KR101897451B1 - Composition for preventing or treating hair-loss - Google Patents

Abstract

The present invention relates to a composition for preventing or treating hair loss comprising plant-derived extracellular endoplasmic reticulum and caffeine.
The composition provided by the present invention is excellent in the effect of promoting hair growth and hair growth, so that it is excellent in prevention and treatment of hair loss, and it is safe without taking side effects even when it is ingested or applied to skin.

Description

[Composition for preventing or treating hair loss]

The present invention relates to a composition for preventing or treating hair loss, which comprises extracellular endoplasmic reticulum derived from a plant juice and caffeine, and is excellent in promoting hair growth and hair growth.

Hair loss is the most important cause of hair loss, but with the recent increase in social stress, the hair loss population is gradually increasing due to westernized eating habits such as environmental pollution and instant food, frequent perm and dyeing, and wrong scalp care.

The hair is maintained by repeating hair growth and hair loss around the cycles of Anagen, Catagen and Telogen. Specifically, there are anagen which grows hair, catagen which is the period of termination of growth and reduction of the size of the moles, talag, which is the period when the papilla stops its activity and keeps its hair on the scalp, Or a generator that generates new hair to depilate old hair.

Hair growth is 3 ~ 5 years for males, 4 ~ 6 years for females, 30 ~ 45 days for degenerative period, and 3 ~ 4 months for rest period. And at the end of the pause, a generator starts to generate new hair.

Hair loss is a normal phenomenon, but normal people have a lot of hair growing, compared with normal hair loss (Alopecia) people are in the off-state hair is a lot of visible hair loss appears.

The characteristic of people who exhibit alopecia is in the miniaturization of hair. As the hair loss progresses, the period of growth is shortened and the hair becomes smaller and smaller. Therefore, in order to treat hair loss, it is important to allow the hair follicle in a resting state to grow rapidly and to shorten the growth period.

Male alopecia are caused by a male hormone called testosterone, which is a hormone that causes male sexiness and develops muscles in adolescence and development of male organs. This testosterone is known as 5-alpha-reductase, (DHT), a hormone that acts on the hair follicle to induce the hair follicle from the growth phase to the regenerator stage, resulting in hair loss. Therefore, in order to treat alopecia caused by these causes, a method of suppressing the production of DHT by 5 alpha-reductase is mainly used.

Female alopecia are caused mainly by a decrease in the amount of estrogen after menopause. Female alopecia is different from the shape of male alopecia, the head is not removed from the front part of the head is mainly hair loss. Female alopecia affects less than men in the association with 5 alpha - reductase. Therefore, drugs that inhibit 5 alpha-reductase inhibition are not effective for postmenopausal women with alopecia. Accordingly, minoxidil or estrogen is mainly used as a therapeutic agent for such alopecia.

Alopecia areata is caused by autoimmune diseases, mental stress, genetic predisposition. It is characterized by the appearance of round or oval hair loss, and the appearance of tofu rings or hairy walls. These alopecia areata are fundamentally different from those of androgenetic alopecia, and different methods of treatment are used, such as a method of treating corticosteroids, minoxidil being applied to the affected part or artificially induced to the affected part.

With respect to such various and complicated hair loss causes, known hair loss prevention products to date have been marketed as products in the market as effective ingredients for promoting blood circulation, inhibiting male hormone function, and strengthening hair follicle function. There is no clear effect yet, and most of the problems are side effects.

It is known that the hair growth promoting effect is caused by side effects among the patients who have been developed and used for promoting blood circulation and have been approved by the US Food and Drug Administration (FDA) 6-Amino-1,2-dihydro-1-hydroxy-2-imino-4-phenoxypyrimidine disclosed in U.S. Patent No. 3,382,247 used as a therapeutic agent is currently used as a therapeutic agent for hair growth. It is also disclosed in U.S. Patent No. 5,215,894 Merck's finasteride was initially used as a treatment for male prostate, but it was also created as a treatment for male alopecia by controlling the dosage in the human body.

However, minoxidil has been reported to have a sticky feel and side effects that cause irritation to the skin. In the case of pinasteride, it is currently used as a preparation for oral administration. However, adverse effects such as sexual dysfunction have been reported depending on its consumption, The effect of 5 alpha-reductase inhibitor on the hair loss is effective only when it is taken steadily, and there is a lot of inconvenience in using the 5 alpha-reductase inhibitor because it is difficult to expect the effect by application and can be expected only by oral administration.

The present invention is intended to provide a composition which is safe and excellent in promoting hair growth and hair growth, and capable of preventing or treating hair loss.

The inventors of the present invention discovered that extracellular vesicles derived from plant juices and caffeine together are excellent in hair growth and hair growth, leading to the present invention.

According to one embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating hair loss comprising plant-derived extracellular vesicles and caffeine as an active ingredient.

In the present invention, the diameter of the extracellular endoplasmic reticulum may be 30-1,000 nm.

In the present invention, the mixing ratio of the extracellular matrix and caffeine is not particularly limited, but is preferably 1: 1 to 10, or 1: 2 to 8, or 1: 3 to 7 or 1: 4 to 6, 4 to 5 weight ratio.

In addition, the content of the extracellular endoplasmic reticulum in the present invention is not particularly limited, but may be preferably 5 to 10 μg / ml.

In the present invention, the content of the caffeine is not particularly limited, but may be in the range of 25 to 50 μM.

According to another embodiment of the present invention, there is provided a cosmetic composition for improving hair loss comprising plant-derived extracellular vesicles and caffeine as an active ingredient.

In the present invention, the diameter of the extracellular endoplasmic reticulum may be 30-1,000 nm.

In the present invention, the mixing ratio of the extracellular matrix and caffeine is not particularly limited, but is preferably 1: 1 to 10, or 1: 2 to 8, or 1: 3 to 7 or 1: 4 to 6, 4 to 5 weight ratio.

In addition, the content of the extracellular endoplasmic reticulum in the present invention is not particularly limited, but may be preferably 5 to 10 μg / ml.

In the present invention, the content of the caffeine is not particularly limited, but may be in the range of 25 to 50 μM.

According to another embodiment of the present invention, there is provided a food composition for improving hair loss comprising extracellular vesicles derived from a plant juice and caffeine as an active ingredient.

In the present invention, the diameter of the extracellular endoplasmic reticulum may be 30-1,000 nm.

In the present invention, the mixing ratio of the extracellular matrix and caffeine is not particularly limited, but is preferably 1: 1 to 10, or 1: 2 to 8, or 1: 3 to 7 or 1: 4 to 6, 4 to 5 weight ratio.

In addition, the content of the extracellular endoplasmic reticulum in the present invention is not particularly limited, but may be preferably 5 to 10 μg / ml.

In the present invention, the content of the caffeine is not particularly limited, but may be in the range of 25 to 50 μM.

According to another embodiment of the present invention, there is provided a method for producing a plant juice comprising the steps of: obtaining a plant juice by juicing a plant using a screw;

Culturing the plant juice by static culture;

Centrifuging the cultured plant juice to recover the supernatant;

Filtering the supernatant through a filtration membrane containing pores having an average pore size of 0.1 to 1.0 mu m to recover the filtrate; And

And mixing the caffeine with the filtrate. The present invention also provides a method for preparing a pharmaceutical composition for preventing or treating hair loss.

In the present invention, the mixing ratio of the filtrate to caffeine is not particularly limited, but is preferably 1: 1 to 10, or 1: 2 to 8, or 1: 3 to 7 or 1: 4 to 6, 5 < / RTI > by weight.

The present invention can further carry out a step of washing plants to remove impurities, if necessary, before juicing the plants.

In addition, in the present invention, it is preferable that the screw uses a low-speed screw having a stirring speed of 20 to 100 rpm to obtain a plant juice containing a large amount of extracellular endoplasmic reticulum excellent in hair growth and hair growth.

In the present invention, the stationary culture may be performed at 20 to 30 ° C for 12 to 48 hours.

In the present invention, the centrifugation may be performed at 100 to 20,000 g for 10 to 120 minutes at least once.

In the present invention, the centrifugation is performed at 100 to 500 g for 10 to 30 minutes, then at 1,000 to 2,000 g for 10 to 30 minutes and then at 5,000 to 20,000 g for 30 minutes to 1 hour. Car.

In the present invention, the mixing ratio of the filtrate to caffeine is not particularly limited, but is preferably 1: 1 to 10, or 1: 2 to 8, or 1: 3 to 7, or 1: 4 to 6, 4 to 5 weight ratio.

In the present invention, the filtration may be performed at least once using a filtration membrane containing pores having an average pore size of 0.2 to 0.5 μm. Preferably, the filtration is firstly performed using a filtration membrane including pores having an average pore size of 0.4 to 0.5 占 퐉, followed by a second filtration using a filtration membrane containing pores having an average pore size of 0.2 to 0.3 占 퐉 , It is possible to obtain a filtrate containing a large amount of extracellular cell bodies excellent in hair growth and hair growth effect.

According to still another embodiment of the present invention, there is provided a method for producing a plant juice, comprising the steps of: obtaining a plant juice by juicing a plant using a screw;

Culturing the plant juice by static culture;

Centrifuging the cultured plant juice to recover the supernatant;

Filtering the supernatant through a filtration membrane containing pores having an average pore size of 0.1 to 1.0 mu m to recover the filtrate; And

And mixing the caffeine with the filtrate. The present invention also provides a method for producing a cosmetic composition for improving hair loss.

The present invention can further carry out a step of washing plants to remove impurities, if necessary, before juicing the plants.

In addition, in the present invention, it is preferable that the screw uses a low-speed screw having a stirring speed of 20 to 100 rpm to obtain a plant juice containing a large amount of extracellular cell body excellent in hair growth and hair growth.

In the present invention, the stationary culture may be performed at 20 to 30 ° C for 12 to 48 hours.

In the present invention, the centrifugation may be performed at 100 to 20,000 g for 10 to 120 minutes at least once.

In the present invention, the centrifugation is performed at 100 to 500 g for 10 to 30 minutes, then at 1,000 to 2,000 g for 10 to 30 minutes and then at 5,000 to 20,000 g for 30 minutes to 1 hour. Car.

In the present invention, the mixing ratio of the filtrate to caffeine is not particularly limited, but is preferably 1: 1 to 10, or 1: 2 to 8, or 1: 3 to 7, or 1: 4 to 6, 4 to 5 weight ratio.

In the present invention, the filtration may be performed at least once using a filtration membrane containing pores having an average pore size of 0.2 to 0.5 μm. Preferably, the filtration is firstly performed using a filtration membrane including pores having an average pore size of 0.4 to 0.5 占 퐉, followed by a second filtration using a filtration membrane containing pores having an average pore size of 0.2 to 0.3 占 퐉 , It is possible to obtain a filtrate containing a large amount of extracellular cell bodies excellent in hair growth and hair growth effect.

According to still another embodiment of the present invention, there is provided a method for producing a plant juice, comprising the steps of: obtaining a plant juice by juicing a plant using a screw;

Culturing the plant juice by static culture;

Centrifuging the cultured plant juice to recover the supernatant;

Filtering the supernatant through a filtration membrane containing pores having an average pore size of 0.1 to 1.0 mu m to recover the filtrate; And

And mixing the caffeine with the filtrate. The present invention also provides a method for producing a food composition for improving hair loss.

The present invention can further carry out a step of washing plants to remove impurities, if necessary, before juicing the plants.

In addition, in the present invention, it is preferable that the screw uses a low-speed screw having a stirring speed of 20 to 100 rpm to obtain a plant juice containing a large amount of extracellular cell body excellent in hair growth and hair growth.

In the present invention, the stationary culture may be performed at 20 to 30 ° C for 12 to 48 hours.

In the present invention, the centrifugation may be performed at 100 to 20,000 g for 10 to 120 minutes at least once.

In the present invention, the centrifugation is performed at 100 to 500 g for 10 to 30 minutes, then at 1,000 to 2,000 g for 10 to 30 minutes and then at 5,000 to 20,000 g for 30 minutes to 1 hour. Car.

In the present invention, the mixing ratio of the filtrate to caffeine is not particularly limited, but is preferably 1: 1 to 10, or 1: 2 to 8, or 1: 3 to 7, or 1: 4 to 6, 4 to 5 weight ratio.

In the present invention, the filtration may be performed at least once using a filtration membrane containing pores having an average pore size of 0.2 to 0.5 μm. Preferably, the filtration is firstly performed using a filtration membrane including pores having an average pore size of 0.4 to 0.5 占 퐉, followed by a second filtration using a filtration membrane containing pores having an average pore size of 0.2 to 0.3 占 퐉 , It is possible to obtain a filtrate containing a large amount of extracellular cell bodies excellent in hair growth and hair growth effect.

As used herein, the term "extracellular < RTI ID = 0.0 > vesicle " refers < / RTI > to a small vesicle of membrane structure that is secreted from various cells and is also defined as a nanobeicle. The extracellular endoplasmic reticulum has a diameter of about 30 to 1,000 nm, preferably an average diameter of 100 to 300 nm, which refers to a follicle that is fused with the plasma membrane and released into the extracellular environment.

As used herein, the term "plant" is understood to include not only mature plants but also plant cells, plant tissues and plant seeds that develop into mature plants.

The kinds of plants that can be used in the present invention are not particularly limited, and for example, food crops including rice, wheat, barley, corn, soybean, potato, wheat, red bean, oats and millet; Vegetable crops including Arabidopsis, cabbage, radish, pepper, garlic, ginger, strawberry, tomato, watermelon, cucumber, cabbage, melon, zucchini, onions, onion, asparagus and carrots; Special crops including ginseng, tobacco, cotton, sesame, sugar cane, beet, perilla, peanut and rapeseed; Fruit trees including apple trees, pears, jujube trees, peaches, sheep grapes, grapes, citrus fruits, persimmons, oranges, grapefruit, plums, apricots, dragon mangoes, blueberries, avocados and bananas; Roses, gladiolus, gerberas, carnations, chrysanthemums, lilies and tulips; And feed crops including raglass, red clover, orchardgrass, alpha-alpha, tall fescue and perennial rice, but preferably asparagus can be used.

In the present specification, the term "plant juice" means a liquid fluid obtained by discharging moisture or the like contained in the inside of a plant by mechanical force or rotational force to the outside. For example, juice, vegetable juice, Juice, and the like, but the present invention is not limited thereto.

In addition, in the present invention, the pharmaceutical composition may further comprise an appropriate carrier, excipient or diluent according to a conventional method. Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium But are not limited to, silicates, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, external preparations, suppositories or sterilized injection solutions, Can be used. More specifically, when formulating the composition, it can be prepared using a diluent or an excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like. The solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations can be prepared by mixing at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. in addition to commonly used diluents such as water and liquid paraffin . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The route of administration of the pharmaceutical composition according to the present invention may be, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, , Sublingual or rectal. Oral or parenteral administration is preferred. The term "parenteral" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical compositions of the present invention may also be administered in the form of suppositories for rectal administration.

The pharmaceutical composition of the present invention may be administered orally or parenterally depending on various factors including the activity of the specific compound used, age, weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination, And the dosage of the pharmaceutical composition may be appropriately selected by a person skilled in the art depending on the condition of the patient, the body weight, the degree of disease, the type of drug, the route of administration and the period of time, and is preferably from 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way. The pharmaceutical compositions according to the present invention can be formulated into pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions.

The food composition of the present invention may be prepared in the form of various foods such as beverages, gums, tea, vitamin complexes, powders, granules, tablets, capsules, confectionery, rice cakes and breads. Since the food composition of the present invention is composed of a plant extract having little toxicity and side effects, it can be safely used for prolonged use even for prophylactic purposes.

When the plant-derived extracellular matrix or the filtrate containing the plant-derived extracellular medium contained in the present invention is included in the food composition, the amount thereof may be added in a proportion of 0.1 to 50% of the total weight.

Here, when the food composition is prepared in a beverage form, there are no particular limitations other than those containing the food composition at the indicated ratios and may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient. That is, natural carbohydrates include monosaccharides such as glucose, disaccharides such as fructose, sucrose and the like and sugar sugars such as polysaccharide, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol can do. Examples of the above-mentioned flavors include natural flavors (such as tau martin and stevia extract (for example, rebaudioside A and glycyrrhizin) and synthetic flavors (for example, saccharine and aspartame).

In addition, the food composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants, pectic acid and its salts, alginic acid and its salts, , a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, a carbonating agent used in a carbonated drink, and the like.

These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0.1 to about 50 parts by weight per 100 parts by weight of the food composition of the present invention.

In addition, the cosmetic composition of the present invention can be used in cosmetics, nutritional lotions, nutritional essences, massage cream, cosmetic bath additives, body lotion, body milk, bath oil, baby oil, baby powder, shower gel, Creams, suntan creams, skin lotions, skin creams, sunscreen cosmetics, cleansing milks, depilatory creams, facial and body lotions, face and body creams, skin whitening creams, hand lotions, hair lotions, cosmetic creams, jasmine Oil, bath soap, water soap, soap, shampoo, hand cleanser, medicinal soap {non-medical use}, cream soap, facial wash, whole body cleanser, scalp cleaner, hair rinse, cosmetic soap, tooth whitening gel, toothpaste And the like. To this end, the composition of the present invention may further comprise a solvent commonly used in the production of a cosmetic composition, or a suitable carrier, excipient or diluent.

For example, water, saline solution, DMSO, or a combination thereof may be used. Examples of the carrier, excipient or diluent include purified water, oil, wax But are not limited to, fatty acids, fatty acid alcohols, fatty acid esters, surfactants, humectants, thickeners, antioxidants, viscosity stabilizers, chelating agents, buffers, lower alcohols and the like. Further, if necessary, it may contain a whitening agent, a moisturizing agent, a vitamin, an ultraviolet screening agent, a perfume, a dye, an antibiotic, an antibacterial agent, and an antifungal agent.

As the oil, hydrogenated vegetable oil, castor oil, cottonseed oil, olive oil, palm oil, jojoba oil and avocado oil may be used. Examples of the wax include wax, wax, carnauba, candelilla, montan, ceresin, liquid paraffin, Can be used.

As the fatty acid, stearic acid, linoleic acid, linolenic acid and oleic acid may be used. As the fatty acid alcohol, cetyl alcohol, octyldodecanol, oleyl alcohol, panthenol, lanolin alcohol, stearyl alcohol and hexadecanol may be used As fatty acid esters, isopropyl myristate, isopropyl palmitate, and butyl stearate may be used. As the surfactant, a cationic surfactant, an anionic surfactant and a nonionic surfactant known in the art can be used, and a surfactant derived from a natural material is preferably used.

In addition, it may contain a hygroscopic agent, a thickening agent, an antioxidant and the like widely known in the field of cosmetics, and the kind and amount thereof are well known in the art.

The composition provided by the present invention is excellent in the effect of promoting hair growth and hair growth, so that it is excellent in prevention and treatment of hair loss, and it is safe without taking side effects even when it is ingested or applied to skin.

FIG. 1 is a graph showing changes in cell survival rate by treatment with asparagus filtrate, caffeine, and a mixture thereof in dermal papilla cells in Experimental Example 1. FIG.
FIG. 2 is a graph showing changes in the degree of expression of PTC-1, VEGF, and CD34 genes by treating aspirin solution, caffeine, and a mixture thereof in dermal papilla cells in Experimental Example 2. FIG.
FIG. 3 is a graph showing changes in the amount of asparagus filtrate, caffeine, and a mixture thereof in the dermal papilla cells in Experimental Example 3 by fluorescence immuno staining.

Hereinafter, preferred embodiments of the present invention will be described. However, the embodiments of the present invention can be modified into various other forms, and the scope of the present invention is not limited to the embodiments described below. Further, the embodiments of the present invention are provided to more fully explain the present invention to those skilled in the art.

Example

In the following experiments, human dermal papilla cells (DPC) were used. In all experiments, the cells were cultured at passage 20 or lower. In culture, the medium was cultured in DMEM (welgene, Korea) lonza) and 1% penicillin-streptomycin (welgene, Korea). The cells were inoculated in the medium and cultured in a 5% CO ₂ incubator at 37 ° C.

[Production Example 1]

Asparagus was washed with distilled water, and liquid juice obtained by squeezing with a low speed screw having a stirring speed of 40 rpm was sifted through a sieve to remove suspended matter. Thereafter, the mixture was allowed to stand at 25 占 폚 for 10 minutes, centrifuged at 2,000 g for 30 minutes to separate the supernatant, and then filtered with a filtration membrane containing 0.45 占 퐉 of polyvinylidene difluoride material. And filtered through a filtration membrane containing 0.22 탆 pores to obtain a filtrate.

[Experimental Example 1] MTS assay

2.5 x 10 ⁴ dermal papilla cells were equally divided into 24-well plates (SPL, Korea). After a stabilization time of 24 hours, the filtrate obtained in Preparation Example 1, caffeine and a mixture thereof were treated for 48 hours. The cells were diluted 1: 5 with the MTS reagent (Promega, USA) and FBS free (1% p / s), and the cells were mixed with 500 μl of each well. After 1 hour and 30 minutes, the absorbance at 490 nm And the change in cell viability was evaluated. The results are shown in FIG. 1 as a graph. However, the positive control group was treated with 10% fetal bovine serum (FBS) and the negative control group was not treated.

As shown in FIG. 1, when the extracts of 10 / / ml of asparagus-containing filtrate and 50 M of caffeine were treated alone, the cell proliferation rates were increased by 24% and 32%, respectively, compared to the negative control, The cell proliferation rate was increased by 47% compared to the negative control group when the filtrate containing the extracellular endoplasmic reticulum was treated with 10 μg / ml of caffeine and 50 μM of caffeine alone.

As a result, synergistic effects can be obtained in improving the growth rate of the papilla cell when the plant-derived extracellular endoplasmic reticulum or caffeine is treated alone by mixing them as in the present invention.

[Experimental Example 2] qPCR (Quantitative Polymerase Chain Reaction) analysis

5 x 10 ⁵ cells / ml of dermal papilla cells were equally divided into 60 mm dishes (SPL, Korea), and the filtrate, caffeine and mixture thereof obtained in Preparation Example 1 were treated. Then, RNA was isolated from the cells using an RNA isolation kit (Quiagen, USA) and a primer for 1 μg of the separated RNA was synthesized using a cDNA synthesis kit (Intronbio, Korea). The primers were used at a concentration of 10 pmol / μl and the degree of expression of PTC-1 (the patched protein-1), VEGF (vascular endothelial growth factor) and CD34 gene was detected by adding 2X Cyber Green Master Mix (Takara, Japan) The results are shown graphically in Fig. However, no treatment was performed for the control group.

As shown in FIG. 2, when the asparagus-derived filtrate containing the extracellular matrix was mixed with caffeine, the expression rates of PTC-1, VEGF and CD34 genes were significantly increased compared with those treated with the caffeine alone, The expression rates of PTC-1, VEGF and CD34 genes were increased by 2.8, 2.3 and 2.2 times, respectively, as compared with the negative control group.

Here, the PTC-1 is a Sonic Hedgehog (Shh) receptor, a gene that regulates morphogenesis and growth of hair follicles, VEGF is a gene related to an angiogenic factor, and CD34 corresponds to a neovascularization marker.

Thus, it can be seen from the above results that the plant-derived extracellular extracellular matrix and caffeine mixed composition according to the present invention are excellent in promoting hair growth and hair growth.

[Experimental Example 3] Immuno-fluorescence staining analysis

5 x 10 ⁴ cells / ml of dermal papilla cells were equally divided into 60 mm dishes (SPL, Korea). After one week, the filtrate, caffeine and mixture thereof obtained in Preparation Example 1 were treated and cultured for 48 hours. Paraformaldehyde (2% paraformaldehyde) was reacted for 10 minutes and then 0.1% Triton X-100 was added to increase cell permeability. In order to reduce nonspecific expression, the cells were reacted with 5% BSA solution for 1 hour and the primary antibody was reacted overnight at 4 ° C in the ratio of 1: 200 (Cytokeratin 15, Ki67, abcam). The secondary antibody conjugated with fluorescence was reacted at a ratio of 1: 200 for 1 hour. Thereafter, a mounting solution mixed with DAPI solution was used for nuclear staining. The resulting photograph is shown in Fig.

As shown in FIG. 3, when the asparagus-derived filtrate containing the extracellular matrix was mixed with caffeine, the expression of Ki67 (proliferation marker) and cytokeratin 15 (expressed in hair follicle) increased .

Thus, it can be seen that the plant-derived extracellular endoplasmic reticulum-caffeine mixed composition according to the present invention significantly influences the proliferation and development of the dermal papilla cells compared with the case of the single treatment.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is clearly understood that the same is by way of illustration and example only and is not to be taken by way of limitation, It is to be understood that the invention may be practiced within the scope of the appended claims.

Claims (30)

A pharmaceutical composition for preventing or treating hair loss comprising extract of asparagus juice containing extracellular vesicles and caffeine as an active ingredient. The method according to claim 1,
Wherein the extracellular endoplasmic reticulum has a diameter of 30 to 1,000 nm. The method according to claim 1,
Wherein the filtrate and caffeine are contained in a weight ratio of 1: 1 to 10. The method according to claim 1,
Wherein the filtrate is contained in an amount of 5 to 10 占 퐂 / ml. The method according to claim 1,
Wherein the caffeine is contained in an amount of 25 to 50 μM. The method according to claim 1,
Wherein the asparagus juice is obtained by extracting asparagus with a screw having a stirring speed of 20 to 100 rpm. delete A cosmetic composition for improving hair loss comprising an extract of asparagus juice containing extracellular vesicles and caffeine as an active ingredient. 9. The method of claim 8,
Wherein the extracellular endoplasmic reticulum has a diameter of 30 to 1,000 nm. 9. The method of claim 8,
Wherein the filtrate and caffeine are contained in a weight ratio of 1: 1 to 10. 9. The method of claim 8,
Wherein the filtrate is contained in an amount of 5 to 10 占 퐂 / ml. 9. The method of claim 8,
Wherein the caffeine is contained in an amount of 25 to 50 μM. 9. The method of claim 8,
Wherein the asparagus juice is obtained by extracting asparagus with a screw having a stirring speed of 20 to 100 rpm. delete A composition for improving hair loss comprising an extract of asparagus juice containing extracellular vesicles and caffeine as an active ingredient. 16. The method of claim 15,
Wherein the extracellular endoplasmic reticulum has a diameter of 30 to 1,000 nm. 16. The method of claim 15,
Wherein the filtrate and caffeine are contained in a weight ratio of 1: 1 to 10. 16. The method of claim 15,
Wherein the filtrate is contained in an amount of 5 to 10 占 퐂 / ml. 16. The method of claim 15,
Wherein the caffeine is contained in an amount of 25 to 50 μM. 16. The method of claim 15,
Wherein the asparagus juice is obtained by extracting asparagus with a screw having a stirring speed of 20 to 100 rpm. delete Extracting asparagus juice using a screw to obtain an asparagus juice;
Culturing the asparagus juice by static culture;
Centrifuging the cultured asparagus juice to recover the supernatant;
Filtering the supernatant through a filtration membrane containing pores having an average pore size of 0.1 to 1.0 mu m to recover the filtrate; And
A method for preparing a pharmaceutical composition for preventing or treating hair loss, comprising mixing caffeine in the filtrate. 23. The method of claim 22,
Wherein the filtrate and caffeine are mixed at a weight ratio of 1: 1 to 10. 23. The method of claim 22,
Further comprising washing the asparagus to remove impurities prior to pouring the asparagus. ≪ RTI ID = 0.0 > 11. < / RTI > 23. The method of claim 22,
Wherein the screw has a stirring speed of 20 to 100 rpm. 23. The method of claim 22,
Wherein the static culture is performed at 20 to 30 DEG C for 12 to 48 hours. 23. The method of claim 22,
Wherein the centrifugation is performed at 100 to 20,000 g for 10 to 120 minutes at least once. 28. The method of claim 27,
The centrifugation is carried out at 100 to 500 g for 10 to 30 minutes, at 1,000 to 2,000 g for 10 to 30 minutes and then at 5,000 to 20,000 g for 30 minutes to 1 hour. Or a pharmaceutically acceptable salt thereof. 23. The method of claim 22,
Wherein the filtration is performed at least once using a filtration membrane containing pores having an average pore size of 0.2 to 0.5 占 퐉. 30. The method of claim 29,
The filtration is performed by first filtering using a filtration membrane containing pores having an average pore size of 0.4 to 0.5 占 퐉 and then performing second filtration using a filtration membrane containing pores having an average pore size of 0.2 to 0.3 占 퐉. A method for the preparation of a pharmaceutical composition for preventing or treating hair loss.

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