JPH05124935A - Hair growth promoter - Google Patents

Abstract

PURPOSE:To provide a hair growth promoter capable of stimulating hair mother cell proliferation leading to contribution to hair growth in terms of hair cycle. CONSTITUTION:The objective promoter giving hair growing activity along with depilation-inhibitory activity through hair mother cell activation, hair growth promotion and depilation-preventive effect, contains, as an active ingredient, platelet-derived growth factor (including PDGF, its effective fragment, its modified product, a modified product from effective fragments of PDGF, or mixture thereof). The PDGF can be isolated from the plasma containing platelet in the blood of animals such as human being, mice or swine (e.g. human platelet) by ion exchange column chromatography, affinity column chromatography, gel filtration and/or dialysis, etc. It is preferable that the amount of the PDGF to be formulated in a cosmetic or hair tonic be 1ng to 1mug/ml (or g) [esp. 10ng to 1mug/ml (or g)].

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a hair growth-promoting agent, and more specifically, it provides hair growth, hair nourishment and hair loss prevention effects by activating hair mother cells, promoting hair growth, and preventing hair loss. It relates to growth promoters.

[0002]

2. Description of the Related Art The mechanism of the hair cycle is roughly as follows. A hair follicle is said to be in the anagen phase during the growth of hair, and when the hair follicles subsequently stop growing and have hair follicles in a contracted state from the anagen phase, the hair follicle is said to be in the telogen phase. When the growing hair follicles enter the resting phase, they undergo a catagen. That is, the hair cycle repeats a growth period, a regression period, and a rest period.

[0003] For example, androgenetic alopecia is not the decrease in the number of hair follicles but the decrease in the size of terminal hairs. The mechanism is that the vascular network of the hair follicle, which gradually decreases in size during repeated hair cycles, atrophies and the plexus dissociates from the hair follicle, but the enzymatic activity involved in energy metabolism causes It is known that there is no qualitative difference. That is, the reason why vellus increases during the development of androgenetic alopecia is that the hair cycle is shortened and the generation of hair follicles is changed at a high resting period. It has been confirmed clinically that this is consistent with the phenomenon of prominence of hair loss before baldness.

Extracts, vitamins, amino acids, hormones and the like extracted from natural products have been used as conventional hair growth, hair nourishing and hair loss preventing agents. For example, capsicum tincture, vitamin E, capronium chloride, senburi and other plant extracts were mixed in various ratios, with the indicators of capillary expansion of the papilla and its duration, and hair growth promotion in animal experiments. Many hair restorers are commercially available in combination with massage.

However, the conventional techniques have no remarkable effect on promoting hair growth and are expected to be more effective. Therefore, an object of the present invention is to provide a hair growth promoter that promotes growth and proliferation of hair mother cells and contributes to hair growth in the hair cycle.

[0006]

[Means for Solving the Problems] In order to solve the above problems, the present inventor has made diligent studies and as a result, has focused on platelet-derived growth factor (PDGF), which is often contained in blood platelets, and has it act on hair cells. However, they have surprisingly found it to be effective against hair growth. PDGF here means
It includes effective fragments of PDGF, modified products of PDGF, modified products of effective fragments of PDGF, and mixtures thereof.

That is, the present invention relates to platelet-derived growth factor (PD
The present invention provides a hair growth promoter containing GF) as an active ingredient. PDGF used in the present invention is present in platelets, a factor that promotes proliferation of fibroblasts, smooth muscle cells, glial cells, etc., from human, mouse, and platelets in blood of animals such as pigs, for example, , Plasma containing human platelets is subjected to any one of ion exchange column chromatography, affinity column chromatography, gel filtration, dialysis, etc.,
Alternatively, it can be isolated by treating with a combination thereof.

More specifically, 1 mM diisopropylfluorophosphate, 0.05 mM
Phenylmethylsulphonyl fluoride
ulfonyl fluoride), 10 mM benzamidine-
HCl, 10 mM EDTA, 0.03 mM L-1-tosylamide-2-
Phenyl-ethyl chloromethyl ketone (L-1-tosylamido
-2-phenyl-ethylchloromethyl ketone), pH 6.1-6.5 at 10
mg / 1 soybean trypsin inhibit
or)), the plasma containing human platelets is allowed to stand overnight, then centrifuged, the supernatant is subjected to gel filtration, and the fraction containing PDGF is boiled for 10 minutes. Centrifuge further and add 0.1M of supernatant.
NaCl, 0.01M sodium phosphate buffer
er) After dialysis against pH 7.4, CM-Sephadex (-Sepha
dex) cation exchange chromatography.
The fraction containing PDGF was added to 0.3 M NaCl, 0.01 M Na-phosphate.
After dialyzing at pH 7.4, Blue-Sepharos
e) It can be isolated by treating with a column [The Journal of Biological Chemistry, 256 , (17), 88.
96-8899, (1981)]. In addition, purified human PDGF is a collaborative, research, and corporation (Collab
commercially available from orative Res. Inc.) may be used.

The isolated and purified PDGF has a molecular weight of 18,000.
It is a relatively large protein with a molecular weight of 34000, in which the A chain of A. and the B chain of 16000 are linked by some S-S bonds. Since the A chain is easily cleaved during the purification process and the molecular weight is reduced to about 11,000, the actual PD
The molecular weight of GF has a wide range of 28,000 to 34,000. Having 10 or more cystines in the molecule, SS
Although forming a bond, it is known to be inactivated by the reduction treatment. In addition, it is a basic peptide having an isoelectric point in the vicinity of pH 9.8 to 10.2 and does not lose its biological activity after heat treatment at 100 ° C for 10 minutes. Since PDGF generally promotes chemotaxis to smooth muscle cells and fibroblasts and has an extremely strong growth promoting action, it is known to act as a wound treatment and a factor in arteriosclerosis.

The PDGF fragment used in the present invention is, for example, a polypeptide and a sugar chain bound by treating the PDGF with trypsin, adding a soybean trypsin inhibitor to stop the reaction, and gel-filtering the reaction solution. Polypeptides are also included. Also, the PDGF ornament used in the present invention,
Alternatively, modified PDGF fragments include, for example, neoglycoprotein, which is a protein to which a monosaccharide or oligosaccharide is covalently bound by chemical synthesis, and the like.

A typical method for preparing neoglycoprotein is an imidator method in which the ε-amino group of the lysine residue of the protein and the N-terminal α-amino group are modified with sugar. The degree of modification with sugar can be freely changed by changing the ratio of 2-imino-2-methoxyethyl-1-thioglycoside to the amino group of protein [protein, nucleic acid, enzyme, 25 (8) , 707-724
(1980)].

In the present invention, PDGF is added to cosmetics or hair in an amount of 1 ng to 10 μg / ml (or g), particularly 10 ng to 1 μg.
It is desirable that the amount of g / ml (or g) be blended in order to exert the effect of the present invention. The hair growth promoter of the present invention includes, in addition to the above essential components, oil, water, a surfactant, a moisturizer,
Alcohol, thickener, fragrance, antioxidant, chelating agent,
It is also possible to mix raw materials usually used as external preparations such as dyes and preservatives.

The dosage form of the present invention is not particularly limited as long as it can be used as an external preparation, but it is particularly preferable to use a coating agent such as an ointment, lotion, tonic, spray, suspension or emulsion. ..

[0014]

The hair growth-promoting agent of the present invention can promote growth and proliferation of hair mother cells and contribute to hair growth in the hair cycle.

[0015]

EXAMPLES The present invention will be described in more detail below with reference to examples.

[0016]

Example 1 The dorsal skin of a newborn C3H mouse was enzymatically separated from the epidermis and dermis using dispase, and hair follicle tissue was separated from the dermis using collagenase. A human PDGF (Collaborative Res.
Inc.) at a final concentration of 50 ng / ml and 500 ng /
Add ³ ml of thymidine to culture D
The proliferation ability of the cells was examined by measuring the NA synthetic activity. As is clear from the results shown in Table 1 below, strong cell division promoting activity was observed. In addition, A: culture solution only,
B: PDGF (50 ng / ml), C: PDGF (50
0 ng / ml), and in Table 1, the activity of A was defined as 100%.

[0017]

[Table 1]

[0018]

Example 2 1. This example is a formulation of a hair nourishing agent suitable for application to the scalp to promote hair growth. W / V% Ethanol 80.00 Glycerin 0.500 Human PDGF (Collaborative Res. Inc .: O.O 1% aqueous solution) 0.05 Dye proper amount Perfume proper amount Purified water proper amount

2. This example is a hair tonic formulation suitable for application to the scalp to promote hair growth. W / V% Salicylic acid 0.100 Propylene glycol 3.00 Glycyrrhetinic acid 0.010 1-Menthol 0.100 Ethanol 50.00 Human PDGF (Collaborative Res. Inc .: O.O1% aqueous solution) 0.05 Perfume Suitable amount Purified water Suitable amount

3. The present embodiment is a formulation of an aerosol type film agent which stimulates the scalp and is effective for hair growth. W / V% Ethylcellulose 8.00 Glycerin 1.00 Mint oil 0.200 Human PDGF (Collaborative Res. Inc .: O.O1% aqueous solution) 0.05 Ethanol 25.75 Propellant (dimethyl ether) 65.00

4. This example is a formulation of lotion to be used topically on the scalp for the purpose of preventing hair loss. W / V% Isopropanol 1.00 Human PDGF (Collaborative Res. Inc .: O.O 1% aqueous solution) 0.05 Ethanol 4.00 Perfume proper amount Purified water proper amount

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Mitsuyuki Hotta 2606-6 Akabane, Kai Town, Haga-gun, Tochigi Prefecture

Claims (1)

[Claims] 1. A hair growth promoter comprising a platelet-derived growth factor as an active ingredient.