Composition comprising betaine enriched yeast ferment extract

Abstract

The present invention relates to a cosmetic and / or dermatological composition comprising a betaine-rich yeast fermentation extract and a physiologically acceptable medium. The invention also relates to a non-therapeutic method using said composition and to the use of said composition for caring for keratin materials.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202411226970.3 (22) Application Date 2024.09.03 (71) Applicant ELC Management LLC Address New York, USA (72) Inventors Li Zhen, Yao Jizong, Zhou Huanjun, Zhang Yan, Wu Yan, Qu Yulan (74) Patent Agency China Patent Agency (Hong Kong) Limited 72001 Patent Attorneys Zhang Qilu, Wang Lunwei (51) Int.Cl. A61K 8 / 9728 (2017.01) A61K 8 / 44 (2006.01) A61Q 19 / 00 (2006.01) A61Q 17 / 04 (2006.01) A61Q 19 / 08 (2006.01) (54) Title of Invention Composition Containing a Betaine-Rich Yeast Fermentation Extract (57) Abstract This invention relates to cosmetic and / or dermatological compositions comprising a betaine-rich yeast fermentation extract and a physiologically acceptable medium. The invention also relates to non-therapeutic uses of the compositions and their use in the care of keratin materials. Claims 1 page Description 11 pages CN 121622515 A 2026.03.10 CN 1 21 62 25 15 A 1. A cosmetic composition comprising a betaine-rich yeast fermentation extract and a physiologically acceptable medium, wherein the betaine-rich yeast fermentation extract comprises proteins, amino acids, peptides, nucleotides, and at least 2000 ppm of betaine. 2. The cosmetic composition according to claim 1, wherein the betaine-rich yeast fermentation extract is present in the cosmetic composition in an amount of 0.01% to 20% by weight, preferably 0.05% to 5% by weight, more preferably 0.1% to 1% by weight relative to the total weight of the cosmetic composition. 3. The cosmetic composition according to any one of the preceding claims, wherein the protein is present in a total amount of 50% to 75% by weight relative to the total weight of the yeast fermentation extract, and the betaine is present in an amount of 0.2% to 4% by weight relative to the total weight of the yeast fermentation extract. 4. The cosmetic composition according to any one of the preceding claims, wherein the betaine-rich yeast fermentation extract is produced by fermenting one or more yeasts selected from *Saccharomyces cerevisiae*, *Pichia pastoris*, *Brasilaria sinensis*, *Candida*, and *Candida albicans* in a culture medium containing at least one betaine-rich carbon source to obtain a yeast ferment, followed by enzymatic hydrolysis of the yeast ferment.5. The cosmetic composition of claim 4, wherein the betaine-rich carbon source comprises 1% to 10% by weight, preferably 2% to 8% by weight, of betaine relative to the total weight of the carbon source on a dry basis. 6. The cosmetic composition of claim 4 or 5, wherein the betaine-rich carbon source is selected from beet juice, beet pulp, beet molasses, beet molasses fermentation waste / fermentation mash, and mixtures thereof. 7. The cosmetic composition according to any one of claims 4 to 6, wherein the yeast is selected from: *Saccharomyces cerevisiae* strain FX-2, with accession number CCTCC NO: M2016418; *Cyclolindnera fabianii* strain C1.8, with accession number CCTCC NO: M2017780; *Saccharomyces boulardii* strain Ang1.27, with accession number CCTCC NO: M2012116; *Wickerhamomyces anomalus* strain C1.7, with accession number CCTCC NO: M2017782; *Saccharomycopsis fibuligera* strain d6.16, with accession number CCTCC NO: M2019570, preferably *Saccharomyces cerevisiae* strain FX-2. 8. A non-therapeutic use for caring for keratin materials, particularly for improving skin hydration, enhancing skin barrier function, and / or resisting or repairing skin damage caused by stressors, the method comprising topically applying a cosmetic composition according to any one of claims 1 to 7 to the keratin material. 9. A non-therapeutic use of a cosmetic composition according to any one of claims 1 to 8 for caring for keratin materials, particularly for improving skin hydration, enhancing skin barrier function, and / or resisting or repairing skin damage caused by stressors. 10. The non-therapeutic use according to claim 8 or according to claim 9, wherein the stressor is UV radiation, chemical and / or osmotic stressor. Claims 1 / 1 page 2 CN 121622515 A Composition comprising a yeast fermentation extract rich in betaine Technical Field

[0001] The present invention relates to the field of cosmetic and / or dermatological compositions. In particular, the present invention relates to cosmetic compositions comprising a yeast fermentation extract rich in betaine and a physiologically acceptable medium. The present invention also relates to non-therapeutic uses of the compositions and the use of the compositions for caring for keratin materials. Background Art

[0002] The skin is one of the most important organs of the human body.Located on the outermost part of the body, it acts as a barrier to protect the human body from adverse effects of the external environment. With the development of modern society, skin cells face continuous osmotic pressure threats due to constant exposure to ultraviolet rays, air pollution, dry environments, chemicals, etc. To address the need to protect human skin cells, various cosmetic or dermatological products have been developed, many of which are used to hydrate the skin, enhance the skin's resistance to stressors, and / or produce other effects. On the other hand, for cosmetic applications, natural ingredients (i.e., ingredients of natural origin) are extremely attractive to consumers. Much effort has been put into developing compositions containing natural ingredients.

[0003] Betaine is a natural osmotic regulator that has been widely used as a humectant in cosmetics. It shuttles in and out of cells to affect their hydration state. Increased intracellular concentration of betaine can promote cell hydration levels and cell resistance to stressors. FR2877843 discloses the use of betaine as an agent to protect skin cells subjected to different types of stress against skin aging and damage.

[0004] It has been reported that yeast fermentation products can be used as active ingredients in cosmetics, possessing skin whitening, spot-removing, moisturizing, antioxidant, and skin conditioning effects. Their natural and safe properties have made yeast fermentation products increasingly popular among consumers. It is expected that the use of betaine and yeast fermentation products can provide further benefits for cosmetic applications.

[0005] Therefore, the object of the present invention is to develop cosmetic or dermatological compositions comprising a yeast fermentation extract rich in betaine, which has good properties for improving skin hydration and / or skin resistance to stressors.

[0006] The present invention relates to a cosmetic or dermatological composition comprising a betaine-rich yeast fermentation extract and a physiologically acceptable medium, wherein the betaine-rich yeast fermentation extract is produced by fermenting one or more yeasts selected from *Saccharomyces cerevisiae*, *Cyberlindnera fabianii*, *Saccharomyces boulardii*, *Wickerhamomyces anomalus*, and *Saccharomycopsis fibuligera* in a culture medium containing at least one betaine-rich carbon source to obtain a yeast ferment, followed by enzymatic hydrolysis of the yeast ferment. Such a yeast fermentation extract is rich in betaine and is superior to conventionally used pure betaine in efficacy and benefit as a cosmetic care active ingredient for improving skin hydration, enhancing skin barrier function, and resisting or repairing skin damage caused by stressors. Ingesting betaine into yeast provides a significant advantage in promoting overall protective effects.

[0007] Therefore, one subject of the present invention is a cosmetic composition comprising a betaine-rich yeast fermentation extract and a physiologically acceptable medium, wherein the betaine-rich yeast fermentation extract comprises at least 2000 ppm of betaine.

[0008] Another subject of the present invention is a non-therapeutic use of a cosmetic composition according to the present invention for caring for keratin materials. Specification 1 / 11 pages 3 CN 121622515 A

[0009] Yet another subject of the present invention is the use of a cosmetic composition according to the present invention for caring for keratin materials, particularly for non-therapeutic use to improve skin hydration, enhance skin barrier function and / or resist or repair skin damage caused by stressors.

[0010] Other advantages of the present invention will become clearer upon reading the following detailed description and examples. Detailed Description

[0011] Detailed embodiments of the present invention are disclosed herein; however, it should be understood that the disclosed embodiments are for illustrative purposes only, and the present invention can be implemented in various forms. Therefore, the specific structural and functional details disclosed herein should not be construed as limiting, but only as a representative basis for teaching those skilled in the art to implement the present invention in various ways.

[0012] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Where the definition of a term in this specification conflicts with the meaning commonly understood by one of ordinary skill in the art to which this invention pertains, the definition set forth herein shall apply.

[0013] The term “keratin material” as used herein is intended to include skin, hair, scalp, or mucous membranes, preferably skin or mucous membranes.

[0014] The term “fermentation” as used herein refers to the process of preparing microbial cells or their metabolites using microorganisms, such as bacteria, yeast, etc.

[0015] The term “betaine-rich yeast fermentation extract” as used herein refers to an extract of the fermentation product of at least one yeast, rich in betaine from the carbon source used in the fermentation process, preferably containing at least 2000 ppm of betaine.

[0016] The term “beauty care benefits” as used herein includes, but is not limited to, one or more of the following: (a) treating, reducing and / or preventing fine lines or wrinkles; (b) minimizing skin pore size; (c) improving skin thickness, plumpness and / or firmness; (d) improving skin suppleness and / or softness; (e) improving skin tone, radiance and / or clarity; (f) improving procollagen and / or collagen production; (g) improving skin texture and / or promoting reorganization; (h) improving skin barrier repair and / or function; (i) treating and / or preventing skin sagging or atrophy; (j) improving the appearance of skin contours; (k) restoring skin radiance and / or brightness; (l) replenishing key nutrients and / or components in the skin; (m) improving the appearance of skin diminished by menopause; (n) improving skin hydration and / or moisture; (o) improving skin's resistance to stressors; and (p) improving skin elasticity and / or firmness.

[0017] As used herein, the term “stressor” refers to a stressor or condition in which skin cells may be exposed, such as ultraviolet radiation, air pollution, dry environments, chemicals, or osmotic stress. Here, osmotic stress refers to a physiological disturbance caused by a sudden change in the concentration of solutes surrounding the cell, resulting in a rapid change in the movement of water across the cell membrane. Under hypertonic conditions, i.e., with high concentrations of salt, substrate, or any solute in the supernatant, water is drawn from the cell by osmosis.

[0018] As used herein, the term “skin cell” refers to the cells that constitute the skin, including but not limited to keratinocytes, melanocytes, fibroblasts, T cells, etc.

[0019] As used herein, the term “physiologically acceptable medium” refers to a fluid, more or less, that may include, but is not limited to, any additives or cosolvents commonly used in the cosmetics industry, as well as adjuvants, humectants, surfactants, emulsifiers, etc., necessary for their formulation, suitable for contact with the skin or mucous membranes without causing toxic or intolerance reactions.

[0020] As used herein, the terms “partial” or “locally” refer to the application or spreading of the cosmetic composition according to the invention onto a healthy area of ​​a keratin material surface.

[0021] Whenever a term is determined by reference range, that range shall be understood to explicitly disclose each of its elements. Specification 2 / 11 pages 4 CN 121622515 A As a non-limiting example, the range 1-10% shall be understood to include 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% and 10%, and all values ​​between 1% and 10%.

[0022] Except as stated in the operational embodiments, or otherwise, all figures indicating the amount of components and / or reaction conditions should be understood to be modified in all cases by the term “about”, which has a meaning conventionally known in the art, such as within ±10% of the figures shown (e.g., “about 10%” means 9%–11%, “about 2%” means 1.8%–2.2%).

[0023] Unless otherwise specifically stated, content, parts and percentages are expressed on a weight basis.

[0024] Betaine-rich yeast fermentation extract (referred to as OsmoshieldB)

[0025] The cosmetic composition according to the invention comprises a betaine-rich yeast fermentation extract, wherein the betaine-rich yeast fermentation extract contains at least 2000 ppm of betaine.

[0026] The term “betaine” as used herein refers to glycine betaine, also known as trimethylglycine or 1-carboxy-N,N,N-trimethylammonium hydroxide inner salt, which has the following structure:

[0027]

[0028] Betaine is an amphoteric electrolyte ion stable at physiological pH, carrying a positive charge on the nitrogen atom and a negative charge on the oxygen atom of the carboxyl functional group. Furthermore, betaine is an amphiphilic compound, with its hydrophilic portion consisting of both positive and negative charges, and its hydrophobic portion consisting of three methyl groups.

[0029] Betaine has a very wide chemical distribution. It is found in bacteria and cyanobacteria, animals and plants, ranging from algae to higher plants.In plants, betaine is mainly found in the following families: Chenopodiaceae (e.g., *Atriplex* sp., *Beta* sp., *Halosarcia* sp., *Maireana* sp., *Rhagodia* sp., *Salicornia* sp., *Salsola* sp., *Sarcocornia* sp., *Sclerolaena* sp., *Spinacia* sp., *Threlkeldia* sp.), Amaranthaceae (e.g., *Amaranthus* sp.), Aviceniaceae (e.g., *Avicennia* sp.), and Poaceae (e.g., *Cymbopogon* sp., *Distichlis* sp., *Enneapogon* sp.). .), *Eragrostis* sp., *Hordeum* sp., *Sorghum* sp., *Spartina* sp., *Themeda* sp., *Triodia* sp., *Triticum* sp., *Zea* sp.), Asteraceae (e.g., *Aster* sp., *Cassinia* sp., *Cratystylis* sp., *Gnaphalium* sp., *Helianthus* sp., *Olearia* sp.), Convolvulaceae (e.g., *Convolvulus* sp., *Cuscuta* sp., *Evolvulus* sp., *Wilsonia* sp.), *Everlastingia* sp. In plants such as Plumbaginaceae (e.g., Plumbago sp., Limonium sp.), Solanaceae (e.g., Lycium sp.), and Fabaceae (e.g., Medicago sp., Trifolium sp.).

[0030] As an amphoteric ion, betaine interacts strongly with surrounding water molecules through hydrogen bonds, giving it important osmotic pressure regulating properties.In environments with limited water content and / or high salinity, certain living organisms synthesize and accumulate betaine. This mobilization of betaine leads to an increase in intracellular osmotic pressure and enhances the cell's water-holding capacity, thereby enabling the organism to maintain its homeostasis and thus promote its adaptation to the environment.

[0031] In addition to the role of betaine as an osmotic regulator, several hypotheses have been proposed that, under the same water conditions or osmotic stress, betaine plays a role in stabilizing the structural integrity of proteins and cell membranes in organisms.

[0032] Betaine used in conventional cosmetics is either extracted directly from sugar beets, for example, purified from sugar beet molasses by chromatographic separation, concentration, crystallization, and drying, or prepared by chemical or enzymatic synthesis.

[0033] The inventors have discovered that by fermenting one or more yeasts selected from *Saccharomyces cerevisiae*, *Pichia pastoris*, *Brasilaria sinensis*, *Candida*, and *Candida albicans* in a culture medium containing at least one betaine-rich carbon source to obtain a yeast ferment, and then enzymatically hydrolyzing the yeast ferment to produce a betaine-rich yeast ferment extract containing proteins, amino acids, peptides, nucleotides, and at least 2000 ppm of betaine, it exhibits superior efficacy and benefits as a cosmetic care active ingredient compared to conventionally used pure betaine, fermentation extracts produced with other substrates or other microorganisms, or combinations thereof, in improving skin hydration, skin barrier function, and / or skin resistance to stressors such as hyperosmotic stressors and oxidative stressors.

[0034] Preferably, the betaine-rich carbon source used to produce the betaine-rich yeast ferment extract according to the present invention is selected from beet juice, beet pulp, beet molasses, beet molasses fermentation waste / fermentation mash, and mixtures thereof. Beet pulp and beet molasses are byproducts of sugar production from beets. Preferably, the betaine-rich carbon source contains 1% to 10% by weight, preferably 2% to 8% by weight, of betaine relative to the total weight of the carbon source on a dry basis.

[0035] Preferably, the yeast used to produce the betaine-rich yeast fermentation extract according to the invention is selected from one or more of Saccharomyces cerevisiae, Pichia pastoris, Saccharomyces boulardii, Candida albicans, and Saccharomyces clavatum, more preferably Saccharomyces cerevisiae. Preferably, the yeast used herein may be selected from those listed in Table 1 below.

[0036] Table 1.

[0037]

[0038] *CCTCC: China Center for Type Culture Collection

[0039] The betaine-rich yeast fermentation extract according to the invention is produced by fermenting one or more of Saccharomyces cerevisiae, Pichia pastoris, Saccharomyces boulardii, Candida albicans, and Saccharomyces clavatum in a culture medium containing at least one betaine-rich carbon source to obtain a yeast ferment, and then enzymatically hydrolyzing the yeast ferment.Preferably, the culture medium comprises: at least one betaine-rich carbon source, ammonia, NaCl, KCl, NH4H2PO4, copper sulfate, biotin, VB1 and VB2. Preferably, the enzyme used for enzymatic hydrolysis comprises at least one of protease, mannanase and / or β-glucanase.

[0040] Preferably, the fermentation method for producing the betaine-rich yeast fermentation extract according to the present invention comprises the following steps: a) shake flask seed culture; b) seed fermentation; c) commercial fermentation; d) enzymatic hydrolysis; and e) drying. Specification 4 / 11 pages 6 CN 121622515 A

[0041] Preferably, the betaine-rich yeast fermentation extract used herein can be produced by a method comprising the following steps.

[0042] a) Shake flask seed culture: Seed culture is performed in a shake flask with a yeast strain to obtain a seed solution.

[0043] Specifically, in step a), a sterile culture medium containing water, 80-120 g / L glucose, 10-30 g / L yeast extract, 0.8-1.2 g / L KH2PO4, and 0.8-1.2 g / L magnesium sulfate, with a pH of 4.5-5.5, is loaded into the bioreactor. Then, at least one yeast strain (e.g., Saccharomyces cerevisiae FX-2 strain, preservation number CCTCC NO: M2016418) is inoculated and cultured at 26℃-35℃ and 150-250 rpm for 16-48 hours to obtain a seed liquid;

[0044] b) Seed fermentation: The seed liquid is inoculated into a seed fermentation tank and seed fermentation is carried out in the presence of at least one betaine-rich carbon source to obtain a seed fermentation broth.

[0045] Specifically, in step b), the shake flask seed liquid is inoculated into a seed fermentation tank containing a culture medium, wherein the culture medium preferably contains: water, 50-150 g / L beet molasses (calculated as reducing sugar, with a betaine content of 2-8% on a dry basis), 0.5-2.0 g / L magnesium sulfate, 10-30 g / L ammonium sulfate, 1-5 g / L NH4H2PO4, 0.004-0.02 g / L copper sulfate pentahydrate, 0.001-0.5 g / L biotin, 0.002-0.2 g / L VB1, and 0.001-0.1 g / L VB2, wherein beet molasses, ammonium sulfate, and NH4H2PO4 are added by fed-batch addition. Preferably, the seed fermentation conditions include: the stirrer speed is controlled at 200-700 rpm, the sterile air flow rate is controlled at 0.5-4 L air / (L fermentation liquid×min), the fermentation temperature is 26-35℃, and the fermentation time is 16-48 hours. After fermentation, the seed fermentation liquid is centrifuged until the yeast concentration is ≥500 g / L;

[0046] c) Commercial fermentation: The seed fermentation liquid is inoculated into a commercial fermentation tank, and commercial fermentation is carried out in the presence of at least one betaine-rich carbon source to obtain commercial fermentation liquid.

[0047] Specifically, in step c), the seed fermentation broth from the seed fermentation tank is inoculated into a commercial fermentation tank containing culture medium. After inoculation, the yeast concentration in the culture medium is 40-70 g / L. The culture medium preferably contains: water, 80-200 g / L beet molasses (calculated as reducing sugar, with a betaine content of 2-8% on a dry basis), 5-30 mL / L 20% ammonia water, 2-20 g / L NaCl, 2-20 g / L KCl, 1-5 g / L NH4H2PO4, 0.001-0.05 g / L copper sulfate pentahydrate, 0.001-0.05 g / L biotin, 0.002-0.2 g / L VB1, and 0.001-0.1 g / L VB2, wherein beet molasses, ammonia water, and NH4H2PO4 are added by fed-batch addition. More preferably, the culture medium consists of the following components: water, 150 g / L beet molasses (calculated as reducing sugar, with a betaine content of 4% on a dry basis), 30 mL / L 20% ammonia water, 6 g / L NaCl, 2 g / L KCl, 3 g / L NH4H2PO4, 0.005 g / L copper sulfate pentahydrate, 0.02 g / L biotin, 0.01 g / L VB1, and 0.002 g / L VB2. Preferably, the commercial fermentation conditions include: agitator speed controlled at 200-700 rpm, sterile air flow rate controlled at 0.5-4 L air / (L fermentation broth × min), fermentation temperature at 28-35℃, and fermentation time at 10-20 hours.

[0048] d) Enzymatic hydrolysis: enzymatically hydrolyze the commercial fermentation broth, and centrifuge to collect the supernatant. Preferably, the enzyme used for enzymatic hydrolysis includes at least one of protease, mannanase, and / or β-glucanase. The enzyme used is 0.1%-1% of the yeast mass. The enzymatic hydrolysis conditions include 40-70°C, pH 5-7, and hydrolysis for 5-20 hours. Preferably, after hydrolysis, the product is heated to 80-100°C for 0.5-2 hours to inactivate the enzyme, and then centrifuged to collect the supernatant.

[0049] e) Drying: The supernatant is filtered and dried to obtain a yeast fermentation extract rich in betaine (0.2-4% betaine on a dry basis).

[0050] In the prior art, due to the nature of the yeast fermentation process, the amount of betaine ingested into the yeast fermentation product is usually very limited (e.g., <1000ppm). However, the yeast fermentation product according to the present invention exhibits a significantly higher level of betaine intake compared to the prior art.

[0051] Physicochemical analysis of the betaine-rich yeast fermentation extract showed that it contained proteins, amino acids, peptides, nucleotides, and betaine, wherein the proteins were present in a total amount of 50% by weight to 75% by weight relative to the total weight of the yeast fermentation extract, for example, about 55.39% by weight, about 68.49% by weight, about 70.42% by weight, or about 73.21% by weight; and the betaine was present in a total amount of at least 0.2% by weight to 4% by weight relative to the total weight of the yeast fermentation extract, for example, about 0.3% by weight, 0.4% by weight, about 0.6% by weight, about 0.8% by weight, about 1% by weight, about 1.2% by weight, about 1.4% by weight, about 1.6% by weight, about 1.8% by weight, about 2% by weight, about 2.2% by weight, about 2.4% by weight, about 2.6% by weight, etc. It is present in amounts of about 2.8% by weight, about 3% by weight, about 3.2% by weight, about 3.4% by weight, about 3.6% by weight, or about 3.8% by weight.

[0052] Preferably, the betaine-rich yeast fermentation extract is present in the cosmetic composition according to the invention in an amount of 0.01% by weight to 20% by weight, preferably 0.05% by weight to 5% by weight, and more preferably 0.1% by weight to 1% by weight relative to the total weight of the composition. For example, a yeast fermentation extract rich in betaine may be present in the cosmetic composition according to the invention in amounts of 0.01 wt%, 0.025 wt%, 0.05 wt%, 0.075 wt%, 0.1 wt%, 0.2 wt%, 0.3 wt%, 0.4 wt%, 0.5 wt%, 0.6 wt%, 0.7 wt%, 0.8 wt%, 0.9 wt%, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, and 20 wt% relative to the total weight of the composition.

[0053] Additional Ingredients

[0054] The cosmetic compositions of the present invention may specifically be aqueous solutions, water-alcohol solutions, or oil solutions; oil-in-water or water-in-oil emulsions or multiple emulsions; aqueous or hydrogels; or colloidal forms. These compositions may also be in the form of creams, suspensions, or powders suitable for application to the skin, mucous membranes, lips, and / or skin appendages. These compositions may be more or less fluid and have the appearance of creams, lotions, emulsions, pastes, or foams. They may also be in solid form, such as sticks, or applied to the skin in aerosol form. In one embodiment, the compositions of the present invention are cosmetic compositions for caring for keratin materials.

[0055] According to various embodiments, the cosmetic compositions of the present invention may include any ingredients or additives commonly used in the cosmetic field, as well as adjuvants necessary for their formulation, such as cosolvents (ethanol, glycerin, benzyl alcohol, humectants, etc.), thickeners, diluents, emulsifiers, antioxidants, colorants, sunscreens, pigments, fillers, preservatives, fragrances, chelating agents, pH adjusters, odor absorbers, essential oils, trace elements, essential fatty acids, surfactants, film-forming polymers, chemical or mineral filters, hydrating agents, or hot water, etc. For example, natural water-soluble polymers may be used, such as polysaccharides or polypeptides, methylcellulose or hydroxypropylcellulose-type cellulose derivatives or synthetic polymers, poloxamer, carbomer, siloxanes, PVA, or PVP.

[0056] A non-exhaustive list of such ingredients can be found in patent application publication number WO 2023 / 141799, the entire contents of which are incorporated herein by reference. Other examples of such additives can be found in the International Dictionary and Handbook of Cosmetic Ingredients (9th edition, 2002).

[0057] Those skilled in the art will carefully select optional additional ingredients and / or their amounts so that the advantageous properties of the cosmetic compositions according to the invention are not, or substantially not, adversely affected by the contemplated additions.

[0058] Those skilled in the art can make various selections of these ingredients to prepare compositions having desired properties, such as consistency or texture. In particular, the ingredients and their amounts are specifically determined according to their specific product / application, such as aerosols, sprays, foams, liquids, lotions, gels, creams, lotions, emulsions, ointments, pastes, powders, makeup, soaps, solid sticks, etc.

[0059] The betaine-rich yeast fermentation extract of the present invention can be used alone or in combination with other active ingredients. For example, the cosmetic compositions of the present invention also contain at least one other active ingredient designed to improve skin physiological function, such as regenerating, anti-aging, anti-wrinkle, thickening, anti-free radical, anti-glycation, moisturizing, antibacterial, antifungal, stratum corneum separation, muscle relaxation, exfoliating and color-correcting ingredients, ingredients that stimulate the synthesis of skin macromolecules or energy metabolism, ingredients that regulate skin differentiation, pigmentation or depigmentation, ingredients that stimulate microcirculation, sunscreens or metalloproteinase inhibitors.

[0060] In one embodiment, in addition to a yeast fermentation extract rich in betaine, the cosmetic composition of the present invention further comprises:

[0061] - sunscreens, UV and infrared shielding agents,

[0062] - anti-free radical ingredients,

[0063] - DHEA (dehydroepiandrosterone),

[0064] - dehydroacetic acid (DHA),

[0065] - natural or synthetic phytosterols,

[0066] - α-hydroxy acids and β-hydroxy acids, silanols,

[0067] - glycosamines, glucosamine, D-glucosamine, N-acetylglucosamine, N-acetyl-D-glucosamine, mannosamine, N-acetylmannosamine, galactosamine, N-acetylgalactosamine,

[0068] - polyphenols, isoflavones, flavonoids, such as grape extract, pine extract, olive extract,

[0069] - lipids, such as ceramides or phospholipids,

[0070] - Animal oils, such as squalene or squalane,

[0071] - Vegetable oils, such as almond oil, coconut oil, castor oil, jojoba oil, olive oil, rapeseed oil, peanut oil, sunflower seed oil, wheat germ oil, corn germ oil, soybean oil, cottonseed oil, alfalfa oil, pumpkin seed oil, evening primrose oil, millet oil, barley oil, rye oil, safflower oil, passion fruit oil, hazelnut oil, palm oil, almond oil, avocado oil, calendula seed oil, ethoxylated vegetable oils, or shea butter,

[0072] The above compounds may be natural, such as plant peptide hydrolysates, or may be synthetic, such as peptide compounds.

[0073] If present, these ingredients may be present in the composition individually in amounts from 0.01% to 80% by weight, in amounts from 20% to 99.99% by weight, or from 30% to 90% by weight, or in total amounts from 40% to 80% by weight.

[0074] Methods and Uses

[0075] While not wishing to be bound by any theory, it is believed that delivering the cosmetic composition according to the invention can provide cosmetic care benefits, such as improving skin hydration, enhancing skin barrier function, resisting or repairing skin damage caused by stressors (e.g., hyperosmolar stressors, oxidative stressors, etc.), thereby resisting signs of epidermal osmotic imbalance caused by stressors, such as fine lines and wrinkles, withering, thinning, dryness, sagging, dullness, or unevenness of the skin.

[0076] Therefore, the invention also relates to non-therapeutic methods for caring for keratin materials, particularly for improving skin hydration, enhancing skin barrier function, and / or resisting or repairing skin damage caused by stressors, comprising the step of topically applying the cosmetic composition of the invention to keratin materials, such as the skin, wherein the stressor is UV radiation, chemical and / or osmotic stressor.

[0077] In one embodiment, the cosmetic composition of the present invention can be applied topically in an effective amount to the neck, face, neckline, back, hands, or body area, such that the composition can provide cosmetic care benefits, particularly improvements in skin hydration, skin barrier function, and / or skin resistance to stressors, wherein the stressors are UV radiation, chemical and / or osmotic stressors.

[0078] In another aspect, the present invention relates to the non-therapeutic use of cosmetic compositions according to the present invention for the care of keratin materials, particularly for improving skin hydration, enhancing skin barrier function, and / or resisting or repairing skin damage caused by stressors, wherein the stressors are UV radiation, chemical and / or osmotic stressors. Specification 7 / 11 pages 9 CN 121622515 A

[0079] In one embodiment, the present invention provides the non-therapeutic use of cosmetic compositions according to the present invention for combating signs of epidermal osmotic imbalance caused by stressors, such as fine lines and wrinkles, withering, thinning, dryness, sagging, dullness, or unevenness of the skin, wherein the stressors are UV radiation, chemical and / or osmotic stressors.

[0080] In one embodiment, the present invention also provides the use of the cosmetic composition according to the invention in the preparation of a medicament for treating signs of skin dehydration or skin inflammation caused by stressors, wherein the stressor is UV radiation, chemical and / or osmotic stressor.

[0081] The following examples are intended to illustrate the invention and not to limit its scope.

[0082] Examples

[0083] The following examples are given to illustrate the invention and should not be construed as limiting the scope of the invention.

[0084] I. Preparation of Fermentation Extracts

[0085] Unless otherwise stated, the raw materials used in this section are commercially available.

[0086] 1. Yeast Fermentation Extract (Osmoshield B)

[0087] An exemplary yeast fermentation extract Osmoshield B was prepared according to the procedure described in the “Detailed Description” section.

[0088] 2. Control Yeast Fermentation Extract (Control)

[0089] A control yeast fermentation extract was prepared according to substantially the same procedure as Osmoshield B, except that sugarcane molasses was used instead of beet molasses used in steps b) and c).

[0090] 3. Lactic acid bacteria fermentation extract

[0091] The lactic acid bacteria fermentation extract was prepared according to the following procedure:

[0092] 1) Preparation of culture medium: The culture medium was prepared according to the following composition: about 1% beet molasses, 10 g / L peptone, 4 g / L yeast powder, 20 g / L glucose, 1 g / L Tween 80, 2 g / L K2HPO4, 5 g / L sodium acetate, 2 g / L triammonium citrate, 0.2 g / L MgSO4·7H2O, 0.05 g / L MnSO4.

[0093] 2) Activation of lactic acid bacteria: The existing *Lactobacillus rhamnosus* strain was activated by placing it in a small amount of culture medium.

[0094] 3) Fermentation: A total of 4 L of fermentation culture medium was prepared, sterilized by high-temperature steam, and then the activated lactic acid bacteria were added to the fermentation culture medium. Fermentation was carried out in a bioreactor at a controlled temperature of 37°C for about 48 hours.

[0095] 4) Centrifugation: The bacterial suspension after fermentation was separated using a centrifuge to obtain bacterial precipitate.

[0096] 5) Cell wall disruption: The bacterial cells were disrupted using a cell wall disruptor, and the soluble components were extracted with water. The mixture was then centrifuged again to obtain the supernatant.

[0097] 6) Freeze-drying: The supernatant was freeze-dried to obtain the lactic acid bacteria fermentation extract.

[0098] II. Determination of Betaine Content in Fermentation Extracts

[0099] The betaine content in fermentation extracts was quantitatively determined by ultra-high performance liquid chromatography (UPLC) using a Waters BEH Amide column (1.7 μm, 100 × 2.1 mm id) connected to a Waters SQD2 mass spectrometer detector. The mobile phase consisted of acetonitrile (from Merck) and formic acid (from Merck) aqueous solution. The gradient flow rate of the mobile phase was set to 0.5 mL / min at 30 °C. The column temperature, injection volume, and detection mass-to-charge ratio (m / z) were set to 30 °C, 1.0 μL, and SIR118 (positive ion mode), respectively. The calibration curve for betaine was linear in the concentration range of 10–800 ppb (r ≥ 0.9900). The betaine content detected in different fermentation extracts is shown in Table 2.

[0100] Table 2.

[0101] Betaine content of fermentation extract Osmoshield B 10932ppm Instruction manual 8 / 11 pages 10 CN 121622515 A Control yeast fermentation extract 80ppm Lactic acid bacteria fermentation extract 2317ppm

[0102] III. Hyperosmolar stress model test

[0103] 1. Cell culture

[0104] Normal human epidermal keratinocytes (NHEK, FC-0007, purchased from Lifeline Cell Technology) were incubated at 37°C, 95% humidity and 5% CO2 in keratinocyte culture medium (DermaLife K Kertinocyte Medium Complete Kit, LL-0007, Lifeline Cell Technology) supplemented with growth factors and 1% penicillin / streptomycin (Thermo Fischer, Waltham, MA, USA). The cells were passaged when they reached 80% confluence.

[0105] 2. Cell treatment

[0106] NHEK cells were seeded in 96-well plates at a density of 3,000 cells / well.After 24 hours of cell adhesion, cells were treated for 48 hours with 100 μL of 300 mOsm medium (primitive cell medium, DermaLife K Keratinocyte Medium Complete Kit) or 450 mOsm medium with / with different concentrations of Osmoshield B. The 450 mOsm medium was prepared by adding 4.39 mg / mL NaCl to the primitive cell medium.

[0107] After treatment, the medium was removed, and the cells were washed twice with Dulbecco phosphate-buffered saline (DPBS), and then incubated for 120 minutes in PBS containing 10% AlamarBlue™ Cell Viability Reagent (Thermo Fisher, Waltham, MA, USA). The fluorescence values ​​of the incubation plates were measured using a Perkin Elmer plate reader (Tecan) at an excitation wavelength of 530 nm / emission wavelength of 615 nm and recorded as relative fluorescence units (RFU). Cell-free blank wells were used as reference blanks.

[0108] 3. Cell viability ratio

[0109] The protective effect can be determined by the following cell viability ratio (%):

[0110] Cell viability ratio (%) = (RFU value 450mOsm with / without Osmoshield B - RFU value blank) / (RFU value 300mOsm - RFU value blank) × 100%

[0111] The results of the hyperosmolar stress model test (in vitro) are shown in Table 3.

[0112] Table 3.

[0113] Group cell viability ratio (%) SD (%) 300mOsm 100 9.913419 450mOsm 74.56 3.734507 450mOsm+0.10%Osmoshield B 82.54 15.46527 450mOsm+0.25%Osmoshield B 93.20 4.965208 450mOsm+0.50%Osmoshield B 95.05 8.889226

[0114] The results of the hyperosmolar stress model experiment show that the yeast fermentation extract supplement rich in betaine can resist cell damage caused by hyperosmolar stressors.

[0115] IV. Loricrin Expression Assay

[0116] The EpiuiKuits reconstructed skin epidermal model (BioCell, Day 8 model, catalog number: PM1011) was adapted overnight in recovery medium (BioCell, EpiRecovery, catalog number: PY3031) at 37°C / 5% CO2 according to the manufacturer's instructions. After equilibration, the EpiuiKuits model was treated with 0.1% OsmoshieldB for 24 hours.After treatment, EpuiKuits skin samples were fixed in 4% formalin for 2 hours, then paraffin-embedded and sectioned. Immunohistochemical staining (IHC) targeting lobelin (LOR) was performed using an anti-lobelin antibody (ab198994, Abcam).

[0117] Images were captured by fluorescence microscopy, and lobelin expression results were extracted from the images. Table 4 shows the relative levels compared to the control group.

[0118] Table 4.

[0119] Group IOD SD Control 1.000 0.123 0.1% Osmoshield B 1.805 0.145

[0120] Lobelin is a major component of the keratinized cell membrane keratin located in the granular layer. The main function of lobelin is to enhance the cell membrane and strengthen its defensive barrier function. Compared with the control group, 0.1% Osmoshield B significantly increased the expression of scleroderma protein by 80% (p<0.005), indicating that yeast fermentation extract can enhance skin barrier function, which plays an important role in skin hydration.

[0121] V. Oxidative Stress Assay

[0122] Normal human epidermal keratinocytes (NHEK, FC-0007, purchased from Lifeline Cell Technology) were seeded in 12-well plates at a density of 1.4 × 10⁵ cells / well and then treated with various active ingredients at the concentrations listed in Tables 5-7 for 24 hours. Then, H₂O₂ (250 μM) (23381-25 mL, Sigma, St. Louis, MO, USA) was added to the cells, followed by incubation for 30 minutes. The cells were then incubated with the reactive oxygen species (ROS) probe DCFH-DA (S0033, Beyotime, Nanjing, China) in the dark for 30 minutes, and the cells were harvested and measured by flow cytometry (Beckman, CA, USA). Data were analyzed using mean fluorescence intensity (MFI). Cell protection rate was calculated using the following formula:

[0123] Protection rate (%) = [(MFIH2O2 - MFI control) - (MFI active ingredient - MFI control)] / (MFIH2O2 - MFI control) × 100%.

[0124] All experiments were repeated three times. The results of the oxidative stress assays are shown in Tables 5-7. Results in the same table represent experiments performed on the same day using the same batch of cells.

[0125] Table 5.

[0126] Average protection rate of each group (%) SD H2O2+Osmoshield B-0.05% 34.87% 0.16% H2O2+Osmoshield B-0.025% 32.24% 2.54% H2O2+Control yeast fermentation extract-0.05% 9.69% 1.73% H2O2+Control yeast fermentation extract-0.025% 5.34% 1.01% H2O2+Pure betaine-0.06% -13.01% 3.47% H2O2+Pure betaine-0.024% 1.05% 0.29% H2O2+Pure betaine-0.012% -0.95% 1.91%

[0127] Table 6.

[0128]

[0129] Table 7.

[0130] Average protection rate of the group (%) SD H2O2+VE-0.05% 34.19% 4.82% H2O2+Osmoshield B-0.05% 39.01% 4.06% Instructions for use 10 / 11 pages 12 CN 121622515 A H2O2+Osmoshield B-0.025% 33.15% 1.92% H2O2+Lactic acid bacteria fermentation extract-0.05% 34.27% 2.81% H2O2+Lactic acid bacteria fermentation extract-0.025% 22.02% 4.04%

[0131] The skin is continuously exposed to oxidative stress induced by reactive oxygen species (ROS) generated by external pro-oxidants. Oxidative stress test results showed that Osmoshield B can resist or repair ROS-mediated oxidative damage. Compared with control yeast fermentation extract, pure betaine, or combinations thereof, Osmoshield B showed significantly improved efficacy in resisting cell damage caused by oxidative stressors. In addition, Osmoshield B also showed superior anti-oxidative stress efficacy compared with commonly used antioxidants VE and lactic acid bacteria fermentation extract.

[0132] The present invention has been explained and described above. In addition, only preferred embodiments have been shown and described in the present invention, but as mentioned above, it should be understood that it can be used in various other combinations, modifications, and environments, and can be changed or modified within the scope of the inventive concept expressed herein, in proportion to the above teachings and / or skills or knowledge in related fields. The embodiments described above are also intended to explain the best mode known to the applicant and to enable others skilled in the art to utilize the disclosure in these or other embodiments, as well as various modifications required for a particular application or use.Specification Page 11 / 11 13 CN 121622515 A ABSTRACT The present disclosure relates to a cosmetic and / or dermatologic composition comprising a betaine enriched yeast ferment extract and a physiologically acceptable medium. The present disclosure also relates to a non-therapeutic method using said composition and use of said composition for caring for keratin material.。

Claims

1. A cosmetic composition comprising a betaine-enriched yeast fermentation extract and a physiologically acceptable medium, wherein the betaine-enriched yeast fermentation extract comprises proteins, amino acids, peptides, nucleotides and at least 2000 ppm of betaine.

2. The cosmetic composition according to claim 1, wherein the betaine-enriched yeast fermentation extract is present in the cosmetic composition in an amount of 0.01 to 20 wt.%, preferably 0.05 to 5 wt.%, more preferably 0.1 to 1 wt.% relative to the total weight of the cosmetic composition.

3. The cosmetic composition according to any one of the preceding claims, wherein the proteins are present in a total amount of 50 to 75 wt.% relative to the total weight of the yeast fermentation extract, and the betaine is present in an amount of 0.2 to 4 wt.% relative to the total weight of the yeast fermentation extract.

4. The cosmetic composition according to any one of the preceding claims, wherein the betaine-enriched yeast fermentation extract is produced by fermenting a yeast selected from one or more of Saccharomyces cerevisiae, Pichia pastoris, Saccharomyces boulardii, Candida, and Saccharomycopsis fibuligera with a culture medium comprising at least one betaine-enriched carbon source to obtain a yeast fermentation, and then enzymatically hydrolyzing the yeast fermentation.

5. The cosmetic composition according to claim 4, wherein the betaine-enriched carbon source comprises betaine in an amount of 1 to 10 wt.%, preferably 2 to 8 wt.% relative to the total weight of the carbon source on a dry basis.

6. The cosmetic composition according to claim 4 or 5, wherein the betaine-enriched carbon source is selected from the group consisting of beet juice, beet pulp, beet molasses, beet molasses fermentation waste / fermentation mash, and mixtures thereof.

7. The cosmetic composition according to any one of claims 4 to 6, wherein the yeast is selected from the group consisting of: Saccharomyces cerevisiae, strain FX-2, having the accession number CCTCC NO: M2016418; Cyberlindnera fabianii, strain C1.8, having the accession number CCTCC NO: M2017780; Saccharomyces boulardii, strain Anqi 1.27, having the accession number CCTCC NO: M2012116, Wickerhamomyces anomalus, strain C1.7, having the accession number CCTCC NO: M2017782; Saccharomycopsis fibuligera, strain d6.16, having the accession number CCTCC NO: M2019570, and preferably Saccharomyces cerevisiae, strain FX-2.

8. A non-therapeutic method for caring for keratin materials, in particular for improving skin hydration, enhancing skin barrier function, and / or resisting or repairing skin damage caused by stressors, comprising the topical application to the keratin materials of a cosmetic composition according to any one of Claims 1 to 7.

9. Use of a cosmetic composition according to any one of Claims 1 to 8 for caring for keratin materials, in particular for improving skin hydration, enhancing skin barrier function, and / or resisting or repairing skin damage caused by stressors.

10. The non-therapeutic method according to Claim 8 or the non-therapeutic use according to Claim 9, wherein the stressors are UV radiation, chemical and / or osmotic stressors.