KR101626319B1 - Composition with the fermentation product of camellia oil or the extract thereof for growing hair or preventing alopecia and the method for prodcution of it - Google Patents

Abstract

The present invention relates to a composition for inhibiting hair growth or alopecia, which comprises a fermented product of a camellia oil or an extract thereof obtained by adding a fat-soluble solvent to a fermentation broth obtained by fermenting a camellia oil with yeast and removing the fat-soluble fraction, Which promotes the proliferation of fibroblasts and dermal papillary cells without side effects and increases the expression of hair growth related genes, thereby exerting an excellent effect in promoting hair growth and preventing hair loss.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing hair growth or alopecia, which contains camellia oil fermented product or an extract thereof, and a composition for preventing hair growth or alopecia, and a method for producing the composition.

The present invention relates to a composition for inhibiting hair growth or alopecia, which contains a fermented product of a camellia oil or an extract thereof obtained by adding a fat-soluble solvent to a fermentation broth obtained by fermenting a camellia oil with yeast, and removing the fat-soluble fraction, More specifically, it is possible to promote the proliferation of fibroblasts and dermal papilla cells without side effects by adding a fat-soluble solvent to the fermentation broth obtained by fermenting the camellia oil with yeast, and then removing the fat soluble fraction, The present invention relates to a composition which can be used for promoting hair growth or preventing hair loss and a method for producing the same.

Hair is an ecologically derived epithelium from the epidermis, and it is formed from the second month of birth. Types of hair are classified into thin hair (Vellus hair) and thick hard hair (terminal hair) depending on their thickness . The normal menstrual cycle of hair consists of growth period, regenerator, rest period, and activity period. Growth period is literally the period of growth from the papillary, 3 to 5 years for males and 4 to 6 years for females. Degenerative growth is stopped and the follicular area is reduced to 30 to 45 days. The dormancy is the period of stay on the scalp as the growth stops, and it is about 3 to 4 months. The activity in the pausing period is the period when hair that has passed the cycle falls and hair loss occurs and new hair is produced.

On the other hand, when the normal menstrual cycle of the hair is broken and the proportion of the rest period is increased, new hair is not produced and hair loss proceeds. The causes of hair loss include genetic, nutritional disorders, endocrine disorders, tissue disorders, hair follicles, tofu compression, dandruff, autoimmunity and local infection (Kim, Minjung, 2005). In addition, it is thought to be due to metabolic abnormalities such as hormones and proteins closely related to hair formation, stress increase, and unbalance of nutrition from westernized eating habits (J. Kor. Soc. Cosm. (2007) p.1366-1375).

Alopecia can be classified into two categories: male pattern baldness, alopecia seniles, and androgenic alopecia, which are common in middle-aged men. Alopecia areata is a common alopecia starting from the frontal and two governments. The second type is called alopecia areata. It usually starts from the occipital area and has a circular or elliptical shape with a clear borderline appearance. It is alopecia.

Hair loss can occur not only in males but also in women, especially in women, which often originate in two governments and there is also alopecia areata. However, it is reported that the incidence is lower than that of men (M. Inaba et al., Androgenetic Alopecia, Springer-Berlag, Tokyo, Japan, 1996).

Currently, prevention and treatment of alopecia are divided into a medical approach and a oriental medicine approach. Medical approaches include pharmacotherapy, surgical methods, nutritional therapy, and diet. Drugs that promote hair growth include minoxidil (MXD) and finasteride, which are approved by the Food and Drug Administration (FDA). However, it has been reported that minoxidil and finasteride have unfavorable side effects such as dermatitis when they are used for a long period of time (Kaufman K.D. (1996) Elsevier Science, 363-365).

Accordingly, there is a need to overcome the problems of the hair loss preventive and therapeutic agent as described above, and to study the hair loss prevention and treatment food or medicament containing no natural side effects and high safety.

On the other hand, camellia oil camellia (Camillia japonica L.) seeds are dried and pulverized, followed by milking or extraction using a solvent. Camellia seeds are usually composed of 9.1 to 11.5% by weight of saturated fatty acids, 85.6 to 89.4% by weight of oleic acid, and 1.3 to 2.9% by weight of linoleic acid, and have been used as hair oil, precision machine oil and edible oil since ancient times.

Pharm. Bull. 45 (12): 2016-2023 (1997)), Epstein-Barr virus inhibitory effect (Chem. Pharm. Bull. (Tokyo) 52 (1): 153-6 (2004)) and ethanol absorption inhibitory effects (Chem. Pharm. Bull. 42 (3): 742-744 (1994)).

However, there has been no previous study on whether the fermented product or its extract after fermentation of camellia oil is effective in promoting hair growth or inhibiting hair loss.

Korean Patent No. 10-0334944 entitled "Composition for treating alopecia of a herbal medicine ingredient" includes a herbal medicine containing a herbal medicine such as astragalus, Angelica gigas, peony, persimmon, alfalfa, chrysanthemum, Alopecia and crustacea, and alopecia areata, bluish white, yellowish white, fenugreek and crustacean water extracts.

It is intended to provide a composition containing a natural oil fermented product having an effect of promoting hair growth or hair loss or an extract thereof, and a method for producing the same, by increasing gene expression related to cell proliferation and hair growth.

A first aspect of the present invention is a hair growth apparatus characterized by comprising a fermented product obtained by removing a fat soluble fraction after adding a fat soluble solvent to a fermentation liquid obtained by fermenting a camellia oil with yeast or an extract thereof, There is provided a composition for promoting or preventing hair loss.

Unlike conventional hair or scalp products using camellia oil, the present invention is characterized in that a fermented product obtained by fermenting a camellia oil with yeast and then removing a fat-soluble fraction that is an oil layer or an extract thereof is used. According to the experiment of the present invention, it was confirmed that such a fermented product or its extract exerts excellent effects on hair growth promotion or hair loss prevention.

On the other hand, in the anti-hair loss or hair growth promoting composition of the present invention, the yeast is preferably of Candida (Candida) recommended lay, and more preferably Candida spring rain coke (Candida bombicola .

Meanwhile, the hair growth promoting or hair loss preventing composition of the present invention may be a cosmetic or scalp cleaner composition.

Meanwhile, in the hair growth promoting or hair loss preventing composition of the present invention, the cosmetic composition may be any one selected from among cream, lotion, skin, cleansing, soap, treatment, and essence.

In addition, the cosmetic composition may contain commonly used ingredients. Examples include water-soluble vitamins, fat-soluble vitamins, polymer peptides, polysaccharides, sphingolipids, seaweed extracts and the like.

The water-soluble vitamin may be any kind of water-soluble vitamin that can be incorporated into cosmetics. Examples of the water-soluble vitamin include vitamin B ₁ , vitamin B ₂ , vitamin B ₆ , pyridoxine, pyridoxine hydrochloride, vitamin B ₁₂ , pantothenic acid, nicotinic acid, , Vitamin C, vitamin H or thiamine hydrochloride, ascorbic acid sodium salt, ascorbic acid-2-phosphate sodium salt, ascorbic acid-2-phosphate magnesium salt and the like. At this time, the water-soluble vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzymatic method, or a chemical synthesis method.

The fat-soluble vitamin may be any kind of fat-soluble vitamin that can be formulated in cosmetics. Examples of the fat-soluble vitamin include vitamin A, carotene, vitamin D ₂ , vitamin D ₃ , vitamin E (d1-? -Tocopherol, Tocopherol vitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenyl ethyl ether, and the like can be used. have. At this time, the fat-soluble vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzyme or a chemical synthesis method.

The polymeric peptide may be any kind of polymeric peptide that can be incorporated into cosmetics. Examples of the polymeric peptide include collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, and keratin. In this case, the polymer peptide can be obtained, for example, by a conventional method such as a purification method from a culture broth of a microorganism, an enzymatic method, or a chemical synthesis method, and purified from a natural product such as a duck of pigs, .

The polymer polysaccharide may be any kind of polymer polysaccharide compoundable in cosmetics. Examples of the polysaccharide may include hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof. At this time, the polymer polysaccharide can be obtained, for example, by purification from a mammal or fish.

The sphingo lipids may be any kind of sphingolipids that can be incorporated into cosmetics. Examples include ceramides, phytosphingosine and sphingoglycolipids. Here, the sphingogly lipids can be purified, for example, from mammal, fish, shellfish, yeast or plants by conventional methods or by chemical synthesis.

The seaweed extract may be any kind of seaweed extract that can be formulated into cosmetics. Examples of the seaweed extract include brown algae extract, red tiger extract, green algae extract, and the like, which are refined from seaweed extract, arginic acid, sodium alginate, potassium alginate .

Meanwhile, in the hair growth promoting or hair loss preventing composition of the present invention, the cosmetic composition may be, for example, a cosmetic composition containing at least one of a preservative component, a moisturizer, an emollient, a surfactant, An antioxidant, a plant extract, a pH adjuster, an alcohol, a pigment, a flavor, a blood circulation, an accelerator, a cold agent, a limiting agent and purified water.

Examples of the above-mentioned sustaining component may include ester-based oil retaining, hydrocarbon-based oil retaining, silicone-based oil retaining, fluoric oil retaining, animal retention and plant preservation. Examples of the ester-based oil include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, Wherein the acid is selected from the group consisting of isostearyl, isostearyl, isostearyl palmitate, octyldodecyl myristate, isosertyl myristate, isobutyl myristate, isobutyl myristate, isopropyl myristate, Tri (2-ethylhexyl) trimethylolpropane, triisostearic acid trimethylolpropane, tetra-2-ethyl (ethyl) But are not limited to, stearic acid, pentaerythritol hexanoic acid, pentaerythritol hexanoate, caprylic acid cetyl laurate, hexyl laurate, myristyl myristate, myristyl myristate, myristyl stearate, Octyldodecyl linoleate, isostearyl linoleate, isostearyl linoleate, isostearyl linoleate, isostearyl linoleate, isostearyl linoleate, isostearyl linoleate, isostearyl linoleate, isocetyl isostearate, Ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di (capryl, capric acid) propylene Glycerin, glyceryl triisostearate, glyceryl triisostearate, glyceryl triisostearate, glyceryl triisostearate, glyceryl triisostearate, glyceryl triisostearate, glyceryl triisostearate, glyceryl triisostearate, glyceryl triisostearate, Octyldodecyl octanoate, octanoate octanoate, octanoate octanoate, octanoate octanoate, octanoate octanoate, octanoate dodecyl neodecanoate, isocetyl isostearate, isostearic acid Polyglycerin isostearic acid ester, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyl lactate, octyldecyl lactate, octyldecyl isostearate, polyglycerin oleic acid ester, polyglycerin isostearic acid ester, Decyl, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, Diisopropyl sebacate, dioctyl sebacate, stearic acid cholesteryl, isostearic acid cholesteryl, hydroxystearic acid cholesteryl, oleic acid cholesteryl, oleic acid dihydrocholesteryl, isostearic acid pitostearyl, oleic acid Phytosteryl, 12-stearoylhydroxystearic acid isosityl, 12-stearoylhydroxystearic acid stearate, Reel, 12-stacking egg may be hydroxy stearic acid rilil source Te.

The hydrocarbonaceous oil may be a hydrocarbonaceous oil such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, floating isoparaffin, polybutene, microcrystalline wax and vaseline.

The silicone-based oil is, for example, selected from the group consisting of polymethyl silicone, methylphenyl silicone, methyl cyclopolysiloxane, octamethyl polysiloxane, decamethyl polysiloxane, dodecamethyl cyclosiloxane, dimethylsiloxane methyl cetyloxysiloxane copolymer, dimethylsiloxane methyl stearoxysiloxane copolymer , Alkyl-modified silicone oil, amino-modified silicone.

The fluorine-based oil may be, for example, a perfluoropolyether.

Examples of the animal or plant oil include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rape oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, It is also possible to use milk, milk, grape seed oil, cottonseed oil, palm oil, cucumber nut oil, wheat germ oil, rice germ oil, shea butter, colostrum colostrum, marker daisy nut oil, Oil, jojoba oil, canned wax, carnauba wax, liquid lanolin, and hardened castor oil.

The moisturizing agent may be, for example, a water-soluble low molecular weight moisturizer, a liposoluble molecular moisturizer, a water-soluble polymer, or a fat-soluble polymer.

Examples of the water-soluble low molecular weight moisturizing agent include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B ), Polypropylene glycol (polymerization degree n = 2 or more), polyglycerin B (polymerization degree n = 2 or more), lactic acid, lactic acid salt.

The lipid-soluble low-molecular moisturizing agent may be, for example, cholesterol or cholesterol ester.

Examples of the water-soluble polymer include carboxyvinyl polymer, polyaspartic acid, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, Lt; / RTI >

The oil-soluble polymer may be, for example, a polyvinylpyrrolidone eicosene copolymer, a polyvinylpyrrolidone hexadecene copolymer, a nitrocellulose, a dextrin fatty acid ester, or a polymeric silicone.

The emollients may be long chain acyl glutamic acid cholesteryl ester, cholesteryl hydroxystearate, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl ester.

The surfactant may be, for example, a nonionic surfactant, an anionic surfactant, a cationic surfactant, or a positive surfactant.

Examples of the nonionic surfactant include self emulsifying monostearic acid glycerin, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbitol POE fatty acid esters, POE glycerin fatty acid esters, POE alkyl ethers, POE fatty acid esters, POE hardened castor oil, POE castor oil, POE POP (polyoxyethylene polyoxypropylene) copolymer, POE POP alkyl ether, polyether- Acid alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids.

Examples of the anionic surfactant include fatty acid soap, alpha-acylsulfonate, alkylsulfonate, alkylallylsulfonate, alkylnaphthalenesulfonate, alkylsulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl phosphate , Alkylamide phosphate, alkyloyl taurine salt, N-acyl amino acid salt, POE alkyl ether carboxylate, alkyl sulfosuccinate, sodium alkylsulfoacetate, acylated hydrolyzed collagen peptide salt, perfluoroalkyl phosphate ester .

The cationic surfactant includes, for example, alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenylbromide Trimethylammonium chloride, benzalkonium chloride, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, and lanolin derivative quaternary ammonium salts.

Examples of the amphoteric surfactant include a carboxybetaine, an amidebetaine, a sulfobetaine, a hydroxysulfobetaine, an amidosulfobetaine, a phosphobetaine, an aminocarboxylate, an imidazoline derivative, an amide Amine type and the like.

Examples of the organic and inorganic pigments include inorganic pigments such as silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, Bengala, clay, bentonite, titanium mica, titanium oxide, bismuth oxychloride, zirconium oxide, Inorganic pigments such as titanium, aluminum oxide, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, chromium oxide, chromium oxide, chromium, and combinations thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene styrene copolymer, silk Organic pigments such as powder, cellulose, CI Pigment Yellow and CI Pigment Orange; And a composite pigment of an inorganic pigment and an organic pigment.

Examples of the organic powder include metallic soap such as calcium stearate; Metal salts of alkyl phosphates such as sodium zinc cetylate, zinc laurylate and calcium lauryl laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc and N-lauroylglycine calcium; Amidosulfonic acid multivalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; Such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylidene, N-alpha-paratyylnitine, N-alpha-lauroyl arginine, Acyl basic amino acids; N-acylpolypeptides such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-aminocaprylic acid, alpha-aminoaurauric acid, and the like; Polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene styrene copolymer, ethylene tetrafluoride.

Examples of the ultraviolet absorber include ultraviolet absorbers such as p-aminobenzoic acid, ethyl p-aminobenzoate, amyl p-aminobenzoate, octyl p-aminobenzoate, ethyleneglycol salicylate, phenyl salicinate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate , Benzyl cinnamate, 2-ethoxyethyl paratomethoxy cinnamate, octyl paratomethoxy cinnamate, mono-2-ethylhexane glyceryl dipyrromethoxy cinnamate, isopropyl paratethoxy cinnamate, diisopropyl diisopropyl cinnamate ester mixture , Urocannic acid, ethyl urocanoate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and salts thereof, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxy Benzophenone, tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianylino-p- (carbo-2'-ethylhexyl- -1,3,5-triazine, 2- (2- Hydroxy-5-methylphenyl) benzotriazole.

Examples of the bactericides include hinokitiol, trichloroacetic acid, trichlorohydroxydiphenyl ether, crohexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, Benzalkonium, photosensitizer 301, mononitro and eicoll sodium, and undecylenic acid.

The antioxidant may be, for example, butylhydroxyanisole, gallic acid propyl, or erosorbic acid.

The pH adjusting agent may be, for example, citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate.

The alcohol may be, for example, a higher alcohol such as cetyl alcohol.

On the other hand, any components that can be added to cosmetics in addition to the above can be compounded within a range that does not impair the objects and effects of the present invention.

In the present invention, the fermented product of the camellia oil or the extract thereof is preferably added in an amount of 0.00001 to 70% by weight, more preferably 0.001 to 3% by weight, based on the total weight of the entire cosmetic composition.

The cosmetic composition of the present invention can be prepared in any form conventionally produced in the art, and examples thereof include a paste, a cream, a gel, a powder, a spray solution, an emulsion, a suspension, a lotion, a soap, Containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, and the like. However, the present invention is not limited thereto.

When the formulation of the cosmetic composition of the present invention is a paste, a cream or a gel, an animal fiber, a plant fiber, a wax, a paraffin, a starch, a tracer, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, Can be used.

When the formulation of the cosmetic composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, Propellants such as carbon, propane / butane or dimethyl ether.

When the formulation of the cosmetic composition of the present invention is in the form of a solution or an emulsion, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan can be used.

When the formulation of the cosmetic composition of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the cosmetic composition of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, methyltaurate, sarcosinate , Fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolenic derivative or ethoxylated glycerol fatty acid ester.

On the other hand, the second aspect of the present invention provides a fermented product obtained by adding a fat-soluble solvent to a fermentation broth obtained by fermenting a camellia oil with yeast, and then removing the fat-soluble fraction or an extract thereof as an active ingredient A pharmaceutical composition for promoting hair growth or preventing hair loss is provided. The pharmaceutical composition of the present invention may be in the form of an oral formulation, an external preparation, a suppository and a sterilized injection solution, preferably in the form of an external preparation.

In the pharmaceutical composition of the present invention, when the oral formulations are solid formulations, they may be, for example, tablets, pills, powders, capsules, starch, calsium carbonate, sucrose or lactose ), And gelatin.

In the pharmaceutical composition of the present invention, when the oral formulation is a liquid preparation, it may be, for example, a suspension, a solution, an emulsion or a syrup, and may contain propylene glycol, polyethylene glycol, vegetable oil, , ≪ / RTI >

In the pharmaceutical composition of the present invention, the external preparation may be formulated into a liquid, cream, paste, or solid form.

On the other hand, the pharmaceutical composition of the present invention is preferably administered at 0.0001 to 100 mg / kg per day, more preferably 0.001 to 10 mg / kg per day. However, the present invention is not limited thereto, and dosage may be different depending on the condition and the weight of the patient, the degree of disease, the drug form, the administration route and the period. In addition, the administration can be administered once a day or divided into several times.

On the other hand, the pharmaceutical composition of the present invention preferably contains the camellia oil fermented product or the extract thereof in an amount of 0.1 to 50% by weight based on the total weight of the composition.

Meanwhile, the pharmaceutical composition of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the production of pharmaceutical compositions. Examples of the carrier include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, But are not limited to, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, Magnesium stearate and mineral can be unique.

In the pharmaceutical compositions of the present invention, the yeast is Candida (Candida) recommended layman, more preferably Candida spring rain coke (Candida bombicola .

On the other hand, the third aspect of the present invention is a method for culturing yeast cells comprising the steps of: (a) inoculating yeast cells into a culture medium containing camellia oil and then culturing; (B) a step of adding the oil-soluble organic solvent to the mixture and stirring the mixture, and then removing the fat-soluble organic solvent fraction layer. The camellia oil fermented extract produced by the production method of the present invention exerts the effect of promoting hair growth or preventing hair loss.

The method for producing the camellia oil fermentation extract of the present invention preferably further comprises the step (c) of removing the fat-soluble organic solvent fraction layer of the step (b) and adding a water-soluble solvent to extract the oil-soluble organic solvent fraction layer. This is because the useful components can be further concentrated and extracted.

In the method for producing the camellia oil fermented extract of the present invention, the yeast of step (a) is preferably Candida , more preferably Candida bombicola .

In the method for producing a camellia oil fermented extract of the present invention, the step (a) is preferably cultivated at a temperature of 20 to 30 DEG C for 3 to 10 days. When the above range is satisfied, the camellia oil fermented extract can be produced with high productivity.

In the method for producing the camellia oil fermented extract of the present invention, the step (a) may preferably include a step of sterilizing the culture medium after the incubation to kill the yeasts present in the culture medium.

In the method for producing the camellia oil fermented extract of the present invention, the step (a) may preferably include a step of culturing and then aging. At this time, the aging is preferably carried out at 15 to 35 DEG C for 3 to 9 months.

In the method for producing the camellia oil fermentation extract of the present invention, the oil-soluble organic solvent of step (b) may be one comprising hexane, ethyl acetate, methylene chloride and ether alone or in admixture.

In the method for producing the camellia oil fermentation extract of the present invention, the step (c) preferably includes a step of removing diol-soluble organic solvent fraction layer, adding diatomaceous earth before extraction with a water-soluble solvent, good. At this time, more preferably, the filtration may be further followed by concentrating the filtrate to prepare a concentrate.

In the method for producing the camellia oil fermented extract of the present invention, the water-soluble solvent of step (c) may be one prepared by mixing water, methanol aqueous solution, aqueous ethanol solution, propylene glycol, butylene glycol, have.

Meanwhile, in the method for producing a camellia oil fermentation extract of the present invention, the method may further include, after the step (c), concentrating the extract obtained by extracting with a water-soluble solvent .

The composition comprising the camellia oil fermented product obtained by removing the oil soluble fraction after adding the oil soluble solvent to the fermentation broth obtained by fermenting the camellia oil of the present invention with yeast or an extract thereof as the active ingredient is a fibroblast and dermal papilla cell Promoting the proliferation of hair growth related genes and increasing the expression of hair growth promotes hair growth and hair loss prevention effect.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

[ Example  1: Production of camellia oil fermented extract]

0.5 g of yeast extract, KH ₂ PO ₄ 0.1 g of MgSO ₄ (7H ₂ O), 0.01 g of CaCl ₂ (2H ₂ O), 0.01 g of NaCl, 0.07 g of peptone, 10 g of glucose and 10 g of camellia oil were mixed with purified water to obtain 1 L By weight. Sterilization 30 minutes at after production, 121 ℃ the mixture 1 L, and the yeast Candida spring rain coke (Candida bombicola ATCC 22214, and cultured at 25 占 폚 for 7 days. The cultured broth was heated to 121 ° C to kill the yeasts, and the sterilized solution was aged at room temperature (15-35 ° C) for 6 months.

After aging, 2 L of hexane was added to the aging solution and stirred, and the hexane oil mixed fraction layer of the upper layer was removed. The lower layer was mixed with a certain amount of diatomaceous earth and stirred. The solution was filtered using a 0.45 μm filter paper, and the filtrate was concentrated under reduced pressure at 65 ° C. to obtain 152 g of a fermented product.

Thereafter, 3 L of 50% ethanol was mixed and extracted, and the resulting extract was concentrated under reduced pressure at a temperature of 60 to 70 캜 to obtain 110 g of a solid.

[ Example  2 to 12: Preparation of camellia oil fermented extract]

The extracts of Examples 2 to 12 were prepared by performing fermentation, aging, and extraction under the same conditions as in Example 1, but under the conditions described in Table 1 below.

Primary oil removal and purification solvent Aging period (month) Extraction solvent Solids (g) Example 2 Hexane 0 Functional ethanol (50% by weight) 120 Example 3 Hexane 3 Functional ethanol (50% by weight) 115 Example 4 Hexane 6 Saturated butanol 85 Example 5 Hexane 6 Function Propylene glycol (50%) 78 Example 6 Hexane 6 Function Butylene glycol (50%) 69 Example 7 Ethyl acetate 0 Functional ethanol (50% by weight) 53 Example 8 Ethyl acetate 3 Functional ethanol (50% by weight) 56 Example 9 Ethyl acetate 6 Functional ethanol (50% by weight) 49 Example 10 Ethyl acetate 6 Saturated butanol 63 Example 11 Ethyl acetate 6 The propylene glycol (50% by weight) 51 Example 12 Ethyl acetate 6  Functional butylene glycol (50% by weight) 49

[Experimental Example 1: Experiment on human fibroblast proliferation]

Hair growth is regulated by the differentiation and proliferation of keratinocytes, melanocytes, and fibroblasts (Hirobe T, Histol Histopathol 1995; 10: 223-237). Therefore, in this Experimental Example, the effects of Examples 1 to 12 on the proliferation of fibroblasts, which are regarded as important factors for hair growth, were evaluated.

MTT assay is an experimental method widely used for cell proliferation and toxicity by measuring the number of living cells. The living cells are soluble in mitochondria and have a yellow salt, MTT (3- [4,5-dimethylthiazole-2-yl] 5-diphenyltetrazolium bromide) is reduced to a blue formazan derivative which is insoluble by succinate dehydrogenase (also called succinate dehydrogenase or mitochondrial dehydrogenase). The resulting formazan derivative is dissolved in a dissolving agent (usually dimethylsulfoxide) and the absorbance is measured.

For the experiment, human fibroblast (ATCC, American Type Culture Collection, Cat. No. CRL-2076) was inoculated into a 24-well plate at a concentration of 5 x 10 ⁴ cells / ml. At this time, IMDM (Iscove's Modified Dulbecco's Medium; GIBCO, USA) containing 10% bovine serum was used as a medium. After 24 hours of inoculation, the medium was replaced with IMDM (Iscove's Modified Dulbecco's Medium; GIBCO, USA) containing 0% bovine serum, 10 μl of the test sample (Examples 1 to 12) was added at a concentration of 100 μg / ml, And incubated for 3 days at 37 ° C in a 5% CO ₂ incubator.

After the incubation, the supernatant was removed and washed again with 200 μl of 5% PBS (phosphate buffered saline). After washing, 1.0 μl of 25 μg / ml MTT solution was added to each well. After 4 hours, MTT was removed. 1.0 ml of DMSO was added to each well. Absorbance was measured at 570 nm.

The survival rate of the cells was calculated using the following equation (1), and the results are shown in Table 2. At this time, Minoxidil, which is known to be effective for proliferation of fibroblasts, was used as a comparative sample.

sample Cell survival rate (%) control 100 Minoxidil 500 μM 115 Example 1 98 Example 2 102 Example 3 106 Example 4 111 Example 5 109 Example 6 112 Example 7 102 Example 8 113 Example 9 126  Example 10 127  Example 11 119  Example 12 121

As a result of the experiment, it was confirmed that the cell proliferation effect was about 20% or more in Examples 9 to 12. This is a result that the cell proliferation effect is similar or higher than that of minoxidil, which is known to be effective for proliferation of fibroblasts.

As a comparative sample, 500 μM (104.46 μg / ml) of minoxidil was used, which showed a 15% cell proliferation effect and a 20% proliferation at 50 μg / ml treatment of camellia oil fermented extract Therefore, camellia oil fermented extract seems to have a better effect on fibroblast proliferation than minoxidil.

[ Experimental Example  2: Dermal papilla cells  Experiment on proliferation]

Dermoid papillary cells are one of the major components of the hair dermal papilla in the hair follicle, and the basic fibroblast growth factor (bFGF), endothelin-1 (ET 1), stem cell factor (SCF), etc.) promote the growth and differentiation of hair matrix cells (Paus R, Foitzik K. Differentiation 2004; 72: 489-511; Fujie T, et al. J Dermatol Sci 2001; 25: 206-212; Botchkarev VA, et al., J Investig Dermatol Symp Proc 2003; 8: 46-55). Therefore, in this Experimental Example, the effects of Examples 1 to 12 on the proliferation of dermal papilla cells, which are regarded as important factors for hair growth, were evaluated.

Dermal papilla cells were inoculated into a 24-well plate at a concentration of 5 × 10 ⁴ cells / ml. At this time, DMEM (Dubelco's Modified Eagle Medium, BRL, USA) containing 10% bovine serum was used. After 24 hours of inoculation, the cells were replaced with DMEM (Dubelco's Modified Eagle Medium, BRL, USA) containing 0% bovine serum and 10 μl of the test sample (Examples 2 to 13) was added at a concentration of 50 μg / Lt; RTI ID = 0.0 > 37 C < / RTI > in a 5% CO ₂ incubator.

After the incubation, the supernatant was removed, and then 200 μl of 5% PBS (phosphate buffered saline) was added. The MTT solution was added at a concentration of 25 μg / ml in an amount of 1.0 ml per well. DMSO was added at a rate of 1.0 ml per well and the absorbance was measured at 570 nm.

The survival rate of the cells was calculated in the same manner as in Equation (1) above, and the results are shown in Table 3. At this time, Minoxidil, which is known to be effective for proliferation of dermal papilla cells, was used as a comparative sample.

sample Cell survival rate (%) control 100 Minoxidil 500 μM 121 Example 1 99 Example 2 102 Example 3 99 Example 4 107 Example 5 108 Example 6 110 Example 7 104 Example 8 113 Example 9 125  Example 10 123  Example 11 117  Example 12 121

As a result of the experiment, it was confirmed that the cell proliferation effect of 20% or more was observed in Examples 9 to 12. This shows a similar level of cell proliferation effect as compared with Minoxidil, which is known to be effective for proliferation of dermal papilla cells.

As a comparative sample, 500 μM (104.46 μg / ml) of minoxidil was used. When the fermented extract of Camellia oil was treated with 50 μg / ml, the fermented extract of Camellia oil showed more than 20% It seems to have excellent effect on proliferation.

[Experimental Example 3: Experiment on human fibroblast proliferation by concentration of extract]

In this experimental example, the proliferation effect of the human fibroblast of Example 9 was measured in the same manner as in Experimental Example 1 by concentration. The results are shown in Table 4 below.

Example Cell survival rate (%)  control 100.00 Example 9 1 占 퐂 / ml 103.21 Example 9 10 [mu] g / ml 119.37 Example 9 100 占 퐂 / ml 125.40  Minoxidil 500 μM 124.70

As a result, the cell proliferation effect was about 20% from 10 ㎍ / ml concentration and the maximum survival rate was 100 ㎍ / ml. This indicates that the optimum concentration of camellia oil fermented extract in the proliferation effect on human fibroblasts is 10 to 100 占 퐂 / ml.

[Experimental Example 4: Experiment on dermal papillary cell proliferation according to concentration of extract]

In this experimental example, the proliferation effect of the dermal papilla cells of Example 9 was measured by the same method as Experimental Example 2, and the results are shown in Table 5 below.

Example Cell survival rate (%) control 100.00 Example 9 1 占 퐂 / ml 106.33 Example 9 10 [mu] g / ml 119.52  Example 9 100 占 퐂 / ml 128.20  Minoxidil 500 μM 126.48

As a result, the cell proliferation effect was about 20% from 10 ㎍ / ml concentration and the maximum survival rate was 100 ㎍ / ml. This indicates that the optimum concentration of camellia oil fermented extract in the proliferation effect on dermal papilla cells is 10 ~ 100 / / ml.

[ Experimental Example  5: Measurement of increased expression of hair-related gene by camellia oil fermented extract]

In this experiment, reverse transcription polymerase chain reaction (RT-PCR) was performed to examine the keratinocyte growth factor (KGF) and vascular endothelial growth factor (VEGF) Respectively.

KGF is one of the fibroblast growth factors that stimulate hair growth and VEGF is a factor promoting neovascularization in the dormant period and is known as a factor that induces new hair growth period (Dermatol. 1: 49-53 1993) 16,17, Biomaterials 24: 2387-2394 (2003)).

The dermal papilla was inoculated in a 100 mm cell culture dish at an 80% density and cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours. At this time, DMEM (Dubelco's Modified Eagle Medium, BRL, USA) containing 10% bovine serum was used. Thereafter, Example 9 was added for each concentration and cultured for 18 hours. 1 ml of Trizol (Invitrogen, USA) was added to the cells and the cells were separated according to the RNA isolation method of Promega.

After separation, RNA was quantitated at 260 nm using an ultraviolet detector and RT-PCR was performed. The primers and reaction conditions were as shown in Tables 6 and 7 below. The RT-PCR was performed using the RT-PCR kit (All-in-one RT-PCR Kit, SuperBio, Korea) .

Keratinocyte factor (KGF) primer sense  5'-GACATGGATCCTGCCAACTT-3 ' Antisense  5'-AATTCCAACTGCCACTGTCC-3 ' Reaction conditions After reverse transcription at 50 ° C for 30 minutes, reverse transcription enzyme was inactivated at 96 ° C for 3 minutes, followed by 35 cycles of 94 ° C for 30 seconds, 58 ° C for 30 seconds, and 72 ° C for 1 minute

Vascular endothelial growth factor (VEGF) primers sense  5'-TCTTCAAGCCATCCTGTGTG-3 ' Antisense  5'-GCGAGTCTGTGTTTTTGCAG-3 ' Reaction conditions After reverse transcription at 50 ° C for 30 minutes, reverse transcription enzyme was inactivated at 96 ° C for 3 minutes, followed by 40 cycles of 94 ° C for 30 seconds, 58 ° C for 30 seconds, and 72 ° C for 1 minute

Beta-actin primer < RTI ID = 0.0 > sense  5'-GAGACCTTCAACACCCCAGCC-3 ' Antisense  5'-GGCCATCTCTTGCTCGAAGTC-3 ' Reaction conditions After reverse transcription at 50 ° C for 30 min, reverse transcription enzyme was inactivated at 96 ° C for 3 min, followed by 25 cycles of 94 ° C for 1 min, 62 ° C for 1 min, and 72 ° C for 1 min.

The results of measurement of keratinocyte factor (KGF) and vascular endothelial growth factor (VEGF) expression are shown in Table 9, corrected by the result of β-actin gene.

mRNA expression level
(%, Corrected for actin) density Control
(%) 1 ug / ml
(%) 10 [mu] g / ml
(%) 100 [mu] g / ml
(%) Keratinocyte cell factor (KGF) 100 134 155 183 Vascular Endothelial Growth Factor (VEGF) 100 104 110 112

As a result, it was confirmed that the effect of increasing the concentration of camellia oil fermented extract was increased.

[Preparation Example 1: Preparation of cosmetic preparation containing camellia oil fermented extract]

In this Example, a hair lotion as a cosmetic composition containing a camellia oil fermented extract was prepared as shown in Table 10 below.

ingredient Production Example (% by weight) Cetostearyl alcohol 2.0 Stearyltriethylammonium chloride 2.0 Hydroxyethyl cellulose 0.5 Phytontholamine 0.1 Example 9 (Camellia oil fermented extract) 10 Spices, coloring 0.5 Purified water balance system 100

On the other hand, as a comparative example, a general cosmetic composition containing no camellia oil fermented extract was prepared as shown in Table 11 below.

ingredient Comparative Example (% by weight) Cetostearyl alcohol 2.0 Stearyltriethylammonium chloride 2.0 Hydroxyethyl cellulose 0.5 Phytontholamine 0.1 Example 9 (Camellia oil fermented extract) - Spices, coloring 0.5 Purified water balance system 100

[ Experimental Example  6: Evaluation of clinical efficacy experiment]

In this Experimental Example, the clinical effects of hair loss prevention and hair growth promotion were measured using the 'formulation containing the camellia oil fermentation extract' of Preparation Example 1 and the 'formulation without the camellia oil fermentation extract' of the comparative example Respectively.

Subjects were male and female adults between the ages of 20 and 50 years, and those who had alopecia or an average daily number of hair loss of more than 100 persons were used. The method of use was 2 ~ 3 times a day (about 2 ml) And to use it for at least 1 month to 5 months depending on the degree of effect confirmation.

To determine the clinical effect, the average daily amount of hair loss, visual evident, and subjective symptom improvement were evaluated according to the following five steps (see Table 12).

Step classification Amount of hair loss Subjective symptoms Visual judgment 5
4
3
2
One Significant reduction
Moderate reduction
Hardness reduction
immutability
Deterioration (increase) A significant improvement
Moderate improvement
Hardness improvement
immutability
worse A significant improvement
Moderate improvement
Hardness improvement
immutability
worse

The final judgment was evaluated as a comprehensive improvement of at least 3.0 points, and the results are shown in Table 13 below.

Step classification Amount of hair loss Subjective symptoms Visual judgment Overall improvement Production Example 1 3.4 3.7 3.8 3.6 Comparative Example 1 2.4 2.4 2.6 2.5

As a result of the experiment, as shown in Table 13, although Comparative Example 1 showed a slight improvement effect, it was difficult to expect effectiveness. However, a moderate improvement effect was shown in the composition formulation (Preparation Example 1) containing the camellia oil fermented extract according to the present invention. Thus, it can be concluded that the composition of the present invention exerts excellent effects in preventing hair loss and promoting hair growth. In addition, no adverse effects were found in all subjects during the period of use in the clinical experiment, and it was found that the composition is safe for human body.

Claims (19)

A fermentation product obtained by fermenting a camellia oil with yeast, a fat-soluble solvent added to the fermentation broth, and then removing the fat-soluble fraction, or an extract thereof,
The yeast is Candida bombicola ATCC 22214,
Wherein the oil-soluble solvent is ethyl acetate.
The method according to claim 1,
The composition may comprise,
Wherein the composition is a cosmetic or scalp detergent.
3. The method of claim 2,
In the cosmetic,
Wherein the composition is one selected from the group consisting of a cream, a lotion, a skin, a cleansing, a soap, a treatment, and an essence.
delete A fermentation product obtained by fermenting a camellia oil with yeast, a fat-soluble solvent added to the fermentation broth, and then removing the fat-soluble fraction, or an extract thereof,
The yeast is Candida bombicola ATCC 22214,
Wherein the oil-soluble solvent is ethyl acetate.
6. The method of claim 5,
The pharmaceutical composition may contain,
A pharmaceutical composition for promoting hair growth or preventing alopecia.
delete (A) inoculating yeast cells in a culture medium containing camellia oil and then culturing them;
(B) after the culturing, adding a fat-soluble organic solvent and stirring, and then removing the fat-soluble organic solvent fraction layer,
The yeast is Candida bombicola ATCC 22214,
Wherein the oil-soluble organic solvent is ethyl acetate.
9. The method of claim 8,
The method for producing the camellia oil fermented extract comprises:
Further comprising the step (c) of removing the oil-soluble organic solvent fraction layer of step (b) and adding a water-soluble solvent to the oil-extracted organic solvent fraction layer to extract the oil-soluble organic solvent fraction layer.
delete 9. The method of claim 8,
The culture of step (a)
And culturing the mixture at a temperature of 20 to 30 占 폚 for 3 to 10 days.
9. The method of claim 8,
The step (a)
Culturing the yeast cells, and sterilizing the culture broth to kill the yeast cells present in the culture broth.
9. The method of claim 8,
The step (a)
Culturing, and aging the camellia oil fermented extract.
14. The method of claim 13,
The above-
Wherein the fermentation is carried out at 15 to 35 占 폚 for 3 to 9 months.
delete The method of claim 8, wherein
The step (b)
After removing the fat-soluble organic solvent fraction layer,
And adding diatomaceous earth, followed by stirring and filtration.
17. The method of claim 16,
The step (b)
Filtrating the filtrate, and concentrating the filtrate to obtain a concentrated product.
10. The method of claim 9,
The water-soluble solvent of step (c)
A method for producing a camellia oil fermented extract, which comprises water, an aqueous methanol solution, an aqueous ethanol solution, propylene glycol, butylene glycol, and glycerin.
10. The method of claim 9,
The method for producing the camellia oil fermented extract comprises:
And (c) concentrating the extract obtained by extracting with a water-soluble solvent after the step (c).