KR20190019999A - Composition for promoting hair growth comprising dphc - Google Patents

Abstract

The present invention relates to a composition for promoting hair growth comprising diphlorethohydroxycarmalol (DPHC) as an active ingredient. The present invention can not only promote hair growth by inducing hair follicle cell proliferation or promoting the activity of hair follicle, but also improve structural stability of skin tissues by increasing an amount of collagen and elastin which are the extracellular matrix.

Description

TECHNICAL FIELD The present invention relates to a composition for accelerating hair growth comprising DPHC (diphlorethohydroxycarmalol)

The present invention relates to a composition for promoting hair growth comprising DPHC (diphlorethohydroxycarmalol). More particularly, it relates to a composition for stimulating hair growth by promoting hair follicle cell proliferation or promoting follicular activity, And to improve the structural stability of the skin tissue by increasing the amount of the hair growth promoting composition.

Depending on the anatomical site, the thickness of the integument varies, and even within one region, the thickness of the epidermis depends on physiological parameters such as the hair cycle . It is also affected by age and various epidermal diseases. The envelope structure of the mouse is different from humans, but it is very useful for understanding the role of various chemicals as a regulator in integument. The stability of the envelope is known to depend on the basal layer of the epidermis and the activity of the skin cells. Rat cortex can be separated into epidermis and dermis (including cutaneous adnexa) and subcutaneous tissue (penniculum carnosum). The hair follicles, sebaceous glands, and exocrine glands found in the dermis are derived from ectoderms. Primary hair follicles are characterized by large sebaceous glands, prominent innervation, and prominent blood supplies. Secondary hair follicles, on the other hand, are small in size and have small sebaceous glands. The sebaceous glands are derived from the follicular isthmus. But the mouse does not have sweat glands. The dermis and subcutaneous tissue are derived from the mesoderm and provide tensile strength, elasticity and flexibility. These are loose connective tissues and have many extracellular matrix (ECM). The subcutaneous tissue is composed of densely packed adipocytes. These adipose tissues regulate regeneration homeostasis of the epidermis, dermis and cutaneous appendages. The ECM of the envelope is also important to maintain tissue integrity.

The ECM of the dermis consists mainly of Type I, III, V, and VI, collagen fibers, reticular and elastic fibers, and dermal ground substance (glycosaminoglycans, proteoglycans, hyaluronic acid, and dermatan sulphate). Collagen and elastin are elastic and elastic skin structures. Fibroblasts produce collagen and elastin substrates that can be activated by glycolic acid and the like.

As you get older, the thickness of the epidermis, dermis, and subcutaneous tissue that is part of your skin becomes thinner, and the extracellular matrix (ECM) that gives your skin elasticity changes. Especially, collagen and elastin, which form the skin connective tissues, are oxidized and lose their functions, so that the skin loses its elasticity and wrinkles are formed and aging proceeds.

As a result, skin aging phenomenon appears as non-homogenization of cells, elimination of elastin, reduction of collagen, etc., and it is still necessary to study a composition capable of improving the skin aging phenomenon.

KR 10-2009-0070895 A

The present invention provides a hair growth promoting composition comprising DPHC (diphlorethohydroxycarmalol) as an active ingredient. More specifically, it relates to a hair growth promoting composition capable of promoting hair growth by promoting hair follicle cell proliferation or promoting hair follicle activity, and also improving the structural stability of skin tissue by increasing the amount of collagen and elastin as extracellular matrix to provide.

The present invention provides a hair growth promoting composition comprising DPHC (diphlorethohydroxycarmalol) as an active ingredient.

In the hair growth promoting composition of the present invention, the DPHC may be derived from Ishige okamurae .

In the composition for promoting hair growth of the present invention, the DPHC is contained in an amount of 0.1 to 10% by weight based on the total weight of the hair growth stimulating composition. When the DPHC is less than 0.1% by weight, it is difficult to exert an expected effect, and when the DPHC is more than 10% by weight, it is difficult to expect further increase of the efficacy.

In the composition for promoting hair growth of the present invention, the composition for promoting hair growth is characterized by inducing proliferation of hair follicle cells or promoting hair follicle activity.

In the composition for promoting hair growth of the present invention, the composition for promoting hair growth is characterized by increasing the amount of collagen and elastin.

The hair growth promoting composition comprising DPHC (diphlorethohydroxycarmalol) according to the present invention as an active ingredient can maintain and improve the health of hair and hair by proliferating cells in the hair follicle, promote the hair follicle activity, There is a maintainable feature. In addition, regardless of the degree of obesity, that is, irrespective of the amount of adipose tissue, it acts on the skin tissue cells and participates in the regulation of the extracellular matrix collagen and elastin levels, thereby improving the structural stability of the skin tissue. In addition, it can contribute to maintaining and improving the elasticity of skin by increasing the expression of genes associated with elastic fibers in hepatic stem cells.

1 is a graph showing the effect of DHPC on cell activity. (a) Cellular activity of human lipid precursor cells (b) Mouse lipid precursor cells (3T3)
FIG. 2 is a graph showing the effect of DHPC on the strength of collagen fibers. FIG.
Figure 3 is a microscopic image of the effect of DHPC on the amount of collagen fibers (Abbreviation: SC, stratum corneau; E, viable epidermis; DS, dermis and subcutis).
4 is a graph showing the effect of DHPC on the strength of elastin fibers.
FIG. 5 is a microscopic image of the effect of DHPC on the amount of elastin fibers (Abbreviation: SC, stratum corneau; E, viable epidermis; DS, dermis and subcutis).
6 is a microscopic image of the histological characteristics of the back skin of the DPHC-treated rats. A, C, E, G and I correspond to the dry mouse group and B, D, F, H and J correspond to the obese mouse group (Abbreviation: SC, stratum corneum; VE, viable epidermis; D, dermis). DHPC concentrations were as follows: A and B, 0 w / v%; C and D, 0.01 w / v%; E and F, 0.1 w / v%; G and H, 1 w / v%; I and J, 10 w / v%.
7 is a graph showing changes in expression levels of elastin and MFAP5 (microfibrillar-associated protein) mRNA according to DPHC treatment. (a) Expression amount of MFAP5 mRNA in a human (b) Expression amount of elastin mRNA in a human (c) Expression amount of elastin mRNA in a mouse
FIG. 8 is a graph (DHPC concentration unit w / v%) showing the effect of DHPC on the proliferation of mouse skin dermal follicular cells. A, normal individuals; B, Obesity object
9 is a graph showing the results of proliferative cell staining by the Ki67 immunohistological method. A, untreated; B, 0 (w / v)% DPHC; C, 0.01 (w / v)% DPHC; D, 10 (w / v)% DPHC; E, negative control
10 is a graph showing the effect of DHPC on the hair follicle activity.

Hereinafter, the present invention will be described in more detail with reference to Examples. The objects, features and advantages of the present invention will be readily understood through the following examples. The present invention is not limited to the embodiments described herein, but may be embodied in other forms. The embodiments described herein are provided to enable those skilled in the art to fully understand the spirit of the present invention. Therefore, the present invention should not be limited by the following examples.

Reagent preparation

All animal experiments were performed in accordance with the Guide to the Care and Use of Laboratory. Animals were maintained at standard conditions under the Sungshin Women's University. Four-week-old C57BL / 6J female mice were purchased from Charles River Laboratories and adapted to experimental conditions for 10 days. The experimental mice were individually housed in a cage and maintained a light / dark cycle (light on at 06:00) of 14 h / 10 h. Obese rats were exposed to a high fat hypercaloric diet (60%). The dry rats group had free access to standard food (Purina). Food consumption and weight gain were measured daily and weekly, respectively. Six weeks after the start of dietary control, the dorsal hairs were cut and DHPC-containing media was applied to the skin as a whole, once a day for 28 days.

Collagen, elastin and tissue staining

In the DHPC-treated area, a skin sample of the back was taken and fixed in 4% buffered paraformaldehyde and embedded in paraffin. Fixed tissue specimens were placed in an automatic infiltration machine ; EG1150H, Leica, Germany) and dehydrated by volatilization of the alcohols. Through a xylene replacement process, tissues were embedded in paraplast (Cat #: P3558 Sigma-Aldrich, USA). The serial section of the sheath sample was then made vertical from the center using a rotary microtome (RM 2245, Leica, Germany) at a thickness of 4 μm. The sections were attached to glass slides and dried in a slide warmer at 42 DEG C for 6 hours. It was then deparaffinized by xylene. The stained tissue was observed with light microscope (Nikon, Japan). The stained tissue was stained with hematoxylieosin (Masson trichrome for collagen fibers, and Verhoff-Van Gieson for elastic fibers). For image analysis, download the freeware image J (Image Institution of Health) v133 and the Color Histogram plug-in from the NIH website (http: // rsbinfonihgov / ij) Respectively.

Other

All measurements were repeated 10 or more times and are expressed as means SD.

Experimental Example 1: Effect of DHPC on the degree of cell activity

The cell activity of human lipid precursor cells and mouse lipid precursor cells (3T3) according to DHPC concentration (μg / ml%) was analyzed using the MTT assays and is shown in FIG. As shown in Fig. 1, it was confirmed that cell activity was increased in all the DHPC-treated groups.

Experimental Example 2: Effect of DHPC on collagen fibers

After the DHPC treatment was performed on the skin of the dry rats and the obese rats, collagen intensity and collagen amount according to the DHPC concentration (w / v%) were analyzed, and the results are shown in FIG. 2 and FIG. The collagen intensity shown in Fig. 2 was analyzed using ImageJ v133 and Color Histogram plug-in. As shown in Fig. 2, when DHPC was treated, it was confirmed that the strength of the collagen fiber was slightly increased. FIG. 3 is a microscopic image analyzed using the Masson trichrome staining method (x100, (insertion, x400)). The collagen fibers were stained blue. As shown in FIG. 3, it was confirmed that as the DHPC treatment concentration was increased, the amount of collagen increased as the concentration of DHPC was increased (Fig. 3) .

Experimental Example 3: Effect of DHPC on elastin fibers

After the DHPC treatment was carried out on the skin of the dry rats and the obese rats, the amount of elastin and the amount of elastin according to DHPC concentration (w / v%) were analyzed, and the results are shown in FIGS. 4 and 5. The collagen intensity shown in FIG. 4 was analyzed using ImageJ v133 and Color Histogram plug-in. As shown in Fig. 4, when DHPC 10 (w / v%) was treated, it was confirmed that the strength of the elastin fiber significantly increased. FIG. 5 shows a microscopic image analyzed using the Verhoff-Van Gieson staining method (x100, (insertion, x400)). The elastin fibers were dyed from blue black to black. Collagen was stained red. The nucleus was stained blue to black (SC, stratum corneau; E, viable epidermis; DS, dermis and subcutis). As shown in FIG. 5, it was confirmed that the amount of elastin increased as the concentration of DHPC treatment increased.

Experimental Example 4: Histological Characterization

Histological characteristics of DPHC - treated rats and obese rats were evaluated by staining the epithelium. In FIG. 6, histological characteristics of the back skin of the DPHC-treated rats were analyzed. 6, A, C, E, G and I correspond to the dry rat group and B, D, F, H and J correspond to the obese mouse group. DHPC concentrations were as follows: A and B, 0 w / v%; C and D, 001 w / v%; E and F, 01 w / v%; G and H, 1 w / v%; I and J, 10 w / v% H-E (hematoxylin-eosin) staining. The collagen was stained with light pink and the muscles were stained with dark pink (SV, stratum corneum, VE, viable epidermis; D, dermis)

As shown in Fig. 6, DPHC increased the thickness of the non-keratinized epidermal layer and increased the size of the nucleated epidermal keratinocytes without increasing the cell layer. The DPHC-treated group showed a diffuse nuclear staining pattern (over DPHC-untreated group) of more than 60% of the epithelial cells (viable epithelium) and cubic morphology. There was no difference in basement membrane between groups. In the dry rat group, hair follicles were located mostly in the dermal matrix of the skin at 0% DHPC, but were found to be located in the dermal adipocytes of the skin in all DPHC-treated groups. On the other hand, most of the hair follicles in the obese rat group were found to be located in the dermal matrix, with or without DPHC treatment.

Experimental Example 5: Skin Elasticity Analysis

The diets containing high fat and high calorie diet were subjected to dietary control for 6 weeks, and DHPC-containing media was applied to the skin once a day for 28 days, The expression levels of elastin and microfibrillar-associated protein (MFAP5) mRNA, which are related to the elasticity of the elastic fibers, were analyzed and the results are shown in FIG. As shown in Fig. 7, the expression of elastin and MFF5 (microfibrillar-associated protein) mRNA was significantly increased in DHPC treatment, especially at 50 (μg / ml)% DHPC, This is a significant effect.

Experimental Example 6: Effect of DHPC on hair cell proliferation

The effect of DHPC on mouse skin follicular cell proliferation was analyzed and the results are shown in FIG. 8 and FIG. As shown in Fig. 8, regardless of the degree of obesity, it was confirmed that DHPC affects hair cell proliferation in the same way. Also, as shown in FIG. 9, it was confirmed that the proliferation regulation of the cells in the hair follicle progressed in a concentration-dependent manner in DPHC in the result of proliferative cell staining by Ki67 immunohistological method.

Experimental Example 7: Effect of DHPC on hair follicle activity

The effect of DHPC on the activity of mouse skin follicles (changes in hair follicle ratio in the hair follicle and the growing stage (Anagen) in the dermal tissue during DPHC treatment) were analyzed and the results are shown in Fig. Mice were treated with DPHC at 0, 001, 01, 1, and 10% (w / v%), respectively, and tissue sections were obtained. The hair follicle stage was analyzed using HE (hematoxylin-eosin) staining method. One-way ANOVA test showed significant differences between the groups. As shown in Fig. 10, when the DPHC treatment was carried out, the ratio of the resting period to the growing period was significantly decreased depending on the concentration. This suggests that DPHC promotes the activity of hair follicles and promotes the formation of hair (Fig. 1). This suggests that DPHC promotes hair follicle development It can be seen that it maintains.

Claims (5)

A composition for accelerating hair growth comprising DPHC (diphlorethohydroxycarmalol) as an active ingredient.
The method according to claim 1,
Wherein said DPHC is derived from Ishige okamurae.
The method according to claim 1,
Wherein the DPHC is contained in an amount of 0.1 to 10% by weight based on the total weight of the composition for promoting hair growth.
The method according to claim 1,
Wherein the composition for promoting hair growth promotes hair follicle cell proliferation or promotes hair follicle activity.
The method according to claim 1,
Wherein the hair growth promoting composition increases the amount of collagen and elastin.

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