KR101721457B1 - Composition for preventing hair loss and promoting hair growth comprising the effective amount of bee venom - Google Patents

Abstract

The present invention relates to a composition for promoting hair growth and preventing alopecia, which comprises bee venom as an active ingredient. The composition according to the present invention contains bee venom as an active ingredient and has an excellent hair growth promoting effect, And can be used as a composition for promoting hair growth and preventing hair loss, which can effectively solve various problems caused by the hair growth promoting agent or the hair loss inhibitor according to the prior art.

Description

[0001] The present invention relates to a composition for promoting hair growth and preventing hair loss comprising bee venom as an active ingredient.

The present invention relates to a composition for preventing, ameliorating or treating hair growth and hair loss comprising bee venom (hereinafter referred to as BV) as an active ingredient.

In addition to the basic function of protecting the skull from the external environment, the hair has a cosmetic function that makes the appearance more beautiful. In addition to the endogenous factors such as genetics and hormone secretion in recent years, the hair loss population is rapidly increasing due to external factors such as stress, drinking or smoking, and the target is limited to middle-aged men and young men and women The trend is spreading.

Hair growth can be divided into anagen phase, catagen phase, and telogen phase. In the growth period, hair grows continuously due to active cell division at the base of the hair follicle, Rapidly shrinks and stops the growth of the hair follicle, and stops the growth of the hair follicle at rest. At this time, a new growth hair is formed at the base of the dormant hair follicle, and the existing dormant hair is torn off by the newly emerged hair of the growing hair or is separated by the mechanical action such as combing.

) 및 트리칼곡실 (Tricalgoxyl

) 성분을 함유한 탈모방지 제품으로서, 폴텐 (Foltene^TM)과 같은 제품들이 시판되고 있기도 하다.Therefore, in order to prevent hair loss effectively and to obtain a wool effect, it is necessary to be able to simultaneously exhibit a growth-inducing effect for promoting the transition from a pausing period to a growing period and a regressive effect for delaying the transition from a growing period to a retrogressive period. However, finasteride and minoxidil are the only drugs that are approved by the United States Food and Drug Administration (FDA) for hair loss treatment. Drugs for hair loss licensed as medicines in Korea include minoxidil, topical and oral hair loss treatments Or a pinasteride ingredient is commercially available. For example, Propecia ( ^TM ), which is marketed by Merck, and Rogaine ( ^TM ), which is marketed by Pfizer, contain minoxidil. In addition, Tricosaccaride, developed by the Italian hair loss specialist company,

) And Tricalgoxyl

), And products such as Foltene ( ^TM ) are also commercially available.

However, in the case of minoxidil, it takes a long time to absorb the skin as well as relying on imports from all over the world for the raw materials, and there is a disadvantage in that the hair loss is recurred when the use is stopped, Propecia has been reported to be prescribed only to adult males, which can lead to decreased number of sperm and sexual dysfunction. Serious problems with existing products are emerging. Moreover, most of the existing hair loss treatment drugs have a strong relationship with the male hormone, thus causing serious problems such as the lack of efficacy in women or the development of malformations. Therefore, in recent years, due to these problems, attention has been focused on natural ingredients for promoting hair growth and preventing hair loss.

Bee venom is a substance consisting of about 40 components, such as polypeptides, enzymes and small molecular weight compounds found in bees' capsules. Natural bee venom contains melittin, apamin, MCD peptide, (Son DJ. Et al., Pharmacology) have been reported to contain major active ingredients such as adolapin, protease inhibitor, phospholipase A2, and hyaluronidase & Therapeutics 2007,115, 246-270). In particular, melittin, which is the main characteristic component of bee venom, is contained in an amount of 40-50%, and phospholipase and hyaluronidase are 10-12% and 1.5-2% , Which is known to exhibit anti-inflammatory and anti-cancer effects. Other amines include histamine, dopamine, and norepinephrine in an amount of less than 1%.

It has been known that bee venom causes allergies, but it has been used in traditional therapies such as degenerative arthritis and acute arthritis because of its analgesic and anti-inflammatory effects. In recent years, there has been a growing interest in the treatment of rheumatoid arthritis (Seo BK et al., Korean Acupuncture Society 2005, 22, 19-32), improvement of Parkinson's disease (Doo AR et al., Brain Research 2012. 1429, 106-115 .; Alvarez- D., PLoS One. 2013, 8, e61700) and anti-dementia, as well as skin whitening (Korean Patent Publication No. 10-2012-0003178) and skin regeneration Japanese Patent Application Laid-Open No. 10-2011-0139486). However, the fact that bee venom is effective in promoting hair growth and preventing hair loss is not known at all in the past.

Accordingly, the present invention provides a composition for promoting hair growth and preventing hair loss containing bee venom as an active ingredient and a method for producing the same, thereby solving various problems of the hair growth promoting agent or the hair loss preventing agent reported in the prior art.

In order to achieve the above object,

A composition for promoting hair growth and preventing hair loss comprising bee venom as an active ingredient.

According to one embodiment of the present invention, the bee venom may be a live bee venom, a dry bee venom, a bee venom extract, a tablet bee venom, or a mixture thereof.

According to another embodiment of the present invention, the content of the bee venom in the composition may be 0.001 to 0.01% by weight.

According to another embodiment of the present invention, the composition may further comprise a carrier component.

According to another embodiment of the present invention, the carrier component may be a mixture of polyethylene glycol, distilled water and ethyl alcohol.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a composition exhibiting excellent hair growth promotion and hair loss prevention effect for both men and women without side effects.

Figure 1 is a graph showing the composition according to the invention containing bee venom at various concentrations (0.001, 0.005, 0.01%) and the wool effect in skin tissue after treatment with minoxidil (2%) as a positive control.
FIG. 2 is a graph showing the effect of the composition according to the present invention containing various concentrations of bee venom (0.001, 0.005, 0.01%) and the growth effect of hair follicles in skin tissue after treatment with minoxidil (2%) as a positive control.
FIG. 3 is a graph showing the effect of increasing the length of hair grown from hair roots after treatment with minoxidil (2%) as a positive control and composition according to the present invention containing bee venom at various concentrations (0.005, 0.01%).
Figure 4 shows the composition according to the invention containing bee venom at various concentrations (0.001, 0.005, 0.01%) after degenerating by treatment with 0.1% dexamethasone, a regressor inducer, and minoxidil (2%) as a positive control. , Which is a graph showing the anti-hair loss effect of regressor.
Figure 5 shows the composition of the present invention containing bee venom at various concentrations (1, 10, 100, 500 ng) and melittin (200 ng), a major component of bee venom, in human hair follicle dermal papilla cells And the cell proliferation effect when treated.

In the present invention, the effect of preventing the hair loss of the bee venom component and promoting the hair growth is confirmed and purified through a predetermined process, followed by mixing with a carrier to prepare a composition, and then the hair loss prevention and hair growth promoting effect of the composition is experimentally confirmed . Specifically, in terms of the effect of preventing hair loss, artificial induction of hair regrowth and treatment of the composition according to the present invention was observed to prevent hair loss. In addition, in terms of hair growth promoting effect, Effect was observed.

As can be seen from the following examples, when the composition according to the present invention was treated, hair loss prevention and hair growth promoting effects were observed depending on the bee venom component content in the composition. This effect was confirmed by using minoxidil 2%) treated group.

Accordingly, the present invention provides a composition for promoting hair growth and preventing hair loss, which contains bee venom as an effective ingredient, in view of the excellent hair loss prevention and hair growth promoting effect of bee venom component.

In the present invention, bee venom is a secreted bee venom that is stored in a bee venom of a bee secreted from a bee's poison and is an animal natural physiologically active substance secreted by external stimuli . The bee venom corresponds to the bee venom of the present invention including, but not limited to, raw bee venom, bee venom extract, bee venom extract, and tablet bee venom, as long as they contain substances secreted from bee venom. In the present invention, bee venom can be purchased commercially, sold, or harvested from nature or cultivated, and unrefined bee venom can be used as a yellow poplar propolis, honey, pollen, It may contain impurities, so it can be purified and used.

The raw bee venom is not dried or purified, and is flavored with a transparent liquid having a directionality and exhibits an acidic reaction. It has a specific gravity of 1.31, an acidity (pH) of 5.2-5.5, an acidic substance of bee venom volatilizes at room temperature % Of the dried matter remains, and most of the dried matter occupies about 75% of the protein or the peptide. In addition, various substances such as potassium, sodium, magnesium, phosphorus, alanine, valine, arginine, tryptophan, methionine, and histidine are contained in trace amounts. A bee venom extract can be obtained by isolating and extracting active ingredients including peptides and the like using various extraction methods from raw bee venom. As a known extraction method, water, a mixed anhydride or water lower alcohol (carbon number 14, methanol, ethanol, propanol, butanol, etc.), a lower alcohol and water, acetone, ethyl acetate and butylene glycol But is not limited thereto.

The bee venom can be obtained by a conventional purification process other than the extraction by the above-mentioned extraction solvent, or by removing the bee venom component by separation using a filtration membrane, separation by various chromatography, Bee venom, and does not limit the purification method.

According to the present invention, the preferred bee venom may be bean gum extract, and may be a butanol extract obtained by successively fractionating bee venom in the order of ethanol, petroleum ether, ethyl acetate and butanol.

When the bee venom extract is a bean-butanol extract obtained by sequentially fractionating the bee venom in the order of ethanol, petroleum ether, ethyl acetate and butanol, specifically bee venom different in allergen induction, specifically bee venom, bee venom, , Bee venom acetone extract, bee venom ethyl acetate extract and ethyl acetate butylene glycol extract. The concentration of hair was increased and the hair regrowth inhibitory effect and hair growth inducing effect were much better than those of bee venom acetone extract, bee venom ethyl acetate extract and ethyl acetate butylene glycol extract. Thus, the hair loss prevention effect and hair growth effect of the bee venom butanol extract according to the present invention were confirmed. In particular, the butanol extract showed an allergen-inducing effect that was twice or more lower than that of the conventional live bean gum, dry bean gum or purified bee gum.

The bee venom in the composition according to the present invention may be 0.001 to 0.01% by weight. In the present invention, the bee venom concentration in the composition is optimized in consideration of the toxicity of bee venom including allergen-inducing effects. When the bee venom concentration in the composition is less than 0.001% by weight, the concentration of the bee venom is too low, However, if it exceeds 0.01% by weight, the toxin itself may be toxic to the human body and cells, resulting in various side effects such as reduction of cell proliferation or induction of inflammation.

In addition to the bee venom as an active ingredient, the composition according to the present invention includes a vehicle component for effectively delivering an effective ingredient to target cells. For example, a mixture of polyethylene glycol, distilled water and ethyl alcohol is used as a carrier , Wherein the ratio of each component in the carrier can be mixed in a volume ratio of 5: 3: 2.

EXAMPLES Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to assist the understanding of the present invention and should not be construed as limiting the scope of the present invention.

Bee venom  Manufacturing and refining

50 g of the crude bee venom (pearl trade) was added to the reaction vessel together with 2.5 L of purified water, and the mixture was stirred at 30 to 35 ° C for 2 hours. The filter paper (ADVANTEC No. 2, pore size 5 탆, thickness 261 탆) was used for filtration in order to primarily remove substances having a large size in the resultant product, and then the filter paper was washed with 100 ml of purified water. The filtered solution was secondary filtered through a 0.45 [mu] m membrane filtration apparatus (Millipore). After filtration, the filter paper was washed with 100 ml of purified water, and the filtrate was lyophilized for 90 to 120 hours to obtain 41 g of tablets.

Beijing  Preparation of extract

100 g of crude bee venom was mixed with 500 ml of a 70% aqueous ethanol solution as an extraction solvent and reacted for 12 hours. The precipitate of the mixed solution was filtered, and the filtrate was concentrated under reduced pressure. Petroleum ether and water were added to the concentrate, followed by layer separation. A petroleum ether layer was obtained. Ethyl acetate was added to the petroleum ether layer to effect reaction. The ethyl acetate layer was separated by layer separation. The obtained ethyl acetate layer was further reacted by adding butanol, followed by layer separation to obtain a butanol fraction layer. The fractions of each step were confirmed to have hair regressor inhibitory effect, and it was confirmed that the butanol fraction had particularly excellent hair regressor inhibitory effect.

Experimental Example  One. Bee venom  Induction effect of hair growth

1-1. Experimental Method

6 week old C57BL / 6 female mice whose skin color was white or pink were used as experimental animals. The experimental group was divided into five groups including a negative control group (including a carrier with no active ingredient) and a positive control group, and six mice were used in each group. The hair was cut using an electric razor, and the test material was applied to the shaved area once a day for 19 days. The negative control group was treated only with a carrier used as a solvent for dissolving the reagents (polyethylene glycol (purified purified gold): distilled water: ethyl alcohol = 5: 3: 2), and a positive control group was treated with minoxidil (Sigma) (W / w), 0.005% (w / w) and 0.01% (w / w) to the carrier, respectively. Respectively. Table 1 summarizes the treatment methods for each experimental group.

Experimental group Processing method Group 1 (negative control, NC) After depilating, a carrier ointment was applied to the skin (carrier ointment was composed of polyethylene glycol: distilled water: ethyl alcohol = 5: 3: 2) Group 2 (positive control, PC) After epilating, 2% (w / w) minoxidil ointment was applied to the skin Group 3 (sample treatment group) After epilating, apply 0.001% (w / w) bee venom to skin Group 4 (sample treatment group) After epilating, apply 0.005% (w / w) bee venom to the skin Group 5 (sample treatment group) After epilating, 0.01% (w / w) bee venom is applied to the skin

1.2 Identification of promoting effect of wool in skin tissue

Each sample was applied to the skin once a day at regular intervals, and after 19 days, the photographs were taken to compare the effect of the wool, and the results are shown in FIG. As can be seen from Fig. 1, the wool effect was apparent in a concentration-dependent manner according to the bee venom treatment, and 0.005% and 0.01% treatment of bee venom was significantly superior to that of minoxidil (2%) used as a positive control group Wool promoting effect.

1.3 Identification of root growth effect

On the last day of the experiment, mice were sacrificed to obtain dermal tissues. After the tissues were fixed with paraffin, tissue slides were prepared and stained with hematoxylin and eosin (H & E) and observed with a microscope (100x). Observation results are shown in Fig. As can be seen in FIG. 2, the growth of hair follicles in the tissue was clearly observed by bee venom treatment, and the effect was dose-dependent. The results of FIG. 2 are consistent with the wool promoting effect observed from the outside of the skin of FIG. 1, and as a result, the bee venom treatment resulted in promoting the wool.

1.4 Identification of Hair Length Growth Effect

After 19 days from the treatment of the sample, the hair was randomly picked and the length was measured. The results are shown in FIG. As can be seen from FIG. 3, it was observed that the length of the hair grows by the bee venom treatment, and the effect thereof is shown to be concentration-dependent, and it is known that the length increases in comparison with the case of the positive control group there was. In conclusion, it was confirmed that the hair-growth promoting effect appears in a concentration-dependent manner in the bee venom-treated group.

Experimental Example  2. Bee venom  Hair regressor inhibition effect

2.1 Experimental Method

6 week old C57BL / 6 female mice whose skin color was white or pink were used as experimental animals. The experimental group was divided into 6 groups including negative control (including carrier with no active ingredient) and positive control group, and 6 mice were used in each group. The hair was removed from the mouse using a depilator, and the test material was applied for 7 days once a day to the depilated area from 7 days after depilation. After 9 days of epilating, 1 ml of 0.1% dexamethasone (regression inducing drug, Sigma) using propylene glycol as a solvent was applied to the depilated area for 5 days once a day. After 16 days, the area where hair growth was maintained in hair removal area was observed. The negative control group was treated with a carrier (polyethylene glycol: distilled water: ethyl alcohol = 5: 3: 2) used as a solvent for dissolving the reagents, and a positive control group was prepared by dissolving minoxidil (Sigma) ), And treated with 0.001% (w / w), 0.005% (w / w) and 0.01% (w / w) of bee venom on the carrier. Table 2 summarizes the treatment methods for each experimental group.

Experimental group Processing method Group 1 (negative control, NC) After 7 days of epilating, the carrier ointment was applied to the skin (carrier ointment was composed of polyethylene glycol: distilled water: ethyl alcohol = 5: 3: 2) Group 2 (dexamethasone treatment) After 7 days of epilating, the carrier ointment was applied to the skin, and after 9 days of epilating, 0.1% (w / w) dexamethasone (Dexamethason) and carrier ointment Group 3 (positive control, PC) From 7 days after epilating, 2% (w / w) minoxidil ointment was applied to the skin Group 4 (sample treatment group) From 7 days after hair removal, 0.001% (w / w) bee venom was applied to the skin. After 9 days, 0.1% (w / w) dexamethasone and 0.001% (w / w) bee venom are applied to the skin. Group 5 (sample treatment group) From 7 days after hair removal, 0.005% (w / w) bee venom is applied to the skin. After 9 days, 0.1% (w / w) dexamethasone and 0.005% (w / w) bee venom are applied to the skin. Group 6 (sample-treated group) From 7 days after hair removal, 0.01% (w / w) bee venom is applied to the skin. After 9 days, 0.1% (w / w) dexamethasone and 0.01% (w / w) bee venom are applied to the skin.

2.2 Identification of Anti-Hair Loss Prevention Effect

As can be seen from Fig. 4, the effect of preventing hair regrowth in the concentration-dependency was observed by the bee venom treatment. In the group treated with dexamethasone only, the hair growth rate was significantly increased in the bee venom treated group, despite dexamethasone treatment. In the bee venom treated group, the hair growth effect was much superior to that of the positive control group, minoxidil treated group.

Experimental Example  3. Bee venom Skin cell  Proliferation effect

3.1 Experimental Method

Dermal papilla cells (DPC) were used to confirm the proliferation of skin-derived cells by bee venom. Cell culture has 10% fetal bovine serum (FBS), 1% penicillin / streptomycin, 1% L- glutamine-containing Dulbecco's modified Eagle's medium (DMEM , Gibco BRL) a was used, 37 ℃, 5% CO ₂ is fed Lt; / RTI > The cultured cells were treated with 1.5 × 10 ⁴ cells / well and cultured for 24 hours. Then, 10% of the medium was removed and MTT (0.5% 3- (4,5- Dimethylthiazol-2-yl) -2,5 diphenyl-2H-tetrazolium bromide, Sigma) was added and incubated for 4 hours. After removing the culture medium, 200 μl of MTT solubilization solution (Sigma) was added and shaken for 10 minutes, and the cell proliferation effect was evaluated by measuring the absorbance at 570 nm using a microplate reader.

3.2 Confirmation of skin cell proliferation effect

As can be seen from FIG. 5, the cell proliferation effect of bee venom treated group was 12%, 19%, and 21% at 10, 100 and 500 ng treatments, respectively, Respectively. On the other hand, 200 ng of melitin, the main component of bee venom, showed 15% cell proliferation effect.

In summary, the composition according to the present invention contains bee venom as an effective ingredient, has an excellent hair growth promoting effect, an anti-hair regrowth prevention effect, and a skin cell proliferation effect, Can be effectively used as a composition for promoting hair growth and preventing hair loss, which can effectively solve various problems caused by an antioxidant.

Claims (5)

A bee venom inhibiting anti-alopecia composition comprising bee venom butanol extract obtained by sequentially fractionating bee venom in the order of aqueous ethanol solution, petroleum ether, ethyl acetate and butanol. delete [2] The composition according to claim 1, wherein the content of the bee venom butanol extract in the composition is 0.001 to 0.01% by weight. The composition of claim 1, wherein the composition further comprises a carrier component. A method for preparing an antiallergic composition having reduced allergenicity, comprising the steps of sequentially fractionating bee venom in the order of ethanol aqueous solution, petroleum ether, ethyl acetate and butanol in order.

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