CN111246868A - Composition for preventing hair loss or promoting hair growth comprising extract of cultured tissue of beans and preparation method thereof - Google Patents

Abstract

The present invention relates to a composition for preventing hair loss or promoting hair growth comprising a bean culture tissue extract and a method for preparing the same, wherein the composition for preventing hair loss or promoting hair growth comprising bean culture tissue can be uniformly mass-produced using one aspect of the present invention, and the composition prepared by the method has excellent effects of preventing hair loss and promoting hair growth, and has characteristics of no side effects unlike conventional alopecia remedies.

Description

Composition for preventing hair loss or promoting hair growth comprising extract of cultured tissue of beans and preparation method thereof

Technical Field

The present invention relates to a composition for preventing hair loss or promoting hair growth comprising an extract of cultured tissue of beans and a method for preparing the same.

Background

Recently, people suffering from alopecia have been increasing and their ages have been decreasing with changes in dietary life, increased stress, and the like, and the number of people suffering from alopecia has been increasing not only in males but also in females.

Among the causes of hair loss, the most common type having a direct relationship with the hypersecretion of male hormones is androgenic alopecia (androgenic alopecias), which is commonly referred to as male pattern baldness (male pattern baldness). If there is androgenic alopecia, healthy hair gradually becomes thinner and shorter, and the hair becomes soft and is easily broken, which is called atrophy (miniturization). The follicle causing atrophy eventually becomes thin and hard to see to the naked eye and becomes short fine hair, resulting in hair loss.

Male pattern alopecia is Dihydrotestosterone (DHT) which is one of male hormones, and thus activated dihydrotestosterone binds to androgen receptor, delays protein synthesis of hair follicle cells, shortens the anagen phase of hair follicles, atrophy hair follicles, and causes hair loss, and in addition, excessive sebum is generated during male hormone-derived alopecia, and as a result, hair loss accompanied by inflammation is also generated on scalp.

Commercially available alopecia remedies such as finasteride (propecia) or minoxidil (minoxidil) have little effect or great side effects, and thus are limited in use according to subjects, and consumers who use them may also resist use. In order to solve the above problems, there is a strong demand for the development of an anti-hair loss agent or a hair restorer which is excellent in the effects of natural materials and the like.

Disclosure of Invention

Technical problem

The subject of the present specification is a composition for preventing hair loss or promoting hair growth and a method for uniformly preparing the same, which utilize natural raw materials while providing no side effects.

Technical scheme

In order to solve the above problems, the present specification provides a composition for preventing hair loss or promoting hair growth, comprising an extract of a cultured bean tissue, which is a cultured bean root or a callus of bean.

In addition, the present specification provides a method for preparing a composition for preventing hair loss or promoting hair growth, which comprises: (1) a step of positioning a soybean culture root in a culture medium in a bioreactor, and proliferating the soybean culture root while maintaining a predetermined air supply amount in the bioreactor; (2) drying the cultured roots subjected to the step (1); (3) step (2), soaking the product obtained in the step (2) into alcohol for extraction; and (4) filtering and drying after the step (3).

Effects of the invention

The present invention can be used to prepare a composition for preventing hair loss or promoting hair growth, which contains bean culture tissue of uniform quality, uniformly and in large quantities, and the composition prepared by the method has significant effects of preventing hair loss and promoting hair growth while using natural raw materials, and thus can be an alternative solution to overcome the disadvantages of the existing products for preventing hair loss or promoting hair growth.

Drawings

FIG. 1 is a diagram of a situation in which bean culture roots are cultured for 4 weeks using an explant technique according to an aspect of the present invention;

FIG. 2 is a graph showing that extracts enhance Wnt/β -catenin activity by concentration according to an aspect of the invention;

fig. 3 is a graph illustrating the effect of extract on the ratio of catagen phase to anagen phase in the follicular cycle according to an aspect of the present invention.

Detailed Description

In this specification, explant (explantation) may be referred to as an explant or in vitro culture, a technique in which a part of an individual animal or plant is mainly isolated and cultured in vitro. Mainly comprises survival in vitro for a certain time and time-consuming culture until a certain change is gradually confirmed. General tissue culture is also a method of in vitro culture. In addition, the in-reactor culture according to an aspect of the present invention is also included in the in vitro culture.

In the present specification, the explant technique includes a case where one or more selected from the group consisting of protoplasts, cells, tissues, organs, embryos, seeds, culture roots, and parts of plant bodies are grown in an artificial culture apparatus containing a medium or the like to which a specific component is added.

In one aspect, the present invention is a composition for preventing hair loss or promoting hair growth, comprising an extract of bean culture tissue, which is bean culture roots or bean callus.

In another aspect, the present invention relates to a method for preventing hair loss or promoting hair growth, comprising administering an extract of a bean culture tissue to an individual, and may relate to a method for preventing hair loss or promoting hair growth, wherein the bean culture tissue is a bean culture root or a bean callus.

In yet another aspect, the present invention relates to the use of an extract of bean culture tissue for the preparation of a composition for preventing hair loss or promoting hair growth, and may relate to the use of an extract of bean culture tissue that is bean culture root or bean callus.

As an example, the bean may be soybean, broad-leaf cranberry (Glycine gracilis), green-kernel black soybean, murraya koenigii, black soybean, green soybean, kidney bean, pinto bean, red bean, small black bean or bean sprout, and may preferably be soybean or broad-leaf cranberry. The variety of the beans is not limited, and on the one hand, the beans can be bean paste, bean curd, bean sprout, rice cooker or green soybean. The varieties for soybean paste and bean curd include Dafeng (Daepung, variety protection application, application-2003-, variety protection application, application-1998-. As the bean sprout products, there are mentioned mythical (Shinhwa, variety name registration No. 01-0003-, application-1998-. The rice cooking materials include Qingzhao (Cheongjakong, breed protection application, application 2001-38), Heugcheng (Heugheong, breed protection application, application 2000-27), Brown (breed protection application, application 2000-25), Xianhukkong (Seongheukkong, breed protection application, application 1999-20), Black Bean and Yipin Black Bean (breed protection application, application 1998-. The species of green soybean products include Dougu (Daol, variety protection application, application-2003-149), Xin Green (variety protection application, application-2001-35), Xin Yi (saeol), Black Yi (geomjeong), Xianliang green (seekyang-putkong), Huayan green (hwaeom-putkong) (variety name registration No. 01-0003-21), and Dayi (keunol). In other aspects of the invention, the beans are preferably of a germinable and pest resistant variety. Examples of the beans include mythic fungus, small round, peace, southwestern, colorful, small green, huhu, Shuangming, multi-element, high yield bean sprouts, Yishan bean sprouts, small white bean sprouts, Guangan, short leaves, and Galaxy. However, beans according to an aspect of the present invention are not limited to the bean variety.

The broad leaf lima beans according to an aspect of the invention are called lentils, lima beans and the like.

As other examples, the extract may be extracted using water, a lower alcohol of C1 to C6, or a mixture thereof as a solvent.

As another example, the lower alcohol may be ethanol.

As other examples, the alcohol concentration may be 60 to 100% (w/v). In one aspect, the alcohol concentration may be 60% (w/v) or more, 70% (w/v) or more, 75% (w/v) or more, 80% (w/v) or more, 85% (w/v) or more, 90% (w/v) or more, or 95% (w/v) or more. On the other hand, the alcohol concentration may be 100% (w/v) or less, 95% (w/v) or less, 90% (w/v) or less, 85% (w/v) or less, 80% (w/v) or less, 75% (w/v) or less, 70% (w/v) or less, or 65% (w/v) or less.

As another example, the extract may contain coumestrol in an amount of 5 to 15 wt% relative to the total weight of the extract. In addition, the composition may contain coumestrol in an amount of 5 to 15% by weight, relative to the total weight of the extract or the composition. In one aspect, the coumestrol may be present in an amount of 5 wt% or more, 7 wt% or more, 8 wt% or more, 9 wt% or more, 10 wt% or more, 12 wt% or more, or 14 wt% or more. On the other hand, the content of coumestrol may be 15 wt% or less, 13 wt% or less, 12 wt% or less, 11 wt% or less, 10 wt% or less, 8 wt% or less, or 6 wt% or less.

As still other embodiments, the preventing hair loss or promoting hair growth may be accomplished by one or more selected from the group consisting of proliferating hair papilla cells, increasing hair follicle stem cell activity, inducing hair follicle anagen phase, maintaining or extending hair follicle anagen phase, and inhibiting transition of hair follicle cycle to catagen phase.

The composition according to one aspect of the present invention increases the activity of hair follicle stem cells and promotes the proliferation of hair papilla cells, thereby enabling the improvement and treatment of alopecia prevention, hair growth promotion, and various types of alopecia (one or more types selected from the group consisting of androgenetic alopecia, traumatic alopecia, alopecia areata, drug-induced alopecia, and alopecia caused by microbial infection).

The hair is a process of periodic changes of regularly repeating growth phase, catagen phase and telogen phase, and each hair is independently repeatedly generated and died. In the anagen phase, the hair bulb cells are surrounded by a papilla composed of stromal cells and are rounded, and the anagen phase is a phase in which cell division is active for the generation of new hair. The catagen is a period in which the growth activity of the hair follicle is static and rapidly atrophies, and the shape of the hair is similar to a stick. The telogen phase is a period in which the cell activity of the hair follicle is completely destroyed, and after 2 to 3 months, the anagen-phase hair newly growing from the base of the telogen phase hair is squeezed out and falls off, or naturally falls off by mechanical action such as combing or washing the hair. Generally, hair loss includes a case where the ratio of hairs in the anagen phase becomes short and the number of hairs in the catagen or telogen phase becomes large in this cycle, resulting in a large number of abnormal hair loss. The composition according to an aspect of the present invention induces, maintains and prolongs the anagen phase of hair follicles, inhibits transition to the catagen phase, and thus can be used for preventing hair loss, promoting hair growth, improvement and treatment of various kinds of hair loss (one or more selected from the group consisting of androgenic alopecia, traumatic alopecia, alopecia areata, drug-induced hair loss, and microbial infection-induced hair loss).

The composition according to an aspect of the present invention increases the transcriptional activity of β -catenin or increases the Wnt/β -catenin activity by means of concentration, thereby regulating the follicular cycle (i.e., inducing the anagen phase and maintaining the anagen phase), and thus, it can be applied to the treatment of alopecia prevention, hair growth promotion, and various kinds of alopecia (one or more selected from the group consisting of androgenetic alopecia, traumatic alopecia, alopecia areata, drug-induced alopecia, and alopecia caused by microbial infection).

As an example, the route of administration of the composition is not limited, and transdermal or external application to the skin may be preferred.

As other examples, the concentration of the extract within the composition may be from 10 to 100ppm (w/v). In one aspect, the input concentration may be 10ppm (w/v) or more, 20ppm (w/v) or more, 25ppm (w/v) or more, 30ppm (w/v) or more, 35ppm (w/v) or more, 40ppm (w/v) or more, 45ppm (w/v) or more, 50ppm (w/v) or more, 60ppm (w/v) or more, 70ppm (w/v) or more, 80ppm (w/v) or more, 90ppm (w/v) or more, or 95ppm (w/v) or more. On the other hand, the input concentration may be 100ppm (w/v) or less, 90ppm (w/v) or less, 80ppm (w/v) or less, 70ppm (w/v) or less, 60ppm (w/v) or less, 50ppm (w/v) or less, 45ppm (w/v) or less, 40ppm (w/v) or less, 35ppm (w/v) or less, 30ppm (w/v) or less, 25ppm (w/v) or less, 20ppm (w/v) or less, or 15ppm (w/v) or less.

As another example, the amount of the extract dosed based on the dosing of the composition may be 5 mg/kg/day to 300 mg/kg/day. On the other hand, the amount to be added may be 5 mg/kg/day or more, 10 mg/kg/day or more, 30 mg/kg/day or more, 50 mg/kg/day or more, 80 mg/kg/day or more, 100 mg/kg/day or more, 120 mg/kg/day or more, 140 mg/kg/day or more, 160 mg/kg/day or more, 180 mg/kg/day or more, 200 mg/kg/day or more, 250 mg/kg/day or more, or 280 mg/kg/day or more. On the other hand, the amount to be added may be 300 mg/kg/day or less, 280 mg/kg/day or less, 250 mg/kg/day or less, 200 mg/kg/day or less, 180 mg/kg/day or less, 160 mg/kg/day or less, 140 mg/kg/day or less, 120 mg/kg/day or less, 100 mg/kg/day or less, 80 mg/kg/day or less, 50 mg/kg/day or less, 30 mg/kg/day or less, 10 mg/kg/day or less, or 8 mg/kg/day or less.

In another aspect, the composition may be a food, pharmaceutical or cosmetic composition.

In yet another aspect, the composition may be a composition for ameliorating or treating one or more selected from the group consisting of androgenic alopecia, traumatic alopecia, alopecia areata, drug-induced alopecia, and alopecia caused by microbial infection.

The food composition according to an aspect of the present invention may be a health food composition in which the amount of the compound to be administered is determined within the level of those skilled in the art, and may vary depending on various factors such as age, health status, complications, and the like of a subject to be administered.

The formulation of the food composition is not particularly limited, and the food composition may be formulated into, for example, tablets, granules, pills, powders, liquids such as drinks, caramel, gels, bars, tea bags, and the like. The food composition of each dosage form can be prepared by selecting and mixing ingredients commonly used in the art according to the dosage form or the use purpose, besides the effective ingredients, and can cause the effect to be improved when being used together with other raw materials.

The composition can be administered by a variety of methods including simple ingestion, drinking, injection, spray, or extrusion.

The health food composition according to one aspect of the present invention may be a health food, and may include not only a health food, but also processed foods in any form, such as various foods including chewing gum, caramel products, candies, frozen sweets, cookies, breads, and the like, beverage products including soft drinks, mineral water, alcoholic beverages, and the like, and functional foods including vitamins, minerals, and the like.

In addition to this, the health food composition according to an aspect of the present invention may include various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salt, alginic acid and its salt, organic acid, protective gel thickener, pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonic acid agent for carbonated beverage, and the like. In addition, the health food composition according to an aspect of the present invention may include natural fruit juice and pulp for preparing fruit juice beverages as well as vegetable beverages. The ingredients may be used alone or in combination. The ratio of the additives is not critical, but generally ranges from 0 to about 50 parts by weight per 100 parts by weight of the composition according to an aspect of the invention.

The cosmetic composition according to an aspect of the present invention is, for example, a composition for skin, finger and foot nails, and/or hair, and may have a form of a soft lotion, an astringent lotion, a nutritive lotion, a nourishing cream, a massage cream, an eye essence, an essence, a cleansing cream, a cleansing foam, a makeup remover, a mask, a powder, a body lotion, a body cream, a body essence, a body cleanser, a hair dye, a shampoo, a hair conditioner (rinse), a hair fixative, a hair tonic, an ointment, a gel, a cream, a patch, a spray, a powder, a skin adhesive type, and the like, but is not limited thereto.

In addition, each dosage form may contain, in addition to the above-mentioned essential components, other components as appropriate by those skilled in the art according to the type or purpose of use of other external preparations.

The cosmetic composition may be provided in all dosage forms suitable for topical use. For example, the following formulations may be provided: a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an aqueous phase in an oil phase, a suspension, a solid, a gel, a powder, a paste, a microprojection, a foam (foam), or an aerosol composition. The composition of the dosage form may be prepared according to methods generally used in the art.

The cosmetic composition according to the present specification may further include functional additives and ingredients included in general cosmetic compositions, in addition to the compound, extract or fraction of the present specification. The functional additive may include a component selected from the group consisting of water-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed juice. The cosmetic composition according to the present specification may preferably include other components that can exert an effect-enhancing effect on the main effect, within a range that does not impair the main effect. In addition, the cosmetic composition according to the present specification may further include a moisturizer, a lubricant, a surfactant, an ultraviolet absorber, a preservative, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cooling agent, or an antiperspirant. The amount of the above-mentioned components can be easily selected by those skilled in the art within a range not to impair the objects and effects of the present specification, and the amount can be 0.001 to 10% by weight, specifically 0.01 to 3% by weight, based on the weight of the whole composition.

According to yet another embodiment, the composition may be a skin external composition. The composition for external skin preparations may be, but is not limited to, compositions for cosmetics, mouth preparations, mouth washes, pharmaceuticals, and external medical supplies. The skin external preparation is not particularly limited in its dosage form.

In addition, the skin external agent composition of the present specification may further include one or more selected from a functional salt (salt) for special purposes and a pH adjuster for adjusting pH. At this time, the salt may be selected from inorganic salts, organic salts and/or organic-inorganic salts for ion blocking, moisture retention, ultraviolet blocking, and the like. Specifically, for example, the salt may be selected from sodium chloride (NaCl), sodium phosphate (Na)₃PO₄) And calcium chloride (CaCl)₂) And the like. The pH adjuster is selected from acids (acids) or bases (bases), and may be selected from the group consisting of hydrochloric acid, sulfuric acid, tartaric acid, citric acid, phosphoric acid, acetic acid, lactic acid, sodium lactate, sodium hydroxide, potassium hydroxide, alkylamines, alkanolamines, ammonia, and the like.

The skin external preparation composition of the present specification may be a cosmetic, pharmaceutical or pharmaceutical external preparation composition, which may further contain a preservative, a stabilizer, a wettable powder or an emulsification promoter, an auxiliary agent such as a salt for adjusting osmotic pressure and/or a buffer, and other therapeutically useful substances. The composition may be formulated as a lotion, cream, ointment, gel, or the like. The composition for external application to the skin is preferably administered transdermally.

The amount of the active ingredient to be administered in the pharmaceutical or parenteral composition can vary depending on the age, sex, weight and the particular disease or condition to be treated, the severity of the disease or condition, the route of administration and the judgment of the prescribing person. It is within the level of the skilled person to target the input amount determined on the basis of said factor.

The pharmaceutical composition according to an aspect of the present invention may be administered orally, non-orally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, etc. The dosage form for oral administration may be, but is not limited to, tablets, pills, soft and hard capsules, granules, powders, fine granules, liquids, emulsions or pellets. The dosage form for non-oral administration may be a solution, a suspension, an emulsion, a gel, an injection, a drop, a suppository (suppository), a patch or a spray, but is not limited thereto. The dosage form may be conveniently prepared by conventional methods in the art, and may further comprise surfactants, excipients, wettable powders, emulsion enhancers, suspending agents, salts or buffers for regulating osmotic pressure, colorants, spices, stabilizers, preservatives or other conventional adjuvants.

The amount of the pharmaceutical composition used or the amount administered according to an aspect of the present invention may vary according to the age, sex, body weight, condition and severity thereof of the subject receiving the administration, the route of administration or the judgment of the prescribing person. It is within the level of skill in the art to target the amount of use determined based on the factors.

In another aspect, the present invention may be a method for preparing a composition for preventing hair loss or promoting hair growth, which comprises: (1) a step of positioning a soybean culture root in a culture medium in a bioreactor, and proliferating the soybean culture root while maintaining a predetermined air supply amount in the bioreactor; (2) drying the cultured roots subjected to the step (1); (3) step (2), soaking the product obtained in the step (2) into alcohol for extraction; and (4) filtering and drying after the step (3).

In another aspect, the present invention may be a method for preparing an extract of bean culture tissue uniformly and in large quantities, comprising: (1) a step of positioning a soybean culture root in a culture medium in a bioreactor, and proliferating the soybean culture root while maintaining a predetermined air supply amount in the bioreactor; (2) drying the cultured roots subjected to the step (1); (3) step (2), soaking the product obtained in the step (2) into alcohol for extraction; and (4) filtering and drying after the step (3).

In one aspect, the method may comprise the step of germinating the legumes in a culture medium prior to step (1). In another aspect, the method may further comprise the step of inducing soybean culture roots from the soybean plant body before the step (1).

In general, since bean extracts cultivated in the open air and extracted by a conventional method are not necessarily ingredients, it is difficult to uniformly mass-produce a composition for achieving a specific purpose. However, according to the method of an aspect of the present invention, external influencing factors and the like are minimized, so that a composition for preventing hair loss or promoting hair growth can be uniformly and mass-produced.

As an example, the bioreactor may be a Stirred Tank Reactor (Stirred Tank Reactor), a Bubble Column Reactor (Bubble Column Reactor), an Air Lift bioreactor (Air Lift Reactor), a fluidized Bed Reactor (fluidized Bed Reactor), a Fixed Bed/Packed Bed Reactor (Fixed/Packed Bed Reactor), or a Tower fermentor (Tower fermentor), and may preferably be a Bubble bioreactor (Bulb Type Bubble bioreactor).

As another example, the medium in the step (1) may be a medium supplemented with IBA (3-indolebutyric acid) and a carbon source. In one aspect, the carbon source may be one or more selected from the group consisting of glucose, fructose, mannose, ribose, arabinose, xylose, galactose, sucrose, cellobiose, trehalose, lactose, raffinose, amylose, Starch (Starch), sorbitol, mannitol, and glycerol.

As another example, the medium in the step (1) may be MS (Murashige and Skoog) medium.

As another example, the pH of the medium when the bean culture roots are placed in the medium may be 4.8 to 6.8. In one aspect, the pH may be 4.8 or more, 5 or more, 5.2 or more, 5.4 or more, 5.6 or more, 5.8 or more, 6.0 or more, 6.2 or more, 6.4 or more, or 6.6 or more. On the other hand, the pH may be 6.8 or less, 6.6 or less, 6.4 or less, 6.2 or less, 6.0 or less, 5.8 or less, 5.6 or less, 5.4 or less, 5.2 or less, or 5.0 or less.

As another example, the density of the bean culture roots when they are placed in a medium may be 2 to 6 g/L. In one aspect, the density can be 2g/L or more, 3g/L or more, 4g/L or more, or 5g/L or more. On the other hand, the density may be 6g/L or less, 5g/L or less, 4g/L or less, or 3g/L or less.

As another example, the proliferation in the step (1) may be performed under dark conditions.

As another embodiment, the step (1) may be performed at 19 to 25 ℃. In one aspect, the temperature can be 19 ℃ or higher, 20 ℃ or higher, 21 ℃ or higher, 22 ℃ or higher, 23 ℃ or higher, or 24 ℃ or higher. On the other hand, the temperature may be 25 ℃ or lower, 24 ℃ or lower, 23 ℃ or lower, 22 ℃ or lower, 21 ℃ or lower, or 20 ℃ or lower.

In one aspect, the air supply amount in the step (1) may be 0.05 to 0.4vvm (air volume/culture volume per min). In one embodiment, the air supply amount may be 0.05vvm or more, 0.08vvm or more, 0.1vvm or more, 0.12vvm or more, 0.14vvm or more, 0.16vvm or more, 0.18vvm or more, 0.2vvm or more, 0.22vvm or more, 0.24vvm or more, 0.26vvm or more, 0.28vvm or more, 0.3vvm or more, 0.32vvm or more, 0.34vvm or more, 0.36vvm or more, or 0.38vvm or more. As another example, the air supply amount may be 0.4vvm or less, 0.38vvm or less, 0.36vvm or less, 0.34vvm or less, 0.32vvm or less, 0.3vvm or less, 0.28vvm or less, 0.26vvm or less, 0.24vvm or less, 0.22vvm or less, 0.2vvm or less, 0.18vvm or less, 0.16vvm or less, 0.14vvm or less, 0.12vvm or less, 0.1vvm or less, 0.08vvm or less, or 0.06vvm or less.

As an example, the proliferation in the step (1) may be performed for 2 to 5 weeks. In one aspect, the proliferation in step (1) may be performed for 2 weeks or more, 3 weeks or more, or 4 weeks or more. On the other hand, the proliferation in the step (1) may be performed for 5 weeks or less, 4 weeks or less, or 3 weeks or less.

As another example, the alcohol concentration in the step (3) may be 60 to 100% (w/v). In one aspect, the alcohol concentration may be 60% (w/v) or more, 70% (w/v) or more, 80% (w/v) or more, 90% (w/v) or more, or 95% (w/v) or more. On the other hand, the alcohol concentration may be 100% (w/v) or less, 95% (w/v) or less, 90% (w/v) or less, 85% (w/v) or less, 80% (w/v) or less, 75% (w/v) or less, 70% (w/v) or less, or 65% (w/v) or less.

In one aspect, the weight ratio of the dried product to alcohol can be 1:50 or less, 1:45 or less, 1:40 or less, 1:35 or less, 1:32 or less, 1:30 or less, 1:25 or less, or 1:22 or less. On the other hand, the weight ratio may be 1:20 or more, 1:22 or more, 1:24 or more, 1:26 or more, 1:28 or more, 1:31 or more, 1:33 or more, 1:35 or more, 1:40 or more, 1:45 or more, or 1:48 or more.

As another embodiment, the extraction time of the step (3) may be 20 to 28 hours. In one aspect, the extraction time may be 20 hours or more, 22 hours or more, 24 hours or more, 25 hours or more, or 26 hours or more. On the other hand, the extraction time may be 28 hours or less, 26 hours or less, 25 hours or less, 24 hours or less, 22 hours or less, or 21 hours or less.

By the method, the bean culture tissue can be harvested in large quantities in an environmentally friendly and uniform manner without using growth promoters or fertilizers, pesticides, etc., thereby preparing the extracts and compositions in large quantities that exert an environmentally friendly and uniform effect.

The structure and effects of one aspect of the present invention will be described in more detail below with reference to examples and the like. However, the following examples and the like are provided for illustrative purposes only to facilitate understanding of one aspect of the present invention, and the scope of the present invention are not limited thereto.

[ example 1 ] culture of Bean culture root

Beans (soybean; Glycine max.) and broad-leaved cranberry (Glycine gracilis) were placed in MS medium (Murashige and Skoog medium, Duchefa, Haarlem, the Netherlands), respectively, and germinated in a reactor. After induction of the cultured roots using the radicle, hypocotyl, cotyledon of the isolated soybean plant (in particular, induction from the radicle of the soybean), the bean-cultured roots were placed in a Bulb-shaped (Bulb Type) bioreactor (generally commercially available, see fig. 1) having an air volume of 3L in an MS medium (Murashige and Skoog medium, Duchefa, the netherlands) supplemented with 4mg/L of IBA (3-indolebutyric acid; indole-3-butyric acid, Sigma-Aldrich; Merck KGaA, Darmstadt, germany) and 30g/L of sucrose, and subcultured and propagated while replacing the fresh medium every 3 weeks. The medium was sterilized at 121 ℃ under 1.2 air pressure for 35 minutes after adjusting the pH to 5.8 with 1N NaOH. The cultured roots were proliferated for 3 weeks, cut into 1 to 1.5cm, inoculated into the medium at a density of 4g/L based on fresh weight, and cultured under dark conditions maintained at 22. + -. 1 ℃. Pesticide, fertilizer and the like are not used in the process. The air supply amount was adjusted to be constant at 0.1vvm (air flow ratio) by an air flow meter (RMA series; Dwyer Instruments, inc.) during the entire period of the culture, and the air supplied to the inside of the bioreactor was supplied to the inside of the bioreactor by an air compressor without using oil after passing through an air condenser capable of condensing compressed air, a filter capable of removing impurities, an air dryer, and the like in order to maintain a constant temperature.

It was confirmed that the use of the bioreactor or the like as described above enabled the production of a bean-cultured root extract of uniform quality all year round, unlike the general bean (derived from seeds or plant bodies) extract cultivated in open field or field (see fig. 1).

[ example 2 ] preparation of extract

After the bean culture roots proliferated for 4 weeks in the bioreactor of example 1 were dried with hot air, the bean culture roots were immersed in 80% (w/v) ethanol so that the weight ratio of the culture roots to the ethanol was 1:30, and the bean culture roots were extracted at room temperature for 24 hours. The extract was filtered through a filter paper and then the solvent was evaporated to dryness to obtain a powder. It was confirmed that the extract contained about 10% by weight of various active substances including coumestrol. Further, it was confirmed by repeating the extraction experiment that the components in the extract were uniformly maintained when the extract was prepared using the cultured roots produced in example 1, and it was confirmed that mass production could be performed by the above-mentioned extract preparation method in combination with subculture or the like.

[ EXAMPLE 1 ] confirmation of Hair follicle Stem cell Activity (Wnt/β -catenin luciferase assay; Wnt/β -catenin luciferase assay)

The extracts were confirmed for viability of hair follicle stem cells using LEADING LIGHT Wnt Reporter Cell Line (Enzo life science Inc.) and ONE-Glo luciferase assay Reporter kit (ONE-Glo luciferase assay Reporter kit, Promega, Madison, USA). Specifically, after mouse embryo fibroblasts (NIH3T3) (WNT) were seeded (seeding) in a 96-well plate (well plate) at 10000 seeds/well, they were cultured for 24 hours (5% CO)₂37 ℃ C.), then the negative control group is not treated at all, and the positive control group is treated with LiCl (lithium chloride; lithiumchloride) for 24 hours, the extracts were treated at the respective concentrations for 24 hours on the experimental groups.

Then, 100 was placed in each well of a 96-well plate

The ONE-Glo reagent (reagent) of (1), after 3 minutes of reaction, luciferase activity was measured by a luminometer (Victor luminometer).

The higher the concentration of the extract, the higher the transcription activity of β -catenin (refer to fig. 2), the higher the treatment of 50ppm (w/v) of the extract showed that Wnt/β -catenin activity was 3.28 times higher than that of the control group, SARE in fig. 2 represents that the extract (soybean adventitious root extract; soybean acquired adjuvant root extract), the maintenance and activation of Wnt signaling pathway (Wnt signaling) through β -catenin pathway were necessary in the hair-inducing activity of hair papilla (dermal papilla), and in addition, in order to maintain the anagen phase in hair papilla cells, Wnt/β -catenin activity increased much due to the treatment of the extract, which increased the activity regulated the hair follicle cycle (maintenance anagen phase and induction anagen phase), so it could be confirmed that the extract of the present invention had the effects of inducing hair, promoting hair growth, preventing alopecia, and the like.

[ Experimental example 2 ] culture of Hair follicle organs and measurement of Hair follicle cycle

For the experiments, for 6 groups of human bodies, 15 hair follicle samples were extracted for each group, and 90 samples were isolated. For isolated hair follicles, 500ul of medium was aliquoted in 24-well plates (Nunc, wissbaden, germany), and 5-6 samples were placed in each well for culture.

As the medium used, Williams E (Gibco, USA) containing 2mM L-glutamine (PAA, Coelbe, Germany), 10. mu.g/ml insulin, 10ng/ml cortisol (hydrocortisone, Sigma, USA), 0.1% amphotericin B (fungizone, Gibco, USA), 10. mu.g/ml streptomycin, and 100U/ml penicillin (Gibco, USA) was used.

The extracts were treated at various concentrations (0, 10, 50ppm) into the hair follicle samples in the medium and the medium was replaced every 2-3 days. The hair follicles were photographed with a microscope and the cycle was classified according to morphological criteria.

The ratio of catagen phase to anagen phase is determined from the hair follicle cycle (HF cycle). In particular, at the transition from the anagen phase to the catagen phase, apoptosis of keratinocytes increased and proliferation of keratinocytes was inhibited, and it was confirmed that only 27 to 36% of hair follicles were transitioned to the catagen phase as a result of administration of the extract, and when 50ppm of the extract was used, the ratio of the catagen phase to the anagen phase was 2 times or more higher than that of the control group after 11 days (fig. 3). That is, when the extract of the present invention was used, it was shown that the anagen phase in the Hair Follicle (HF) cycle was significantly prolonged (in fig. 3, MNX was minoxidil (minoxidil) and SARE was the extract of the present invention (soybean adventitious root extract)). From this, it was confirmed that the extract of the present invention has hair growth promoting and alopecia preventing effects. In addition, since beans which are natural plants are used, it has been confirmed that the side effects of the conventional therapeutic agents can be solved.

[ dosage form example 1 ] Soft capsules

A soft capsule was prepared by mixing 10mg of the extract of example 2, 80-140mg of L-carnitine (L-carnitine), 180mg of soybean oil, 2mg of palm oil, 8mg of a vegetable hardened oil, 4mg of yellow beeswax and 6mg of lecithin, and filling 1 capsule with the mixture according to a conventional method.

[ dosage form example 2 ] tablets

10mg of the extract of example 2, 200mg of galactooligosaccharide, 60mg of lactose and 140mg of maltose were mixed, granulated by a fluidized bed dryer, and then 6mg of sugar ester (sugar ester) was added, and tabletted by a tabletting machine to prepare tablets.

[ dosage form example 3 ] drink preparation

After 5mg of the extract of example 2, 10g of glucose, 0.6g of citric acid and 25g of liquid-phase oligosaccharide were mixed, 300ml of purified water was added, and 200ml of the mixture was filled in each bottle. After filling into the bottle, sterilizing at 130 deg.C for 4-5 s to prepare beverage.

[ dosage form example 4 ] injection

An injection was prepared by a usual method using 20mg of the extract of example 2, an appropriate amount of distilled water for injection, and an appropriate amount of pH adjuster.

[ dosage form example 5 ] health food

A health food was prepared from 5mg of the extract of example 2, 70. mu.g of vitamin A acetate, 1.0mg of vitamin E, 0.13mg of vitamin B1, 0.15mg of vitamin B2, 60.5mg of vitamin B, 0.2. mu.g of vitamin B12, 10mg of vitamin C, 10. mu.g of biotin, 1.7mg of nicotinamide, 50. mu.g of folic acid, 0.5mg of calcium pantothenate, 1.75mg of ferrous sulfate, 0.82mg of zinc oxide, 25.3mg of magnesium carbonate, 15mg of monopotassium phosphate, 55mg of calcium hydrogen phosphate, 90mg of potassium citrate, 100mg of calcium carbonate, and 24.8mg of magnesium chloride according to a usual method.

The composition ratio of the vitamin and inorganic substance mixture is a composition obtained by mixing components relatively suitable for health foods, but the mixing ratio may be arbitrarily modified, and the components may be mixed according to a general health food preparation method and then a health food composition may be prepared according to a general method.

[ dosage form example 6 ] cream preparation

Using 0.2 wt% of the extract of said example 2, 2.00 wt% of polyethylene glycol monostearate, 5.00 wt% of self-emulsifying glyceryl monostearate, 4.00 wt% of propylene glycol, 6.00 wt% of squalene, 6.00 wt% of tri-2-ethylhexanetriol, 1.00 wt% of sphingolipids, 7.00 wt% of 1, 3-butanediol, 5.00 wt% of beeswax, the remaining amount of purified water, a cream formulation was prepared. In addition, an ointment is prepared by adjusting the amount of purified water and adjusting the viscosity.

Claims (24)

1. A composition for preventing hair loss or promoting hair growth, comprising an extract of a bean-cultured tissue, which is a bean-cultured root or a bean callus.

2. The composition of claim 1, wherein the bean is a soybean or a broad leaf cranberry.

3. The composition as claimed in claim 1, wherein the extract is extracted using water, a lower alcohol of C1 to C6, or a mixture thereof as a solvent.

4. The composition of claim 3, wherein the lower alcohol is ethanol.

5. The composition of claim 3, wherein the lower alcohol concentration is 60 to 100% (w/v).

6. The composition according to claim 1, wherein the composition comprises from 5 to 15% by weight of coumestrol, relative to the total weight of the composition.

7. The composition according to claim 1, wherein the preventing hair loss or promoting hair growth is accomplished by one or more selected from the group consisting of proliferating hair papilla cells, increasing hair follicle stem cell activity, inducing hair follicle anagen phase, maintaining or prolonging hair follicle anagen phase, and inhibiting transition of hair follicle cycle toward catagen phase.

8. The composition of claim 1, wherein the preventing hair loss or promoting hair growth is accomplished by increasing the transcriptional activity of β -catenin or increasing the activity of Wnt/β -catenin.

9. The composition of claim 1, wherein the composition is administered by a route selected from transdermal or external dermal administration.

10. The composition of claim 1, wherein the concentration of the extract within the composition is 10 to 100ppm (w/v).

11. The composition according to claim 1, wherein the input amount of the extract based on the input of the composition is 5 mg/kg/day to 300 mg/kg/day.

12. The composition of claim 1, wherein the composition is a food, pharmaceutical or cosmetic composition.

13. A method for uniformly and massively preparing the composition for preventing hair loss or promoting hair growth according to claim 1, which comprises:

(1) a step of positioning a soybean culture root in a culture medium in a bioreactor, and proliferating the soybean culture root while maintaining a predetermined air supply amount in the bioreactor;

(2) drying the cultured roots subjected to the step (1);

(3) step (2), soaking the product obtained in the step (2) into alcohol for extraction; and

(4) and (3) filtering and drying after the step (3).

14. The method of claim 13, wherein the bioreactor is a bulb bubble bioreactor.

15. The method according to claim 13, wherein the culture medium in the step (1) is a culture medium supplemented with 3-indolebutyric acid and a carbon source.

16. The method according to claim 13, wherein the medium in the step (1) is MS medium.

17. The method according to claim 13, wherein, in the step (1), the medium pH at which the bean-cultured roots are placed in the medium is 4.8 to 6.8.

18. The method according to claim 13, wherein, in the step (1), the density of the bean-cultured roots when placed in the medium is 2 to 6 g/L.

19. The method according to claim 13, wherein the proliferation in step (1) is performed under dark conditions.

20. The method according to claim 13, wherein the (1) step is performed at 19 to 25 ℃.

21. The method according to claim 13, wherein the air supply amount of the (1) step is 0.05 to 0.4 vvm.

22. The method of claim 13, wherein the proliferation of step (1) is performed for 3 to 5 weeks.

23. The method according to claim 13, wherein the alcohol concentration in the step (3) is 60 to 100% (w/v).

24. The method according to claim 13, wherein the extraction time of the step (3) is 20 to 28 hours.

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