JP3218282B2 - Epidermal keratinization accelerator, cosmetic, and bath agent - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a keratinization accelerator, cosmetic and bath agent which are expected to improve rough skin and prevent aging.

[0002]

2. Description of the Related Art The stratum corneum in the skin epidermis plays a series of physiological roles such as moisturizing properties of the skin and physical protection of living bodies, and plays an important role in vital activities. However, the formation of a healthy stratum corneum is often hindered in so-called rough skin with keratinization failure.

[0003] Age-related changes in the skin surface include clinical dryness of the skin. When the rate of keratinization of the epidermal cells decreases with aging, the water retention ability of the skin and the amount of water loss that the skin originally possesses increase, and as a result, a rough skin state is induced.

[0004] In order to prevent such rough skin and the accompanying aging and maintain healthy skin, it is conceivable to administer certain components to normalize skin troubles. Heretofore, as a main method for the above purpose, a method has been adopted in which a moisturizing component is administered to prevent the skin from drying out and give it moisture.

[0005] However, the above-mentioned conventional method generally replenishes water on the keratinous surface or partially replenishes moisturizing components, and the effect is only temporary, and sufficient moisture is maintained inside the skin. (Toshiyuki Takemura:
Pharmacia, 28, 1 (1992)). Therefore,
It was desired to solve this problem.

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

[0007] Further, when a PAF antagonist is applied to the skin as a medicinal product, the anti-inflammatory action (edema, dermatitis, etc.) targeting skin neutrophils, eosinophils, basophils, etc., is not limited. Was what I expected (Merlos M et al .:
Br. J. Pharmacol., 104, 990 (1991), La.
vaud P and others: J. Invest. Dermatol., 97, 101
(1991)).

[0008]

DISCLOSURE OF THE INVENTION The present inventors have made intensive studies in view of such circumstances, and as a result, have found that a PAF antagonist exhibits an effective keratinization-promoting action and normalizes epidermal keratinization. The present invention has been completed,
An object of the present invention is to provide an epidermal keratinization accelerator, a cosmetic, and a bathing agent, which are expected to improve rough skin and prevent aging.

[0009]

The above objects can be attained by an epidermal keratinization accelerator comprising an antagonist to PAF, a cosmetic containing the epidermal keratinization accelerator, and a bath preparation.

[0010]

DESCRIPTION OF THE PREFERRED EMBODIMENTS PAF antagonists used in the present invention include CV-3988, CV-6209, 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, Octyloni
Umbromide ( octylonium bromide ) , chazlenone
Known compounds such as non-lipid antagonists such as ( kadsurenone ) (Muneo Takatani: The latest medicine, 45, 46)
3 (1990)).

[0011] These PAF antagonists can be used in plant extracts and microbial cultures containing them, for example, BN-5.
Ginkgo biloba extract, including the 2020 and related substances ginkgolides (ginkgolides) (Braquet PG addition: Blood Vessel
s, 16, 558 (1985)), kazurenone ( kadsure
none ) containing Spodoptera brilliosa stem extract (Shen TY)
Others: Proc. Natl. Acad. Sci. USA, 82, 672 (1
985)), goboushi and forsythia containing lignans, alnicolides of sesquiterpene lactones ( arni
colide ) (Iwakami S and others)
Chem. Pharm. Bull., 40, 1196 (1992)) and Arnica (Toshi Mikawa: Plant Cell Engineering, 16, 118 (1)
994)) It can be used in the form of an extract or the like.

The epidermal keratinization promoter of the present invention refers to a PAF antagonist itself and may be in any form, but may contain water, alcohol, or other excipients in some cases.

The form of the cosmetic or bath containing the epidermal keratinization promoter of the present invention is not particularly limited.
Ointments, powders, emulsions, lotions, and other common skin external preparations may be used.

The content of the PAF antagonist in the cosmetic amount of the present invention is 0.0000001 to 5 based on the total amount of the composition.
% By weight (hereinafter referred to as%), particularly preferably 0.0000
1-0.005% is preferable. The amount of the plant extract or microbial culture solution containing the PAF antagonist varies depending on the content of the PAF antagonist. For example, 5% by weight of PAF
The Ginkgo biloba dry extract containing antagonists (ginkgolides (ginkgolides)), based on the total composition,
0.000001% or more is preferable, and especially 0.0001%
~ 0.5% is preferred. When the PAF antagonist of the present invention, or a plant extract or a microorganism culture containing the PAF antagonist is used 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. In particular, it is preferable that the amount is 0.00001 to 0.005%. Cosmetics or baths containing these PAF antagonists can be mixed with other components commonly used in cosmetics and baths.

[0015]

The present invention will be described in more detail with reference to the following examples. Prior to the examples, a method for evaluating an epidermal keratinization promoter will be described. The effect of the present invention on keratinization of cells can be evaluated by culturing human epidermal cells and measuring the keratinocyte-forming ability. In addition, the preparation method and measurement method of the reagent used for each test are as follows.

Epidermal keratinization promotion test (a) Cultured epidermal cells As human normal epidermal keratinocytes, a commercially available product (purchased from Cascade Biology) was used.

(B) Medium for Cell Culture The medium is based on MCDB153HAA medium (purchased from Wako) and contains hydrocortisone (0.5 μm).
M), ethanolamine (0.1 mM), phosphoethanolamine (0.1 mM), insulin (5 μg / m
l) and EGF (epidermal growth factor: 10 ng / m
The K-GM medium to which 1) was added was used. During cell growth culture, BPE (Cattle pituitary extract, Cascad)
e Biology) was added in an amount of 2 μl per 1 ml of the medium.

(C) Preparation of Hepes buffer 7.15 g of Hepes (manufactured by Dojindo Laboratories Inc.)
1.8 g of glucose (Kanto Kagaku), 0.22 g of potassium chloride (Kanto Kagaku), 7.7 g of sodium chloride (Kanto Kagaku), disodium hydrogen phosphate dodecahydrate (Kanto Kagaku) ) Dissolve 0.27g in purified water and add 1N
After adjusting to pH 7.4 with aqueous sodium hydroxide solution, 1 l
Up.

(D) Trypsin solution 0.025% trypsin (manufactured by Sigma) and 0.01
A Hepes buffer containing 5% disodium ethylenediaminetetraacetate (Kanto Chemical) was prepared.

(E) Trypsin reaction stopping solution Soybean trypsin inhibitor (hereinafter abbreviated as SBTI, manufactured by Sigma) is dissolved (0.2%) in Hepes buffer prepared in (c), and the trypsin reaction is stopped as an SBTI solution. It was used for.

(F) Cell culture The number of normal human epidermal cells was increased to 1000 in K-GM medium.
0 cells / ml, inoculate 1 ml each on a 24-well collagen-coated plate (manufactured by Falcon), and give 95% (V
/ V) 37% in an atmosphere of air-5% (V / V) carbon dioxide.
The culture was allowed to stand at 4 ° C. for 4 days. The medium was changed every two days.

The culture supernatant was removed by suction, and 1 ml of a K-GM medium containing an epidermal keratinization promoter containing a PAF antagonist at each concentration was added to each dish. The dish was incubated 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 keratinocyte-forming ability The culture supernatant was removed by suction, washed twice with 0.5 ml of Hepes buffer, and treated with 0.15 ml of trypsin solution to remove cells. 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 were subjected to 1% SDS (sodium lauryl sulfate) / 20 mM DTT (@thiothreitol).
It was dissolved in the solution and heat-treated at 90 ° C. for 3 minutes. After heat treatment,
The number of insolubilized cells was measured, and the keratinization ratio was determined from the ratio to the number of cells before heat treatment.

Test Example 1 (Keratinization-Promoting Effect of Epidermal Keratinization Promoter Containing PAF Antagonist) The final concentration of the aqueous solution of the epidermal keratinization accelerator containing the PAF antagonist prepared in Examples 1 to 6 below was 0, 0.03,
The keratinization ratio of epidermal cells when added to a human epidermal cell culture system to be 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 used in an amount of 1000 m of distilled water.
and filtered through a 0.22 μm membrane (Millipore) membrane with a filtration hole diameter of 0.22 μm to give an aqueous solution of WEB 2086 (concentration 1).
(0 μM, 0.0004% by weight).

Example 2 SM-10661 (SM-10661) was used in the same manner as in Example 1 except that 4.8 mg of SM-10661 (Funakoshi) was used instead of WEB2086 and dissolved in 100 ml of distilled water.
(200 μM, 0.0048% by weight) was obtained.

Example 3 BN-5202 was prepared in the same manner as in Example 1 except that 4.08 mg of BN-52020 (Sigma) was used instead of WEB2086 and dissolved in 100 ml of distilled water.
0 (100 μM, 0.0041%) was obtained.

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

Example 5 An aqueous solution of CV-66985 (5 μM, 0.00M) was prepared in the same manner as in Example 1 except that 2.9 mg of U-66985 (Funakoshi) was used in place of WEB2086.
029%).

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

Example 7 (Cream) The following hydrophilic component was heated to 80 ° C. in a hot water bath, and gradually added to the mixed lipophilic component with stirring. This was stirred with a homogenizer to sufficiently emulsify and disperse each component, and then gradually cooled while stirring to obtain a WEB2086-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. 7 g isopropyl myristate 6.0 g isopropyl monostearate 1.3 g polyethylene alkyl ether phosphoric acid 2.3 g glycerin monostearate 2.0 g butyl paraoxybenzoate 0.1 g

Examples 8 to 12 (Cream) Instead of the epidermal keratinization agent consisting of the WEB 2086 aqueous solution of Example 1, an epidermal keratinization agent consisting of the SM-10661 aqueous solution of Example 2 and BN-52020 of Example 3 Epidermal keratinization accelerator comprising an aqueous solution, CV-39 of Example 4
Epidermal keratinization accelerator consisting of 88 aqueous solution, U-
Each of the creams (Examples 8 to 12) was prepared in the same manner as in Example 7 except that the epidermal keratinization accelerator consisting of the aqueous solution 66985 and the FR-49175 aqueous solution of the skin in Example 6 were used. ) Got.

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

Examples 14 to 19 (Lotion) Instead of the epidermal keratinization promoter consisting of the WEB 2086 aqueous solution of Example 1, an epidermal keratinization promoter consisting of the SM-10661 aqueous solution of Example 2, and BN-52020 of Example 3 Epidermal keratinization accelerator comprising an aqueous solution, CV-39 of Example 4
Epidermal keratinization accelerator consisting of 88 aqueous solution, U-
Epidermal keratinization accelerator comprising aqueous solution 66985, Example 6
An epidermal keratinization accelerator comprising an aqueous solution of FR-49175;
And Ginkgo biloba leaf extract (Nippon Powder Chemical Co., Ltd.) 1
Except for using each of 00 mg, each lotion (Examples 14 to 19) was obtained in the same manner as in Example 13.

Examples 20 to 23 (bath agents) Bath agents were prepared with the following compositions.

[0038]

[Table 2]

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

[0040]

According to the present invention, it is apparent that an epidermal keratinization accelerator, a cosmetic, and a bathing agent, which are expected to improve rough skin and prevent aging, can be provided.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61K 35/78 A61K 35/78 X A61P 17/00 A61P 17/00 (56) References JP-A-9-208483 (JP, A JP-A-9-110674 (JP, A) JP-A-9-110673 (JP, A) JP-A-8-245410 (JP, A) JP-A-6-336417 (JP, A) JP-A-7-107 2868 (JP, A) JP-A-7-2841 (JP, A) JP-A-7-277947 (JP, A) JP-A-6-293652 (JP, A) Proc. Natl. Acad. Sc i. , Vol. 82, No. 3, (1985) p. 672-676 Chem. Pharm. Bull. , Vol. 40, no. 5, (1992) p. 1196-1198 (58) Fields investigated (Int. Cl. ⁷ , DB name) A61K 45/00 A61K 7/00 A61K 7/48 A61K 7/50 A61K 35/78 A61P 17/00 CA (STN) EMBASE (STN) MEDLINE (STN)

Claims (3)

(57) [Claims] 1. An epidermal keratinization promoter comprising an antagonist to platelet activating factor. 2. The method according to claim 1, further comprising an antagonist to platelet activating factor.
No, cosmetic, characterized in that it contains a Secco Sui extract. 3. An anti- platelet activating factor antagonist.
Free, bath agent characterized by containing the Secco Sui extract.