WO2016137284A1 - Composition for promoting hair growth - Google Patents

Abstract

The present invention relates to a composition for promoting hair growth, the composition containing, as an active ingredient, at least one selected from the group consisting of a Rheum undulatum extract, a Nardostachys chinensis extract, and organic solvent fractions thereof. The use of the composition of the present invention can lead to a hair growth promoting effect.

Description

Hair growth promoting composition

The present invention was made by the task number 113011-3 under the support of the Ministry of Agriculture, Food and Rural Affairs, the research and management agency of the project is the Ministry of Agriculture, Food and Rural Affairs, the research project name is "High Value Food Technology Development Project," and the project title is "Betacatenin Signal." Hair Loss Prevention Inducing Activity to Explore Nontoxic Food and Herbal Ingredients and Development of Functional Skin Care Products for Wool ”, The Supervising Organization is Natural Endotech Co., Ltd., and the period of research is 2013. 07. 16 ~ 2016. 07. 15.

This patent application is filed with the Korean Patent Application No. 10-2015-0028320 filed with the Korean Patent Office on February 27, 2015 and the Korean Patent Application No. 10-2016-0023500 filed with the Korean Patent Office on February 26, 2016. Priority is claimed, the disclosures of which are incorporated herein by reference.

The present invention relates to a composition for preventing hair loss or promoting hair growth comprising at least one component selected from rhubarb extract and extract persimmon.

Many causes and mechanisms of alopecia are unknown. The main causes are reportedly related to male hormones and steroid hormones. For example, the cause of androgenetic alopecia is androgenic hormone balance, which has been reported to be the main cause of androgenetic alopecia (androgen) (J. Shapiro et al., Hair regrowth.Therapeutic agents, Dermatol. Clin. 16 (2), 341-356, 1998). In addition, a paper presented that stress is a significant cause of alopecia. On the other hand, studies to determine the cause of baldness and hair loss, has recently been suggested that the impairment of kidney function is a direct cause of hair loss.

Attempts have been made to develop materials that can prevent hair loss or promote hair growth. This effort, however, was an unfinished development that almost resulted in failure or only partially succeeded. There are several anti-hair loss or hair growth drugs that have been certified by the US Food and Drug Administration (FDA), but they do not achieve satisfactory hair loss or hair growth.

On the other hand, Wnt protein is known that beta-catenin (β-catenin) plays an important role in hair follicle formation, and is involved in the development and growth of hair. Wnt protein is a signaling protein involved in hair development and growth. Wnt signaling leads to betacatenin to form hair follicles and plays an important role in hair loss treatment [1].

In addition, 5α-reductase is an enzyme that converts testosterone, a male hormone, to DHT (Dihydtotestosterone), and is known to inhibit hair loss and promote hair growth by inhibiting the action of 5α-reductase.

US Patent Publication No. 2005-260152 discloses that the garlic component dissolved in a liquid carrier including water, polysorbate and auxiliary ingredients exerts a hair growth action. US Patent Publication No. 2005-239697 discloses a composition for promoting hair growth comprising an actin-binding peptide as an active ingredient.

WO 2005-063279 discloses that IL (interleukin) -15 is associated with promoting hair growth or preventing hair loss. WO 2005-053763 discloses a method for promoting hair growth comprising transplanting the dermal papilla or cultured dermal papilla cells into the skin.

Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

The present inventors earnestly researched to develop a hair growth promoting composition comprising a natural extract component. As a result, by clarifying that the hair growth promoting effect can be obtained by using at least one component selected from rhubarb extract and persimmon extract, the present invention has been completed.

Accordingly, it is an object of the present invention to provide a composition for preventing hair loss or promoting hair growth.

Another object of the present invention is to provide a method for preventing hair loss or promoting hair growth.

Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.

According to an aspect of the present invention, the present invention provides a composition for preventing hair loss or promoting hair growth comprising any one or more selected from the group consisting of rhubarb extract, extract persimmon extract and each organic solvent fraction as an active ingredient.

The present inventors earnestly researched to develop a hair growth promoting composition comprising a natural extract component. As a result, it was found that the hair growth promoting effect can be obtained by using at least one component selected from rhubarb extract and extract of persimmon.

As used herein, the term “hair loss” refers to a state in which the hair ratio in the growth phase decreases, the number of hairs in the degenerative or resting phase increases, and the number of abnormal hair dropouts increases compared to the average number of hair dropouts in a normal state.

"Rhubarb" used as an active ingredient of the present invention is a medicinal herb made from the root of rhubarb, which is a perennial herb of the genusaceae. It is made using Rheum palmatum L., Rheum tanguticum Maximowicz, Rheum officinale Baillon, and Rheum stem of the same plant. When you take rhubarb, you have diarrhea that removes the poison and heat in your body. Cleans the intestines, promotes metabolism, and treats febrile constipation and bloating. It is also effective for headache, congestion, sore throat, constipation, nosebleeds, bleeding, and boils. It is also used for diuresis and edema. Its pharmacological action is known to promote colon movement, fever, lowering body temperature, promoting bile secretion, reducing blood clotting time, antibacterial, diuretic, and liver function.

In addition, "Nardostachys" used as an active ingredient of the present invention is a medicinal herb (Nardostachys chinensis Batalin), leaf persimmon (Nardostachys jatamansi DC.) And medicinal herbs made using the roots and roots of the same plant. It is effective in relieving abdominal pain and is effective for blotch, blemishes, malnutrition accompanied by swelling, tooth decay, and hemorrhoids.

The rhubarb extract and the extract of persimmon are various extraction solvents such as (a) water, (b) anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), (c) the lower alcohol A mixed solvent of water and water, (d) acetone, (e) ethyl acetate, (f) chloroform, (g) 1,3-butylene glycol and (h) butyl acetate can be obtained as an extraction solvent. Most preferably, the extract of the present invention is obtained using water as the extraction solvent. On the other hand, it will be apparent to those skilled in the art that the extract of the present invention can obtain an extract having substantially the same effect using not only the extraction solvent but also other extraction solvents.

In addition, the present invention may include an organic solvent fraction for the primary extract as an active ingredient. As used herein, the term “fractions” refers to secondary extracts obtained by extracting the above-mentioned primary extract with at least one organic solvent selected from the group consisting of hexane, ethyl acetate, dichloromethane, n-butanol, petroleum ether, benzene, and chloroform. Means. More specifically, the primary extract is prepared in a powder state by distillation under reduced pressure, which is water and any one selected from the group consisting of hexane, ethyl acetate, dichloromethane, n-butanol, petroleum ether, benzene and chloroform. Fractions may be obtained in such a way that the organic solvent layer is recovered by putting the organic solvents together.

In an embodiment of the present invention, the powder obtained by distillation of the primary extract of each component under reduced pressure (a) was subjected to layer separation using the same amount of water and hexane, and the hexane layer was recovered to obtain a hexane fraction, (b ) After recovering the hexane fraction, the same amount of ethyl acetate was added to the remaining water layer to separate the layers, and the ethyl acetate layer was recovered to obtain an ethyl acetate fraction. (C) After recovering the ethyl acetate fraction, the same amount of dichloromethane was added to the remaining water layer. Methane was added to separate the layers, and the dichloromethane layer was recovered to obtain a dichloromethane fraction. (D) Finally, the same amount of butanol was added to the remaining water layer to separate the layers, and the butanol layer was recovered to recover the butanol fraction. Got it. Fractions of the present invention may be obtained by using hexane, ethyl acetate, dichloromethane, and butanol sequentially as in one embodiment of the present invention described above, but is not limited thereto, and ethyl acetate, dichloromethane, or butanol In the case of the fractions, similarly to the hexane fractions, ethyl acetate, dichloromethane, or butanol may be recovered with the same amount of water with respect to the primary extract powder after the first layer separation.

As used herein, the term “primary extract” is a term used to distinguish a “fraction” from the meaning of a secondary extract. The term “a) water, (b) anhydrous or water containing 1 to 4 carbon atoms is described above. Lower alcohols (methanol, ethanol, propanol, butanol, etc.), (c) a mixed solvent of the lower alcohols with water, (d) acetone, (e) ethyl acetate, (f) chloroform, (g) 1,3-butyl The extract extracted for the first time using solvents, such as len glycol and (h) butyl acetate, as an extraction solvent is shown. Extracts of the present invention include those prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

Any one or more selected from the group consisting of rhubarb extract, extract of persimmon and extract of each of the organic solvents, which are the active ingredients of the present invention, exhibit high winch / beta-catenin signal induction promoting activity and 5α-reductase inhibitory effect (see : Figure 2-5).

In one embodiment of the present invention, the organic solvent fraction of the present invention is an ethyl acetate fraction of rhubarb, dichloromethane fraction, butanol fraction, obtained by sequentially fractionating rhubarb or persimmon fragrance with hexane, ethyl acetate, dichloromethane, butanol, water At least one selected from the group consisting of hexane fraction, ethyl acetate fraction, butanol fraction and water fraction of persimmon flavor. The ethyl acetate fraction, dichloromethane fraction, butanol fraction, hexane fraction, ethyl acetate fraction, butanol fraction and water fraction of rhubarb of the present invention may be specifically prepared according to, for example, Preparation Example 1-2 on the present specification. have. Rhubarb ethyl acetate fraction, dichloromethane fraction, butanol fraction, persimmon hexane fraction, ethyl acetate fraction, butanol fraction and water fraction show very good wint / betacatenin signaling inducing activity and / or 5α-reductase inhibitory effect. (See FIG. 5).

In one embodiment of the present invention, the active ingredient of the present invention is a single rhubarb extract, a persimmon fragrance extract, or a combination of rhubarb and persimmon fragrance extract. Rhubarb single extract, sweet persimmon extract, and rhubarb and persimmon extract were found to exhibit the same level of efficacy as commercially available hair regrowth minoxil in mouse hair growth test (see Fig. 6).

In one embodiment of the invention, the composition of the present invention is a food composition.

When the composition of the present invention is prepared as a food composition, as an active ingredient, as well as rhubarb extracts, persimmon extract, as well as components commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings And flavoring agents. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

For example, when the food composition of the present invention is prepared with a drink, in addition to the rhubarb and persimmon extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, jujube extract, licorice extract, and the like may be further included. have.

In one embodiment of the invention, the composition of the present invention is a cosmetic composition.

The components included in the cosmetic composition of the present invention include components commonly used in cosmetic compositions in addition to rhubarb extract and extract of persimmon as active ingredients, and are commonly used for example, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings. Phosphorus aids, and carriers.

The composition for improving skin condition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactants -Can be formulated with, but not limited to, cleansing, oils, powder foundations, emulsion foundations, wax foundations, sprays, and the like. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

In one embodiment of the invention, the composition of the invention is a pharmaceutical composition.

When the composition of the present invention is made into a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be oral or parenteral administration, and may preferably be an external composition for skin applied by topical application by parenteral administration, more preferably by application.

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. The dosage of the pharmaceutical composition of the present invention is in the range of 0.001-80 mg / kg on an adult basis. In addition, in the case of external preparations, it is recommended to continue the application for one month to three times a day in an amount of 1.0 to 3.0 ml on an adult basis. However, the dosage does not limit the scope of the present invention.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulation may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of extracts, powders, powders, granules, tablets or capsules, and may further comprise dispersants or stabilizers.

According to another aspect of the present invention, the present invention requires a composition for preventing hair loss or promoting hair growth comprising any one or more selected from the group consisting of rhubarb extract, extract persimmon extract and each organic solvent fraction as an active ingredient It provides a method for preventing hair loss or promoting hair growth comprising administering to a subject.

“Administration” in the methods of the invention may be oral or parenteral administration, preferably parenteral administration, more preferably topical application by application.

Suitable dosages in the methods of the present invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of the patient, food, time of administration, route of administration, rate of excretion and response to response. Can be. Suitable dosages for the methods of the invention are in the range of 0.001-80 mg / kg on an adult basis. In addition, in the case of external preparations, it is recommended to continue the application for one month to three times a day in an amount of 1.0 to 3.0 ml on an adult basis. However, the dosage does not limit the scope of the present invention.

In one embodiment of the present invention, the organic solvent fraction of the present invention is an ethyl acetate fraction of rhubarb, dichloromethane fraction, butanol fraction, obtained by sequentially fractionating rhubarb or persimmon fragrance with hexane, ethyl acetate, dichloromethane, butanol, water At least one selected from the group consisting of hexane fraction, ethyl acetate fraction, butanol fraction and water fraction of persimmon flavor.

In one embodiment of the present invention, the active ingredient of the present invention is a single rhubarb extract, a single extract of persimmon fragrance, or a complex extract of rhubarb and persimmon fragrance.

In one embodiment of the invention, the administration of the invention is a topical application to the skin.

The method of the present invention relates to a method of using the composition for preventing hair loss or hair growth, which is another aspect of the present invention described above, and the overlapping content will be omitted in order to avoid excessive complexity of the present specification.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a composition for preventing hair loss or promoting hair growth.

(b) The present invention provides a method for preventing hair loss or promoting hair growth.

(c) The composition and method of the present invention can effectively prevent hair loss and promote hair growth.

(d) Since the active ingredient of the composition of the present invention is a component of a natural product, it can be safely applied to food, cosmetics, medicine and the like.

Figure 1 shows the manufacturing process of the methanol extract and its fractions of rhubarb, persimmon.

Figure 2 shows the results of searching for Wnt / Beta-catenin signaling induction activity in each single extract. In FIG. 2, the term “DW” refers to distilled water, “VPN” refers to valproic acid, and “LiCl” refers to lithium chloride.

FIG. 3 shows the results of the Wnt / beta-catenin signaling induction activity by treating the fractions of each of the solvent layers of rhubarb and persimmon scent prepared in Preparation Example 1-2 at a concentration of 50 μg / ml. Indicates. The term “Hex” in FIG. 3 represents a hexane fraction, “EA” represents an ethyl acetate fraction, “DCM” represents a dichloromethane fraction, “BuOH” represents a butanol fraction, and “W” represents a water fraction.

Figure 4 shows the result of confirming the inhibitory effect on 40-50% 5α-reductase in the rhubarb, persimmon extract extracted in Preparation Example 1-1. In FIG. 4, the term “EB” denotes an enzyme blank containing only a buffer, and “Liv” denotes a reaction result of adding a mouse liver extract.

5 shows the results of confirming the inhibitory effect on 5α-reductase by adding rhubarb and persimmon fractions prepared in Preparation Example 1-2 at a final concentration of 3.3 mg / ml.

Figure 6 shows the results of evaluating the effect of the extract of Preparation Example 1 on the mouse hair growth effect. The term “persimmon” in FIG. 6 refers to rhubarb and persimmon mixed extract.

7 is treated with a test substance for 14 days once a day, after hair removal of the mouse to evaluate the hair growth efficacy of the extract prepared in Preparation Example 1-1, the day of hair removal, 3, 7, 10, 13, And a photograph of the 15-day hair growth site.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example

Preparation Example 1-1: Preparation of Rhubarb, Persimmon Scent, and Complex Extracts thereof

To each 50 g of rhubarb (domestic, Chinese herbal medicine market), persimmon scent (Chinese, Chinese herbal medicine market) or a mixture of two kinds of rhubarb and two kinds of persimmon scent, 500 ml to 600 ml of distilled water was added and refluxed once at 100 ° C for 6 hours. After cooling, the mixture was filtered through a filter paper (Whatman No 4. filter paper) to remove solids. Thereafter, the filtered extract was completely dried using a reduced pressure concentrator, and then dissolved in distilled water at an appropriate concentration to carry out cell experiments, and ethanol was used for dissolution of the animal test sample.

Preparation Example 1-2: Preparation of Rhubarb, Methanol Extract of Persimmon Scent, and Fractions thereof

After grinding 50 g of rhubarb and 50g each, 500-600 ml of methanol was added and refluxed for 4 hours in a 65 ° C. hot water bath. The total methanol extract obtained above was completely removed with a solvent under reduced pressure concentrator (Rotavapor R-215, Buchi), hexane (Hexane) and distilled water (DW) were added to each 200 ml to separate the layers to recover the hexane layer. 200 ml of ethyl acetate was added to the remaining water layer, and the layers were separated in the same manner to recover the ethyl acetate layer. 200 ml of dichloromethane was added to the residual water layer, and the layers were separated to recover the dichloromethane layer. 200 ml of butanol (n-Butanol) was added to the residual water layer, and the layers were separated to recover a butanol layer and a water layer. Each fractional layer was concentrated under reduced pressure to completely remove the solvent, and then dissolved in DMSO at an appropriate concentration and used in the experiment. The overall manufacturing process is as shown in FIG. 1.

Example 1 Evaluation of Wnt / beta-catenin Signal Induction Promoting Activity in Extracts of Rhubarb and Persimmon Scent

In relation to hair growth and hair growth, we investigated the Wnt / beta-catenin signaling induction activity of the extract prepared in Preparation Example 1-1. To this end, HEK-293T (human embryonic kidney cell), which is easy to incubate and transform, is transformed with pCMV renilla vector by transforming TOPFlash, which is a Wnt receptor vector, or FOPFlash, which is a control vector, with a pCMV renilla vector. Used for evaluation test. Specifically, HEK-293T cells were cultured in DMEM medium with 5% CO ₂ , 37 in 10% fetal bovine serum (FBS), 100 unit / ml penicillin and 100 μg / ml streptomycin. Cultured in an incubator at ℃ ℃ conditions, transformed with TOPFlash plasmid, FOPFlash plasmid, pCMV / renilla plasmid. After 12 hours, transformed HEK-293T cells were plated in a 96-well plate for tissue culture. After 24 hours, the extract was treated with a 200 ug / ml concentration, and then incubated for 48 hours. Incubated at. Cells were lysed according to the standard method of the Dual Luciferase Assay System (Promega, USA) to measure their activity. The activity of Renilla luciferase was used to standardize the transformation efficiency.The ratio of TOP / renilla activity was divided by the ratio of FOP / renilla activity, and then the distilled water control value of the comparative group was converted to 1 and the relative TOP / FOP activity ratio was Was obtained. As a positive control, lithium chloride (LiCl) and valproic acid, which are known to activate the Wnt signaling system, were used at 50 μM and 5 μM, respectively. As a result of investigating Wnt / beta-catenin signaling induction activity in each single extract, the value of TOP / FOP in rhubarb extract was increased to 1.5-2.0 compared to the distilled water control as shown in FIG. In particular, the value of TOP / FOP increased more than 2.0 in the extract of the Persimmon scent, indicating that the wink / betacatenin signal-inducing activity was shown.

Example 2 Evaluation of Wnt / beta-catenin Signal Induction Promoting Activity in Each Fraction of Rhubarb and Persimmon Scent

As a result of exploring Wnt / beta-catenin signaling induction activity by treating the fractions of each of the solvent layer prepared in Preparation Example 1-2, each concentration of 50 g / ml of rhubarb. As can be seen, the ratio of TOP / FOP in the dichloromethane (DCM) and butanol (n-BuOH) fractions of rhubarb increased 1.5-2 times compared to the DMSO control, indicating that the wink / betacatenin signaling-inducing activity was observed. .

Example 3: Confirmation of 5α-reductase inhibitor in each extract of rhubarb and persimmon

5α-reductase is an enzyme that converts testosterone to more active Dihydrotestosterone (DHT), which is closely related to hair loss mechanism due to a decrease in wint / betacatenin signaling due to excess DHT. In order to confirm the inhibitory effect on the 5α-reductase of the distilled water extracts of rhubarb and persimmon scent prepared in Preparation Example 1-1, 5α-reductase obtained from mouse liver microsomes by the extract treatment was testosterone. The degree to which the process of decreasing the amount was suppressed was measured. To obtain 5α-reductase of liver microsomes, anesthetized livers of 10-week-old BL / 6 male mice were anesthetized and excised, and the 5α-reductase in liver was sonicated through sonication in lysis buffer. After extraction, the supernatant obtained by centrifugation was used for the experiment, the rest was stored at 80 ℃. For the experiment, a mixture of phosphatase buffer, testosterone, and NaDPH necessary for 5α-reductase reaction was mixed, and the final concentration of 5 mg / ml was obtained by lysis buffer and extract extracted with 5α-reductase. After the addition, the reaction was carried out at 37 ° C. for 180 minutes, and dichloromethane was added to the reaction product to terminate the reaction. As the internal control group, propyl-p-hydroxybenzoate was added to correct the amount of testosterone, and high performance liquid chromatography was performed to evaluate testosterone reduction and the content of the internal control group. , HPLC) was used. As the conditions for the analysis, the stationary phase column was a C18 reverse phase column, and the mobile phase solvent was set to 65:35 ratio of methanol and ultrapure distilled water, and the sample injection amount was 20 μl. Was evaluated at 245 nm using a UV detector. When the value obtained from the enzyme-enzyme blank control group containing the buffer only was 100%, the relative testosterone / internal control ratio of the reaction result of the mouse liver extract (Liv) was calculated, and the inhibitory activity was compared with the result of the herbal extract. (inhibition activity) was calculated and the specific method is shown in Equation 1 below.

(Calculation 1)

[{Sample treatment (testosterone area / internal control area)}-{enzyme only treatment (testosterone area / internal control area)} / Treatment (testosterone area / internal control area)}] * 100 (%)

 Treatment of rhubarb and persimmon extracts prepared in Preparation Example 1-1 showed inhibition of 5α-reductase reaction of 40-50%, confirming hair loss prevention efficacy (see FIG. 4).

Example 4 Identification of 5α-reductase Inhibitors in Each Fraction of Rhubarb and Persimmon Scent

As a result of adding rhubarb and persimmon fragrance fraction prepared in Preparation Example 1-2 at a concentration of 3.3 mg / ml to confirm the inhibitory effect on 5α-reductase (see FIG. 5), rhubarb butanol fraction and persimmon hexane fraction Inhibitory effect of 5α-reductase was apparent in.

Example 5: Mouse hair growth test

For hair growth efficacy test, 6-week-old male C57BL / 6 specific pathogen free mouse (Coatec) was purchased and bred in a stainless steel breeding box [165 W x 240 L x 145 H (mm)]. The animals were housed as less than 3 animals / boxes during the acclimation period and 1 animals / boxes during the administration and observation periods. The experimental animals were freely supplied with rodent feed (2918C, Harlan Co. Ltd., USA) and sterilized filtered drinking water, and the animal breeding room had a temperature of 23 ± 3 ℃, a humidity of 55 ± 15%, a ventilation frequency of 10-20 times, 12 It is managed in an environment that automatically turns on at intervals of time. Healthy animals that did not find any abnormalities during the acclimatization period were anesthetized at the end of the anesthesia at the end of the anesthesia by using an electric clipper for hair removal. Tea depilation was performed. After depilation, only individuals with uniform dorsal skin color were selected and used for the experiment, and groups were divided based on weight. Animal experimental group was set to excipient control group coated only with excipients (ethanol), each of the rhubarb, the single extract group of persimmon scent prepared in Preparation Example 1-1, and two complex extract groups of rhubarb, persimmon fragrance, as a positive control group Minoxyl 3% (modern drug) group was added. From the day after hair removal, excipients and extracts were applied to the hair removal site once a day for a total of 14 days, 200 μl each, and treated constantly to soak naturally. A total of six pictures were taken on the back of the mouse, such as 0 (the day of epilation) and 3, 7, 10, 13, and 15 days (the day of autopsy) after the administration of the test substance. It was used to measure the area of hair growth (%) using. In order to evaluate the hair growth efficacy of the extract prepared in Preparation Example 1-1, the results obtained by quantifying the area of hair growth (%) on the 15th day of sample application are shown in FIG. 6, and the hair growth photograph of each group is shown in FIG. 7. . After hair removal, hair growth of 60-100% occurred according to the extract on the 15th day of application of the sample. At 4% concentration, equivalent efficacy was found to be higher than that of the positive control group, 3%.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

references

Ruth Schmidt-Ullrich et al., Molecular principles of hair follicle induction and morphogenesis, BioEssays 27: 247-261 (2005).

Claims (11)

1. Anti-hair loss or hair growth promoting composition comprising any one or more selected from the group consisting of rhubarb extract, extract persimmon extract and each organic solvent fraction as an active ingredient.
2. According to claim 1, wherein the organic solvent fraction is rhubarb or ethyl acetate fraction of rhubarb obtained by sequentially fractionating the fragrance or scent of hexane, ethyl acetate, dichloromethane, butanol, water, dichloromethane fraction, butanol fraction, hexane fraction of , Ethylacetate fraction, butanol fraction and water fraction, characterized in that any one or more selected from the group consisting of.
3. The composition of claim 1, wherein the active ingredient is rhubarb single extract, persimmon single extract, or rhubarb and persimmon complex extract.
4. The composition of claim 1 wherein the composition is a food composition.
5. The composition of claim 1, wherein the composition is a cosmetic composition.
6. The composition of claim 1, wherein the composition is a pharmaceutical composition.
7. The composition according to claim 6, wherein the composition is an external composition for skin.
8. Preventing hair loss comprising administering to the subject in need thereof a hair loss preventing or hair growth promoting composition comprising any one or more selected from the group consisting of rhubarb extract, extract of persimmon and each organic solvent fraction as an active ingredient Or hair growth promoting method.
9. According to claim 8, wherein the organic solvent fraction of rhubarb or ethyl acetate fraction of rhubarb obtained by sequentially fractionating the rhubarb or persimmon fragrance with hexane, ethyl acetate, dichloromethane, butanol, water, dichloromethane fraction, butanol fraction, hexane fraction of the , Ethyl acetate fraction, butanol fraction and water fraction, any one or more selected from the group consisting of.
10. 9. The method according to claim 8, wherein the active ingredient is rhubarb single extract, persimmon single extract, or rhubarb and persimmon complex extract.
11. 9. The method of claim 8, wherein said administration is a topical application to the skin.

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