KR101629503B1 - Composition for promoting the growth of hair - Google Patents

Abstract

The present invention relates to a composition for promoting hair growth, which comprises Tetramethyl pyrazine, Madecassoside or a mixture thereof as an active ingredient. The composition according to the present invention can accelerate the production of VEGF, increase the nutritional supply of the hair follicle, and improve the hair cycle, thereby promoting hair growth.

Description

[Technical Field] The present invention relates to a composition for promoting hair growth,

The present invention relates to a hair growth promoting composition, and more particularly, to a composition capable of promoting the production of VEGF, increasing nutritional supply of hair root, and improving hair growth through hair growth promoting effect.

VEGF is a cytokine, a factor that promotes the differentiation and growth of endothelial cells in blood vessels ( Science 1989; 246, 1306-1309). Cells that have experienced a lack of oxygen usually produce Hypoxia Inducible Factor (HIF), which promotes the production and release of VEGF. The epidermal cells that recognize this will proceed with angiogenesis. In addition, VEGF also extends blood vessels and enhances the permeability of capillaries.

In humans, VEGF is made from a single gene through alternative splicing and about five types of VEGF are known. Three of them (VEGF ₁₂₁ , VEGF ₁₄₅ , and VEGF ₁₆₅ ) are secreted from the cells produced, while the two functions in the form bound to the cells ( Biochem . Biophys . Res. Commun . 2000 : 270 , 709-713). These expressed VEGF bind to receptors such as VEGFR-1, VEGFR-2, and VEGFR-3, and the activated receptors act as trans-membrane kinases (C ancer J. 1998; 4 , 209-217).

VEGF expression was observed in various cells in the structure of the hair follicle. In particular, 46 kDa of VEGF corresponding to VEGF ₁₆₅ was produced in DPC (Dermal papilla cell) ( Arch. Dermatol . Res. 1998; 290 : 661-668). In addition, outer root sheath keratinocyte and dermal pailla fibroblast were also observed ( Br. J. Dermatol . 1998; 138, 407-411)

The effects of VEGF on hair can be found in various studies. The occurrence of angiogenic action in normal skin is limited only to the case of skin injury. However, in the process of hair growth, an angiogenic action takes place in the hair follicle, and it is actively observed when it is changed from telogen to anagen ( J. Invest. Dermatol . 2000; 114 (5) 909- 196). In addition, inhibition of hair growth by angiogenesis inhibitors such as TNP 407 indicates the importance of VEGF in hair formation. In addition to angiogenesis, VEGF plays an important role in the metabolism of DPC (Dermal papilla cell), which may play a role in the synthesis of collagen and in the synthesis of lipids, thereby contributing to the formation of hair ( Arch. Dermatol . Res. 1996 288 : 469-473, Eur ., J. Dermatol ., 1996; 6 : 365-368). Was observed that the expression of VEGF in a further reduced hair loss model (J. Invest Dermatol 1995;.. 104 18S-20S), representative of minoxidil hair restorer has also been reported to increase the expression of VEGF (British J. Dermatol 1998;. 138 407-411).

The effect of increased expression of VEGF on hair was confirmed in transgenic mice overexpressing VEGF, where the distribution of blood vessels around the hair follicles was increased and the thickness of the hair was increased by about 30% ( The Journal of Clinical Investigation 2001; 107 (4) 409-417).

Therefore, the development of a material that increases the expression level of VEGF is essential for not only suppression of hair loss but also for promoting hair growth. However, the search for a material capable of regulating the production of VEGF externally is insufficient.

 The Journal of Clinical Investigation 2001; 107 (4) 409-417

The present invention provides a composition which promotes the production of VEGF, promotes hair growth, and improves hair damage.

In order to accomplish the above object, the present invention provides a process for preparing a compound represented by the following formula (1): Tetramethyl pyrazine represented by the following formula (1), Madecassoside represented by the following formula (2) A composition for accelerating the production of VEGF (Vascular Endothelial Growth Factor), a composition for promoting hair growth, or a composition for improving hair damage.

≪ Formula 1 >

(2)

The tetramethyl pyrazine used in the present invention is also called Chuanxiongqin in China is known as an active ingredient of the soil Cnidium (ligusticum chunaxiong), it has been used in the treatment of angina pectoris (Proc. Chin. Acad. Med . Sci. Peking Union Me. Coll 1989; 4 (4): 224-8). Madecasone according to the present invention is an active substance extracted from Centella asiatica, which increases the synthesis of wound treatment.

The present invention is based on the surprising discovery that these conventionally used compounds can promote the production of VEGF, help in hair growth, and improve hair damage.

In the composition according to the present invention, the total content of the active ingredient is preferably 0.00001 to 10% by weight based on the total weight of the composition.

The composition for promoting VEGF production, for promoting hair growth or for treating hair damage according to the present invention can be produced in the form of medicines, cosmetics and the like. Such medicaments and cosmetics may include pharmaceutically or cosmetically acceptable excipients or additives. The compositions of the present invention may be administered alone or in admixture with any convenient vehicle, excipient, etc., and such dosage forms may be single-dose or multiple-dose formulations.

The formulation of the composition of the present invention is not particularly limited, but is preferably in the form of an external preparation for skin such as semi-solid and liquid preparations. Examples of the semi-solid preparation include a cream, a lotion, an emulsifier, a liniment, and the like, but the present invention is not limited thereto, and can be produced by adding a suitable colorant, a flavoring agent, a stabilizer, a tackifier, a surfactant, and the like. Examples of the liquid preparation include water, alcohols, solutions such as solutions of propylene glycol, suspensions, emulsions and the like, but not limited thereto, and they can be prepared by adding appropriate coloring agents, flavoring agents, stabilizers, tackifiers and the like.

For example, the external preparation for skin may be a suitable pharmaceutically or cosmetically acceptable substrate such as vaseline, stearyl alcohol and the like; A suitable pharmaceutically or cosmetically acceptable surfactant such as polysorbate, sorbitan sesquioleate, and the like; Suitable pharmacologically or cosmetically acceptable moisturizing agents such as glycerin; A suitable pharmaceutically or cosmetically acceptable solvent; And a conventional external preparation for skin preparation in which a flavoring agent, a coloring agent, a stabilizer, a tackifier and the like are homogeneously mixed.

The composition according to the present invention has excellent VEGF expression increasing effect, promotes hair growth without side effects, and can improve hair damage.

FIG. 1 is a diagram showing a recombinant vector by cloning a VEGF promoter region into a pGL3 vector.
FIG. 2 is a graph showing the activity of the VEGF promoter after transfecting the vector of FIG. 1 into a HaCat cell and treating tetramethylpyrazine or madecassoside, respectively.
FIG. 3 is a photograph showing the expression of VEGF gene by RT-PCR after treating HaCat cells with tetramethylpyrazine or madecassoside, respectively.
FIG. 4 shows the result of applying a composition containing tetramethylpyrazine or madecassoside to a test site for a period of 8 days and then measuring the thickness of the hair using a microscope. And the growth rate was compared with the thickness of the comparative example.

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.

In the following examples of the present invention, Sample 1 refers to Tertamethyl pyrazine (TMP), and Sample 2 refers to Madecassoside, each purchased from Shaanxi Shipyard Biological Co.,

<Experimental Example 1> VEGF transcription promoter activity increase test

To investigate the degree of transcriptional activation of VEGF, a reporter vector was prepared to show the promoter activity of VEGF (Biochemical and Biophysical Research Communications 270, 709-713 (2000)). For this purpose, the VEGF promoter region (-2350 to +101) was amplified from human genomic DNA (PCR). The amplification product was cloned into pGL3vector (Figure 1). This vector was transfected into Hacat cells. Hacat cells were inoculated into 24-well plates and cultured for 24 hours. Cells grown in 70-80% were transfected with Serum-free DMEM and then transfected with pFila-luc vector and pGH control vector using Effecten. Samples 1 or 2 were treated after 5 hours. As a comparative example, the test substance solvent (DMSO) was treated. After incubation for 16 hours, the cells were washed with PBS, and the activity was measured using Promega's luciferase assay system (FIG. 2). The results are summarized in Table 1 below.

Control group (Sample 1)
5 ppm (Sample 1)
20 ppm (Sample 2)
5 ppm (Sample 2)
20 ppm 100% 125% 154% 122% 145%

As shown in Table 1, tetramethylpyrazine or madecassoside according to the present invention was able to activate the VEGF promoter in a concentration-dependent manner.

<Experimental Example 2>

Hacat cells were inoculated into 6 well plates. After 24 hours, the cells grown at 70-80% were replaced with serum-free DMEM and further incubated for 5 hours. Sample 1 or 2 was treated at a given concentration after the time. As a comparative example, the test substance solvent (DMSO) was treated. After further incubation for 16 hours, the cells were washed with PBS and RNA was extracted using RNAeasy kit. The amount of RNA was then standardized using UV absorbance and quantified using RT-PCR (Figure 3). The primers used for RT-PCR of VEGF are as follows (Biochemical and Biophysical Research Communications 270, 709-713 (2000)).

For: 5'-TCGGGCCTCCGAAACCATGA-3 '

Rev: 5'-CCTGGTGAGAGATCTGGTTC-3 '

The primer used for beta-actin rt-PCR was Bioneer N-1083, the sequence of which is as follows.

For: 5'-GTGGGGCGCCCCAGGCACCA-3 '

Rev: 5'-CTCCTTAATGTCACGCACGATTC-3 '

The experimental results are shown in Table 2 below.

Control group (Sample 1)
5 ppm (Sample 1)
20 ppm (Sample 2)
5 ppm (Sample 2)
20 ppm 100% 180% 320% 175% 315%

As shown in Table 2, tetramethylpyrazine or madecassoside according to the present invention activated the expression of the VEGF gene in a concentration-dependent manner.

&Lt; Production Examples 1-1 and 1-2 and Comparative Example 1 >

Preparation Examples of compositions containing Sample 1 or 2 were prepared as shown in Table 3 below.

(Unit: wt%) Production Example 1-1 Production Example 1-2 Comparative Example 1 Sample 1 5 - - Sample 2 - 5 - ethanol 35 35 35 Twin 60 0.5 0.5 0.5 Purified water to 100 to 100 to 100

<Experimental Example 3> Evaluation of hair growth promoting effect

The hair growth promoting effect of each sample was measured. The composition of Comparative Example 1, Preparation 1-1 or Preparation 1-2 was applied to each group of C75BL / 6 rats for 8 days. After that, the hairs were randomly collected, and the thickness of the hairs for preventing hair loss and for the hair growth was measured by a microscope. Table 4 shows the rate of increase in the thickness of hair based on the comparative example.

Production Example 1-1 Production Example 1-2 Growth rate 24.5% 27.6%

As shown in Table 4, it was found that the composition containing the sample 1 or 2 had better hair growth promoting effect than the comparative example 1.

Claims (3)

A composition for improving hair growth or hair damage, which comprises Madecassoside represented by the following formula (2) as an active ingredient, and the total content of the effective ingredient is 0.00001 to 10% by weight based on the total weight of the composition.
(2)

2. The composition of claim 1, wherein the composition promotes the production of VEGF (Vascular Endothelial Growth Factor). delete

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