Composition for improving hair health comprising latex rubber powder and mint extract

Abstract

Disclosed is a composition for improving hair health, which comprises a latex rubber powder and a mint extract. The present invention relates to a hair growth and hair growth promotion composition which comprises a latex rubber powder and a mint extract as active ingredients, and which can promote the proliferation of hair papilla cells, enhance the expression of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1) and beta-catenin (beta-catenin), which are related to hair growth and hair growth promotion, and improve the hair growth and hair growth promotion. Furthermore, hair growth can be promoted in an animal model, the hair density is improved, and the long-term hair growth proportion is improved.

Description

Technical Field

[0001] The present invention relates to a compound for improving hair health, comprising latex rubber powder and mint extract, and in particular to a compound for promoting hair growth and inhibiting hair loss. Background Art

[0002] Hair is formed in hair follicles. There are about 5 million hair follicles in an adult, which are distributed on the entire body except the palms and soles of the feet. One million hair follicles are found on the head, and 100,000 of them are found on the scalp (Bulletin of Food Technology 2009, 22(4):651-663). Scalp hair accounts for about 20% of the hair on the human body. It not only plays a physiological function such as sweat evaporation and body temperature regulation, but also protects the head from impact damage by cushioning the head. In addition, because modern people pay attention to the aesthetic tendency of appearance, it plays a very important social function for individuals to show their personality and image (Clin Dermatol 2001, 19(2):161-166).

[0003] Although the exact cause of hair loss has not been fully identified so far, the viewpoints that have been put forward include the inhibition of proliferation or decreased function of dermal papilla cells related to hair cycle regulation, abnormal hair cycle due to the action of androgen, abnormal changes in the hair cycle due to decreased scalp blood flow, the effects of drugs such as anticancer agents, mental stress, physical stimulation and environmental pollution (J Invest Dermatol, 1999, 113: 873-877; Mol Cell Endocrinol, 2002 198: 89-95; J. Dermatol. Sci. 2011, 62: 154-159).

[0004] Dermal papilla cells are a type of specialized fibroblasts that occur in the hair bulb. They can interact with various types of epithelial cells in the hair follicles and are known to play an important role in the formation of hair follicles, hair regeneration, and hair growth. Therefore, the growth, proliferation, and apoptosis of dermal papilla cells are an important target for developing drugs to maintain the growth phase of hair and inhibit hair loss (Physiol. Rev. 81: 449-494; J. Dermatol. Sci. 2011, 62: 154-159; FASEB J. 2008, 22: 1725-1736).

[0005] Insulin-like growth factor 1 (IGF-1) can promote the growth of hair papilla cells and inhibit apoptosis of hair papilla cells, thus playing a certain role in promoting hair growth. As a substance produced in the dermal papilla, insulin-like growth factor 1 (IGF-1) is a representative growth factor that helps hair growth by promoting the differentiation of keratinocytes (Ann Dermatol. 2012; 24 (1): 26-31).

[0006] Vascular endothelial growth factor (VEGF) is an angiogenic factor that improves blood circulation by promoting the growth of vascular endothelial cells, thereby enhancing hair growth and differentiation of hair papilla cells. Therefore, it is also used as an indicator factor in the screening of hair loss inhibitory or hair growth inducing substances (Korean J Dermatol. 1999; 37(1): 23-30; Exp Cell Res, 2012 Aug 15; 318(14): 1633-40).

[0007] Cell proliferation is closely related to the cell cycle consisting of G0 / G1, S and G2 / M steps, and extracellular factor / β-catenin (Wnt / β-catenin) signaling affects the cell cycle as described above. In addition, extracellular factor / β-catenin (Wnt / β-catenin) signaling also plays an important role in hair growth and hair regeneration (Dev.Biol.2005, 185:82-91; Mol.Cell.Biol.2005, 25:9063-9072; J.Immunol.2001, 166:4713-4720).

[0008] The most widely used drugs for the treatment of hair loss are substances that inhibit the activity of type 2 5α-reductase, such as minoxidil preparations (see U.S. Pat. Nos. 4,139,619 and 4,596,812) and finasteride (U.S. Pat. Nos. 5,547,957, 5,760,046 or Korean Patent No. 375083).

[0009] Because hair loss treatment requires long-term treatment, it is crucial to reduce side effects. People are working hard to extract new hair growth ingredients or materials from natural substances that have relatively few side effects on the human body.

[0010] The invention discloses a hair health improving compound comprising mastic gum (Pistacia lentiscus) powder and mentha piperite extract, which has the effects of promoting hair growth and inhibiting hair loss. Summary of the invention

[0011] An object of the present invention is to provide a hair health improving compound comprising latex rubber powder and peppermint extract.

[0012] In order to achieve the above object, the present invention adopts the following technical means:

[0013] As shown in the following embodiments and test examples, the present invention can confirm that the complex of latex rubber powder and peppermint extract can promote the proliferation of hair papilla cells and enhance the expression of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1) and β-catenin, which are related to hair growth and hair growth promotion, and can promote hair growth, increase hair density and increase the growth phase ratio of hair in animal models.

[0014] The present invention provides a compound based on the test results as described above. The compound is a food compound for improving hair health, which contains latex rubber powder and peppermint extract as active ingredients. The hair health improvement is to promote hair growth or inhibit hair loss.

[0015] Among them, preferably, the peppermint extract is an extract of water, a lower alcohol having a carbon number of 1 to 4, or a mixed solvent of water and a lower alcohol having a carbon number of 1 to 4.

[0016] Furthermore, the present invention also provides a pharmaceutical composition for promoting hair growth or inhibiting hair loss, which comprises a complex of latex rubber powder and peppermint extract as an active ingredient.

[0017] Among them, preferably, the peppermint extract is an extract of water, a lower alcohol having a carbon number of 1 to 4, or a mixed solvent of water and a lower alcohol having a carbon number of 1 to 4.

[0018] Furthermore, the present invention also provides a cosmetic composition for promoting hair growth or inhibiting hair loss, which comprises a complex of latex rubber powder and mint extract as an effective ingredient.

[0019] Among them, preferably, the peppermint extract is an extract of water, a lower alcohol having a carbon number of 1 to 4, or a mixed solvent of water and a lower alcohol having a carbon number of 1 to 4.

[0020] In the present specification, "mint extract" refers to an extract obtained by leaching the leaves, stems, fruits, seeds or a mixture of the mint object, i.e., mint, using a solvent such as water, a lower alcohol having a carbon number of 1 to 4 (such as methanol, ethanol and butanol), dichloromethane, ethylene, ether, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butanediol, propylene glycol or a mixed solvent of the above solvents, and an extract obtained by using a supercritical extraction solvent such as carbon dioxide and pentane, or a fraction obtained by fractionating the extract. As an extraction method, any method such as cold soaking, convection, heating, supercritical radiation and supercritical extraction can be applied in consideration of the polarity of the active substance, the degree of extraction and the degree of preservation. The extract after fractionation includes the fraction obtained by suspending the extract in a specific solvent, mixing it with a solvent of different polarity, and letting it stand, and the fraction obtained by adsorbing the crude extract onto a chromatographic column filled with silica gel, etc., using a hydrophobic solvent, a hydrophilic solvent, or a mixed solvent thereof as a mobile phase. In addition, the extract also includes a concentrated liquid extract or solid extract from which the extraction solvent has been removed by freeze drying, vacuum drying, hot air drying, and spray drying. Preferably, it refers to an extract obtained by using water, ethanol, or a mixed solvent of the two as the extraction solvent, and preferably refers to a 30-80% (v / v) ethanol extract.

[0021] In the present specification, the "active ingredient" means an ingredient that can exhibit the desired activity alone or can exhibit activity together with a carrier that has no activity itself.

[0022] In this specification, the term "hair loss" should be understood to include all symptoms classified as hair loss in the industry, regardless of direct or indirect causes. Specifically, it includes all symptoms of hair loss caused by factors such as poor blood circulation, excessive sebum secretion due to excessive production of dihydrotestosterone, decreased scalp function due to peroxides or bacteria, aging, genetic factors, stress, and the combined effects of the above factors.

[0023] As long as the composite of the present invention can exhibit hair growth promotion or hair loss inhibition activity, it can contain any amount (effective amount) of the active ingredient according to its use, dosage form, and purpose of mixing. Generally speaking, the effective amount can be determined in the range of 0.001% by weight to 50.0% by weight based on the total weight of the composite. Here, "effective amount" refers to the amount of the active ingredient contained in the composite of the present invention that can exhibit the desired function and pharmacological effect such as hair growth promotion when the composite of the present invention is administered to its applicable object, i.e., mammals, preferably humans, during the administration period under the advice of medical experts. The effective amount as described above can be determined by experiments within the general ability of relevant practitioners.

[0024] In order to enhance or strengthen the hair growth promoting or hair loss inhibiting effect or to improve the convenience of taking or ingesting the complex of the present invention by adding similar activities such as blood pressure regulating activity, in addition to the active ingredients, any compound or natural extract whose safety has been verified in the relevant industry and is known to have the corresponding activity may be added. The above-mentioned compounds or extracts may include compounds or extracts disclosed in legal documents such as the pharmacopoeia of various countries (in Korea, the "Pharmacopoeia of the Republic of Korea"), the public codes of health functional foods of various countries (in Korea, the "Standards and Specifications for Health Functional Foods" announced by the Food and Drug Administration), compounds or extracts licensed according to laws of various countries for regulating the manufacture and sale of pharmaceuticals (in Korea, the "Pharmaceutical Affairs Act"), and compounds or extracts that have obtained independent functional certification according to laws of various countries for regulating the manufacture and sale of health functional foods (in Korea, the "Health Functional Food Related Laws").

[0025] For example, the compounds or extracts described above may include ingredients with the function of alleviating hair loss symptoms, such as dexpanthenol, biotin, L-menthol, and zinc pyrithione, which are listed in the "Standards and Test Methods for Functional Cosmetics" announced by the Food and Drug Administration in relation to the Korean Cosmetics Act; and ingredients with the function of regulating blood pressure, such as γ-aminobutyric acid (GABA) powder derived from L-glutamic acid, bonito oligopeptides, natto bacteria culture powder, mycelium peptide complex, salmon peptide, olive leaf extract, sardine peptide, casein hydrolyzate, coenzyme Q10, grape seed enzyme decomposition extract powder, and seaweed oligopeptides, which are listed in the "Standards and Specifications for Health Functional Foods" related to the Korean Health Functional Food Act or have obtained blood pressure regulating functionality certification separately.

[0026] A specific form of the complex of the present invention may be a food composition.

[0027] The food composition applicable to the present invention can be manufactured into any form, for example, it can be manufactured into beverages such as tea, juice, carbonated drinks, electrolyte drinks, processed milk such as milk, yogurt, chewing gum, rice cakes, Korean cakes, bread, biscuits, noodles, etc., and health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, sticks, etc.

[0028] In addition, the food composition of the present invention can be classified into any product type that meets the implementation regulations when manufacturing and circulating in terms of the classification in law and function. For example, it can be the health functional food that meets the "health functional food related law" of Korea, or it can be the biscuits, beans, teas, beverages, special purpose foods etc. of each food type on the food code ("food standards and specifications" of the food and drug department notice) that meets the "food sanitation law" of Korea.

[0029] In addition to its effective ingredients, the food composition applicable to the present invention may also contain food additives. Food additives can generally be understood as substances added to food for mixing or soaking in order to manufacture, process or preserve food. Because they are taken with food every day and for a long time, their safety should be guaranteed. In the food additive code of various countries' laws (the "Food Sanitation Law" in South Korea) for regulating the manufacture and circulation of food, restrictive provisions are made for food additives that can guarantee their safety in terms of ingredients or functions. In the Korean Food Additive Code (the "Food Additive Standards and Specifications" announced by the Food and Drug Administration), the classification of food additives in terms of ingredients is defined as chemical synthetic products, natural additives and mixed preparations, and the classification of the above-mentioned food additives in terms of functions is defined as sweeteners, flavoring agents, preservatives, emulsifiers, acidulants, thickeners, etc.

[0030] The role of sweeteners is to give food a suitable sweet taste. Natural or synthetic sweeteners can be used in the food composition applicable to the present invention. Preferably, natural sweeteners are used. As natural sweeteners, sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, maltose, etc. can be used.

[0031] The effect of flavoring agent is to improve taste or aroma, and natural or synthetic flavoring agent can be used. Preferably, it is advisable to use natural flavoring agent. When using natural flavoring agent, the purpose of nutritional enhancement can be achieved in addition to improving flavor. As natural flavoring agent, flavoring agent extracted from apple, lemon, citrus, grape, strawberry, peach, etc. or flavoring agent extracted from green tea leaves, polygonatum, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, etc. can be used. In addition, flavoring agent extracted from ginseng (red ginseng), bamboo shoots, aloe juice, ginkgo, etc. can also be used. Natural flavoring agent can be a liquid concentrate or a solid extract. In some cases, synthetic flavoring agent can be used, and as synthetic flavoring agent, ester, alcohol, aldehyde or terpene can be used.

[0032] As preservative, calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, ethylenediaminetetraacetic acid (EDTA) etc. can be used, and as emulsifier, gum arabic, carboxymethyl cellulose, xanthan gum, pectin etc. can be used, and as acidulant, citric acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid and acetic acid etc. can be used. Acidulant, except the purpose of improving taste, can also be added in food composition with the purpose of suppressing microbial proliferation until reaching suitable acidity. As thickener, suspending agent, sedimentation agent, gel forming agent, swelling agent etc. can be used.

[0033] The food composition to which the present invention is applied may contain, in addition to the food additives described above, physiologically active substances or minerals which are well known in the industry and can ensure their stability as food additives for the purpose of supplementing and strengthening functionality and nutrition.

[0034] As the physiologically active substances as mentioned above, catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, vitamin B12, tocopherol, dibenzoylthiamine, etc. can be used, and as minerals, calcium preparations such as calcium citrate, magnesium preparations such as magnesium stearate, iron preparations such as ferric citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc. can be used.

[0035] In the food composition applicable to the present invention, the above-mentioned food additives can be added in an appropriate amount to achieve the purpose of addition according to the product type.

[0036] For other food additives that may be included in the food composition applicable to the present invention, please refer to the food code or food additive code of each country.

[0037] Another specific form of the composition of the present invention may be a pharmaceutical composition.

[0038] The pharmaceutical composition applicable to the present invention may contain a pharmaceutically acceptable carrier in addition to its active ingredient, and may be manufactured into an oral dosage form or a non-oral dosage form according to the administration route by a general method known in the industry. The administration route may be any appropriate route including a topical route, an oral route, an intravenous route, an intramuscular route, and direct absorption through mucosal tissues, or a combination of two or more routes may be used.

[0039] Pharmaceutically acceptable carriers for different administration routes or dosage forms are well known in the industry. For details, please refer to the pharmacopoeias of various countries including the Korean Pharmacopoeia.

[0040] When the pharmaceutical composition of the present invention is manufactured into an oral dosage form, it can be manufactured into a dosage form such as powder, granules, tablets, pills, sugar-coated tablets, capsules, liquids, gels, syrups, suspensions, and thin slices together with an appropriate carrier by a method known in the industry. At this time, examples of appropriate carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, and wheat starch, celluloses such as cellulose, methyl cellulose, ethyl cellulose, sodium carboxymethyl cellulose, and hydroxypropyl methyl cellulose, polyvinyl pyrrolidone, water, methyl paraben, propyl paraben, magnesium stearate, mineral oil, malt, gelatin, talc, polyols, vegetable oils, ethanol, glycerol, etc. When the formulation is carried out, appropriate binders, lubricants, disintegrants, colorants, diluents, etc. can be included as needed. As suitable binders, starch, magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinyl pyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, etc. can be used, as lubricants, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silicon dioxide, talc, stearic acid or its magnesium salt and calcium salt, polyethylene glycol, etc. can be used, and as disintegrants, starch, methylcellulose, agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt can be used. In addition, as diluents, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc. can be used.

[0041] When the pharmaceutical composition of the present invention is manufactured into a non-oral dosage form, it can be formulated in the form of injection, transdermal administration, nasal absorption agent and suppository together with an appropriate carrier by a method known in the industry. As an appropriate carrier for formulation in the form of injection, an aqueous isotonic solution or suspension can be used, specifically, a phosphate buffered saline (PBS) containing triethanolamine or sterile water for injection, 5% dextrose and other isotonic solutions can be used. When it is formulated in the form of a transdermal administration, it can be formulated in the form of an ointment, cream, emulsion, gel, external liquid, powder, liniment, aerosol and the like. When the formulation is in the form of a nasal inhaler, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and when the formulation is in the form of a suppository, as its carrier, it can be used such as Witepsol, Tween 61, polyethylene glycols, cocoa butter, lauric acid glyceride, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, etc.

[0042] The specific preparation method of the pharmaceutical composition is well known in the industry, and reference may be made to the literature [Remington's Pharmaceutical Sciences (19th edition, 1995)], etc. The above literature shall be regarded as a part of this specification.

[0043] The preferred dosage of the pharmaceutical composition of the present invention may be in the range of 0.001 mg / kg to 10 g / kg per day, preferably in the range of 0.001 mg / kg to 10 g / kg per day, depending on the patient's condition, weight, sex, age, severity of the patient, and route of administration. Administration may be performed once or multiple times per day. In any aspect, the dosage as described above should not be construed as limiting the scope of the present invention.

[0044] Another specific form of the composition of the present invention may be a cosmetic composition.

[0045] When the composition applicable to the present invention is defined as a cosmetic composition, the above-mentioned cosmetic composition can also adopt any product type in terms of its use and legal classification, and specifically can be functional cosmetics with uses such as hair loss relief, hair growth promotion, skin problem improvement and atopic dermatitis improvement, non-functional general cosmetics, etc. In terms of product form, any product form can also be adopted, specifically, it can adopt product forms such as solutions, suspensions, emulsions, pastes, gels, creams, emulsions, powders, soaps, shampoos, facial cleansers containing surfactants, oils, powder bases, emulsion bases, wax bases, sprays, etc. In terms of specific product forms, it can be skin softening lotions, nutritional lotions, nutritional creams, massage creams, essences, eye creams, facial cleansing creams, facial cleansing foams, facial cleansing waters, facial masks, sprays or powder dosage forms, etc.

[0046] The cosmetic composition applicable to the present invention may contain, in addition to its active ingredients, ingredients commonly used in cosmetic compositions, such as stabilizers, solubilizers, surfactants, vitamins, pigments, fragrances, and other general auxiliary agents and carriers.

[0047] When the dosage form of the present invention is a paste, cream or gel, as a carrier component, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone resin, bentonite, silicon dioxide, talc or zinc oxide can be used.

[0048] When the dosage form of the present invention is a powder or a spray, lactose, talc, silicon dioxide, aluminum hydroxide, calcium silicate or polyamide powder can be used as a carrier component. In particular, when the dosage form is a spray, a propellant such as chlorofluorocarbons, propane / butane or dimethyl ether can also be added.

[0049] When the dosage form of the present invention is a solution or an emulsion, as a carrier component, a solvent, a solubilizer or an emulsifier can be used, specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol, fatty acid esters of sorbitan, etc. can be used.

[0050] When the dosage form of the present invention is a suspension, as the carrier component, liquid diluents such as water, ethanol or polyethylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitan esters, polyoxyethylene sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, etc. can be used.

[0051] When the dosage form of the present invention is a facial cleanser containing a surfactant, as the carrier component, fatty alcohol sulfates, fatty alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazoline derivatives, methyl taurates, sarcosinates, fatty acid amide ether sulfates, alkylamide betaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glyceryl fatty acid esters can be used.

[0052] The cosmetic composition applicable to the present invention can be manufactured according to the manufacturing method of the cosmetic composition generally used in the industry, except that it contains its effective ingredients that exhibit hair growth promoting activity and the like.

[0053] As described above, the present invention can provide a composition for promoting hair growth or inhibiting hair loss using a complex of latex rubber powder and peppermint extract.

[0054] The composition of the present invention can be formulated into foods, medicines, hair cosmetics, and the like. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 as well as Figure 2 It is an assessment of the proliferation degree of dermal papilla cells.

[0056] Figure 3 It is the antioxidant activity evaluation result.

[0057] Figure 4 It is the Western blot result of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1) and β-catenin.

[0058] Figure 5 It is the result of the evaluation of the growth index in animal models.

[0059] Figure 6 is the result of the assessment of hair density in animal models.

[0060] Figure 7 is an assessment of the hair anagen phase ratio in an animal model. DETAILED DESCRIPTION

[0061] Next, the present invention will be described in conjunction with embodiments and test examples. However, the scope of the present invention is not limited by the following embodiments and test examples.

[0062] <Example 1> Preparation of a composite of latex rubber powder and mint extract

[0063] As the latex rubber powder, the latex rubber was put into a pulverizer and pulverized, and then sieved with an AMKCO mechanical sieve to be homogenized.

[0064] As a mint extract, 4 L of 50% ethanol was added to 200 g of dried mint and then extracted at 50° C. for 6 hours to obtain a mint extract. Next, the filtered extract was concentrated under reduced pressure at 50° C. or less and freeze-dried to produce a powdered extract.

[0065] The latex rubber powder and the mint extract composite (FHH-MG) was prepared by mixing the latex rubber powder and the mint extract at a weight ratio of latex rubber powder:mint extract=7:3.

[0066] <Test Example 1> Evaluation of Hair Loss Inhibition or Hair Growth Promotion Effect

[0067] 1. In vitro efficacy evaluation

[0068] 1.1 Assessment of cell proliferation rate

[0069] This test is used to confirm the appropriate concentration that does not produce cytotoxicity when the cells are treated with the test product (prepared in Example 1). It is a method for measuring the amount of surviving cells using water-soluble tetrazolium salts (WST, water soluble tetrazolium salts), which is suitable for cytotoxicity assessment, cell viability and cell division ability, etc. Water-soluble tetrazolium salts (WST) are highly sensitive water-soluble tetrazolium salts that are easily soluble in water, and formazan is generated by reacting with dehydrogenases in cells. The more surviving cells are, the more formazan is generated, thereby presenting a higher absorbance, by which the degree of toxicity of the cells or the promotion of cell proliferation can be judged.

[0070] Human follicle dermal papilla cells (PromoCell) were cultured in a culture medium to which fetal calf serum, bovine pituitary extract, basic fibroblast growth factor and insulin were added at 37°C and 5% CO2.

[0071] The cultured cells were plated in a 96-well plate at a rate of 5 × 10 3 The cells were divided and treated with the test product when the cells reached 80% or more and then cultured for another 24 hours. Then, a water-soluble tetrazolium salt (WST) substrate solution (CCK-8, Dojindo) was added to the culture medium and cultured at 37°C for 2 hours. The absorbance (OD) at 450nm was measured using a microplate reader (VARIOSKAN LUX, Thermo Fisher Scientific). Based on the measured absorbance (OD) value, the cell proliferation rate was calculated as a percentage compared to the untreated group.

[0072] The results are as follows Figure 1 as well as Figure 2 shown. Figure 1 The results are as follows: The latex rubber powder of Example 1 and the single substance and the composite (FHH-MG) of the peppermint extract were treated at 200 ppm and cultured for 24 hours, 48 ​​hours and 72 hours. Figure 2 The results are shown when the cells were treated with the complex (FHH-MG) at 62.5 ppm, 125 ppm, and 250 ppm and cultured for 24 hours, 48 ​​hours, and 72 hours.

[0073] See also Figure 1 It was confirmed that the cell proliferation rate at 48 hours and 72 hours of culture was further promoted when the complex (FHH-MG) was used for treatment compared with the control group, compared with the single substance of latex rubber and peppermint extract.

[0074] See also Figure 2The complex (FHH-MG) can promote the proliferation of dermal papilla cells compared with the control group in proportion to the treatment concentration and culture time.

[0075] exist Figure 1 as well as Figure 2 Minoxidil, used as a positive control group, can also promote the proliferation of dermal papilla cells compared with the control group.

[0076] 1.2 Evaluation of antioxidant efficacy

[0077] 1.2.1 Evaluation of superoxide dismutase activity (SOD activity)

[0078] This test adopts the Marklun method (Eur. J. Biochem., 47: 469-474, 1974) for measuring superoxide dismutase (SOD) activity by observing the inhibition of pyrogallol autoxidation.

[0079] Superoxide dismutase (SOD) is an antioxidant enzyme that protects organisms from the effects of reactive oxygen by converting oxygen radicals that are harmful to cells into hydrogen peroxide, which is then further converted into harmless water molecules and oxygen molecules by catalase.

[0080] The cultured human dermal papilla cells were cultured at 5×10 4 The cells were divided into 6-well plates and treated with H2O2 (400 μM) to induce oxidative stress when the cells reached 80% or more. The cells were then treated with the test product and cultured for another 24 hours. The culture medium was then obtained and centrifuged at 2,000 × g for 10 minutes, and the supernatant excluding cell debris was collected for testing.

[0081] The superoxide dismutase activity (SOD activity) was measured using a superoxide dismutase kit (SODkit, BIOMAX). The stock solution was used as the culture medium, and the test was performed according to the attached test method, and the absorbance (OD) was measured using a microplate reader (VARIOSKAN LUX, Thermo Fisher Scientific). Based on the measured absorbance (OD) value, the superoxide dismutase activity (SOD activity) was calculated according to the method provided by the manufacturer.

[0082] The results are as follows Figure 3 See Figure 3 It was observed that the superoxide dismutase activity (SOD activity) of the hair papilla cells, which was reduced by H2O2, increased in a concentration-dependent manner compared with the control group by treatment with the complex (FHH-MG). In addition, it was confirmed that the activity was further increased when the complex (FHH-MG) was treated at 125ppm and 250ppm compared with the positive control group, minoxidil.

[0083] 1.2.2 Evaluation of catalase activity (CAT activity)

[0084] This test adopts the Aebi method (Methods in enzymology, Academic press. 105: 121-126, 1984) which confirms the activity of catalase (CAT) by measuring the decrease in absorbance due to the decomposition of hydrogen peroxide.

[0085] Catalase (CAT) is a representative antioxidant enzyme, and is known to be involved in the decomposition reaction of hydrogen peroxide (two molecules of H2O2) generated by superoxide dismutase (SOD) into one molecule of oxygen and two molecules of water, as well as the decomposition reaction of reactive oxygen species (ROS).

[0086] The cultured human dermal papilla cells were divided into 6-well plates at 5×104 cells per well and treated with H2O2 (400 μM) to induce oxidative stress when the cells reached 80% or more. The cells were then treated with the test product and cultured for another 24 hours. The culture medium was then obtained and centrifuged at 2,000×g for 10 minutes, and the supernatant except for cell debris was collected for testing.

[0087] A catalase kit (CAT kit, BIOMAX) was used for the catalase activity (CAT activity). The stock solution was used as the culture medium, and the test was performed according to the attached test method, and the absorbance (OD) was measured using a microplate reader (VARIOSKAN LUX, Thermo Fisher Scientific). Based on the measured absorbance (OD) value, the superoxide dismutase activity (CAT activity) was calculated according to the method provided by the manufacturer.

[0088] The results are as follows Figure 3 shown.

[0089] See also Figure 3 It was observed that the catalase activity (CAT activity) of the hair papilla cells, which was reduced by H2O2, increased in a concentration-dependent manner compared with the control group by treatment with the complex (FHH-MG). In addition, it was confirmed that the CAT activity was further increased when the complex (FHH-MG) was treated compared with the positive control group, minoxidil.

[0090] 1.3 Evaluation of hair growth stimulating factor protein production

[0091] This test is a method for quantifying the production of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1), and β-catenin protein, which are hair growth stimulating factors, by reacting with an enzyme-labeled antibody. The production of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1), and β-catenin protein by the test product was measured in human dermal papilla cells.

[0092] The cultured human dermal papilla cells were divided into 6-well plates at 5×104 cells, and each well was treated with the test product when the cells reached 80% or more, and then cultured for another 24 hours. Next, the culture medium was obtained and centrifuged at 2,000×g for 10 minutes, and the supernatant excluding cell debris was collected for the test.

[0093] Human VEGF ELISA kit (Abeam) was used for the quantification of vascular endothelial growth factor (VEGF) protein, Human IGF-1 ELISA kit (Abeam) was used for the quantification of insulin-like growth factor 1 (IGF-1) protein, and Human β-catenin ELISA kit (MyBioSource) was used for the quantification of β-catenin protein. The stock solution was used as the culture medium, and the test was performed according to the attached test method. The absorbance (OD) was measured using a microplate reader (VARIOSKAN LUX, Thermo Fisher Scientific), and the measured value was substituted into the regression equation obtained by the standard curve, and then multiplied by the dilution factor to calculate the protein concentration.

[0094] The results are as follows Figure 4 shown.

[0095] See also Figure 4 It was observed that the insulin-like growth factor 1 (IGF-1) protein increased in a concentration-dependent manner compared with the control group by treatment with the complex (FHH-MG), and further increased compared with the positive control group, minoxidil. In addition, the vascular endothelial growth factor (VEGF) and β-catenin proteins increased by treatment with the complex (FHH-MG) compared with the control group, and further increased compared with the positive control group, minoxidil.

[0096] Statistical analysis

[0097] The statistical analysis was performed using IBM SPSS statistics 25.0 program, and the significance of the experimental group and the control group was confirmed based on the assumption of a mean difference of 5% (p<0.05).

[0098] After the normality test, the significance was confirmed by independent sample T-test (parametric method) according to whether the normality was satisfied or not.

[0099] 2. In vivo efficacy evaluation

[0100] 2.1 Animal husbandry and experimental composition

[0101] Male C57BL / 6 mice aged 5 weeks were purchased from ORIENT BIO and acclimated for 1 week. The experimental animals were fed with experimental animal feed freely under the conditions of 24℃±0.5℃, 55%-65% humidity, and automatic light and dark adjustment in a 12-hour cycle. All experimental procedures and steps were approved by the Institutional Animal Care and Use Committee of Yonsei Medical Center (approval number: 2023-0017) and performed in accordance with ethical regulations in the specific pathogen-free (SPF) area of ​​the Yonsei University Avison Biomedical Research Center (ABMRC).

[0102] As a negative control group, distilled water, a solvent in which the sample was dissolved, was used, and as a positive control group, minoxidil (Sigma, US) was used as a hair loss treatment agent. After the mice that had undergone the acclimation period were anesthetized by inhalation of isoflurane, the back of the mice was depilated for the first time using an animal clipper. After the depilatory agent was applied to the depilatory area and left for 3 minutes, the fine hair remaining on the skin was removed for the second time. After 24 hours of skin stabilization after depilation, the 6-week-old mice with resting hair on the back side of the depilated back were weighed and divided into 5 groups of 6 each, and then orally fed with 100 μl of the test product, "FHH-MG (100, 150, 200 mg / kg)" 5 times a week at a specific time within 3 weeks. In order to confirm the effect of drug administration on body weight changes, body weight was measured once a week. 7 days and 21 days after the start of the experiment, they were anesthetized and killed using CO2.

[0103] 2.2 Determination of hair growth index

[0104] In order to evaluate hair growth by measuring the hair growth index within 3 weeks after the start of the experiment, a scanner (Aura X Pro; ZUR) was used to take photos once a week. The changes in hair growth in the depilatory back area of ​​the mice were observed with the naked eye, and the degree of hair growth was judged as 0-19% (1-1.5 points), 20-39% (2-2.5 points), 40-59% (3-3.5 points), 60-79% (4-4.5 points), and 80-100% (5 points). The average value of the judgment scores evaluated by experienced researchers in a blind test was calculated.

[0105] The results are as follows Figure 5 See Figure 5It was confirmed that the administration of the complex (FHH-MG) from the 7th day to the 14th day increased the hair growth index compared with the control group. In addition, the hair growth index of minoxidil was also increased compared with the control group.

[0106] 2.3 Hair density determination

[0107] In order to measure the hair density based on the number of hair follicles, skin tissues of mice that had been orally administered for 7 days were removed and fixed with 10% formalin for more than 24 hours to make paraffin blocks and paraffin embedded slides. The deparaffinized tissues were stained with hematoxylin (104302; Merck) solution for nuclei and then washed with running water. Then, the cytoplasm was stained with eosin (230251; Sigma) solution and washed with water. The completely dehydrated tissues were then fixed with mounting solution. The number of hair follicles in the dermis and subcutaneous was measured in the tissues magnified 400 times using an optical microscope (BX43F; OLYMPUS).

[0108] The results are as follows Figure 6 See Figure 6 It was confirmed that the administration of the complex (FHH-MG) increased hair density compared to the control group. In addition, minoxidil also increased hair density compared to the control group.

[0109] 2.4 Determination of growth period ratio

[0110] To measure the growth phase ratio of hair follicles, skin tissues of mice that had been orally administered for 7 days were removed and fixed with 10% formalin for more than 24 hours to make paraffin blocks and paraffin embedded slides. The deparaffinized tissues were stained with hematoxylin (104302; Merck) solution for nuclei and then washed with running water. Then, the cytoplasm was stained with eosin (230251; Sigma) solution and washed with water. The completely dehydrated tissues were then fixed with mounting solution. The subcutaneous hair follicle ratio was measured in the whole hair follicle book of the tissue magnified 400 times using an optical microscope (BX43F; OLYMPUS).

[0111] The results are as follows Figure 7 See Figure 7 It was confirmed that the administration of the complex (FHH-MG) increased the growth period ratio compared to the control group. In addition, the growth period ratio of minoxidil was also increased compared to the control group.

[0112] 2.5 Statistical analysis

[0113] The statistical analysis was performed using IBM SPSS statistics 25.0 program, and the significance of the experimental group and the control group was confirmed based on the assumption of a mean difference of 5% (p<0.05).

[0114] After the normality test, the significance was confirmed by independent sample T-test (parametric method) according to whether the normality was satisfied or not.

Claims

1. A composite, characterized in that: The compound is a food compound for improving hair health, which contains latex rubber powder and mint extract as effective ingredients. The hair health improvement is to promote hair growth or inhibit hair loss.

2. The composite according to claim 1, characterized in that: The mint extract is an extract of water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent of water and a lower alcohol having 1 to 4 carbon atoms.

3. A pharmaceutical composition for promoting hair growth or inhibiting hair loss, characterized in that: Contains a complex of latex rubber powder and mint extract as active ingredients.

4. The composition according to claim 3, characterized in that: The mint extract is an extract of water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent of water and a lower alcohol having 1 to 4 carbon atoms.

5. A cosmetic composition for promoting hair growth or inhibiting hair loss, characterized in that: Contains a complex of latex rubber powder and mint extract as active ingredients.

6. The composition according to claim 5, characterized in that: The mint extract is an extract of water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent of water and a lower alcohol having 1 to 4 carbon atoms.

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