Composition for alleviating hair loss and promoting hair growth, containing heat-treated lactic acid bacterium Rimosilactobacillus fermentum LM1020

Abstract

The present application relates to heat-treated Lactobacillus fermentum LM1020 having an anti-hair loss and hair growth promoting effect, and more particularly, to heat-treated Lactobacillus fermentum LM1020 having an anti-hair loss and hair growth promoting effect by promoting the proliferation of dermal papilla cells of hair follicles and a composition containing the same. The heat-treated Lactobacillus fermentum LM1020 of the present application and the composition containing the same promote the proliferation of dermal papilla cells, are excellent in the inhibitory activity of 5α-reductase-1, and have an anti-hair loss and hair growth promoting effect by promoting the expression of FGF7, FGF10, and EGF, which are a kind of growth factors.

Description

[Technical Field]

[0001] This application relates to heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020, which has the effect of preventing hair loss and promoting hair growth, and more specifically, to heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020, which promotes the proliferation of hair follicle dermal papilla cells and has the effect of preventing hair loss and promoting hair growth, and to compositions containing the same. [Background technology]

[0002] Hair loss refers to a condition where hair is absent in areas where it should normally be present, generally meaning the shedding of mature hair on the scalp. While hair loss was historically considered a male-only problem, the number of women suffering from hair loss is increasing in modern society, and the age range at which it occurs is gradually becoming younger. As the number of hair loss patients increases, research into the causes of hair loss, treatment methods, and therapeutic agents is attracting attention.

[0003] Currently, minoxidil and finasteride (Propecia) are cited as examples of the most effective drugs for preventing or treating hair loss. However, minoxidil affects hair all over the body, causing it to grow thicker and denser, and it also lowers blood pressure, so people with low blood pressure should refrain from using it. Furthermore, since the drug is metabolized in the liver, people with liver problems should also refrain from using it. Propecia is a male hormone, so it is prohibited for women to take it, and it is known to cause side effects such as loss of libido and sexual dysfunction. Recently, depression and low mood have been reported in patients who have been administered it, and these have been added as precautions. Therefore, there is a need to develop a hair loss treatment that has fewer side effects and is more effective than minoxidil and finasteride.

[0004] On the other hand, microbial resources, unlike petroleum and water, are classified as sustainable resources because they can be reproduced, and they have very high potential for research and industrial application because they utilize the unique characteristics of microorganisms adapted to diverse environments. In particular, components derived from microbial strains that have been inactivated by heat treatment or other means but still provide health benefits are called postbiotics. Postbiotics have functionalities such as immunomodulation, improvement of skin condition, antioxidant and inflammation regulation, and because they are inactivated, they have advantages over probiotics in terms of stability, safety, economy, and storability.

[0005] The skin surface and hair follicle openings are areas rich in microbial communities and exhibiting strong immune activation, demonstrating a strong correlation between immune privileges essential for the hair cycle and microbial communities. Changes in the microbiome or absorption of microbial metabolites in hair follicles are associated with inflammatory responses in the hair follicles, including the regulation and homeostasis of skin immune responses. Since reducing inflammation in the hair follicles increases the likelihood of hair regrowth, managing the microbiome in localized areas of the scalp is crucial for both hair loss and hair regrowth.

[0006] Therefore, products utilizing microorganisms are being developed for use in the rapidly increasing number of hair loss patients, with the aim of preventing hair loss, promoting hair growth, and stimulating hair development. One example is the Republic of Korea Published Patent No. 10-2017-0038462. However, research into the precise mechanisms of hair growth regulation by dermal papilla cells and new regulatory substances is still insufficient.

[0007] Therefore, the present inventors studied hair loss prevention or hair growth promotion compositions that can promote the proliferation of hair follicle dermal papilla cells (hDPCs, human dermal papilla cells). As a result, they confirmed that heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020 (KCCM12918P) promotes the proliferation of hair follicle dermal papilla cells, and thus completed the present invention. [Overview of the Initiative] [Problems that the invention aims to solve]

[0008] The present invention aims to provide a composition containing heat-treated lactic acid bacteria rimosilactobacillus fermentum LM1020 (KCCM12918P) that has the effect of preventing hair loss and promoting hair growth.

[0009] However, the problems that this application aims to solve are not limited to those mentioned above, and other problems not mentioned should be clearly understandable to an average engineer from the following description. [Means for solving the problem]

[0010] The first aspect of this application provides a cosmetic composition for hair papilla cell proliferation, comprising heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 (KCCM12918P) as an active ingredient.

[0011] A second aspect of the present application provides a food composition for hair papilla cell proliferation, comprising heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 (KCCM12918P) as an active ingredient.

[0012] A third aspect of this application is to provide a pharmaceutical composition for the treatment of hair loss, comprising heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 (KCCM12918P) as an active ingredient. [Effects of the Invention]

[0013] The heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 and compositions containing it promote the proliferation of dermal papilla cells, exhibit excellent inhibitory activity of 5α-reductase-1, and have hair loss prevention and hair growth promotion effects by promoting the expression of growth factors such as FGF7, FGF10, and EGF. [Brief explanation of the drawing]

[0014] [Figure 1] This figure shows the results of comparing the effects of heat-treated lactic acid bacteria on promoting hair papilla cell proliferation. [Figure 2]It is a figure showing the results of comparing the effects of heat-treated lactic acid bacteria on promoting the growth of papilla cells. [Figure 3] It is a figure showing the results of comparing the effects of different concentrations of heat-treated lactic acid bacterium Lactobacillus fermentum LM1020 on promoting the growth of papilla cells. [Figure 4a] It is a figure showing the change in the expression level of CDK2 in papilla cells after treatment with heat-treated lactic acid bacterium Lactobacillus fermentum LM1020. [Figure 4b] It is a figure showing the change in the expression level of CDK4 in papilla cells after treatment with heat-treated lactic acid bacterium Lactobacillus fermentum LM1020. [Figure 4c] It is a figure showing the change in the expression level of CDK6 in papilla cells after treatment with heat-treated lactic acid bacterium Lactobacillus fermentum LM1020. [Figure 4d] It is a figure showing the change in the expression level of cyclin B1 in papilla cells after treatment with heat-treated lactic acid bacterium Lactobacillus fermentum LM1020. [Figure 4e] It is a figure showing the change in the expression level of cyclin D1 in papilla cells after treatment with heat-treated lactic acid bacterium Lactobacillus fermentum LM1020. [Figure 4f] It is a figure showing the change in the expression level of cyclin E1 in papilla cells after treatment with heat-treated lactic acid bacterium Lactobacillus fermentum LM1020. [Figure 5a] It is a figure showing the results of confirming the gene expression level of 5α-reductase in human scalp tissue after treatment with heat-treated lactic acid bacterium Lactobacillus fermentum LM1020. [Figure 5b] It is a figure showing the results of confirming the gene expression level of FGF7 (Fibroblast growth factor 7) in human scalp tissue after treatment with heat-treated lactic acid bacterium Lactobacillus fermentum LM1020. [Figure 5c]This figure shows the results of examining the gene expression level of FGF10 (Fibroblast growth factor 10) in heat-treated Lactobacillus rimosilactobacillus fermentum LM1020 in human scalp tissue. [Figure 5d] This figure shows the results of examining the gene expression level of EGF (Epidermal Growth Factor) in heat-treated Lactobacillus rimosilactobacillus fermentum LM1020 in human scalp tissue. [Figure 6] This figure shows the results of comparing the hair papilla cell growth promoting effects of ermenthol, salicylic acid, dexpanthenol and mixtures thereof (LSD), heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020, and the ermenthol, salicylic acid, and dexpanthenol complex with heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020. [Figure 7a] This figure shows the change in the ratio of Staphylococcus and Lactic acid bacteria (Operational Taxonomy unit) in the scalp of participants before and after using a product containing heat-treated lactic acid bacteria rimosilactobacillus fermentum LM1020. [Figure 7b] This figure shows the changes in the Shannon index of Staphylococcus and Lactic acid bacteria against the overall microbiome in participants who used a product containing heat-treated lactic acid bacteria rimosilactobacillus fermentum LM1020, before and after use. [Figure 7c] This figure shows the changes in the Shannon index of Staphylococcus and Lactic acid bacteria in the scalp relative to total Lactic acid bacteria (LAB) in participants who used a product containing heat-treated lactic acid bacteria rimosilactobacillus fermentum LM1020 before and after use. [Figure 7d]This figure shows the changes in the major microorganisms on the scalp of participants before and after using a product containing heat-treated lactic acid bacteria, Limosilactobacillus fermentum LM1020. [Figure 7e] This figure shows the changes in the major microorganisms on the scalp of participants before and after using a product containing heat-treated lactic acid bacteria, Limosilactobacillus fermentum LM1020. [Modes for carrying out the invention]

[0015] In the following, embodiments of the present application will be described in detail with reference to the attached drawings, so that they can be easily implemented by a person with ordinary skill in the art to which the present application pertains. However, the present application can be embodied in various different forms and is not limited to the embodiments described herein. Furthermore, in order to clearly illustrate the present application, parts unrelated to the description have been omitted from the drawings, and similar parts throughout the specification are denoted by similar reference numerals.

[0016] Throughout the specification of this application, when a member is described as being "on top of" another member, this includes not only cases where the member is in contact with another member, but also cases where there is yet another member between the two members.

[0017] Throughout this specification, when a part “includes” a component, this means, unless otherwise stated, that it may include other components rather than excluding them. Throughout this specification, terms of degree such as “about” and “substantially” are used in the sense of, or close to, the numerical values ​​of the manufacturing and material tolerances inherent to the meaning referred to, where such values ​​are presented, and are used to prevent unscrupulous infringers from unfairly exploiting disclosures that refer to precise or absolute numerical values ​​to aid in understanding this application. Throughout this specification, terms of degree such as “~step” or “~step” do not mean “~step for.”

[0018] Throughout the specification of this application, the term “these combinations” as used in the Markush expression means one or more mixtures or combinations selected from the group of components described in the Markush expression, and means including one or more selected from the group of components.

[0019] Throughout the specification of this application, the phrase "A and / or B" means "A or B, or A and B."

[0020] The following describes in detail the embodiment and examples of the present application with reference to the attached drawings. However, the present application is not limited to these embodiment and examples and drawings.

[0021] The first aspect of this application provides a cosmetic composition for hair papilla cell proliferation, comprising heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 (KCCM12918P) as an active ingredient.

[0022] The term "heat-treated lactic acid bacteria" as used throughout the specification of this application refers to dead cells of the Limosilactobacillus fermentum LM1020 strain that have been inactivated by heat treatment.

[0023] The term "dead bacterial cells," as used throughout this specification, is the opposite of "live bacteria," and refers to a form in which live bacteria and metabolites obtained by fermentation are rendered unable to grow through heat treatment or other means. Dead bacterial cells may contain cytoplasm, cell walls, antimicrobial active substances such as bacteriocins, polysaccharides, organic acids, etc. Products utilizing the aforementioned dead bacterial cells have higher stability compared to live bacterial products, particularly excellent heat resistance, and high stability against the external environment, making them easier to store and allowing for a longer distribution period than existing live bacterial products. Furthermore, given the strengthening of regulations on the use of antibiotics, there is high potential for use as an alternative, and because only a handful of companies have seriously entered the production of dead bacterial cell products, the market potential and growth potential are very high.

[0024] In one embodiment of the present invention, the component derived from the rimosilactobacillus fermentum LM1020 strain may be derived from fermented dough, but is not limited thereto.

[0025] In one embodiment of the present application, the heat-treated lactic acid bacterium Rimosilactobacillus fermentum LM1020 may have a saturated fatty acid:unsaturated fatty acid:cyclic fatty acid ratio of 0.1-2:0.25-5:0.03-0.6, preferably 1:2.5:0.3, but is not limited thereto.

[0026] In one embodiment of the present invention, the heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 may increase the expression of CDK2, CDK4, cyclin B1, and cyclin D1.

[0027] In one embodiment of the present invention, the heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 may decrease the expression level of the 5α-reductase-1 gene and increase the expression levels of the FGF7, FGF10, and EGF genes.

[0028] In one embodiment of the present invention, the cosmetic composition for hair papilla cell proliferation may further contain ermenthol, salicylic acid, and dexpanthenol.

[0029] In one embodiment of the present invention, when the heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 of the present invention was treated together with a complex of elmenthol, salicylic acid, and dexpanthenol, the effect of promoting hair papilla cell growth was superior to that of using the heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 alone.

[0030] In one embodiment of the present invention, the heat-treated lactic acid bacterium Limosilactobacillus fermentum LM1020 is 1 × 10 4 cells / mL ~ 1 × 10 8It may be cells / mL, preferably 1 × 10 6 cells / mL ~ 1 × 10 7 It may be cells / mL, and more preferably 1 × 10 6 cells / mL or 1 × 10 7 The value may be in the format of cells / mL, but is not limited to this.

[0031] In one embodiment of the present application, the cosmetic composition for hair papilla cell proliferation may further contain purified water, ethanol, alanine / histidine / lysine polypeptide copper HCl, polyacrylate crosspolymer-6, coconut acid, proline, tea tree oil, glycerin, peptide, butylene glycol, 1,2-hexanediol, green tea extract, lavender flower extract, quince extract, and ethylhexanediol, and the cosmetic composition for hair papilla cell proliferation may also increase the microbial diversity and lactic acid bacteria diversity of the scalp.

[0032] The second aspect of this application provides a food composition for hair papilla cell proliferation, comprising the heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 (KCCM12918P) as an active ingredient. Content overlapping with the first aspect of this application also applies to the food composition of the second aspect.

[0033] As used throughout the specification of this application, the term "food" includes all foods in the ordinary sense, such as meats, sausages, bread, chocolates, candies, snacks, confectionery, pizzas, ramen and other noodle products, gums, dairy products including ice cream, various soups, beverages, teas, energy drinks, alcoholic beverages, vitamin complexes, functional foods, and health foods.

[0034] The term "health functional food" as used throughout the specification of this application means a food manufactured and processed using raw materials or components that have functional properties useful to the human body, as defined in Act No. 6727 on Health Functional Foods. "Functionality" means obtaining effects useful for health purposes, such as regulating nutrients or physiological effects on the structure and function of the human body.

[0035] The food product of this invention can be manufactured by methods commonly used in the industry, and during such manufacturing, raw materials and components commonly added in the industry may be added. Furthermore, the dosage form of the food product is not limited as long as it is a dosage form recognized as a food product. The food composition of the present invention can be manufactured in various dosage forms, and unlike general pharmaceuticals, it is made from food as a raw material, so it has the advantage of not having side effects that may occur with long-term use of pharmaceuticals, and it is highly portable, so the food product of the present invention can be taken as an auxiliary agent to enhance the effect of improving the intestinal environment.

[0036] The term "health food" refers to foods that have a more active effect on maintaining or promoting health compared to general foods, while "health supplement food" refers to foods intended to supplement health. Depending on the context, the terms "health functional food," "health food," and "health supplement food" may be used interchangeably. Specifically, the term "health functional food" refers to foods in which the heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 of this application is added to food ingredients such as beverages, teas, spices, gums, and confectionery, or manufactured in the form of capsules, powders, or suspensions, and which, when consumed, produce specific health effects. However, unlike general medicines, it uses food as a raw material, and therefore has the advantage of not having the side effects that can occur with long-term use of medicines.

[0037] Because the food composition of this invention can be consumed on a daily basis, it is expected to have a high effect in improving depression and can therefore be used very effectively.

[0038] The food composition may further contain a physiologically acceptable carrier, but the type of carrier is not particularly limited, and any carrier commonly used in the art can be used.

[0039] Furthermore, the food composition may contain additional ingredients commonly used in food compositions that can improve smell, taste, appearance, etc. For example, it may contain vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. It may also contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr). It may also contain amino acids such as lysine, tryptophan, cysteine, and valine.

[0040] Furthermore, the food composition may also contain food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder and high-grade bleaching powder, sodium hypochlorite, etc.), antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), colorants (tar dyes, etc.), color fixatives (sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, potassium bitartrate, etc.), fortifiers, emulsifiers, thickeners, coating agents, gum bases, antifoaming agents, solvents, and improvers. The additives may be selected according to the type of food and used in appropriate amounts.

[0041] The heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 of this application may be added as is, used together with other foods or food components, or used appropriately by conventional methods. The amount of active ingredient mixed may be appropriately determined according to its intended use (prevention, health, or therapeutic treatment). Generally, when manufacturing food or beverages, the food composition of the present invention may be added to the food or beverage in an amount of 50 parts by weight or less, specifically 20 parts by weight or less. However, when ingested for long-term purposes for health and hygiene, the content may be below the above range, and there are no safety concerns, so the active ingredient may be used in amounts above the above range.

[0042] One example of the food composition of the present invention is that it may be used as a health beverage composition, in which case it may contain various flavorings or natural carbohydrates as additional ingredients, as in ordinary beverages. The natural carbohydrates mentioned above may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; or sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The proportion of the natural carbohydrates is generally about 0.01 to 0.04 g per 100 mL of the health beverage composition of the present invention, and specifically, it may be about 0.02 to 0.03 g.

[0043] In addition to the above, the health beverage composition may also contain various nutrients, vitamins, electrolytes, flavorings, colorings, pectin, salts of pectin, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonation agents. Furthermore, it may contain fruit pulp for producing natural fruit juice, fruit juice beverages, or vegetable beverages. These components may be used individually or in combination. The proportion of such additives is not particularly important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health beverage composition of the present invention.

[0044] The food composition of the present application may contain heat-treated lactic acid bacteria rimosilactobacillus fermentum LM1020 of the present application in various weight percentages, as long as it can exhibit the effect of preventing hair loss and promoting hair growth. Specifically, the food composition may contain heat-treated lactic acid bacteria rimosilactobacillus fermentum LM1020 of the present application in an amount of 0.00001 to 100% by weight or 0.01 to 80% by weight relative to the total weight of the food composition, but is not limited thereto.

[0045] In one embodiment of the present invention, the food composition may be a health functional food composition.

[0046] The third aspect of this application provides a pharmaceutical composition for the treatment of hair loss, comprising heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 (KCCM12918P) as an active ingredient. The content that overlaps with the first and second aspects also applies to the pharmaceutical composition of the third aspect of this application.

[0047] In one embodiment of the present application, the pharmaceutical composition may be formulated by conventional methods into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, topical preparations, suppositories, or sterile injection solutions for use, but is not limited thereto.

[0048] In one embodiment of the present application, when formulating the pharmaceutical composition, it may be compounded using commonly used fillers, bulking agents, binders, wetting agents, disintegrants, or diluents or excipients such as surfactants, but is not limited to these.

[0049] In one embodiment of the present invention, the solid preparation for oral administration includes tablets, pills, powders, granules, or capsules, and such solid preparations may be compounded by mixing the component derived from the bacterial strain with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, or gelatin. In addition to mere excipients, lubricants such as magnesium stearate or talc may also be used, but are not limited thereto.

[0050] In one embodiment of the present invention, the liquid formulation for oral administration includes suspensions, oral solutions, emulsions, syrups, etc., and may include, but is not limited to, various excipients such as water and liquid paraffin, which are commonly used diluents, as well as humectants, sweeteners, fragrances, and preservatives.

[0051] In one embodiment of the present invention, the preparation for parenteral administration may include, but is not limited to, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. For example, the non-aqueous solvent or suspension may include, but is not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. For example, the suppository may include, but is not limited to, witepsol, macrogol, tween 61, cocoa butter, lauric butter, and glycerol gelatin.

[0052] The pharmaceutical composition relating to one embodiment of the present application may be a pharmaceutical composition or a quasi-drug composition.

[0053] The term "quasi-drug" as used throughout this specification refers to articles used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases in humans or animals, which have a milder effect than pharmaceuticals. For example, according to the Pharmaceutical Affairs Law, quasi-drugs are articles other than those used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases in humans and animals, and products that have a mild or no direct effect on the human body.

[0054] The quasi-drug composition of this application may be manufactured in a dosage form selected from the group consisting of body cleansers, disinfectants, detergents, kitchen detergents, cleaning detergents, toothpaste, mouthwashes, wet wipes, detergents, soaps, hand washes, hair washes, hair softeners, humidifier fillers, masks, ointments, and filter fillers, but is not limited thereto.

[0055] In one embodiment of the present application, the pharmaceutical composition may be administered in a pharmacokinetically effective amount, where the term "pharmacokinetically effective amount" means an amount sufficient to treat or prevent a disease with a reasonable benefit / risk applicable to medical treatment or prevention, and the level of the effective dose may be determined by the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's drug sensitivity, the time of administration, route of administration and excretion rate of the composition of the present invention used, the duration of treatment, factors including drugs used in combination with or concurrently with the composition of the present invention used, and other factors well known in the medical field. The pharmaceutical composition of the present application may be administered alone or in combination with components known to exhibit therapeutic effects for known intestinal diseases. Taking all of the above factors into consideration, it is important to administer an amount that provides the maximum effect with the minimum amount without side effects.

[0056] In one embodiment of the present application, the dosage of the pharmaceutical composition may be determined by a person skilled in the art, taking into consideration the purpose of use, the severity of the disease, the patient's age, weight, sex, medical history, or the type of substance used as the active ingredient. For example, the pharmaceutical composition of the present invention may be administered at a dose of about 0.1 ng to about 1,000 mg / kg per adult, preferably 1 ng to about 100 mg / kg. The frequency of administration of the composition of the present application is not particularly limited thereto, but may be administered once a day or divided into several doses. The aforementioned dosage or frequency of administration does not limit the scope of the present application in any way.

[0057] The pharmaceutical composition of this application is not limited to the present invention, but may be administered via routes such as intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, intrapulmonary administration, or intrarectal administration, depending on the purpose. However, in the case of oral administration, it may be administered in an unformed dosage form, and since the heat-treated lactic acid bacterium limosiractobacillus fermentum LM1020 may be denatured or degraded by gastric acid, the oral composition may be administered orally in a dosage form that is coated with the active agent or protected from degradation in the stomach, or in the form of an oral patch. Furthermore, the composition may be administered by any device that allows the active substance to move to target cells.

[0058] The present invention will be described in more detail below with reference to the embodiments of this application, but the embodiments described below are for illustrative purposes only to aid in understanding this application, and the content of this application is not limited to the embodiments described below. [Examples]

[0059] Example 1. Comparison of the effect of heat-treated lactic acid bacteria on hair papilla cell proliferation. Hair papilla cells were purchased from PromoCell, at a rate of 5000-10000 cells / cm². 2The cells were inoculated into flasks at the specified density and then cultured. Dermal papilla cells were cultured in a dermal papilla cell culture medium prepared by adding growth factors (40 μL / mL fetal calf serum, 4 μL / mL bovine pituitary extract, 1 ng / mL fibroblast growth factor, 5 μg / mL insulin) to basal media, a commercial culture medium, and cultured at 37°C while maintaining a CO2 concentration of 5%. Dermal papilla cells were used for subculturing and experiments once they reached 80-90% confluency. Dermal papilla cells used in the study were stored in liquid nitrogen for 2-5 generations, and up to 10 generations were used in experiments.

[0060] The effect of heat-treated lactic acid bacteria on promoting hair papilla cell proliferation was confirmed by observing viable cells under a microscope via trypan blue staining. Hair papilla cells that had reached 80-90% confluency were inoculated into a 24-well plate at a concentration of 100,000 cells / well and cultured for 24 hours, after which 1 × 10⁶ cells were cultured. 7 The increase in the number of cells compared to the control group was confirmed by directly counting the heat-treated lactic acid bacteria at a concentration of cells / mL using a microscope. The heat-treated lactic acid bacteria used to compare the hair papilla cell proliferation-promoting effect were Lactobacillus plantarum LP1001, Lactococcus lactis LP1009, Lacticaseibacillus rhamnosus LP1011, Lacticaseibacillus paracasei LP1014, Bifidobacterium animalis lactis LP1017, Rimosilactobacillus fermentum LP1016, Rimosilactobacillus fermentum LM1020, Lactobacillus acidophilus LP1060, and Lactobacillus gasseri LP1065.

[0061] The comparison revealed that none of the heat-treated lactic acid bacteria promoted the proliferation of hair papilla cells. Of the nine types of heat-treated lactic acid bacteria, only Lactobacillus paracasei LP1014, Lactobacillus acidophilus LP1060, Rimosilactobacillus fermentum LP1016, and Rimosilactobacillus fermentum LM1020 were able to promote the growth of hair papilla cells, while the other heat-treated lactic acid bacteria actually inhibited their growth (Figure 1).

[0062] Although Rimosilactobacillus fermentum LP1016 and Rimosilactobacillus fermentum LM1020 belong to the same genus and species, LP1016 originated from kimchi, while LM1020 originated from fermented dough, thus differing in their origins. Research results showed that even when heat-treated using the same method, heat-treated Rimosilactobacillus fermentum LP1016 derived from kimchi improved hair papilla cell proliferation by 3.7%, while heat-treated Rimosilactobacillus fermentum LM1020 derived from fermented dough improved hair papilla cell proliferation by 24.9%, demonstrating a significant difference.

[0063] Example 2. Cellular fatty acid composition of heat-treated lactic acid bacteria Cellular fatty acids are precursors of intracellular membranous organelles, including cell membranes, and are known to play various roles within the bacterial cell. The internal standard (ISTD) was selected as methyl undecanoate, which is not commonly found in nature. It was prepared at a concentration of 10,000 μg / mL using HPLC-grade n-hexane as the solvent, and then used in research while being stored at -20°C and diluted as needed. To extract fatty acids from heat-treated lactic acid bacteria, 25 μl of the internal standard was added, followed by 200 μl of a chloroform and methanol 2:1 (v / v) mixture. Then, 300 μl of 0.6 M hydrochloric acid methanol solution was added for fatty acid derivatization, homogenized for 120 seconds, and heated at 85°C for 60 minutes, after which it was allowed to cool to room temperature. 1 mL of n-hexane was added again, homogenized for 120 seconds, and left at room temperature for 60-120 minutes before the supernatant was used for analysis. Fatty acids were analyzed using gas chromatography / mass spectrometry electron ionization analysis, with helium as the mobile phase and a DB-FastFAME column.

[0064] As a result, it was confirmed that the fatty acid composition of the cells of heat-treated lactic acid bacteria limosilactobacillus fermentum LP1016 and LM1020, which belong to the same genus and species but have different strain origins, differs from each other (Table 1). In particular, linoleate was present only in heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020, and behenic acid was present only in heat-treated lactic acid bacteria limosilactobacillus fermentum LP1016. Among the fatty acids, oleate was present in approximately 2.4 times more of heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020, and lactobacillic acid was present in approximately 2.1 times more of heat-treated lactic acid bacteria limosilactobacillus fermentum LP1016.

[0065] The two strains also had significantly different ratios of saturated fatty acids to unsaturated fatty acids in their cell bodies. For Limosilactobacillus fermentum LP1016 derived from kimchi, the ratio of saturated fatty acids:unsaturated fatty acids:cyclic fatty acids was 1:1:0.6, while Limosilactobacillus fermentum LM1020 derived from fermented dough showed a ratio of 1:2.5:0.3. The composition of each fatty acid was different, and it was confirmed that the characteristics of the overall fatty acids also showed significant differences.

[0066]

Table 1

[0067] Example 3. Comparison of the effect of heat-treated lactic acid bacteria on promoting the growth of hair papilla cells Through comparison with strains (Lactobacillus casei paracasei KCTC14004BP and Limosilactobacillus fermentum KCCM11910P) known to be effective in preventing hair loss, promoting hair growth, or promoting hair regrowth, the effect of Limosilactobacillus fermentum LM1020, a heat-treated lactic acid bacterium, on the proliferation of hair papilla cells was verified (Figure 2).

[0068] The strains used for comparison were all heat-treated at a high temperature of 121°C for 15 minutes to be inactivated.

[0069] After treating hair papilla cells (10,000 cells / well) with heat-treated lactic acid bacterium at a concentration of 2.5×10 7 cell / mL, they were cultured as in Example 1, and the growth-promoting effect of the hair papilla cells was calculated using the following formula.

[0070] The growth rate of hair papilla cells cultured alone was compared with the growth rate when heat-treated lactic acid bacteria were added to confirm the hair papilla cell growth promoting effect of heat-treated lactic acid bacteria. The growth rate of hair papilla cells cultured using the above method without any additives was calculated, as well as the growth rate of hair papilla cells when heat-treated lactic acid bacteria Lactobacillus paracasei KCTC14004BP, heat-treated lactic acid bacteria Rimosiractobacillus fermentum KCCM11910P, and heat-treated lactic acid bacteria Rimosiractobacillus fermentum LM1020 were added. The hair papilla cell growth promoting effect of heat-treated lactic acid bacteria was calculated using the following formula and compared. All tests were repeated three times, and the average of each result was calculated and compared.

[0071] Hair papilla cell growth promoting effect (%) = (BA) ÷ A × 100 A = Hair papilla cell growth rate in the untreated group B = Growth rate of hair papilla cells when treated with heat-treated lactic acid bacteria

[0072] As a result, 2.5 × 10 7 At a concentration of cells / mL, heat-treated lactic acid bacterium rimosilactobacillus fermentum KCCM11910P actually inhibited the growth of hair papilla cells, while heat-treated lactic acid bacterium lacticaseibacillus paracasei KCTC14004BP promoted the growth of hair papilla cells, but showed a cell growth rate more than twice as low as that of the heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 of the present invention.

[0073] Example 4. Effect of heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 on hair papilla cell proliferation at different concentrations. In Examples 1 and 2, heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 showed the best growth-promoting effect on dermal papilla cells. This study investigated whether it could promote the proliferation of dermal papilla cells even in conditions where growth factors were deficient. Dermal papilla cells were inoculated into a 96-well plate and cultured for 24 hours. Then, the culture medium in the wells was replaced with a growth factor-deficient medium, and cultured for an additional 24 hours. Subsequently, heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 was added at different concentrations, and the growth of dermal papilla cells was checked after 24 hours. The growth-promoting effect on dermal papilla cells was confirmed by dissolving the formazan produced while culturing at 37°C with 0.5 mg / mL of thiazolyl blue tetrazolium bromide (MTT) in DMSO and measuring the absorbance at 570 nm.

[0074] As a result, heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 can proliferate dermal papilla cells even under culture conditions deficient in dermal papilla cell growth factor, 1 × 10⁻⁶ 3 cells / mL ~ 1 × 10 5 Within the concentration range of cells / mL, hair papilla cell growth increases in a concentration-dependent manner, 1 × 10⁻⁶ 6 At high concentrations of cells / mL or higher, the hair papilla cell proliferation effect showed a similar trend regardless of concentration. The hair papilla cell proliferation promoting effect was 1 × 10⁻⁶ 7 The cell / mL ratio was the most impressive, showing a 156% growth-promoting effect compared to the untreated group (Figure 3).

[0075] Example 5. Increased protein expression levels associated with hair papilla cell proliferation. In order to confirm the mechanism by which heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020, which showed the best hair papilla cell growth promoting effect in Examples 1 and 2, promotes hair papilla cell proliferation, we measured the expression levels of proteins related to hair papilla cell proliferation.

[0076] Dermal papilla cells are fibroblasts derived from the mesoderm, and their proliferation is regulated by cell cycle proteins such as cyclin, CDKs (cyclin-dependent kinases), and CDK inhibitors. Cells exist in either the resting or proliferating phase, and growing cells go through the cell cycle of G1 phase, S phase (DNA synthesis phase), G2 phase, and M phase (mitosis phase), ultimately dividing into two daughter cells and proliferating.

[0077] CDK2 (Cyclin-dependent kinase 2) is a kinase involved in cell cycle regulation. It is essential for meiosis but not for mitosis. It regulates the activation of cyclin BCDK1 as a mechanism to control the timing of entry into mitosis and meiosis by controlling the subsequent activation of cyclin B / CDK1 through phosphorylation. CDK2 activity is maximally expressed in the S and G2 phases of cell division. CDK4 (Cyclin-dependent kinase 4) is required for the transition from the G1 to the S phase of the cell cycle. CDK6 (Cyclin-dependent kinase 6) promotes the G1 / S transition and is involved in the initiation and maintenance of the cell cycle during cell differentiation. Its main function is to prevent cell proliferation and regulate cell differentiation in a negative way. Cyclin B1 is a regulatory protein involved in mitosis and is a cell cycle regulatory transcript expressed during the G2 / M phase of the cell cycle.

[0078] Cyclin B1 plays a role in determining mitosis in cells, and after being activated and forming the cyclin B1-CDK1 complex, it promotes various mechanisms such as early mitosis.

[0079] Cyclin D1 is one of the target genes of the Wnt / β-catenin signaling pathway, which plays an important role in processes such as hair growth, stem cell regulation, and cell proliferation, and it influences cell division throughout the entire cell division cycle of dermal papilla cells.

[0080] Cyclin E1 may induce the early G1 phase of the cell division cycle in dermal papilla cells. Cyclin E1 promotes the transition from G1 to S phase, initiating DNA synthesis. Cyclin E1 is expressed most maximally at the G1 / S stage of the cell cycle.

[0081] To measure the protein expression levels of dermal papilla cells, 4 × 10⁶ dermal papilla cells were placed in a 6-well microplate. 5 After injecting at a concentration of cells / mL, the cells were cultured for 24 hours. Then, the dermal papilla cell culture medium was replaced with a basic medium (a medium from which growth factors for dermal papilla cells, such as insulin, have been removed), and cultured for an additional 24 hours. Subsequently, the cells were cultured at different concentrations (final concentration: heat-treated lactic acid bacteria rimosilactobacillus fermentum LM1020 1 × 10⁶). 6 , 1 x 10 7 and 1 × 10 8 The dermal papilla cell culture medium was replaced with a medium treated with (cell / mL), and dermal papilla cells were cultured together with heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020 at different concentrations for 24 hours to induce proliferation of the dermal papilla cells. After the culture was complete, Pro-Prep TM Proteins from dermal papilla cells were extracted using lysis buffer (Intron, South Korea). The extracted proteins were analyzed by measuring protein quantity using a BCA protein assay, and protein expression levels were determined by performing Western blotting using the Protein Simple Jess system (Jess; Protein Simple, USA). Electrophoresis, blocking, primary and secondary antibody reactions were carried out within the Jess capillary cartridge, and expression levels were confirmed using ECL detection reagent.

[0082] As a result, it was confirmed that heat-treated lactic acid bacterium limosiractobacillus fermentum LM1020 increased the expression of cyclin and CDK proteins, which are necessary for the growth cycle of dermal papilla cells, and promoted dermal papilla cell growth (Figures 4a to 4f). Although the degree of protein expression affecting dermal papilla cell proliferation differed depending on the treatment concentration of heat-treated lactic acid bacterium limosiractobacillus fermentum LM1020, the expression of CDK2, CDK4, cyclin B1, and cyclin D1 increased, while the expression of CDK6 and cyclin E1 tended to decrease (Table 2).

[0083] In particular, heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 increased the expression level of cyclin B1 by up to 3.9 times (Figure 4d), which means that during the same growth period, a greater number of cells reached the G2 / M phase of the dermal papilla cell division cycle compared to the control group (untreated), and a greater increase in the number of dividing dermal papilla cells after DNA replication.

[0084] Furthermore, when treated with heat-treated lactic acid bacterium rimosiractobacillus fermentum LM1020, the expression of cyclin D1 protein, which affects cell division throughout the entire cell division cycle of dermal papilla cells, increased by up to 2.4 times (Figure 4e). This indicates that heat-treated lactic acid bacterium rimosiractobacillus fermentum LM1020 formed a cyclin D1-CDK4 complex within dermal papilla cells, inducing the transition from the G1 phase of cell division to the S phase where DNA is replicated, thereby promoting the growth of dermal papilla cells.

[0085] On the other hand, after treatment with heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020, the expression levels of cyclin E and CDK6, which induce the early G1 phase of the cell division cycle in dermal papilla cells, were slightly decreased or maintained at levels similar to those before treatment (Figures 4c and 4f). This means that dermal papilla cells treated with heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020 progressed more frequently through the G1 phase, the initial stage of the cell division cycle, to the G2 / M phase.

[0086] [Table 2]

[0087] Example 6. Measurement of hair loss prevention and hair growth effects using human-derived tissue. 6-1) Culture of Explant Tissue Human scalp tissue provided for research purposes (IRB No. 4-2021-1524; Yonsei University College of Medicine, Severance Medical Center Bioethics Committee) was washed several times with PBS to remove residual impurities, and then prepared in 1cm x 1cm sections.

[0088] Heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020 were added to a scalp tissue culture medium in a 1x10⁶ format. 8 cells / mL or 1 × 10⁶ 9 After dilution to a concentration of cells / mL, 50 μL was applied to human scalp tissue. The negative control group was treated with 50 μL of scalp tissue culture medium instead of heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020. The cultures were incubated in the culture medium at 37°C and 5% CO2, with the culture medium being replaced and the treatment substance applied at 24-hour intervals. 72 hours after application, subcutaneous, epidermal, and dermal tissues were separated from each tissue sample and used for testing.

[0089] 6-2) Real-Time Polymerase Chain Reaction (RT-PCR) The gene expression levels of hair loss and hair growth-related factors (5α-reductase-1, FGF7, FGF10, EGF) were confirmed using real-time polymerase chain reaction. The gene expression level of 5α-reductase-1 was examined in subcutaneous tissue isolated from human scalp tissue, while the gene expression levels of FGF7, FGF10, and EGF were examined in epidermal and dermal tissue.

[0090] 5α-reductase-1, one of the factors associated with hair loss, is an enzyme mainly distributed in dermal papilla cells, sebaceous glands, epidermis, and keratinocytes of hair follicles. It acts with testosterone to produce DHT (dihydrotestosterone), which is a major cause of hair loss, and it is known that a decrease in 5α-reductase-1 gene expression in human dermal papilla cells is involved in preventing hair loss (H Rastegar et al., 2015).

[0091] FGF7 (fibroblast growth factor type 7) is a growth factor produced by fibroblasts and secreted to keratinocytes, which promotes the proliferation of epithelial and epidermal cells. It is known to promote hair follicle cell differentiation, induce proliferation of the vascular system of the dermal papilla, and increase the amount of extracellular matrix, thereby maintaining hair follicles in the growth phase. FGF10 (fibroblast growth factor type 10) is known to contribute to the initial formation of cells by inducing resting hair follicles into the growth or proliferation phase, and is a factor that promotes hair growth (Sole Cho et al., 2016).

[0092] EGF is well known as a growth factor involved in hair follicle maturation and has been studied to promote the proliferation of dermal papilla cells through the activation of the notch mechanism (Zhang et al., 2016).

[0093] After the testing was completed, the tissue was pulverized using TissueLyser II (Qiagen), and total RNA was extracted using TRIzol Reagent (Invitrogen). The extracted total RNA was then used to synthesize cDNA using RNA to cDNA EcoDry™ Premix (Oligo dT) (Clontech). Real-time polymerase chain reaction was carried out using synthesized cDNA, Taqman Fast Advanced Master Mix (Applied Biosystems), and Taqman primers for each target (SRD5A1 (5-α-reductase-1):Hs00971645_g1, FGF7:Hs00940253_m1, FGF10:Hs00610298_m1, EGF:Hs01100002_m1;Applied Biosystems). Relative quantitative analysis of each gene was performed using the housekeeping gene GAPDH (Hs02786624_g1, Applied Biosystems).

[0094] During cDNA amplification, the amplification amount is monitored in real time, and the threshold cycle (Ct) value, which is the intersection of the amplification curve and the threshold line, is obtained. Based on this value, the relative mRNA expression level (RQ) can be determined, allowing for the confirmation of the relative mRNA expression level of the target gene. RQ was calculated using the following formula.

[0095] RQ=2 -△△Cт △△Cт=△Cт(treatment)-△Cт(control) △Cт=Cт(target gene)-Cт(housekeeping gene) -Treatment (experimental group): Treatment of test product -Control (control group): Negative control group -Target gene: SRD5A1 (5α-reductase-1), FGF7, FGF10, EGF -Housekeeping gene:GAPDH

[0096] 6-3) Statistical analysis Statistical analysis was performed using the IBM SPSS statistics 25.0 program, and the significance of the difference between the experimental group and the control group was confirmed using a hypothetical mean difference of 5% (p<0.05). After testing for normality, significance was confirmed using an independent sample t-test (parametric method) based on whether normality was satisfied.

[0097] 6-4) Research results 5α-reductase, known to induce hair loss, was significantly reduced at all concentrations, regardless of the concentration of heat-treated lactic acid bacterium Rimosilactobacillus fermentum LM1020 (p<0.05). FGF7, FGF10, and EGF, which induce hair growth and hair development, were significantly reduced in the high-concentration treatment group (1 × 10⁻⁶). 9 The cell / mL ratio increased significantly (p<0.05). Therefore, it was confirmed that heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 is useful in preventing hair loss and promoting hair growth.

[0098] Example 7. Functional ingredients that help alleviate hair loss symptoms and the hair loss prevention effect of heat-treated lactic acid bacteria rimosilactobacillus fermentum LM1020. The effects on dermal papilla cell growth were investigated when using ermenthol, salicylic acid, and dexpanthenol—active ingredients commonly used in functional cosmetics to alleviate hair loss symptoms—in combination with heat-treated lactic acid bacterium Rimosilactobacillus fermentum LM1020. Dermal papilla cells were inoculated into a 96-well plate and cultured for 24 hours. Then, the culture medium in the wells was replaced with a growth factor-deficient medium, and cultured for an additional 24 hours. Subsequently, heat-treated lactic acid bacterium Rimosilactobacillus fermentum LM1020, ermenthol, salicylic acid, and dexpanthenol individually or mixed in a ratio of 3:2:6:2—and the effects on dermal papilla cell growth were investigated.

[0099] After calculating the growth rate of dermal papilla cells cultured alone and the growth rate of dermal papilla cells treated individually or in combination with menthol, salicylic acid, dexpanthenol, and heat-treated lactic acid bacteria rimosilactobacillus fermentum LM1020, the dermal papilla cell growth promoting effect of each treatment substance was calculated using the following formula, and the effects were compared. All tests were repeated three times, and the average of each result was calculated and compared.

[0100] Hair papilla cell growth promoting effect (%) = (BA) ÷ A × 100 A = Hair papilla cell growth rate in the untreated group B = Growth rate of hair papilla cells after treatment with elmenthol, salicylic acid, dexpanthenol, and heat-treated lactic acid bacterium rimosilactobacillus fermentum LM1020

[0101] The results showed that, among ermenthol, salicylic acid, and dexpanthenol, which are widely known to have hair loss mitigating effects, ermenthol and salicylic acid inhibited the growth of dermal papilla cells. Dexpanthenol promoted the growth of dermal papilla cells, but its effect was lower than that of heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020 at all concentrations. On the other hand, when heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020 was treated with a complex of ermenthol, salicylic acid, and dexpanthenol, it was confirmed that the complex's inhibitory effect on dermal papilla cell growth was overcome, and the growth of dermal papilla cells was promoted. To understand these synergistic effects in detail, the predicted effect was calculated using Colby's formula, and the predicted value was compared with the measured value to verify whether a synergistic effect exceeding the predicted value could be achieved.

[0102] Colby's formula Predicted value = (A + B) - (A × B / 100) (Source: SR Colby, "Calculating Synergistic and Antagonistic Responses of Herbicide Combinations", Weeds 1967, 15, 20-22)

[0103] As a result, a mixture of ermenthol, salicylic acid, and dexpanthenol in a ratio of 3:2:6:2 inhibited the growth of hair papilla cells, but when this mixture was treated together with heat-treated lactic acid bacteria limosiractobacillus fermentum LM1020, the growth of hair papilla cells was promoted. Furthermore, when this mixture was treated together with heat-treated lactic acid bacteria limosiractobacillus fermentum LM1020, the growth of hair papilla cells was promoted. 6 or 1 × 10 7 When treated with cells / mL, the hair papilla cell growth promoting effect was superior to that of heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 when used alone.

[0104] [Table 3]

[0105] Manufacturing Example 1: Development of a liquid formulation containing heat-treated lactic acid bacterium Rimosilactobacillus fermentum LM1020. In Example 5, a synergistic effect was confirmed in the hair loss alleviation functional cosmetic. We developed a liquid formulation that can be sprayed onto the scalp for even application, containing ermenthol, salicylic acid, and dexpanthenol, which are active ingredients in the hair loss alleviation functional cosmetic, along with heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020. The developed liquid formulation contains 0.3% ermenthol, 0.26% salicylic acid, 0.2% dexpanthenol, and 5% heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020. In addition, purified water, ethanol, alanine / histidine / lysine polypeptide copper HCl, polyacrylate crosspolymer-6, coconut acid, proline, tea tree oil, glycerin, peptide, butylene glycol, 1,2-hexanediol, green tea extract, lavender flower extract, quince extract, ethylhexanediol, and other additives.

[0106] Example 8. Verification of hair loss reduction effect through human application testing. The human clinical trial was conducted by Global Medical Research Center Co., Ltd., and was approved by the Research Ethics Committee (Approval Number: GIRB-21029-ET). The trial involved 25 Korean men and women aged 18-54, of whom two dropped out.

[0107] All participants applied the product from Manufacturing Example 1 evenly to their scalp before going to bed for six months. Visits were conducted at 0, 8, 16, and 24-week intervals to evaluate product adaptation, hair density measurement using phototrichograms, visual evaluation by experts, and effectiveness questionnaires from participants (satisfaction with hair growth, satisfaction with hair loss reduction, and satisfaction with the bangs hairline).

[0108] The results of the study examining the participants' adaptation to the product showed that "0" participants used the product less than 80% of the prescribed number of uses. Therefore, all participants used the test product as prescribed, applying it to their scalp once a day before bedtime, for a total of 24 weeks.

[0109] The efficacy of the test product was verified by measuring hair density (phototrichogram), taking photographs and visual evaluations by experts, and evaluating the effectiveness in terms of user satisfaction. To measure hair density, the hair loss area to be evaluated was measured in 1 cm². 2 After removing hair in a circular area, a 1mm diameter dot was tattooed, and the area was photographed using a hair density meter. Hair density was measured four times, every eight weeks, with the tattoo as the reference point (1cm). 2 The number of hairs within the circle was measured.

[0110] As a result, participants in the study experienced a sustained increase in hair density compared to before use of the test product, and after 16 weeks, the increase in hair density was statistically significant compared to before use (p<0.05).

[0111] [Table 4]

[0112] Visual evaluation by experts was performed by photographing the test areas: the crown of the head (90°) and the hairline (45°). Two experts performed the visual evaluation and rated the changes on a 7-point scale. As a result, it was found that the participants experienced a reduction in hair loss symptoms that was visible to the naked eye while using the test product, and that hair regrowth was progressing. The degree of agreement in the visual evaluations between researchers was confirmed using the ICC (Intraclass Correlation Coefficients) method to verify the precision of the research results. The intraclass correlation coefficient for the visual evaluations between researchers was 0.916, which is close to 1, indicating perfect agreement.

[0113] [Table 5]

[0114] After using the product, participants evaluated its effectiveness using a 7-point scale (-3 points: significantly worse, -2 points: worse, -1 point: slightly worse, 0 points: no change, 1 point: slightly better, 2 points: better, 3 points: significantly better). The results showed that participants who used the test product for a total of 24 weeks were satisfied with hair growth, reduced hair loss, and their bangs hairline, and satisfaction tended to increase with longer usage periods.

[0115] [Table 6]

[0116] [Table 7]

[0117] [Table 8]

[0118] Example 9. Changes in the scalp microbial environment before and after use of a product containing heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020. The scalp microbial environment was compared before and 24 weeks after using the product described in Manufacturing Example 1. Scalp microorganisms were collected using sterile skin-microbiome collection swabs and immediately sent to a microbiological analysis laboratory while maintaining a low temperature using hexane transport medium. For the analysis of scalp microorganisms, a library was prepared after sequencing and 16S metagenome analysis was performed for comparison.

[0119] As a result, the participants in the study showed a fourfold increase in scalp lactic acid bacteria after using the product of Production Example 1 (Figure 7a). Furthermore, the diversity of scalp microorganisms also improved, with total scalp microbial diversity increasing by 60% (Figure 7b) and scalp lactic acid bacteria diversity increasing by 176% (Figure 7c). After using the product of Production Example 1, there was a tendency for Staphylococcus caprae, a microorganism that decreases in people with alopecia areata, to increase (Figures 7d, 7e). Therefore, it was confirmed that applying the heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020 to the scalp can change the scalp microbial environment and prevent hair loss symptoms.

[0120] The above description of the present application is illustrative, and a person with ordinary skill in the art to which the present application pertains should understand that it can be easily modified into other specific forms without altering the technical idea or essential features of the present application. Therefore, the above-described embodiments should be understood to be illustrative in all respects and not limiting. For example, each component described as a single type may be implemented in a dispersed manner, and similarly, components described as dispersed may be implemented in a combined form.

Claims

1. It contains heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020 (KCCM12918P) as an active ingredient. A cosmetic composition for hair papilla cell proliferation, further comprising ermenthol, salicylic acid, and dexpanthenol.

2. The heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 is derived from fermented dough, as described in claim 1, for the cosmetic composition for hair papilla cell proliferation.

3. The heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 has a ratio of saturated fatty acids:unsaturated fatty acids:cyclic fatty acids of 0.1 to 2:0.25 to 5:0.03 to 0.6, wherein this is the cosmetic composition for hair papilla cell proliferation according to claim 1.

4. The heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 increases the expression of CDK2, CDK4, cyclin B1, and cyclin D1, as described in claim 1 of the cosmetic composition for hair papilla cell proliferation.

5. The heat-treated lactic acid bacterium Limosilactobacillus fermentum LM1020 reduces the expression level of the 5α-reductase-1 gene and increases the expression levels of FGF7, FGF10, and EGF genes, as described in claim 1, for the cosmetic composition for hair papilla cell proliferation.

6. The concentration of the heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 in the cosmetic composition for hair papilla cell proliferation is 1 × 10 ６ cell / mL or 1 × 10 ７ A cosmetic composition for hair papilla cell proliferation according to claim 1, wherein the concentration is cell / mL.

7. The cosmetic composition for hair papilla cell proliferation according to claim 1, further comprising purified water, ethanol, alanine / histidine / lysine polypeptide copper HCl, polyacrylate crosspolymer-6, coconut acid, proline, tea tree oil, glycerin, peptide, butylene glycol, 1,2-hexanediol, green tea extract, lavender flower extract, quince extract, and ethylhexanediol.

8. The cosmetic composition for hair papilla cell proliferation according to claim 7, wherein the cosmetic composition for hair papilla cell proliferation increases the microbial diversity and lactic acid bacteria diversity of the scalp.

9. It contains heat-treated lactic acid bacteria limosilactobacillus fermentum LM1020 (KCCM12918P) as an active ingredient. A pharmaceutical composition for the treatment of hair loss, further comprising elmenthol, salicylic acid, and dexpanthenol.

10. The aforementioned heat-treated lactic acid bacterium, limosilactobacillus fermentum LM1020, is derived from fermented dough, as per claim 9, the pharmaceutical composition for treating hair loss.

11. The aforementioned heat-treated lactic acid bacterium, limosilactobacillus fermentum LM1020, has a ratio of saturated fatty acids:unsaturated fatty acids:cyclic fatty acids of 0.1 to 2:0.25 to 5:0.03 to 0.6, as described in claim 9, for the treatment of hair loss.

12. The heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 increases the expression of CDK2, CDK4, cyclin B1, and cyclin D1, as described in claim 9 of the pharmaceutical composition for treating hair loss.

13. The heat-treated lactic acid bacterium limosilactobacillus fermentum LM1020 reduces the expression level of the 5α-reductase-1 gene and increases the expression levels of FGF7, FGF10, and EGF genes, as described in claim 9, for the treatment of hair loss.

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