JPH0657652B2 - Skin aging prevention cosmetics - Google Patents

Description

Description: FIELD OF THE INVENTION The present invention relates to a skin anti-aging cosmetic (a skin cosmetic used for preventing skin aging).

More specifically, the present invention relates to a skin cosmetic having excellent skin aging prevention effects (rough skin improving effect, keratin improving effect, turnover speed increasing effect, beautiful skin effect, etc.).

(Prior Art) Aged skin is dry and non-smooth rough skin in which keratinocyte exfoliation phenomenon is observed. It is said that aging skin has a slow turnover speed, and that the turnover speed increases when the skin has an antiaging effect.

The present applicant has previously found and proposed that vitamin E orotate and γ-aminobutyric acid-based compounds enhance the skin function by the peripheral vasodilatory action of the skin and have an antiaging effect (Japanese Patent Publication No. No. 52-2979 and Japanese Patent Publication No. 58-26726).

In addition, it was found and submitted that diisopropylamine dichloroacetate enhances the skin function by the skin tissue activating action and similarly has the skin aging preventing effect. (JP-A-53-196528).

However, all of these effects are slow-acting, and the effect appears after 6 months in the case of cream and after 3 months in the case of lotion, which is not sufficiently satisfactory and leaves room for improvement.

DISCLOSURE OF THE INVENTION The inventors of the present invention have conducted extensive studies as an improvement of this difficulty, and as a result, in the skin cosmetic base, the epidermal growth factor (Ep
idermal Growth Factor, hereinafter abbreviated as EGF),
When blended with at least one compound selected from γ-aminobutyric acid, vitamin E orotate, and diisopropylamine dichloroacetate, the synergistic effect of the two causes an excellent anti-aging effect on skin (improves rough skin, keratin) It has been found that a rapid-acting anti-aging cosmetic composition is obtained, which is excellent in improving effect and has an effect of accelerating turnover speed) within a very short period of 1-2 months after the start of use, and lasting. The present invention has been completed.

(Object of the Invention) The present invention provides a skin anti-aging cosmetic composition having an excellent anti-aging effect on skin (excellent effect on rough skin, effect on improving keratin, fast turnover speed, beautiful skin effect, etc.). There is.

(Structure of the Invention) The present invention comprises a skin cosmetic base comprising EGF and at least one compound selected from γ-aminobutyric acid, vitamin E orotate, and diisopropylamine dichloroacetate to prevent skin aging. Cosmetics.

(Detailed Description of Configuration) The EGF according to the present invention is widely used in the submandibular gland, thyroid, pancreas, kidney, plasma, saliva, urine of mammals such as humans, mice, rats, rabbits, horses, cows and sheep. It is present in various organs or body fluids, but especially in the mouse submandibular gland (0.3-1 μg / m
l), a kind of peptide hormone present in human urine (25 to 250 ng / ml) at high concentration. [Refer to "Cell Growth Factors", pp. 20-30, Asakura Shoten Publishing Co., Ltd., 1984] EGF is obtained by extraction and purification from the mouse submandibular gland or human urine described above. It can also be extracted from fermentation products. For example, the method of extracting and purifying EGF from the submandibular gland of a mouse is as shown in the following experimental examples. [Journal of Biological Chemistry, Volume 237, 1552-
1562 (J, Biol, Chem, Vol 1237; 1555-
1562)] (Experimental Example-1) 200 g of distilled water was added to 22 g of the submandibular gland collected from 150 male adult mice (body weight 25 g or more), and the mixture was crushed with a homomixer and then heated at a temperature of 0 to 3 ° C. Centrifuge for minutes to obtain the extract.

An antibiotic, streptomycin, is added to the extract to adjust the PH value to 6.8 to 7.1, the mixture is allowed to stand overnight at a temperature of 0 ° C., and then centrifuged for 5 minutes to obtain an extract.

To this extract, 100 g of solid ammonium sulfate was added, and the mixture was left standing at a temperature of 0 ° C. for 30 minutes and then centrifuged to obtain a precipitate.

A solution obtained by suspending this precipitate in 15 ml of distilled water was dialyzed in distilled water (the distilled water of 2 was exchanged 5 times in 24 hours), then allowed to stand in a boiling water bath for 5 minutes, and then cooled to 10
The extract is obtained by centrifuging for minutes.

After dialyzing this extract for 24 hours in the same manner as in the above, using a carboxymethylcellulose column, a 0.01 molar sodium acetate buffer solution having a PH value of 4.2 was used as an effluent to give a fraction having a high absorbance at 280 nm. Collect. Then, after dialyzing for 24 hours in the same manner as in the above section, it was dried under reduced pressure to obtain 6.5 mg of the target mouse EGF.

The mouse EGF obtained in the above (Experimental Example-1) has asparagine and arginine at both ends, as shown in the following formula.
It is a peptide consisting of 3 amino acids and has a molecular weight of 6.
045, the isoelectric point (PI) was 4.6, and the extinction coefficient (EC) was 30.9. It was also confirmed that there were three SS bonds in the molecule.

(Mouse EGF Amino Acid Primary Structure) Asn-Ser-Tyr-Pro-Gly-Cys-Pro-Ser-Ser-Tyr-
Asp-Gly-Tyr-Cys-Leu-Asn-Gly-Gly-Val-Cys-
Met-His-Ile-Glu-Ser-Leu-Asp-Ser-Tyr-Thr-
Cys-Asn-Cys-Val-Ile-Gly-Tyr-Ser-Gly-Asp-
Arg-Cys-Gln-Thr-Arg-Asp-Leu-Arg-Trp-Trp-
Glu-Leu-Arg The three letters in the formula are abbreviations for amino acids as shown in Table 2.

(Experimental Example-2) 100 g of urinary protein was obtained by concentrating 10 Kl of human urine by extraction with acetone as a solvent by a conventional method.

100 g of this urinary protein was suspended in 400 ml of distilled water to obtain a suspension, and this suspension was replaced with the extract obtained in (Experimental Example-1) Human EGF (5.5 mg) was obtained in the same manner as in the steps (1) to (3).

The human EGF obtained in the above (Experimental Example-2) is a peptide consisting of 53 amino acids whose both ends are asparagine and arginine, and has a molecular weight of 6,201, as shown in the following formula. Isoelectric point (PI) is 4.
5, and it was also confirmed that there were three SS bonds in the molecule.

(Human EGF amino acid primary structure) Asn-Ser-Asp-Ser-Glu-Cys-Pro-Leu-Ser-His-
Asp-Gly-Tyr-Cys-Leu-His-Asp-Gly-Val-Cys-
Met-Tyr-Ile-Glu-Ala-Leu-Asp-Lys-Tyr-Ala-
Cys-Asn-Cys-Val-Val-Gly-Tyr-Ile-Gly-Glu-
Arg-Cys-Gln-Tyr-Arg-Asp-Leu-Lys-Trp-Trp-
GLu-Leu-Arg Regarding the three letters in the formula, in addition to those shown in Table 2, Lys
Is lysine and Ala is alanine.

It is known by the present inventors that EGF promotes proliferation and differentiation of various cells even in a small amount such as about 1 ng by an experiment using tissue culture [see the above-mentioned "cell growth factor"]. Note that each of the 1 μg / ml aqueous solutions of mouse and human EGF was applied to the upper arm of 30 women complaining of rough skin twice a morning and evening for 3 consecutive months, and as a result, an improving effect was observed.

It is a compound known to the skin-activating components γ-aminobutyric acid, vitamin E orotate and diisopropylamine dichloroacetate according to the present invention, and each of them is described in detail in the above publication.

In the cosmetic of the present invention, at least one of EGF and γ-aminobutyric acid, vitamin E orotate, and diisopropylamine dichloroacetate is directly added to a base such as cream, emulsion, lotion, or a compound thereof. Is dissolved in an oil phase component, an aqueous phase component, a solvent such as alcohol, and the like to be mixed, and then subjected to treatments such as emulsification, mixing, dispersion, dissolution and solubilization. The above-mentioned γ-aminobutyric acid, vitamin E orotate, and diisopropylamine dichloroacetate mixed in cosmetics are stable and absorbed into the skin to enhance the skin function by the peripheral vasodilatory effect and prevent wrinkles on the skin. In addition to providing a smooth and moist skin, it has an excellent effect of preventing skin aging (effects of improving rough skin and keratin, and accelerating turnover speed) synergistically with EGF in a short time. , Lasting, etc. can have remarkable effects.

The amount of EGF compounded is based on the total amount of the cosmetic of the present invention.
0.0001 to 0.1% by weight (hereinafter abbreviated as wt%), γ-
Similarly, the amount of aminobutyric acid is 0.01 to 1 wt%, the amount of vitamin E orotate is 0.01 to 1 wt%, and the amount of diisopropylamine dichloroacetate is 0.01 to 1%.
It may be 1.0 wt%. Even if the upper limit of each of these blending amounts is exceeded, the effect commensurate with the blending amount exceeding that cannot be expected, and if the blending amount is less than the lower limit, the object of the present invention cannot be achieved.

In addition to the above, a colorant, a fragrance, an antiseptic, a surfactant, a pigment, an antioxidant and the like can be appropriately added to the skin cosmetic of the present invention within the range where the object of the present invention is achieved.

Examples will be described below.

The methods for measuring the turnover speed of the stratum corneum, the measurement test method for improving rough skin, the measurement test method for improving stratum corneum, and the sensory test described in Examples are as follows.

(1) Method for measuring turnover speed of stratum corneum 5w of dansyl chloride, which is the foundation of fluorescent color, in white petrolatum
Make an ointment containing t% and apply it to the skin of the forearm of the subject 24
Apply occluded for a time and allow dansyl chloride to permeate the stratum corneum. After that, apply the test sample to the same site twice a day (morning and evening) and examine the fluorescence of dansyl chloride every day.
The number of days until the fluorescence disappeared was defined as the turnover speed of the stratum corneum of the skin. The normal turnover rate of the stratum corneum of the skin is 14 to 16 days, but in aged skin, the turnover speed extends to around 18 days. On the other hand, when the anti-aging effect appears, it is shortened to around 12 days.

(2) Test method for measuring rough skin improving effect The application effect for 20 consecutive weeks was examined for 20 middle-aged and elderly subjects who have rough skin on their lower legs. About 1 g of cream was applied to the left lower leg test site of the test subject twice a day, and the skin condition before and after the test was measured according to the criteria in Table 3. The lower right leg was not coated with the sample and served as a control.

Table 3 Judgment criteria for skin dryness −: Normal ±: Minor dryness, no desquamation +: Dryness, mild desquamation ++: Dryness, moderate desquamation +++: Dryness, remarkable desquamation The determination results of the test site before and after the test and the control site are shown. In comparison, when the skin dryness is improved by two or more stages (for example, + → −,
++ → ±) is defined as “valid”, a case of improvement by 1 level is defined as “slightly valid”, and a case of no change is defined as “invalid”. The judgment result was shown by the number of subjects who were “effective” and “somewhat effective”.

(3) Measurement test method for keratin improvement (increased keratinocyte anti-exfoliation) effect Scotch tape (Nichiban Mending Tape) on the skin of the test area before and after the start of the rough skin improvement measurement test described above
The state of keratinocytes adhered to the tape when adhered was examined by a scanning electron microscope in detail, the skin keratinocyte anti-peelability was separated according to the criteria in Table 4, and the effect of improving keratin was determined.

Table 4 Criteria for keratin improvement effect (increased keratinocyte anti-exfoliation) Evaluation point 1: No scale is recognized 〃: Small scale is scattered 〃 3: Small to medium scale is remarkable 〃 4: Large scale is remarkable When the difference between the evaluation point of the test site and that of the control site after continuous application for 4 weeks was 2 points or more, it was defined as "effective", 1 point was defined as "moderately effective", and 0 point was defined as "ineffective". The judgment result was shown by the number of subjects who were “effective” and “somewhat effective”.

(4) Sensory test (Beautiful skin effect test) Female test subject who complains of rough skin, fine wrinkles, dry skin, etc. (35
The sample was applied to 20 people (up to 55 years old) twice a day (morning and evening) for 3 consecutive months, and the effect after 1, 2 and 3 months was evaluated. The evaluation results were "wet skin,""smoothskin," and "tension on skin" for each item of wettability, smoothness, and elasticity of skin. The number of people is shown.

Examples 1 to 6 and Comparative Examples 1 to 5 [Skin cream] The mouse and human EGFs obtained in Experimental Examples 1 and 2 above,
Γ-aminobutyric acid, vitamin E orotate, diisopropylamine dichloroacetate, which are skin activating ingredients,
Each skin cream was prepared by blending as described in the table, and the above-mentioned tests were carried out.

In addition, γ-aminobutyric acid is abbreviated as GABA, vitamin E orotate is abbreviated as VEOT, and diisopropylamine dichloroacetate is abbreviated as DADA.

(1) Composition (2) Preparation method Each of the components (A) and (B) was dissolved by heating at 80 ° C, mixed, and cooled to 30 ° C with stirring to prepare each skin cream. Incidentally, (C) component γ-aminobutyric acid and diisopropylamine dichloroacetate are dissolved in the (B) component,
Vitamin E orotate was dissolved in the component (A).

(3) Characteristics Table 5 shows the results of various tests conducted on each skin cream.

As shown in Table 5, the skin creams containing only the EGF or the skin activating component of Comparative Examples 1-5 did not provide sufficient effects in various properties, and thus E of Examples 1 to 6 of the present invention were not obtained.
A skin cream containing at least one of GF, GABA, VEOT, and DADA showed remarkable effects in various properties, and a sensory test showed excellent skin-beautifying effect 1-2 months after application of the sample.

Examples 7 to 10 and Comparative Example 6 [Skin lotion] Similar to Example 1, each skin lotion was prepared using the following raw material components, and the results of various properties are shown in Table 6.

Raw material components, ethanol 10 wt%, glycerin 5
wt%, polyoxyethylene sorbitan monooleate (solubilizing agent) 0.2 wt%, preservative 0.01 wt%, perfume 0.01 wt%, and a suitable amount of colorant purified water as the remaining amount to make the total amount 100 wt% A skin lotion was prepared according to a conventional method (Comparative Example 6). Next, this comparative example 2
To the same ingredients as the skin lotion, add the ingredients shown in Table 6 and add purified water as the remaining amount to make the total amount 1
The amount was adjusted to 00 wt% and skin lotions were prepared in the same manner (Examples 7 to 10).

As shown in Table 6, the skin lotions of Examples 7 to 10 of the present invention showed excellent effects in various characteristics, and the amount of EGF was 0.0001 to 0.1 wt% to achieve the object of the present invention. obtain.

(Effects of the Invention) As described above, the present invention clearly provides a skin anti-aging cosmetic composition which is excellent in rough skin improving effect, keratin improving effect, and effect of accelerating turnover speed, and also has a remarkable skin beautiful effect. Is.

Claims (1)

[Claims] 1. A skin anti-aging cosmetic composition comprising an epidermal growth factor and at least one compound selected from γ-aminobutyric acid, vitamin E orotate, and diisopropylamine dichloroacetate in a skin cosmetic base. .