Cosmetic composition for skin improvement

Abstract

The present invention relates to a skin-improving cosmetic composition comprising a polysaccharide, a yeast extract, and a fermentation product of a strain having probiotic properties as effective ingredients, harmless to humans and having a skin condition-improving function, the cosmetic composition being provided for use in improving a microbial flora on the skin, for skin soothing, for skin wrinkle improvement, for skin elasticity improvement, for scalp soothing, for scalp oil improvement, for hair loss prevention, or for hair growth improvement.

Description

[0001] This application is a divisional application. The international application number of the original application is PCT / KR2021 / 001717, the international application date is February 9, 2021, the Chinese national application number is 202180012425.2, the entry date into the Chinese national phase is August 2, 2022, and the invention title is "Cosmetic composition for skin improvement comprising polysaccharides, yeast extract and fermentation products of strains with probiotic characteristics as active ingredients". Technical Field

[0002] This invention relates to a topical composition for skin improvement, comprising polysaccharides, yeast extracts, and ferments of strains with probiotic properties as active ingredients. Background Technology

[0003] As living standards improve, people are not only concerned about health, but also paying more and more attention to beauty. As a result, people generally strive to have beautiful skin, leading to the emergence of various cosmetic compositions for skin management.

[0004] From this perspective of beauty, hair plays a crucial role. Hair is a fine, keratinized structure that grows from the surface of the skin. This not only cushions against external impacts but also protects the body from external stimuli such as direct sunlight, cold, friction, and danger, allowing harmful heavy metals like arsenic, mercury, and zinc to be excreted from the body. Furthermore, hair plays an increasingly important role in modern cosmetic applications.

[0005] Various factors, primarily changes in diet or increased internal and external stress, can lead to a deterioration in hair condition, including hair loss. Since the scalp and hair are usually interdependent, the condition of the scalp is one of the most important factors affecting hair (Int.J.Trichology.,2018Nov-Dec;10(6):262-270).

[0006] On the other hand, topical skin care compositions that primarily focus on scalp and hair management come in various forms in terms of raw materials, appearance, and function. In most cases, artificial compounds are added to kill harmful bacteria in the skin or to regulate oil content. While such artificial compounds have excellent functional advantages, they can sometimes irritate the skin and, in the long run, become a cause of skin damage.

[0007] Therefore, there is a need to develop a composition that improves upon the problems of previous topical skin-applied compositions, is harmless to the human body, and can effectively improve skin condition. Summary of the Invention

[0008] The problem that the invention aims to solve

[0009] This invention was developed to solve the problems of the prior art as described above. The purpose of this invention is to provide a cosmetic composition for skin improvement that contains plant-derived polysaccharides, yeast-derived extracts, and fermented strains with probiotic properties as active ingredients, thereby achieving the effect of increasing beneficial bacteria or reducing harmful bacteria on the skin.

[0010] Methods for solving problems

[0011] This invention provides a cosmetic composition for skin improvement containing polysaccharides, yeast extract derived from microorganisms, and fermented products derived from microorganisms, i.e., fermented products of strains with probiotic characteristics, as active ingredients.

[0012] The term "probiotics" as used in this invention refers to microorganisms that exist in the human body and have beneficial effects on human health, including dead bacterial cells, extracts, and fermentation products of these microorganisms. "Prebiotics" refers to foods used to promote the reproduction, growth, and activity of these microorganisms.

[0013] The term "microbial colony" used in this invention refers to a group of microorganisms that exist normally in a specific part of the human body and affect human health.

[0014] In this invention, the fermentation product of the strain with probiotic characteristics cannot achieve the meaningful effect of increasing beneficial bacteria or reducing harmful bacteria to improve skin condition. However, it has been confirmed that the composition including the fermentation product of the above strain and probiotics, i.e., polysaccharides and other probiotics, i.e., extracts derived from yeast, exhibits prebiotic properties that promote the growth and reproduction of beneficial bacteria. Therefore, the above composition is provided as a composition for improving scalp or skin condition.

[0015] In this invention, the polysaccharide is selected from one or more of the group consisting of inulin, β-glucan, and maltodextrin. Such polysaccharides are beneficial in enhancing immunity and reducing oxidative stress in cells by increasing the activity of immune cells in the human body.

[0016] Inulin, a polysaccharide used as an energy storage agent in many plants such as chicory, wheat, onion, banana, garlic, and asparagus, is generally obtained from the roots or rhizomes of these plants. When applied to the skin, inulin can protect prebiotic microorganisms present in the skin as an antibacterial agent. Inulin is represented by the following chemical formula 1 (C... 6n H 10n+2 O 5n+1 It includes a glucose moiety at the end of the chain and repeating fructose moiety in between.

[0017] [Chemical Formula 1]

[0018]

[0019] The number n mentioned above can be from 2 to 60, but is not limited to this. Inulin in this invention can be obtained from chicory root, but can also be purchased and used from domestic and international markets.

[0020] Beta-glucan is a polysaccharide formed in the cell walls of grains, bacteria, and fungi. When applied to the skin, beta-glucan enhances the antioxidant activity of skin cells, prevents wrinkles, improves UV protection, wound healing, and moisturizes. The beta-glucan represented by the following chemical formula 2 includes repetitive glucose moieties linked at the 1,3 positions of the beta.

[0021] [Chemical Formula 2]

[0022]

[0023] The number n mentioned above can be from 2 to 100, but is not limited to this. In this invention, β-glucan is obtained by extraction from mushroom fruiting bodies, but it can also be purchased and used from domestic and international markets.

[0024] Maltodextrin is a polysaccharide formed through the partial hydrolysis of plant starch. When applied to the skin, maltodextrin enhances the antioxidant activity of skin cells. The maltodextrin represented by the following chemical formula 3 includes a repetitive glucose moiety with glycosidic bonds at the alpha 1,4 positions.

[0025] [Chemical Formula 3]

[0026]

[0027] The number n can be 2 to 20, but is not limited thereto. In this invention, corn or wheat starch can be obtained by hydrolysis, but it can also be purchased from domestic or international markets.

[0028] The yeast extract in this invention includes extracts obtained by solvent extraction of yeast, components obtained by hydrolyzing yeast cells, yeast culture media containing both yeast and culture medium, extracts obtained from the aforementioned components or culture media, filtrates from which yeast is filtered out of the aforementioned components or culture media, and diluted or dried products of the aforementioned components or extracts. For example, the yeast extract is a component obtained by extracting yeast using pharmaceutically permissible organic solvents or by hydrolyzing yeast cells using enzymes and then separating the extract using a solvent.

[0029] In this invention, the yeast extract is selected from one or more yeast extracts derived from beer and yeast extracts derived from truffles.

[0030] The yeast extract derived from beer is obtained by culturing yeast of the genus *Saccharomyces* (corresponding to ascomycetes) or by culturing purchased yeast of the same species, and then extracting the sterilized culture. Preferably, the yeast is *Saccharomyces cerevisiae*. *Saccharomyces cerevisiae* is also known as baker's yeast and is used in the manufacture of alcoholic beverages, primarily beer, and bread. The yeast extract derived from beer is obtained by drying the isolated yeast after filtering the beer following wort fermentation in a beer-making process using *Saccharomyces cerevisiae*. The yeast extract derived from beer is non-fermentable and contains a large amount of carbohydrates, proteins, nucleic acids, B vitamins, and minerals such as phosphorus. B vitamins have skin health and hair condition-improving effects. In this invention, the yeast used in the beer-making process is isolated, cultured, and then sterilized; however, it can also be purchased and used from domestic and international markets.

[0031] Yeast extract derived from truffles (matsutake mushrooms) is an extract obtained by isolating and culturing yeast from microorganisms present in truffles, or by culturing purchased yeast of the same species found in truffles and then sterilizing the culture. Yeast extract derived from truffles contains a large amount of β-glucan, including small amounts of B vitamins. The extract can be obtained by the methods illustrated above in this invention, but it can also be purchased and used from domestic and international markets.

[0032] The strains possessing probiotic characteristics in this invention are selected from one or more anaerobic strains belonging to the group consisting of *Lactobacillus* and *Bifidobacterium*. Therefore, the fermentation products of strains possessing probiotic characteristics include, but are not limited to, the fermentation metabolites of the aforementioned strains.

[0033] The fermentation product of this invention includes not only fermented substances obtained from the strain, but also culture media containing the strain and the culture medium, fermentation product obtained by filtering the strain from the aforementioned culture medium, fermentation product obtained by sterilizing the strain from the aforementioned culture medium and then filtering it, the aforementioned fermentation product or extracts extracted from culture media containing these, dilutions or dried products of the aforementioned fermentation product or extracts, and soluble products obtained by breaking down the bacterial cells of the strain. Preferably, the fermentation product refers to fermentation soluble products and fermentation filtrates.

[0034] For example, fermentation products include fermentation lysates and fermentation filtrates. Fermentation lysates are the product obtained after fermentation with anaerobic strains, followed by the inactivation of the bacteria using conditions such as heat, pH, enzymes, and pressure. Fermentation lysates include not only fermentation metabolites but also various useful components from the strain. Fermentation filtrates are the supernatant obtained after fermentation with anaerobic strains, through separation methods such as filtration to remove the dominant microorganisms of that strain.

[0035] Lactobacillus strains are Gram-positive bacteria distributed throughout the natural environment, including the human body, plants, and soil. Lactobacillus strains produce lactic acid through the fermentation of six-carbon sugars. As a metabolic byproduct in the human body, lactic acid or hydrogen peroxide produced by Lactobacillus strains inhibits the growth of harmful bacteria, primarily pathogenic bacteria.

[0036] The *Lactobacillus* strains in this invention include homolactic fermentation strains that ferment one 6-carbon sugar molecule into two lactic acid molecules, and heterolactic fermentation strains that generate lactic acid molecules, ethanol, acetic acid, carbon dioxide, etc., from 6-carbon sugar molecules. For example, *Lactobacillus* strains may include *Lactobacillus acidophilus*, *Lactobacillus delbrueckii*, *Lactobacillus helveticus*, *Lactobacillus salivarius*, *Lactobacillus casei*, *Lactobacillus plantarum*, *Lactobacillus savei*, *Lactobacillus brevis*, *Lactobacillus buchneri*, *Lactobacillus fermentum*, *Lactobacillus reuteri*, *Lactobacillus rhamnosus*, *Lactobacillus paracasei*, *Lactobacillus Johnsonii*, *Lactobacillus bulgaricus*, etc., but are not limited to these. Preferably, the *Lactobacillus* strain is at least one of *Lactobacillus acidophilus*, *Lactobacillus casei*, *Lactobacillus plantarum*, *Lactobacillus fermentum*, *Lactobacillus paracasei*, and *Lactobacillus bulgaricus*. More preferably, the *Lactobacillus* strain is at least one of *Lactobacillus acidophilus*, *Lactobacillus plantarum*, *Lactobacillus fermentum*, and *Lactobacillus paracasei*. Most preferably, the *Lactobacillus* strain is at least one of *Lactobacillus acidophilus* and *Lactobacillus plantarum*.

[0037] Therefore, in this invention, the fermentation product of the Lactobacillus strain includes the fermentation metabolite of the above-mentioned strain with a relative 6-carbon sugar content, preferably the fermentation lysate from which the above-mentioned strain has been killed.

[0038] Bifidobacterium strains are Gram-positive bacteria found throughout the natural environment, including human skin, food, and soil. They are heterolactic fermentation strains that produce lactic acid, acetic acid, formic acid, and other acids through the unique fermentation metabolism of a 6-carbon sugar, fructose-6-phosphate phosphoketolase.

[0039] The Bifidobacterium strains used in this invention include, for example, *Bifidobacterium animalis*, *Bifidobacterium bifidum*, *Bifidobacterium breve*, *Bifidobacterium dentium*, *Bifidobacterium longum*, and *Bifidobacterium pseudolongum*, but are not limited thereto. Preferably, the Bifidobacterium strain is at least one of *Bifidobacterium animalis*, *Bifidobacterium bifidum*, and *Bifidobacterium longum*.

[0040] The fermentation products of Bifidobacterium strains in this invention include fermentation metabolites of the above-mentioned strains with a relative 6-carbon sugar content, preferably fermentation lysates from strains that have been inactivated.

[0041] One embodiment of the present invention provides a skin-improving cosmetic composition comprising inulin, β-glucan, maltodextrin, a yeast extract derived from beer, a yeast extract derived from truffles, lactobacillus ferment lysate, or bifidobacteria ferment lysate as active ingredients.

[0042] Preferably, the cosmetic composition comprises inulin, β-glucan, maltodextrin, a yeast extract derived from beer, a yeast extract derived from truffles, lactobacillus ferment lysate, and bifidobacteria ferment lysate in the same ratio. For example, the cosmetic composition comprises inulin, β-glucan, maltodextrin, a yeast extract derived from beer, a yeast extract derived from truffles, lactobacillus ferment lysate, and bifidobacteria ferment lysate in a weight ratio of a:b:c:d:e:f:g. Here, the distribution of a to g is 1 to 10, preferably a to g are each 1 to 5, and most preferably a to g are all 1.

[0043] An embodiment of the cosmetic composition for skin improvement of the present invention comprises, relative to 100% by weight, 0.00001% to 10% by weight of one or more active ingredients selected from the group consisting of inulin, β-glucan, maltodextrin, yeast extract derived from beer, yeast extract derived from truffles, lactobacillus ferment lysate, and bifidobacteria ferment lysate. Preferably, it comprises 0.0001% to 1% by weight, more preferably 0.0005% to 0.5% by weight, and most preferably 0.001% to 0.1% by weight.

[0044] The term "skin improvement" used in this invention includes not only improvements in skin condition as observed by the naked eye, but also factors that have a direct and indirect impact on skin health.

[0045] Specifically, the skin-improving cosmetic composition of the present invention is a composition for improving the microbial flora inhabiting the skin. Improving the microbial flora includes promoting the growth of beneficial bacteria present on the skin, thereby increasing beneficial bacteria, or inhibiting the growth of harmful bacteria, thereby reducing harmful bacteria.

[0046] Specifically, the skin-improving cosmetic composition of the present invention is any one of a skin-soothing composition, a skin wrinkle-improving composition, and a skin elasticity-improving composition. Skin soothing includes preventing or inhibiting skin inflammation and other skin impairments or normalization. Skin wrinkle improvement or elasticity improvement includes preventing or inhibiting damage or volume reduction of the dermis caused by natural causes such as aging and external environmental causes such as ultraviolet radiation and oxidative stress.

[0047] Specifically, the skin-improving cosmetic composition of the present invention is applied to the scalp. For example, the scalp-improving cosmetic composition is any one of a scalp-soothing composition, a scalp-oil-reducing composition, a hair loss prevention composition, and a hair growth improvement composition. Skin soothing refers to preventing or suppressing inflammation on the scalp. Scalp-oil-reducing means reducing the amount of oil secreted when there is excessive oil secretion on the skin. Hair loss prevention or hair growth improvement includes increasing the activity of human hair papilla cells in the scalp or inhibiting inflammatory factors on the cells that make up the skin or increasing the rate of collagen biosynthesis in the cells that make up the skin.

[0048] Preferably, "skin improvement" refers to increasing the number of beneficial bacteria present on the skin and reducing harmful bacteria. The beneficial and harmful bacteria are distinguished based on the effect of the strain on human skin health; for example, they may be strains of the genus Staphylococcus, but are not limited to this.

[0049] For example, strains of Staphylococcus spp., such as Staphylococcus epidermidis, which are epidermal Staphylococci, do not act as pathogens on normal skin. Instead, they produce antimicrobial peptides against other pathogens or have the effect of inhibiting the development of some cancers, and therefore can be classified as beneficial bacteria.

[0050] For example, strains of the genus Staphylococcus, such as Staphylococcus aureus (also known as Staphylococcus faecalis), can cause various skin infections by existing on the skin surface or in pores, and may also cause disease on normal skin; therefore, they are classified as harmful bacteria. In the case of the scalp, harmful bacteria include Aspergillus niger, which produces mycotoxins and causes inflammation of the scalp.

[0051] As described later, the composition of one embodiment of the present invention provides a skin-improving effect by increasing beneficial bacteria and the like, thereby improving the flora in the human body.

[0052] Human dermal papilla cells, located in the middle and outer part of hair follicles on the scalp, play a crucial role in controlling hair formation and the hair growth cycle. Excessive sebum production on the scalp can trigger dermatitis, with inflammatory factors inducing an inflammatory response against scalp cells. Collagen, a protein found in the extracellular matrix that forms the binding tissue, can lead to decreased collagen biosynthesis in the scalp, resulting in dermal deterioration and consequently, shrinkage of hair follicles.

[0053] As described below, the composition of one embodiment of the present invention can provide the following effects: increase the activity of specific cells; reduce skin oiliness; inhibit the production of inflammatory factors in skin cells; and improve the skin by increasing the rate of collagen biosynthesis.

[0054] The term "cosmetic composition" as used in this invention refers to a composition provided by direct application to the skin or by spraying, etc. The term "provided" as used in this invention means the composition of this invention being delivered to a target tissue using methods conventional in the art.

[0055] The skin-improving cosmetic composition of the present invention can be in the form of lotion, emulsion, moisturizing lotion, nourishing cream, hydrating cream, eye cream, serum, cosmetic ointment, spray, gel, mask, sunscreen, liquid foundation, foundation cream, powder, facial cleanser, facial cleanser milk, facial cleanser oil, facial cleanser, soap or shower gel.

[0056] In the case of the scalp or hair, the skin improvement cosmetic composition of the present invention can be in the form of hair growth tonic, hair conditioner, hair essence, hair lotion, hair nourishing lotion, shampoo, hair conditioner, hair treatment liquid, hair cream, hair nourishing cream, hair moisturizing cream, hair massage cream, hair wax, hair spray, hair mask, hair nourishing mask, shampoo bar, shampoo milk, hair oil, hair dryer, hair preservation treatment, hair dye, hair wave agent, hair bleaching agent, hair gel, hair conditioner, hair dye, hair moisturizer, mousse or hair spray.

[0057] When the composition of the present invention is formulated into a liquid, the composition of the present invention includes a carrier for the above-mentioned active ingredient. The carrier may be, for example, water, ethanol, castor oil, glycerol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol oil, glycerol aliphatic esters, polyethylene glycol, or fatty acid esters such as sorbitan, but is not limited thereto. These may be used alone or in combination of two or more.

[0058] When the compositions of the present invention are formulated as pastes, creams, or gels, the compositions of the present invention include a carrier for the above-mentioned active ingredients. Such carriers are, for example, animal oils, vegetable oils, waxes, paraffin wax, starch, tragacanth gum, cellulose derivatives such as hydroxyethyl, polyethylene glycol, silicon, bentonite, silica, talc, zinc oxide, hexadecyl stearyl alcohol, or stearyl triethylammonium chloride, but are not limited thereto. These can be used alone or in combination of two or more.

[0059] When the compositions of the present invention are formulated as powders or sprays, the compositions of the present invention may include a carrier for the above-mentioned active ingredients. Such carriers are, for example, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powders, but are not limited thereto. These can be used alone or in combination of two or more. When the compositions of the present invention are formulated as sprays, they may also include a propellant such as chlorofluorocarbons, propane, butane, or dimethyl ether.

[0060] When the composition of the present invention is formulated as a soap, the composition of the present invention includes a carrier of the above-mentioned active ingredients. The carriers are, for example, alkali metal salts of fatty acids, fatty acid half-esters, fatty acid protein hydrolysates, ethyl thiophosphate, lanolin derivatives, fatty alcohols, vegetable oils, glycerin, sugars, etc., but are not limited thereto. These can be used alone or in combination of two or more.

[0061] The compositions of the present invention also include additives commonly used in cosmetic compositions. These additives include, but are not limited to, purified water, surfactants, moisturizers, lower alcohols, chelating agents, bactericides, antioxidants, preservatives, pigments, and fragrances. These can be used alone or in combination of two or more.

[0062] This invention provides the use of one or more of the following: polysaccharides, yeast extracts, and fermented strains possessing probiotic characteristics. Specifically, this invention provides the use of one or more of the following in a skin-improving cosmetic composition: polysaccharides, yeast extracts, and fermented strains possessing probiotic characteristics. More specifically, this invention provides the use of one or more of the following in a method for manufacturing a skin-improving cosmetic composition: polysaccharides, yeast extracts, and fermented strains possessing probiotic characteristics.

[0063] In addition, the present invention provides the use of any one or more of the following in skin improvement methods, skin microbial flora improvement methods, skin soothing methods, skin wrinkle improvement methods, skin elasticity improvement methods, scalp soothing methods, scalp oiliness improvement methods, hair loss prevention methods, or hair growth improvement methods.

[0064] In addition, the present invention provides a method for improving skin by treating a subject with a cosmetic product comprising any one or more of polysaccharides, yeast extracts and ferments of strains with probiotic characteristics, a method for improving the microbial flora inhabiting the skin, a method for soothing the skin, a method for improving skin wrinkles, a method for improving skin elasticity, a method for soothing the scalp, a method for improving scalp oiliness, a method for preventing hair loss, or a method for improving hair growth.

[0065] The content of all ingredients included in this invention does not exceed the specifications stipulated by each country. Preferably, the ingredients mentioned in the cosmetic composition of this invention are included within the range not exceeding the maximum usage specified in the "Cosmetic Safety Technical Specifications" designated by the Chinese government.

[0066] Invention Effects

[0067] The skin-improving cosmetic composition of the present invention comprises polysaccharides as prebiotics, extracts from yeast as probiotics, and fermentation products of yeast strains. Compared with the application of prebiotics or probiotics alone, it achieves excellent effects in promoting the growth of beneficial bacteria in the skin and inhibiting harmful bacteria. Such a cosmetic composition is harmless to the human body and improves skin condition by improving the skin's flora.

[0068] Specifically, the skin-improving cosmetic composition of the present invention improves scalp health and hair condition by increasing the activity of human hair papilla cells.

[0069] In addition, the skin-improving cosmetic composition of the present invention can improve skin condition by inhibiting the production of inflammatory factors in skin cells.

[0070] In addition, the skin-improving cosmetic composition of the present invention increases the rate of collagen biosynthesis in skin cells, thereby improving the condition and health of the skin.

[0071] In addition, the skin-improving cosmetic composition of the present invention removes foreign matter and oil from the skin without side effects, thereby improving the oil-water balance of the skin surface. Attached Figure Description

[0072] Figure 1 These are scalp photographs of the shampoo according to an embodiment of the present invention, taken before (a) application and after (b) one application.

[0073] Figure 2 These are scalp photographs of the shampoo according to an embodiment of the present invention, taken before (a) application and after (b) one application.

[0074] Figure 3 These are scalp photographs of the shampoo according to an embodiment of the present invention, taken before (a) application and after (b) one application. Detailed Implementation

[0075] The objectives, specific advantages, and novel features of the present invention will become clearer with reference to the following detailed description and preferred embodiments. However, these embodiments are merely illustrative and the scope of the invention is not limited to these embodiments. Furthermore, in describing the present invention, detailed descriptions of relevant prior art are omitted where such detailed descriptions would obscure the essence of the invention.

[0076] Manufacturing example: A complex comprising polysaccharides, yeast extract, and fermented products of strains possessing probiotic properties. Regarding the inulin (Beneo), β-glucan (Quegen Biotech Co., Ltd.), maltodextrin (samyang), and beer-derived yeast extract (activon) used in the following examples, commercially available products were used. The truffle-derived yeast extract was an extract obtained from yeast isolated from truffles, cultured at 37°C for 48 hours, and then sterilized. The Lactobacillus fermentation lysate and Bifidobacterium fermentation lysate were extracts obtained from Lactobacillus and Bifidobacterium (Chr. Hansen) cultures, cultured at 37°C for 48 hours, and then sterilized.

[0077] In Example 1, a complex was manufactured by mixing 10g of inulin, a yeast extract derived from beer, and a Lactobacillus ferment lysate. In Example 2, a complex was manufactured by mixing 10g of β-glucan, a yeast extract derived from beer, and a Lactobacillus ferment lysate. In Example 3, a complex was manufactured by mixing 10g of maltodextrin, a yeast extract derived from beer, and a Lactobacillus ferment lysate. In Example 4, a complex was manufactured by mixing 10g of inulin, a yeast extract derived from truffles, and a Bifidobacterium ferment lysate. In Example 5, a complex was manufactured by mixing 10g of β-glucan, a yeast extract derived from truffles, and a Bifidobacterium ferment lysate. In Example 6, a complex was manufactured by mixing 10g of maltodextrin, a yeast extract derived from truffles, and a Bifidobacterium ferment lysate. In (Manufacturing Example 7), a complex was manufactured by mixing 10g each of inulin, β-glucan, maltodextrin, yeast extract derived from beer, yeast extract derived from truffles, lactobacillus ferment lysate, and bifidobacteria ferment lysate.

[0078] Example: The active ingredients include polysaccharides, yeast extract, and fermentation products of strains possessing probiotic properties. Cosmetic composition

[0079] Example 1-1. Shampoo composition

[0080] A shampoo composition comprising the components shown in Table 1 below was manufactured.

[0081] [Table 1]

[0082]

[0083] Examples 1-2. Shampoo Composition

[0084] A shampoo composition having the composition shown in Table 2 below was manufactured.

[0085] [Table 2]

[0086]

[0087] Example 2. Hair care liquid composition

[0088] A hair care liquid composition having the composition shown in Table 3 below was manufactured.

[0089] [Table 3]

[0090]

[0091] Example 3. Hair growth tonic

[0092] A hair growth tonic composition having the composition shown in Table 4 below was manufactured.

[0093] [Table 4]

[0094]

[0095] Example 4. Shower Gel

[0096] A shower gel composition having the composition shown in Table 5 below was manufactured.

[0097] [Table 5]

[0098]

[0099] Example 5. Moisturizing Lotion

[0100] A moisturizing lotion composition having the composition shown in Table 6 below was manufactured.

[0101] [Table 6]

[0102]

[0103] Experiment Example 1. Confirming the effect on the reproduction of beneficial bacteria

[0104] To confirm the effectiveness of the compound in the manufacturing example regarding the proliferation of beneficial bacteria, *Staphylococcus epidermidis* was selected as the beneficial bacteria, and *Staphylococcus aureus* was selected as the harmful bacteria. Before use, these bacteria were stored at -80°C. Multiple 100 mL TSB (tryptophan broth) media were prepared, and 500 μL of each bacterial stock was inoculated into each medium. The cultures were then incubated at 37°C with rotation at 210 rpm for 17 hours. Afterward, 4 mL of the culture was inoculated into 40 mL of TSB and subcultured at 37°C with rotation at 210 rpm for 8 hours.

[0105] After 8 hours, the optical density (OD) value was adjusted to 0.65, and the initial OD value was measured after inoculating the complexes prepared according to Manufacturing Examples 1 to 7 above at a concentration of 1%. Subsequently, the complexes were cultured at 37°C and rotated at 210 rpm for 16 hours, and the OD value was measured again. For each culture medium, the difference between the OD value measured after 16 hours and the initial OD value was divided by the initial OD value for standardization, and the results were evaluated.

[0106] As comparative examples, products inoculated at 1% concentrations with inulin, β-glucan, maltodextrin, yeast extract derived from beer, yeast extract derived from truffles, Lactobacillus fermentation lysate, and Bifidobacterium fermentation lysate were used. An untreated control group was used.

[0107] The inoculation results of the above manufacturing examples and comparative examples were compared with the untreated group, and the average increase or decrease of beneficial and harmful bacteria were converted into percentages. The results are shown in Table 7 below.

[0108] [Table 7]

[0109] distinguish Beneficial bacteria (S. epidermidis) Harmful bacteria (S. aureus) Lactobacillus fermentation lysate -9.90 21.42 Bifidobacterium fermentation lysate -4.69 26.00 Yeast extract derived from beer 2.48 16.37 Yeast extract derived from truffles 3.39 14.95 Inulin 0.29 -0.84 β-glucan -0.26 -0.78 maltodextrin 0.14 1.43 Manufacturing Example 1 5.51 -2.46 Manufacturing Example 2 4.79 -2.88 Manufacturing Example 3 3.88 -2.31 Manufacturing Example 4 7.18 -2.49 Manufacturing Example 5 6.07 -2.71 Manufacturing Example 6 5.99 -2.19 Manufacturing Example 7 15.54 -7.47

[0110] As shown in Table 7, although inulin, β-glucan and maltodextrin are polysaccharides with prebiotic properties, they did not show a significant effect on increasing beneficial bacteria and decreasing harmful bacteria when used alone. In the case of fermented lysates with probiotic properties, they showed a decrease in beneficial bacteria and an increase in harmful bacteria when used alone.

[0111] In contrast, the complexes of Manufacturing Examples 1 to 7 of the present invention, compared with comparative examples using polysaccharides, yeast extracts, or fermentation lysates alone, exhibit significantly superior effects in increasing beneficial bacteria and reducing harmful bacteria. When polysaccharides, acting as prebiotics, are used in combination with yeast extracts and fermentation lysates, acting as probiotics, the effects of increasing beneficial bacteria and reducing harmful bacteria are further enhanced. In particular, Manufacturing Example 7 exhibits the most superior effect regarding the increase in beneficial bacteria and the reduction in harmful bacteria compared to the other manufacturing examples.

[0112] Experimental Example 2. Confirmation of the activity of human dermal papilla cells

[0113] To confirm the effect of the complex on the human dermal papilla cells, 4-8 passages of human dermal papilla cells (Promocell, C-12071) were used as the cell line, and dermal papilla cell growth media (Promocell, C-26501) was used as the initial culture medium.

[0114] Cells were seeded at a level of 3,000 to 6,000 cells per well in 96-well plates and cultured for 24 hours at 37°C and 5% CO2. The culture medium was then replaced with DMEM medium supplemented with 0.1% FBS (fetal bovine serum) and cultured for 24 hours at 37°C and 5% CO2. The culture medium was then replaced with DMEM medium supplemented with 0.1% FBS, containing the complexes from Examples 1 to 7, and cultured for 48 hours at 37°C and 5% CO2. Each well was treated with 10 μl of CCK-8 solution, and the optical density at 450 nm was measured after 1 hour of culture at 37°C.

[0115] The optical density readings of each well after 48 hours were standardized by dividing the initial reading by the initial reading, and the average values ​​are shown in Table 8 below. As a control group, 1 μM minoxidil was applied. For significant differences, p-values ​​were plotted assuming equal dispersion (p < 0.05).

[0116] [Table 8]

[0117]

[0118] As shown in Table 8, the complexes of Manufacturing Examples 1 to 7 of the present invention have the effect of improving the activity of human dermal papilla cells. In particular, Manufacturing Example 7 has the best effect of improving the activity of human dermal papilla cells compared with the other manufacturing examples.

[0119] Experimental Example 3. Confirmation of inhibition of inflammatory factors

[0120] As an inflammatory factor in cells, to confirm the effect of the complex in the manufacturing example on NO production, mouse Raw264.7 cell line was used as the initial culture medium, which was a culture medium with NaOH added to Complete RPMI (RPMI:FBS:antibiotic = 10:1:0.1) and the final pH was adjusted to 8.5.

[0121] After culturing at 37°C and 5% CO2 for 24 hours, Raw264.7 cells were seeded into 24-well plates prepared with DMEM medium when they appeared to float on the surface of the culture medium, and cultured at 37°C and 5% CO2 for another 24 hours. Subsequently, the culture medium was replaced with serum-free DMEM medium, and the complexes from Preparation Examples 1 to 7 were diluted to the following concentrations and treated with 1 μg / mL LPS. After culturing at 37°C and 5% CO2 for 24 hours, the supernatant was mixed with Griessreagent reagent (Sigma-Aldrich) at a 1:1 ratio. The NO production inhibition function was evaluated by absorbance, and the average values ​​are shown in Table 9 below.

[0122] NO generation inhibition performance (%) = {1 – (NO generation with the addition of the complex / NO generation in the untreated group)} × 100

[0123] As a control group, the NO inhibitor L-NMMA1 (N G -Methyl-L-arginine acetate salt, N G 20 μg / mL of 2-methyl-L-arginine acetate.

[0124] [Table 9]

[0125]

[0126] As shown in Table 9, the complexes of Manufacturing Examples 1 to 7 of the present invention exhibit inhibitory properties against the production of the inflammatory factor NO in cells. In particular, Manufacturing Example 7 demonstrates the best NO production inhibition performance compared to the other manufacturing examples.

[0127] Experiment Example 4. Confirming the collagen biosynthesis rate

[0128] Using human dermal fibroblasts as subjects, the effect of the manufactured complex on collagen biosynthesis in these cells was confirmed. Dermal fibroblasts were cultured in complete DMEM medium at 37°C and 5% CO2 for 72 hours. Afterwards, they were seeded into 24-well plates and cultured at 37°C and 5% CO2 for 24 hours. The manufactured complex was then treated and cultured at 37°C and 5% CO2 for 48 hours. The supernatant was separated, and the collagen synthesis effect was confirmed by absorbance using a procollagen type I C-peptide (PIP) EIA kit. The collagen biosynthesis growth rate was evaluated as follows, and the average values ​​are shown in Table 10 below.

[0129] Collagen biosynthesis growth rate (%) = {(Absorbance of the treated group – Absorbance of the untreated group) / Absorbance of the untreated group} × 100

[0130] The control group used the cytokine TGF-β 10 ng / mL.

[0131] [Table 10]

[0132]

[0133] As shown in Table 10, the complexes of Manufacturing Examples 1 to 7 of the present invention were confirmed to have the effect of enhancing collagen biosynthesis in fibroblasts. In particular, Manufacturing Example 7 showed the best effect of enhancing collagen biosynthesis in fibroblasts compared with the other manufacturing examples.

[0134] Experimental Example 5. Confirming whether scalp oiliness has improved.

[0135] The effect of using the shampoo composition of Example 1-1 on improving scalp oiliness was confirmed. Twenty-two adult women aged 20 to 50 were selected as subjects, and their scalps were chosen as the experimental sites. During this experiment, subjects were prohibited from using other shampoos, conditioners, hair serums, and hair treatments such as dyeing or perming.

[0136] The subjects were sedated for 30 minutes in a constant temperature and humidity chamber at 22±2℃ and 50±5% before the experiment was conducted. All experiments were performed in the aforementioned constant temperature and humidity chamber. After thoroughly wetting their hair and scalp with lukewarm water, the subjects applied an equal amount of the shampoo composition of Example 1-1 evenly to their hair and scalp, massaged it in, and then rinsed thoroughly with running water.

[0137] The evaluation of scalp oiliness before and after shampoo use was conducted using a sebumeter (SKIN-O-MAT, Cosmomed GmbH) and an electron microscope (KongPC Camera, Bomtech). A probe cartridge with an attached oil-absorbing strip was placed on the top of the scalp of all subjects, and contact was applied with the same pressure for 30 seconds to ensure sufficient oil absorption. The cartridge was then inserted into the sebumeter to measure the sebum level. Furthermore, the top of the scalp of all subjects was observed under the same illumination conditions using an electron microscope at 300x magnification.

[0138] The statistical analysis in this experiment was performed using SPSS 17.0 for Windows. Table 11 below shows the sebum levels (μg / cm³) before and after one application of the composition of Examples 1-1. 2 The results showed that the maximum detectable sebum amount using a sebumeter was 350 μg / cm³. 2 ).

[0139] [Table 11]

[0140] Before use After one use average 200.41 32.05 Standard deviation 37.03 14.87

[0141] Table 12 below shows the results of paired t-test analysis (*p<.05, **p<.01, ***p<.001) of the improvement rate (%) of sebum mass after one use of the composition of Example 1-1 and whether the change was significant: Improvement rate (%) = {(value after one use – value before use) / value before use} × 100

[0142] [Table 12]

[0143] Improvement rate of sebum content p-value After one use 84.01 <![CDATA[.001 *** ]]>

[0144] In addition, the results of consultations regarding the individual scalp condition of the subjects and changes in scalp condition before and after using the composition of Example 1-1 once are shown in Tables 13 and 14 below (N = total frequency = number of subjects = 22).

[0145] [Table 13]

[0146]

[0147] [Table 14]

[0148]

[0149] Additionally, consultations were conducted regarding whether any adverse reactions occurred on the scalp and hair when using the compositions of the examples, and the results are shown in Table 15 below (0: none, 1: slight, 2: moderate, 3: severe).

[0150] [Table 15]

[0151] Adverse reactions After one use Adverse reactions After one use Erythema (turning red) 0 Stinging pain (pain) 0 Edema (swelling) 0 burning sensation 0 Dandruff (keratin) 0 stiff 0 itch 0 Scratching 0

[0152] As shown in Tables 11 and 12, after applying the shampoo of Example 1-1 of the present invention once, the amount of sebum was improved by more than 80% compared with the previous application, and the effect of improving scalp oiliness was excellent. This result is consistent with the situation shown in Tables 13 to 15, in which more than 90% of the subjects answered that there were no side effects and that it had the effect of cleansing and removing sebum.

[0153] Figures 1 to 3 These are magnified photographs showing the observation results of three subjects before (a) application and after (b) one application of the shampoo in Example 1-1 of the present invention, thereby confirming its excellent cleansing and sebum removal effects.

[0154] Experimental Example 6. Confirming whether the scalp flora has improved.

[0155] The effects of using the shampoo composition of Example 1, the hair care liquid composition of Example 2, and the hair growth liquid composition of Example 3 on improving scalp flora were confirmed. Twenty-four adult women with an average age of 45 years were selected as subjects, and their scalps were chosen as the experimental sites. During this experiment, subjects were prohibited from using other shampoos, hair conditioners, hair serums, and hair treatments such as dyeing or perming.

[0156] The subjects were placed in a constant temperature and humidity chamber at 20-25°C and 40-60% humidity for 30 minutes before the experiments were conducted. All experiments were performed in the aforementioned constant temperature and humidity chamber. After thoroughly wetting their hair and scalp with slightly warm water, the subjects applied the same amount of the shampoo compositions of Examples 1-1 and 1-2 and the hair care liquid composition of Example 2 to their hair and scalp once a day, massaged it in, and then rinsed thoroughly with running water. Additionally, the subjects applied the same amount of the hair growth tonic composition of Example 3 to their scalp once a day, massaged it in, and then dried it directly without rinsing.

[0157] After the experiment, without applying shampoo or water to the hair and scalp for 30 minutes in the aforementioned constant temperature and humidity room, the hairline on the top of the head was fixed and the scalp was moved up and down 40 times with a cotton swab to collect microorganisms, which were then compared and analyzed with the microorganisms collected before the experiment.

[0158] The following statistical analysis was performed using SPSS in this experiment. The results were considered to have an improvement effect when the significance probability p < 0.05 was within the 95% confidence interval. As statistical analysis methods, the paired-samples t-test was used when using parametric methods, and the Wilcoxon signed-rank test was used when using non-parametric methods. Table 16 shows the results of ANOVA performed at the genus level before and after using the composition of the example.

[0159] [Table 16]

[0160] Genus p-value Before use After one week of use After 2 weeks of use Aspergillus p<0.01 2.21 2.01 0.78 Fusarium p<0.01 1.58 1.49 0.28 Penicillium p<0.01 1.12 0.99 0.21 Cladosporium p<0.01 1.03 0.98 0.19 Mucor p<0.01 0.66 0.71 0.2 Alternaria p<0.01 0.6 0.56 0.13 Kluyveromyces p<0.01 0.66 0.54 0.06 genus Clitocybe p<0.01 0.6 0.57 0.032 genus *Sarocladium* p<0.01 0.53 0.55 0

[0161] After using the compositions of the examples for two weeks, the flora of nine genera present on the scalp changed statistically significantly. As shown in Table 16, the Saccharomyces genera, which helps improve scalp moisturization and elasticity, began to increase statistically significantly after one week of using the compositions of Examples 1 to 3 of the present invention. The Aspergillus, Fusarium, and Penicillium genera, which cause fungal diseases or dermatitis, decreased statistically significantly after two weeks of use, thus confirming a significant improvement in the flora on the scalp.

[0162] Experimental Example 7. Confirming whether the skin flora has improved.

[0163] The skin flora improvement effects of using the shower gel composition of Example 4 and the body lotion composition of Example 5 were confirmed. Twenty-two adult women aged 20 to 60 years were selected as test subjects, and the skin on their forearms was chosen as the test site. The subjects were prohibited from using other skincare products during this experiment.

[0164] The subjects were left unwashed for more than 8 hours before the experiment was conducted. The subjects were then placed in a constant temperature and humidity chamber (20-25°C, 40-60% humidity) for 30 minutes before the experiment was performed. All experiments were conducted within the aforementioned constant temperature and humidity chamber. The subjects wetted the skin of their forearms with slightly warm water, then washed with the shower gel composition of Example 4, and evenly applied the moisturizing lotion composition of Example 5.

[0165] After the experimental period, microorganisms were collected from the skin of the forearm by moving a cotton swab up and down 40 times, and then compared with the microorganisms collected before the experiment. The statistical processing was the same as in Experiment 6 above. Table 17 shows the results of dispersion analysis (ANOVA) performed at the genus level before and after using the composition of the example.

[0166] [Table 17]

[0167] Genus pvalue Before use After one week of use After 2 weeks of use Ehrlichia p<0.001 7.43 7.54 2.62 Sphingomonas p<0.001 3.19 3.22 2.29 Pseudomonas p<0.001 1.56 1.5 3.31 Acetobacter p<0.001 2.65 2.7 0.95 Cutibacterium p<0.001 1.22 1.17 3.07 Streptococcus p<0.001 1.13 0.96 1.93 Akkermansia p<0.001 1.55 1.56 0.85 Bacteroides p<0.001 1.37 1.41 0.69 Staphylococcus p<0.001 0.72 0.72 1.81 Enterococcus p<0.001 0.93 0.92 0.80

[0168] After using the compositions of the examples for two weeks, a statistically significant change was confirmed in the bacterial flora of 10 genera present on the skin. As shown in Table 17, after using the compositions of Examples 4 and 5 of the present invention for two weeks, the spp. of Staphylococcus and Cutibacterium significantly increased, while the spp. of Ehrlichia and Sphingomonas, which cause infections, significantly decreased after two weeks of use, thereby achieving a statistically significant improvement in the bacterial flora on the skin. In addition, the diversity of the bacterial flora (β-diversity) was tested, and the results showed a statistically significant increase, confirming a statistically significant improvement in the bacterial flora on the skin.

[0169] The present invention is not limited to the embodiments described above. As another embodiment, it may include a combination of the above embodiments or a combination of at least one of the above embodiments and known technologies.

[0170] The present invention has been specifically described above through specific embodiments. However, this is only for the purpose of specific description of the present invention. The present invention is not limited thereto, and those skilled in the art can make modifications or improvements within the technical concept of the present invention.

[0171] Simple variations and modifications of this invention are all included in the scope of this invention. The specific scope of protection of this invention can be clearly understood through the appended claims.

Claims

1. Use of a truffle-derived yeast extract in the manufacture of a skin-improving cosmetic composition that achieves the effect of increasing beneficial bacteria and reducing harmful bacteria in the skin, characterized in that, The composition comprises, relative to 100% by weight, 0.00001% by weight to 10% by weight of polysaccharides, the truffle-derived yeast extract, and Bifidobacterium fermentation lysate as active ingredients. The polysaccharides mentioned above are selected from one or more of the group consisting of inulin, β-glucan and maltodextrin.

2. The use according to claim 1, characterized in that, The above-mentioned cosmetic composition is a composition for improving the microbial flora inhabiting the skin.

3. The use according to claim 2, characterized in that, The above cosmetic composition promotes the growth of beneficial bacteria residing on the skin or inhibits the growth of harmful bacteria.

4. The use according to claim 3, characterized in that, The above-mentioned beneficial bacteria are Staphylococcus epidermidis (ATCC 14984) S.epidermidis ), The above-mentioned harmful bacteria are Staphylococcus aureus (ATCC 6538) and Escherichia coli (ATCC 8739). S.aureus ) 5. The use according to claim 1, characterized in that, The above-mentioned cosmetic composition is any one of the following: a composition for soothing the skin, a composition for improving skin wrinkles, and a composition for improving skin elasticity.

6. The use according to claim 1, characterized in that, The skin mentioned above refers to the scalp.

7. The use according to claim 6, characterized in that, The above-mentioned cosmetic composition is any one of the following: scalp soothing composition, scalp oil improvement composition, hair loss prevention composition, and hair growth improvement composition.

8. The use according to claim 6, characterized in that, The above-mentioned cosmetic composition is a composition for improving the microbial flora inhabiting the scalp.

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