KR20150087659A - Composition for inhibiting growth of body hair comprising coumarin as an effective ingredient - Google Patents

Abstract

The present invention relates to a composition for inhibiting body hair growth including coumarine as an effective ingredient and, more specifically, to a cosmetic or pharmaceutical composition for inhibiting body hair growth including 0.0005 to 5 wt% of coumarine compared with total weight of the composition. The composition of the present invention effectively inhibits growth of body hair by inhibiting propagation of outer rood sheath cells by activating a promotor of TGF- β1 and increasing expression of TGF- β1. The composition of the present invention has no side effect even being applied to the body by not having toxicity, and is useful in reducing and removing hair of local parts.

Description

[0001] The present invention relates to a composition for inhibiting hair growth comprising coumarin as an active ingredient,

The present invention relates to a composition for inhibiting hair growth comprising coumarin as an active ingredient, and more particularly, to a cosmetic composition for inhibiting hair growth and containing a coumarin as an active ingredient and a pharmaceutical composition for inhibiting hair growth.

Human hair has about 100,000 to 150,000 hairs, and each hair has a different growth period. The hair growth cycle consists of three stages: the anagen stage where hair grows most actively, the catagen stage where hair starts to degenerate, and the telogen stage where hair growth stops or rests. .

Among the factors regulated by hair follicles in the hair follicle, transforming growth factor-β (TGF-β) inhibits hair follicle growth and hair follicle growth 2003). It has been shown that TGF-beta is a potent inhibitor of TGF-beta, and that it is involved in the induction of metastasis (Tsuji Y, Denda S, Soma T, Raftery L, Momoi T, Hibino T. A potential suppressor of TGF-beta delays in hair follicles, J Investig Dermatol Symp Proc 2003: 8 (1): 65-68). In other words, TGF-β is expressed in the dermal papilla cells of the hair follicle by the male hormone, causing the apoptosis of the hair follicle stromal cells and the follicular keratinocyte and inducing the growing hair follicle as a regressor, whereby the dormant hair follicle is increased, (Lee, Young-Joo, Lee, Joo-young, Clinical diagnosis and management of male androgenetic dehumanizer, Journal of Korean Society of Hairdressing, 2007: 13 (2): 799-810).

Hair or body hair has been functioning as one of the important defenses of the body that protects the organs of the body from external stimuli. Now, except for the hair, the function and role as the defensive means are becoming increasingly irrelevant.

In particular, when viewed from an aesthetic point of view, it is desirable for women to have no hair on the contrary. Accordingly, various methods are used to remove hairs. Examples of methods for removing hairs include a method in which hairs are removed or cut off by mechanical means using an electric razor, an epilator, an ultrasonic wave, or a laser, a method using physical means such as an adhesive tape, There are various methods such as depilatory methods, and a material containing a specific protease is used as a permanent depilator.

However, hair removal methods using mechanical means or physical means to pull hair removal have painful disadvantages, and chemical means using conventional hair removal cream or wax may cause skin damage due to chemical components . In addition, the conventional methods are still insufficient in terms of suppression of hair growth, have an effect in a short period of time, and have a disadvantage in that hair removal treatment must be performed again after a certain period of time has elapsed due to hair growth.

SUMMARY OF THE INVENTION Accordingly, the present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide a composition for inhibiting hair growth, which is safe for human body but can effectively inhibit hair growth, And to provide a method for inhibiting hair growth.

In order to solve the above problems, the present invention provides a cosmetic composition for inhibiting hair growth comprising coumarin as an active ingredient. The present invention also provides a pharmaceutical composition for inhibiting hair growth comprising coumarin as an active ingredient.

In the cosmetic or pharmaceutical composition for inhibiting hair growth, the coumarin may be represented by the following formula (1), but is not limited thereto.

The present inventors have researched to provide a composition for inhibiting hair growth which can safely protect the human body and effectively inhibit hair growth in order to solve the problems of the hair removal agent by the conventional chemical and physical means. As a result, coumarin activates the promoter of human TGF-beta1 (Transforming growth factor beta 1) gene and increases the expression of human TGF-beta1 gene, thereby inhibiting proliferation of outer appearance root cells and effectively inhibiting hair growth And confirmed the present invention.

Hereinafter, the present invention will be described in more detail.

In one embodiment of the present invention, the cells were treated with coumarin in Human Hair Outer Root Sheath Cells (SCIENCELL, USA) and then stained with trypan blue (Sigma, USA) And inhibited proliferation (Example 1).

In another embodiment of the present invention, luciferase assay is carried out after treating coumarin with a vector obtained by combining a promoter of human TGF-? 1 and luciferase with lipofectamine, and as a result, -β1 promoter (Example 2).

In another embodiment of the present invention, the relative expression of TGF-beta1 is detected by treatment with coumarin in Follicle Dermal Papilla (Promocell, Germany), which is known to express human TGF-beta1, using reverse transcription polymerase chain reaction As a result of the comparison, it was confirmed that coumarin increases the expression of human TGF-β1 (Example 3). Increasing the expression of human TGF- beta 1 preferably means increasing the expression of human TGF- beta 1 at the transcription level.

In addition, it was confirmed that coumarin was manufactured into ointment for external use or cream formulation in an embodiment of the present invention, and when it was applied to human skin, excellent hair growth inhibitory effect was shown (Examples 4 and 5).

In view of the above-described embodiments of the present invention, the coumarin of the present invention has an effect of reducing the appearance and development of the root hair, and thus can be effectively used as an effective ingredient of the hair growth inhibitor.

The content of the coumarin contained in the cosmetic composition or the pharmaceutical composition may be 0.0005% by weight to 5% by weight based on the total weight of the composition. When the concentration of the coumarin is less than 0.0005% by weight based on the total weight of the composition, it is difficult to obtain the effect. When the concentration exceeds 5% by weight, there is a problem in formulation stability.

The coumarin contained as an active ingredient in the cosmetic or pharmaceutical composition may contain coumarin hydrate, coumarin derivative, a solvent or a stereoisomer thereof in the range of the same effectiveness. The coumarin may be chemically synthesized or extracted directly, or a commercially available coumarin may be purchased and used. When the coumarin is separated from a natural substance, it includes all the extracts of natural products or fractions thereof, as long as it contains coumarin.

The term "hairs" used in the present invention refers collectively to hairs distributed in the body. The distribution of the hairs is not particularly limited. However, for the purpose of the present invention, it is preferable that the hairs are distributed in a body part which causes harm to aesthetics due to occurrence, growth, or excessive distribution of hairs such as armpits, arms, It can mean hair.

The term "hair growth inhibition" used in the present invention includes not only suppressing hair growth or growth, slowing hair growth rate, but also reducing hair thickness or length.

The coumarin of the present invention may preferably be added to an external preparation for skin hair growth inhibition, but is not limited thereto. The external skin preparation for hair growth inhibition may be a cosmetic composition or a pharmaceutical composition. When formulating a skin external preparation for hair growth inhibition with a cosmetic or a medicinal agent, it may contain known dermatologically acceptable excipients which act as a carrier for the active ingredient. When formulated as a medicament, the contents disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA can be referred to. When formulating into cosmetics, International cosmetic ingredient dictionary, 6th ed., The cosmetic, Toiletry and Fragrance Association , Inc., Washington, 1995, which is incorporated herein by reference in its entirety. Which are incorporated herein by reference.

Specifically, the cosmetic or pharmaceutical composition may be formulated into a form such as a liquid, oil, cream, ointment, stick, pack, pasta, powder, gel or spray as an external preparation for skin, And the like. Preferably, for the purposes of the present invention, the cosmetic or pharmaceutical composition may be formulated as a semi-solid preparation, such as an external ointment, lotion, and the like.

As the external preparation for skin, the cosmetic composition or the pharmaceutical composition may contain a cosmetically or pharmaceutically acceptable carrier, diluent, adjuvant, colorant, stabilizer, flavor, interfacial agent, oil, moisturizer, alcohol, , An antioxidant, a pH adjuster or an ultraviolet screening agent.

In addition, the pharmaceutical composition of the present invention may be formulated into various formulations suitable for oral administration or parenteral administration.

Formulations for oral administration of the pharmaceutical compositions may be in the form of troches, lozenges, tablets, aqueous suspensions, oily suspensions, prepared powders, granules, emulsions, hard capsules, soft capsules, syrups or elixirs But is not limited thereto.

Binding agents such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin to formulate the pharmaceutical composition for oral administration; Excipients such as dicalcium phosphate and the like; Disintegrating agents such as corn starch or sweet potato starch; Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax may be used. However, it is not limited thereto, and sweeteners, Fragrances, syrups, and the like may also be used. Furthermore, in the case of capsules, in addition to the above-mentioned substances, liquid carriers such as fatty oils and the like may further be used.

Examples of the parenteral preparation for the pharmaceutical composition include, but are not limited to, injections, suppositories, respiratory inhalation powders, aerosol preparations for spray, ointment, powder for application, oils or creams.

In order to formulate the pharmaceutical composition for parenteral administration, a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized preparation or a external preparation may be used. Examples of the non-aqueous solution and the suspension include propylene glycol, polyethylene glycol, Vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used.

More specifically, when the pharmaceutical composition is formulated into an injection, the pharmaceutical composition of the present invention is mixed with a stabilizer or a buffer in water to prepare a solution or suspension, which is then mixed with an ampoule or a vial Unit dosage form. Further, when the pharmaceutical composition of the present invention is formulated into an aerosol formulation, a propellant or the like may be formulated together with the additive such that the water-dispersed concentrate or the wet powder is dispersed.

When the pharmaceutical composition is formulated into an ointment, cream, or the like, it may be mixed with an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, Can be used as a carrier.

The route of administration and the mode of administration of the pharmaceutical composition of the present invention may be independent of each other and are not particularly limited in the method and may be arbitrarily administered and administered as long as the pharmaceutical composition can reach the desired site It is possible to follow the method. The pharmaceutical composition may be administered orally or parenterally. The pharmaceutical composition of the present invention may preferably be administered parenterally, and more preferably, it may be administered in a manner such that it is applied to the skin.

Examples of the parenteral administration method include a method of applying, spraying, or inhalation the composition to a diseased site, Intraperitoneal, intramuscular, transdermal or subcutaneous administration may also be used, but are not limited thereto.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the age, sex, weight, symptom, degree of disease, drug form, administration route and period of time of the subject, but can be appropriately selected by those skilled in the art. However, for a desired effect, the pharmaceutical composition of the present invention may be administered at 0.01 to 1000 mg / day. The administration can be done once a day, or divided into several times. In addition, the dose may be increased or decreased according to age, sex, weight, degree of disease, administration route, or the like. Accordingly, the dosage is not limited in any way to the scope of the present invention.

The present invention provides a hair growth inhibition method comprising applying a cosmetic composition or a pharmaceutical composition containing coumarin as an active ingredient to the skin. The present invention also provides a method for inhibiting hair growth comprising oral or parenteral administration of a pharmaceutical composition containing coumarin as an active ingredient.

The present invention also provides a composition for promoting TGF-β1 gene expression comprising coumarin represented by the above formula (1) as an active ingredient.

The composition comprising coumarin as an active ingredient according to the present invention promotes the activation of the TGF-beta1 promoter and the production of TGF-beta1, and can effectively inhibit hair growth by inhibiting the proliferation of the appearance root cultured cells. The composition of the present invention is not toxic and has no side effects even when it is applied to a human body, and is useful for removing or reducing hairs in a local area.

1 is a diagram showing a recombinant vector obtained by cloning a human TGF-beta promoter region into a pGL3 vector.
FIG. 2 is a graph showing the relative expression of human TGF-β1 after treatment with coumarin in dermal papilla cells using reverse transcription PCR (RT-PCR).

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.

The coumarin used in the experimental examples of the present invention was purchased from Xi'an Guanyu Bio-tech Co., Ltd.

Experimental Example  One: Coumarin Apparent cells  Confirming the proliferation inhibiting effect

In this example, coumarin was treated with Human Hair Outer Root Sheath Cells (SCIENCELL, USA), and cells were stained with trypan blue (Sigma, USA) The number of cells was measured to observe changes in cell division.

First, a uniform number of cells were plated on a 96-well plate, stabilized, treated with coumarin at 20 ppm, and further incubated for 3 days. The cells were then stained with trypan blue and the number of cells was measured. DHT (dihydrotestosterone) treated with 100 nM was evaluated as a positive control. Relative cell viability of cells treated with coumarin without treatment was analyzed as follows. The results are shown in Table 1 below.

Relative cell viability = (compound treated group) / (negative control group) X 100

sample Negative control group
(No treatment) Positive control group
(DHT treatment) Coumarin
(coumarin) Survival rate (%) 100 63 60

As shown in Table 1 above, the survival rate of the appearance-induced myocytes decreased in the group treated with coumarin compared to the negative control group and the positive control group.

Experimental Example  2: In dermal papilla cells  human TGF -β1 promoter activity

A reporter vector was prepared to examine the degree of promoter activity of human TGF-? 1. For this purpose, the promoter region (-1,132 to +271) of human TGF-beta 1 was amplified from human genomic DNA using PCR. The primers used at this time are shown in Table 2.

order SEQ ID NO: Forward 5'-GGGAAGCTTTGGAAGGATCCTTAGCAGGGG-3 ' One Reverse 5'-GGGTCGACCGCGGAGGGAGGTGGGA-3 ' 2

The amplified DNA was cut with HindIII and SalI restriction enzymes and cloned into pGL3 vector expressing firefly luciferase (Fig. 1). The prepared plasmid vector and a plasmid always expressing Renilla luciferase were used in a promoter assay in combination with a DNA mixture in a ratio of 40: 1. Human dermal papilla cells (Promocell, Germany) were cultured in a DMEM medium containing 10% fetal bovine serum for 18 hours, and then the resulting vector, in which the promoter of human TGF-β1 and luciferase were combined with the cultured cells, And injected into cells via transfection using lipofectamine.

After 5 hours of incubation, the cells were washed twice with DMEM medium containing no serum, and treated with coumarin at 20 ppm in DMEM medium and cultured for 18 hours. After 18 hours of treatment, the cells were recovered, washed with cold phosphate buffer (PBS), and the cells were lysed using a cell lysis buffer. Then, only the supernatant was extracted using a centrifugal separator and subjected to luciferase assay (Promega, USA) Fluorescence luminescence was measured and analyzed with a Perkin Elmer Victor 3. The negative control group was untreated and the positive control group was treated with only 100 nM of DHT. The growth rate was analyzed as follows, and the results are shown in Table 3.

Growth rate (%) = (Compound treated group) / (Negative control group) X 100

sample Negative control group
(No treatment) Positive control group
(DHT treatment) Coumarin
(coumarin) Growth rate (%) 100 135 145

As shown in Table 3, coumarin according to one embodiment of the present invention activates the promoter of human TGF-? 1. The measured value is the value obtained by correcting the firefly luciferase value with the Renilla luciferase value.

Experimental Example  3: In dermal papilla cells  human TGF -β1 increase

In this experiment, Human Hair Outer Root Sheath Cells (SCIENCELL, USA) was used to investigate the effect of increasing expression of human TGF-β1 in coumarin dermal papilla cells. A uniform number of cells were spread on a 6-well plate and stabilized before treatment with coumarin. After culturing for 48 hours after the treatment, the RNA was synthesized with cDNA and the relative content of human TGF-? 1 was measured. The changes in human TGF-β1 were determined by reverse transcription-polymerase chain reaction (RT-PCR) in the presence of TGF-β1 mRNA sequences from human genomic DNA. The primers used at this time are shown in Table 4 below.

order SEQ ID NO: TGF-β1 (forward) 5'-GGGACTATCCACCTGCAAGA-3 ' 3 TGF-β1 (reverse direction) 5'-CCTCCTTGGCGTAGTAGTCG-3 ' 4 GAPDH (forward direction) 5'-cgagatccctccaaaatcaa-3 ' 5 GAPDH (reverse direction) 5'-ttcacacccatgacgaacat-3 ' 6

As a result of the experiment, it was confirmed that the amount of human TGF-β1 mRNA increased in the dermal papilla cells by treatment with coumarin. The results were analyzed using the Comparative C _T (ΔΔC _T ) Experiment, and the internal control was performed using GAPDH. The results are shown in FIG.

Coumarin  Manufacture of ointment for external skin containing

(Example 1) containing coumarin and a skin external ointment containing no coumarin (Comparative Example 1) were prepared with the composition shown in Table 5 below.

Composition Example 1 (% by weight) Comparative Example 1 Coumarin 0.5 - Diethyl sebacate 8 8 Nonsense 5 5 Polyoxyethylene oleyl ether phosphate 6 6 Sodium benzoate Suitable amount Suitable amount vaseline to 100 to 100

Coumarin  Manufacture of cream containing

A cream containing coumarin (Example 2) and a cream containing no coumarin (Comparative Example 2) were prepared with the composition shown in Table 6 below.

Composition Example 2 (% by weight) Comparative Example 2 Coumarin 0.2 - Stearic acid 15.0 15.0 ethanol 1.0 1.0 Potassium hydroxide 0.7 0.7 glycerin 5.0 5.0 Propylene glycol 3.0 3.0 antiseptic Suitable amount Suitable amount incense Suitable amount Suitable amount Purified water to 100 to 100

Experimental Example  4: Investigation of hair removal effect

Twenty women aged 25 to 35 years old who were healthy to cream of Example 2 and Comparative Example 2 were subjected to the following hair removal effect.

The subjects were applied with the cream of Comparative Example 2 without coumarin on the left leg and the cream of Example 2 containing 0.2% coumarin on the right leg twice a day for three months. After three months, the effects of hair removal were evaluated by questionnaire and image analysis. The subjects' questionnaires were compared with those before use with regard to changes in hair thickness and growth retardation, and the three stages of no improvement, slight improvement, and significant improvement were determined.

sample No improvement Slight improvement A significant improvement Comparative Example 2 18 One One Production Example 2 6 7 7

As can be seen from Table 7, when the cream of Example 2 according to the present invention is applied to the skin, it can be seen that the effect of hair removal due to the change in hair thickness and growth delay is excellent.

Example  5: Body image analysis

For image analysis of body hair, subjects shaved their legs four days before evaluation. Evaluation was carried out before and after the experiment. The results of measurement of hair growth of Example 2 and Comparative Example 2 by image analysis are shown in Table 8 by averaging the reduction rate with respect to the hair length before the experiment.

Example 2 Comparative Example 2 Before experiment 1.00 1.00 after 2 weeks 0.76 (24% reduction) 1.00

As can be seen from Table 8, when the cream of Example 2 containing coumarin according to the present invention is applied to the skin, it can be seen that the hair growth delay effect is remarkably exerted.

Claims (10)

A cosmetic composition for inhibiting hair growth, comprising coumarin as an active ingredient. The cosmetic composition for suppressing hair growth according to claim 1, wherein the coumarin is a compound represented by the following formula (1).
[Chemical Formula 1]

The cosmetic composition for suppressing hair growth according to claim 1, wherein the coumarin is 0.0005 to 5% by weight based on the total weight of the composition. A pharmaceutical composition for inhibiting hair growth comprising coumarin as an active ingredient. 5. The pharmaceutical composition for suppressing hair growth according to claim 4, wherein the coumarin is a compound represented by the following formula (1).
[Chemical Formula 1]

The pharmaceutical composition according to claim 4, wherein the coumarin is 0.0005 to 5% by weight based on the total weight of the composition. The composition according to any one of claims 1 to 4, wherein the composition is any one of external preparations for skin selected from the group consisting of liquid, oil, cream, ointment, stick, pack, pasta, powder, Wherein the hair growth inhibitory composition is a hair growth inhibitory composition. The composition according to claim 7, wherein the composition is a cosmetic or pharmaceutical acceptable carrier, diluent, adjuvant, colorant, stabilizer, flavor, interfacial agent, oil, humectant, alcohol, thickener, antioxidant, And an ultraviolet screening agent. The composition for suppressing hair growth according to any one of claims 1 to 3, A method for inhibiting hair growth, comprising applying the composition of any one of claims 1 to 6 to the skin of a subject in need of hair growth inhibition. 1. A composition for promoting TGF-? 1 gene expression comprising coumarin represented by the following formula (1) as an active ingredient.
[Chemical Formula 1]

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