JP3159419B2 - Cosmetics - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to the synthesis of collagen proteins.
Related to cosmetics that promote smoothness .

[0002]

2. Description of the Related Art Compared with ordinary protein turnover (metabolism), the turnover rate of collagen is extremely slow, and even under physiological conditions, the turnover rate of collagen further decreases with aging. It is known. The rate of turnover of collagen is determined by the rate of degradation and the rate of synthesis of collagen. Such a decrease in the rate of turnover associated with aging leads to an increase in the cross-linked structure of collagen (aged cross-linking). Involved in the formation.
The increase in hard-to-decompose / hard-to-extract hard collagen impairs the function of collagen involved in proliferation, differentiation and migration as a cell scaffold, reduces cell activity, and further reduces collagen turnover. (Hyundai Chemistry, December, p. 36, 1990)
Year).

[0003] In order to prevent such a decrease in the rate of collagen turnover due to aging, the activity of collagenase, a key enzyme of collagen metabolism, is stimulated to promote the decomposition and the rate of collagen synthesis is increased. New collagen needs to be supplied.

[0004] Ascorbic acid derivatives, estradiol, testosterone and the like are known as those which activate the ability of cells to synthesize collagen. They increase the rate of collagen synthesis by stimulating the collagen gene or releasing the rate-limiting process after protein synthesis.

However, even when the above-mentioned collagen synthesis promoter is used, the amount of synthesized collagen is actually insufficient.

[0006]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to provide a cosmetic composition which smoothly promotes the synthesis of collagen protein.
To provide a fee .

[0007]

SUMMARY OF THE INVENTION The above-mentioned object is achieved by the following method.
, Proline and alanine in three molar ratios
This is achieved by a cosmetic characterized by containing 3: 2: 1 .

Glycine, proline,
Each amino acid molar ratio of alanine, the strongest effect of nearby, promoting synthesis to the amino acid composition ratio of collagen 3: 2: 1
(Glycine Proline: Alanine) Ru der (see Test Example 2).

A collagen synthesis promoter can be prepared from these amino acids and used as a supplement in a cell system to which a collagen synthesis promoter has been added. May be used.

As the collagen synthesis promoting substance used in the present invention, those generally known as collagen synthesis promoting substance can be used. However, when applied to the skin as anti-aging cosmetics, the presence of dermal fibroblasts is considered. Since it has a large effect on the dermis layer (connective tissue), low molecular substances such as ascorbic acid and its derivatives, estrogen, and testosterone are desirable.

The collagenase production promoter used in the present invention includes silk peptide (Japanese Patent Application No. 3-59752),
Serine, N-methylserine (JP-A-4-1130),
Sulfate, ammonium and nitrate (Japanese Patent Application No. Hei 4-3556)
No. 52), ethanolamine derivatives (JP-A-3-294)
222), pentoxifylline and the like. A collagen metabolism activator containing these collagenase production promoter and collagen synthesis promoter can promote both synthesis and decomposition of collagen.

The above-mentioned collagen synthesis promoter and collagen metabolism activator are formulated into various dosage forms such as a liquid preparation, a solid preparation and a semi-solid preparation by blending them with known components which are generally used according to the purpose of use. Preferred compositions include ointments, gels, creams, sprays, patches, lotions, powders, granules, tablets and the like.

For example, the above-mentioned collagen synthesis promoter may be used in combination with hydrocarbons such as petrolatum, higher alcohols such as stearyl alcohol, lower alkyl esters of higher fatty acids such as isopropyl myristate, animal fats and oils such as lanolin, and glycerin. By mixing with surfactants such as polyhydric alcohols, glycerin fatty acid esters and polyethylene glycol monostearate, inorganic salts, waxes, resins, water and, if necessary, preservatives such as methyl parahydroxybenzoate and butyl paraoxybenzoate, cosmetics And can manufacture pharmaceuticals.

The content of the collagen synthesis promoter at that time varies depending on the dosage form, but is preferably 0.001 to 2% by weight (as amino acid solids), more preferably 0.011% by weight, based on the total amount of the applied composition. １1% by weight. The content of the collagen synthesis promoting substance such as an ascorbic acid derivative is preferably 0.01 to 10% by weight, more preferably 0.1 to 3% by weight. However, those that are diluted at the time of use, such as bath salts, can be added in a further increased amount. Furthermore, when used as an adjunct to cell lines that contain pre collagen synthesis-promoting substances are, for 10 ⁵ U / ml of cell lines, a collagen synthesis promoter solution 30mM contained as for example amino acids, 0.5 to 2. What is necessary is just to add about 0 volume%.

[0015]

Industrial Applicability The cosmetic of the present invention acts on fibroblasts to enhance the ability of collagen synthesis-promoting substances such as ascorbic acid derivatives to synthesize collagen proteins.

[0016]

EXAMPLES Hereinafter, the present invention will be described in detail with reference to examples. Prior to the description, a method for testing the ability of the present invention to promote collagen synthesis will be described.

(Test Method for Measuring Collagen Synthesis-Promoting Ability) The cell density of a normal human fibroblast cell line [CCD45SK (ATCC CRL 1506) collected from the skin of a Caucasian woman] was adjusted to 10% by volume of fetal bovine serum (hereinafter abbreviated as FBS). The mixture was adjusted to 1 × 10 ⁵ cells / ml in a MEM medium containing the cells, and 0.4 ml of each was inoculated into a 24-well plate (4 × 10 ⁴ cells / well). Cultured.

The MEM medium was prepared by adding 0.12% by weight of sodium bicarbonate to the minimum essential medium 10-101 manufactured by Dainippon Pharmaceutical Co., Ltd.

After 24 hours, the culture was aspirated off and the cells were washed twice with MEM medium supplemented with a final concentration of 0.6% by volume of FBS, and then 400 μl of disodium ascorbate sulfate at a final concentration of 500 μg / ml. And MEM medium supplemented with 0.6% by volume of FBS.

A collagen synthesis accelerator solution was sterilized by filtration through a nitrocellulose membrane having a pore size of 0.2 μm (manufactured by Advantech Toyo, DISMIC-25), and then added to each well to a final concentration of 0.5 to 2% by volume.

After culturing under the same conditions for 24 hours, β-aminopropionitrile was added to a final concentration of 50 μg / ml and tritium-L-proline to a final concentration of 1 μCi / ml, and the cells were further cultured for 24 hours (culture test solution). .

Quantitative determination of collagen production: The collagen production was measured by the radioactivity incorporated into the collagen fraction, which was resistant to pepsin and fractionated according to the salt concentration-dependent solubility, from the above culture test solution. Collagen fractionation by pepsin treatment and salt concentration was performed according to the method of Webster et al. (Analytical Biochemistry, p. 220, 1979).
Year).

[0023]sample1-3 The three amino acids of glycine, proline and alanine are described in Table 1.
And mixed in a molar ratio of 1: 1: 1 (sample
1), 1: 2: 3 (sample2), 3: 2: 1 (sample3)
A collagen synthesis promoter was obtained.

[0024]

[Table 1] Displays the amount of each amino acid in g when mixed

In the cell addition experiment, samples 1 to 3 were used.
279.29 mg of collagen synthesis promoter, 28
6.31 mg and 272.29 mg were dissolved in water so that the final volume of each was 100 ml to prepare a collagen synthesis promoter solution having a total amino acid concentration of 30 mM.

(Test Example 1) A solution of the collagen synthesis promoter of Sample 1 was added to a final concentration of 0.5 to
Table 2 shows the results of the above-mentioned test conducted by adding 1.5% by volume to a cell culture system containing disodium ascorbic acid sulfate. As Comparative Example 1, a group was prepared in which no collagen synthesis accelerator solution was added (only disodium ascorbic acid sulfate).

[0027]

[Table 2] Mean ± standard deviation (n = 4)

As can be seen from Table 2, the collagen synthesis promoter of Sample 1 exhibited a strong collagen synthesis promoting activity in a dose-dependent manner.

(Test Example 2) Collagen synthesis promoters of Samples 1 to 3 were dissolved in water to prepare a collagen synthesis promoter solution having a total amino acid concentration of 30 mM, and a final concentration of 1% by volume, disodium ascorbate sulfate was used. Table 3 shows the results of the same test as in Test Example 1 performed by adding the compound to a cell culture system containing. As Comparative Example 2, no collagen synthesis promoter solution was added (only disodium ascorbic acid sulfate).
Groups were provided.

[0030]

[Table 3] Mean ± standard deviation (n = 3)

As can be seen from Table 3, Samples 1 to 3 all enhance the collagen synthesis promoting ability of ascorbic acid sulfate disodium, but the collagen synthesis promoter of Sample 3 is the strongest. Had the ability to promote.

From the above results, it was found that the collagen synthesis promoter used in the present invention acts on fibroblasts to increase the effect of the ascorbic acid derivative which is a collagen synthesis promoter.

The following is a formulation example of a composition to which the above-mentioned collagen synthesis promoter or collagen metabolism activator is applied.

Example 1-3 mg of the collagen synthesis promoter of the ointment sample 3 and the following hydrophilic component were mixed by heating to 80 ° C in a hot water bath.
The mixture was gradually added to the following mixture of lipophilic components which had been heated. Next, a homogenizer (TOKUSYUKIKA KOGYO
Vigorous stirring (2500 rpm) for 2 and a half minutes to sufficiently emulsify and disperse each component.
An ointment containing 0.003% by weight of a collagen synthesis promoter in g was obtained.

“Hydrophilic component” (g) Magnesium ascorbic acid phosphate ester 0.03 Methyl paraoxybenzoate 0.1 Propylene glycol 6.7 Purified water 44.1

"Lipophilic component" Squalane 4.7 White petrolatum 24.0 Stearyl alcohol 8.7 Isopropyl myristate 6.0 Polyethylene glycol monostearate (45E.O.) (trade name NIKKOL MYS-45, Nippon Surfactant Industries, Ltd.) 1.3 Dipolyoxyethylene alkyl ether phosphoric acid (2E.O.) (trade name: NIKKOL DDP-2, manufactured by Nippon Surfactant Industries, Ltd.) 2.3 Glycerin monostearate 2.0 Paraoxybenzoate Butyl acid 0.1 0.1---------------------------100

Example 2 -Lotion (Preparation of Collagenase Production Accelerator) 56 g of refined silk raw material was dissolved in 400 ml of a 60% by weight aqueous solution of calcium chloride by heating. At this time, 160 ml of ethyl alcohol was added to facilitate dissolution. Next, this was dialyzed for 24 hours and concentrated to obtain 312 ml of an aqueous solution of 18% by weight of solubilized fibroin.

235 ml of concentrated sulfuric acid was slowly added to 235 ml of the mixture, and the mixture was heated at 60 ° C. for 6 hours. Then, water was added thereto to make a total volume of 1.8 liters, left overnight at room temperature, neutralized with sodium hydroxide while cooling with ice, and filtered to obtain a silk peptide solution (concentration: 235% by weight).

(Preparation of Collagen Metabolism Activator) The silk peptide solution obtained above and the collagen synthesis promoter of Sample 3 were blended according to the following table to prepare a lotion (a lotion containing a collagen metabolism activator).

Weight% Sample 3 collagen synthesis promoter 1.0 Ascorbic acid phosphate magnesium salt 0.03 Collagenase production promoter (silk peptide solution) 0.1 Ethanol 10.0 Lactate 0.3 Sodium citrate 1 Glycerin 2.0 Preservatives, flavors and surfactants Appropriate amount Purified water Remaining amount----------------------------------------------------------------------------- ---- 100% by weight

Example 3 -Lotion% by weight Collagen synthesis promoter of Sample 3 0.001 Ethanol 10.0 Lactic acid 0.3 Sodium citrate 0.1 Glycerin 2.0 Preservative, flavor and surfactant Appropriate amount Purified water residue Amount 100% by weight-----------------------------------------

Example 4 -Lotion% by weight Collagen synthesis promoter of sample 3 2.0 Ascorbic acid sulfate disodium salt 0.01 N-methyl-L-serine 2.0 N-methylethanolamine 3.0 Ethanol 10 0.0 Lactic acid 0.3 Sodium citrate 0.1 Glycerin 2.0 Preservatives, flavors and surfactants Appropriate amount Purified water Remaining amount -------------- ------ 100% by weight

Example 5 3 g of the collagen synthesis promoter of the bath preparation sample 3 and the following components were mixed to obtain 100 g of a bath preparation as a collagen metabolism activator.

Perfume 0.1 g Organic pigment 10 mg Sodium bicarbonate 14.9 g Sodium sulfate 50.0 g Ammonium sulfate 35.0 g Ascorbic acid sulfate disodium salt 0.3 g

The bath agent is diluted about 3000 times at the time of use.

Example 6 -Cream The components (C) shown in the following table were uniformly mixed and dissolved at about 80 ° C., added to the components (A) which had been uniformly mixed and dissolved at about 80 ° C., and emulsified. About 50
The component (B), which was uniformly mixed and dissolved at a temperature of ° C, was added, and the mixture was cooled to about 30 ° C to prepare the mixture.

(A) Compounding amount (%) Squalane 10.0 Olive oil 10.0 Solid paraffin 5.0 Cetanol 4.0 Sorbitan monostearate 2.0 Polyoxyethylene sorbitan monostearate (20E.O.) 0 (B) Collagen synthesis promoter of sample 3 0.5 Ascorbic acid sulfate disodium salt 2.0 N-methyl-L-serine 2.0 Purified water 25.5 (C) Glycerin 5.0 Methyl paraben 0.1 Purified water balance

──────────────────────────────────────────────────続 き Continued on front page (58) Field surveyed (Int. Cl. ⁷ , DB name) A61K 7/00

Claims (3)

(57) [Claims] 1. A cosmetic comprising three amino acids of glycine, proline and alanine in a molar ratio of 3: 2: 1. 2. The method according to claim 1, further comprising: ascorbic acid sulfate.
The cosmetic according to claim 1, which contains. 3. The method according to claim 1, wherein the ascorbic acid sulfate is Asco.
3. The cosmetic according to claim 2, which is a disodium salt of rubic acid sulfate.
Fees.