JPH09301881A - Epidermis cornification promoting agent, cosmetic and bathing agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an epidermis cornification promoting agent, a cosmetic and a bathing agent acting on epidermis and expected to be effective for improving roughened skin and preventing the aging of the skin. SOLUTION: A roughened skin improving effect, etc., can be achieved by an epidermis cornification promoting agent composed of an antagonistic substance against a platelet activation factor(PAF) and a cosmetic and a bathing agent containing the epidermis cornification promoting agent.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an epidermal keratinization accelerator, cosmetics, and bath agents, which are expected to improve rough skin and prevent aging.

[0002]

BACKGROUND OF THE INVENTION The stratum corneum in the epidermis of the skin plays a series of physiological roles such as moisturizing ability of the skin and physical protection of the living body, and plays an important role in life activities. However, the formation of a healthy stratum corneum is often hindered in so-called rough skin accompanied by keratinization failure.

[0003] The age-related changes in the skin surface include clinically dry skin. When the rate of keratinization of epidermal cells decreases with aging, the water-retaining ability originally possessed by the skin and the amount of water vaporization increase, resulting in a rough skin condition.

[0004] In order to prevent such rough skin and aging associated with it and maintain healthy skin, it is considered that the skin trouble is normalized by administering a certain component. Heretofore, as a main method for the above-mentioned purpose, a method of preventing a dry state of the skin and moisturizing it by administering a moisturizing component has been taken.

However, the above-mentioned conventional method generally replenishes water on the surface of the corneum or partially replenishes the moisturizing component, and the effect is only temporary, and sufficient moisture is maintained inside the skin. Cannot be given (Toshiyuki Takemura:
Pharmacia, 28, 1 (1992)). Therefore,
A solution to this problem was desired.

On the other hand, Platelet Activator
ating factor; hereinafter abbreviated as PAF) is being developed for pharmaceutical applications such as asthma, and is known to suppress physiological activities of PAF such as platelet aggregation inhibitory action, airway stenosis inhibitory action, and blood pressure lowering action. (Terazawa Michio: Protein Nucleic Acid Enzyme, 36, 184 (199
1)). However, the action of PAF on epidermal cells is not well known, and PAF is slightly synthesized in human epidermal cells (Michel L et al .: J. Invest. Dermato.
l., 95, 576 (1990)), and the presence of PAF receptors in epidermal cells (Travers JB et al .: J. Inve.
st. Dermatol., 105, 816 (1995)) was only reported.

Also, when a PAF antagonist is applied to the skin as a medicinal use, the anti-inflammatory action (edema, dermatitis, etc.) that targets skin neutrophils, eosinophils, basophils, etc. It was what I expected (Merlos M et al .:
Br. J. Pharmacol., 104, 990 (1991), La.
vaud P et al .: J. Invest. Dermatol., 97, 101
(1991)).

[0008]

DISCLOSURE OF THE INVENTION The present inventors have conducted extensive studies in view of such circumstances, and as a result, found that a PAF antagonist has an effective keratinization promoting action and normalizes keratinization of the epidermis, The present invention has been completed,
Its purpose is to provide an epidermal keratinization accelerator, a cosmetic and a bath agent, which are expected to improve rough skin and prevent aging.

[0009]

The above-mentioned object is achieved by an epidermal keratinization promoting agent comprising an antagonist to PAF, and a cosmetic and a bath agent containing the epidermal keratinization promoting agent.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION PAF antagonists used in the present invention include CV-3988, CV-6209 and U-6.
6985, R-74717, SIR63-441, ON
Lipid antagonists such as O-6240, WEB2086,
BN-52020, FR-49175, SM-1066
1, L-652731, PCA-4248, octylonium
Known compounds such as non-lipid antagonists such as bromide and kadsurenone are mentioned (Muneo Takatani: The latest medicine, 4
5, 463 (1990)).

Further, these PAF antagonists are plant extracts and microbial cultures containing them, such as BN-52.
Ginkgo biloba extract containing 020 and related substances (ginkgolides) (Braquet PG and others: Blood Vessels, 16, 558)
(1985)), and the extract of Spodoptera scutella containing kadsurenone (Shen TY et al .: Proc. Natl. Acad. Sci.
USA, 82, 672 (1985)), burdock and forsythia containing lignans, and gypsum (Iwakami S et al .: Chem. Pharm. Bull., 40, 1196 (199) containing arnicolides of sesquiterpene lactones.
2)) and Arnica (Mikawa Ushio: Plant Cell Engineering, 16, 1)
18 (1994)) extract and the like.

The epidermal keratinization promoting agent of the present invention refers to a PAF antagonist itself and its form is not limited, but it may contain water, alcohol or other excipients depending on the case.

The form of the cosmetic or bathing agent containing the epidermal keratinization promoting agent of the present invention is not particularly limited, and examples thereof include an aqueous solution, an aqueous alcohol solution, an emulsion,
The form of a general external preparation for skin such as ointment, powder, emulsion or lotion can be mentioned.

The content of the PAF antagonist in the cosmetic of the present invention is 0.0000001-5 based on the total amount of the composition.
% By weight (hereinafter referred to as%) is preferable, and particularly 0.0000
1 to 0.005% is preferable. The blending amount of the plant extract or the microorganism culture solution containing the PAF antagonist varies depending on the content of the PAF antagonist. For example, 5% by weight PAF
Ginkgo biloba leaf dry extract containing antagonist (ginkgolides) is 0.000001% based on the total amount of the composition.
The above is preferable, and 0.0001 to 0.5% is particularly preferable. When using the PAF antagonist of the present invention, or a plant extract or a microbial culture containing the PAF antagonist as a bathing agent, the content in the bath water at the time of use is preferably 0.0000001 to 5% as the PAF antagonist. , Especially 0.
It is preferable to mix it so as to be 0,001 to 0.005%. The cosmetics or bath agents containing these PAF antagonists can be blended with other components usually used in cosmetics and bath agents at the same time.

[0015]

The present invention will be described in more detail with reference to the following examples. Prior to the examples, a method for evaluating the epidermal keratinization promoting agent will be described. The effect of the present invention on keratinization of cells can be evaluated by culturing human epidermal cells and measuring the corneal membrane forming ability. The methods for preparing and measuring the reagents used in each test are as follows.

Epidermal keratinization promotion test (a) Cultured epidermal cells Human normal epidermal keratinocytes were commercially available products (purchased from Cascade Biology).

(B) Cell culture medium The medium is MCDB153HAA medium (purchased from Wako), and hydrocortisone (0.5 μm) is used as a base.
M), ethanolamine (0.1 mM), phosphoethanolamine (0.1 mM), insulin (5 μg / m
1) and EGF (epithelial cell growth factor: 10 ng / m)
The K-GM medium to which 1) was added was used. Also, during cell culture, BPE (bovine pituitary extract, Cascad
(purchased from e Biology) was added in an amount of 2 μl per 1 ml of the medium.

(C) Preparation of Hepes buffer solution Hepes (manufactured by Dojindo Laboratories Ltd.) 7.15 g,
Glucose (Kanto Chemical Co., Ltd.) 1.8 g, potassium chloride (Kanto Chemical Co., Ltd.) 0.22 g, sodium chloride (Kanto Chemical Co., Ltd.) 7.7 g, disodium hydrogen phosphate dodecahydrate (Kanto Chemical Co., Ltd.) ) 0.27 g was dissolved in purified water to give 1N
After adjusting the pH to 7.4 with an aqueous solution of sodium hydroxide, 1 liter
Up.

(D) Trypsin solution 0.025% trypsin (manufactured by Sigma) and 0.01
A Hepes buffer solution containing% disodium ethylenediaminetetraacetate (manufactured by Kanto Chemical Co., Inc.) was prepared.

(E) Trypsin reaction termination solution Soybean trypsin inhibitor (hereinafter abbreviated as SBTI, manufactured by Sigma) was dissolved in the Hepes buffer prepared in (c) (0.2%) to terminate the trypsin reaction as an SBTI solution. Used for.

(F) Cell culture The number of normal human epidermal cells was determined to be 1000 in K-GM medium.
It was adjusted to 0 cells / ml and seeded on a 24-well collagen-coated plate (manufactured by Falcon Co., Ltd.) in an amount of 1 ml each, and 95% (V
/ V) air-5% (V / V) carbon dioxide gas atmosphere, 37
The cells were cultivated at static temperature for 4 days. The medium was changed every two days.

The culture supernatant was removed by aspiration, and 1 ml of K-GM medium containing the epidermal keratinization promoting agent containing each concentration of PAF antagonist was added to each dish. This dish was statically cultured at 37 ° C. for 6 days in an atmosphere of 95% (V / V) air-5% (V / V) carbon dioxide gas. The medium was changed every two days.

(G) Measurement of Corneal Membrane Forming Ability After removing the culture supernatant by suction and washing twice with 0.5 ml of Hepes buffer, the cells were treated with 0.15 ml of trypsin solution and detached, and then 0 The reaction was stopped with 0.35 ml of SBTI solution, and the number of cells in each well was counted with a hemocytometer. After counting the number of cells, the remaining cells are treated with 1% SDS (sodium lauryl sulfate) / 20 mM DTT (dithiothreitol).
It was dissolved in the liquid and heat-treated at 90 ° C. for 3 minutes. After heat treatment,
The number of insolubilized cells was counted, and the keratinization rate was calculated from the ratio with the number of cells before heat treatment.

Test Example 1 (Effect of keratinization of epidermal keratinization promoting agent containing PAF antagonist) An aqueous solution of epidermal keratinization promoting agent containing PAF antagonist prepared in Examples 1 to 6 below was used as a final concentration. 0, 0.03,
The keratinization rate of epidermal cells when added to the human epidermal cell culture system at 0.1, 0.3, 1, 3, or 10 μM was measured (Table 1). As a result, it was found that keratinization of human epidermal cells was promoted depending on the concentration.

[0025]

[Table 1]

Example 1 4.1 mg of WEB 2086 (synthesized according to the method described in JP-A-61-176591) was added to 1000 m of distilled water.
l, and filtered with a membrane having a pore size of 0.22 μm (manufactured by Millipore) to obtain an aqueous solution of WEB2086 (concentration: 1).
An epidermal keratinization promoter consisting of 0 μM, 0.0004% by weight) was obtained.

Example 2 SM-10661 was prepared in the same manner as in Example 1 except that 4.8 mg of SM-10661 (Funakoshi Co., Ltd.) was used in place of WEB2086 and dissolved in 100 ml of distilled water.
An epidermal keratinization promoter consisting of the above aqueous solution (200 μM, 0.0048% by weight) was obtained.

Example 3 BN-5202 was prepared in the same manner as in Example 1 except that BN-52020 (Sigma Co.) 4.08 mg was used in place of WEB2086 and dissolved in 100 ml of distilled water.
An epidermal keratinization promoter consisting of 0 aqueous solution (100 μM, 0.0041%) was obtained.

Example 4 An aqueous solution of CV-3988 (10 μM, 0.00 μM) was prepared in the same manner as in Example 1 except that 5.9 mg of CV-3988 (Cosmo Bio) was used instead of WEB2086.
059% by weight) was obtained.

Example 5 An aqueous solution of CV-66985 (5 μM, 0.005) was prepared in the same manner as in Example 1 except that 2.9 mg of U-66985 (Funakoshi Co., Ltd.) was used instead of WEB2086.
029%) was obtained.

Example 6 An aqueous solution of FR-49175 (10 μM, 10 μM, was used in the same manner as in Example 1 except that 3.57 mg of FR-49175 (Sigma) was used instead of WEB2086.
An epidermal keratinization promoter consisting of 0.00036%) was obtained.

Example 7 (Cream) The following hydrophilic component was heated to 80 ° C. in a water bath and gradually added to the mixed lipophilic component described below with stirring. This was stirred with a homogenizer to sufficiently emulsify and disperse each component, and then gradually cooled with stirring to obtain a WEB 2086-containing cream.

"Hydrophilic component" Methyl paraoxybenzoate 0.1 g Propylene glycol 6.7 g Epidermal keratinization promoter of Example 1 44.1 g "Lipophilic component" Squalane 4.7 g White petrolatum 24.0 g Stearyl alcohol 8. 7g Isopropyl myristate 6.0g Isopropyl monostearate 1.3g Polyethylene alkyl ether phosphate 2.3g Glycerin monostearate 2.0g Butyl paraoxybenzoate 0.1g

Examples 8 to 12 (cream) Instead of the epidermal keratinization promoter consisting of the WEB2086 aqueous solution of Example 1, the epidermal keratinization promoting agent consisting of the SM-10661 aqueous solution of Example 2 and BN-52020 of Example 3 were used. Epidermal keratinization promoter consisting of aqueous solution, CV-39 of Example 4
Epidermal keratinization promoter consisting of 88 aqueous solution, U-of Example 5
Each cream (Examples 8 to 12) was prepared in the same manner as in Example 7, except that the epidermal keratinization promoter composed of the 66985 aqueous solution and the epidermal keratinization promoter composed of the FR-49175 aqueous solution of Example 6 were used. ) Got.

Example 13 (Lotion) 1 ml of the epidermal keratinization promoter of Example 1 was added to the following composition to obtain 100 g of lotion. Ethanol 10.0 g Lactic acid 0.3 g Sodium citrate 0.1 g Glycerin 2.0 g Preservatives, fragrances and surfactants Appropriate amount Purified water Remaining amount

Examples 14 to 19 (lotion) Instead of the epidermal keratinization promoter consisting of the WEB2086 aqueous solution of Example 1, a skin keratinization promoting agent consisting of the SM-10661 aqueous solution of Example 2 and BN-52020 of Example 3 were used. Epidermal keratinization promoter consisting of aqueous solution, CV-39 of Example 4
Epidermal keratinization promoter consisting of 88 aqueous solution, U-of Example 5
Epidermal keratinization promoter consisting of 66985 aqueous solution, Example 6
Epidermal keratinization promoter consisting of FR-49175 aqueous solution,
And ginkgo biloba leaf extract (Japan Powder Chemicals Co., Ltd.) 1
Lotions (Examples 14 to 19) were obtained in the same manner as in Example 13, except that 00 mg was used.

Examples 20 to 23 (Bathing Agent) Bathing agents having the compositions shown below were prepared.

[0038]

[Table 2]

Preparation method: Each component was mixed to prepare a bathing agent. In addition, this bath agent is diluted about 3000 times at the time of use.

[0040]

EFFECTS OF THE INVENTION It is apparent that the present invention can provide an epidermal keratinization promoting agent, a cosmetic, and a bath agent, which are expected to improve rough skin and prevent aging.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI technical display location A61K 45/00 A61K 45/00

Claims (3)

[Claims] 1. An epidermal keratinization promoting agent comprising an antagonist to a platelet activating factor. 2. A cosmetic containing the epidermal keratinization promoter according to claim 1. 3. A bath salt containing the epidermal keratinization promoter according to claim 1.