The composition of claim 1, wherein the extract is present in an amount of 0.001% to 10% based on the total weight of the composition.

10. The composition of claim 1, wherein the composition increases the expression of at least one selected from the group consisting of type i collagen alpha 1, fibrin 1, and elastin.

11. The composition according to claim 1, which is prepared in at least one formulation selected from the group consisting of a lotion, a smoothing lotion, a toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrient solution, a massage cream, a nutrient cream, a moisturizing cream, a hand cream, a foundation, an essence, a nutrient essence, a film, a soap, a cleansing foam, a cleansing milk, a cleansing cream, a body lotion, a body cleaning solution, a suspension, a gel, a powder, a stick, a pack or a sheet pack, or a spray composition.

12. A food composition for improving skin beauty comprises a fermented Panax notoginseng root extract obtained by fermenting Panax notoginseng root with a strain of Aspergillus coronarius as an effective ingredient.

13. A method for preparing a fermented Panax notoginseng root extract, comprising:

fermenting Notoginseng radix with Monascus coronarius strain to obtain fermented Notoginseng radix; and

contacting said fermented notoginseng root with a C1-C6 alcohol, water or a mixture thereof.

14. The method according to claim 13, wherein the strain is Aspergillus papulosus GF8 strain (accession No: KCCM 12432P).

15. The method of claim 13, wherein the fermentation is a solid fermentation.

16. The method of claim 13, wherein the fermentation is performed for 1 to 10 days.

17. The method of claim 13, wherein the fermentation is performed at 25 ℃ to 35 ℃.