JP4452203B2 - Whitening cosmetics - Google Patents

Description

  The present invention relates to a serine protease activity inhibitor. In particular, the present invention relates to a serine protease activity inhibitor having an excellent antagonistic action against the activity of serine protease related to trypsin.

  Conventionally, whitening cosmetics have been screened for ingredients that have the action of inhibiting melanin production or reducing the produced melanin pigment in order to prevent or improve skin darkening due to ultraviolet rays and skin pigmentation such as spots and freckles. Has been formulated. For example, ascorbic acid, cysteine, and derivatives thereof, placental extracts, extracts from plants and algae are used.

  However, ascorbic acid, cysteine, and derivatives thereof are susceptible to redox reactions and are unstable. In addition, placental extracts, extracts from plants and algae are accompanied by an unpleasant odor and coloring when whitening cosmetics are incorporated in an effective amount. Further, the whitening effect is not sufficient, and there are many problems.

Recently, attempts have been made to suppress skin pigmentation by suppressing melanosome phagocytosis by keratinocytes as described in Patent Document 1. Melanin is produced in melanosomes among melanocytes, which are phagocytosed by keratinocytes. This causes melanin to be distributed throughout the epidermis. By using a trypsin inhibitor, the melanosome phagocytosis by keratinocytes is suppressed and the diffusion of melanin is prevented. It did not exhibit a sufficient effect as a cosmetic.

Serine protease is a protease having a serine residue in the active site, and representative ones include trypsin, chymotrypsin, thrombin, plasmin, elastase, etc., and Patent Documents 2 and 3 describe serine protease inhibitors. However, none of them are related to anti-aging effects and rough skin improvement effects, and no content relating to whitening is disclosed or suggested.
JP-T-2001-502360 JP-A-11-124325 JP 2000-327555 A

An object of the present invention is to solve the above-mentioned problems, have a strong skin melanin production inhibitory action, and inhibit the dispersion of the produced melanin to keratinocytes, thereby causing skin irritation and skin sensitization. It is to provide a whitening cosmetic that is highly effective and has no safety problems.

As a result of intensive studies in view of the above circumstances, the present invention considers that it is effective to inhibit the activity of serine protease, and as a result of examining serine protease activity inhibitory action for a wide variety of substances, Thr-Arg, Leu Serine protease inhibitor containing one or more compounds selected from -Leu-Arg, Thr-Lys-Pro-Arg, Tyr-Ile-Gly-Ser-Arg, Arg ethyl ester and melanin production inhibition By containing an agent, it has been found that it has an extremely high whitening effect, and the present invention has been completed.

That is, by activating protease activator receptor-2 (PAR-2) on the cell membrane of keratinocytes by serine protease, incorporation of melanosomes into keratinocytes is activated. The serine protease activity inhibitor of the present invention inhibits the activation of PAR-2 on the keratinocyte cell membrane, thereby suppressing the incorporation of melanosomes into keratinocytes and preventing the diffusion of melanin into the stratum corneum. By preventing the diffusion of melanin, the skin color is not darkened. In addition, whitening cosmetics that have a strong whitening effect by reducing the amount of melanin produced by using a melanin production inhibitor that is a material that directly inhibits melanin production in melanocytes, such as an ascorbic acid derivative, are provided. It is done.

As described in detail above, the present invention has a very high whitening effect and high safety by blending a melanin production inhibitor and a serine protease inhibitor, and thus can be widely applied to cosmetics.

Hereinafter, the present invention will be described in detail. Commercially available reagents are used for Thr-Arg, Leu-Leu-Arg, Thr-Lys-Pro-Arg, Tyr-Ile-Gly-Ser-Arg, and Arg ethyl ester showing serine protease inhibitory action used in the present invention be able to. It is also possible to extract and purify from various proteins that are randomly decomposed by hydrolysis and enzymatic degradation.

Melanin production inhibitors used in the present invention are ellagic acid and its derivatives and their salts, endothelin antagonists, ascorbic acid and its derivatives and their salts, glutathione and their derivatives and their salts, cysteine and its derivatives and their salts In addition, commercially available reagents can be used for resorcin and derivatives thereof and salts thereof, hydroquinone and derivatives thereof and salts thereof. Various solvents from plants containing a large amount of these compounds, such as water; lower monohydric alcohols such as methyl alcohol and ethyl alcohol; liquid polyhydric alcohols such as glycerin, propylene glycol, and 1,3-butylene glycol; acetone Extraction using one or more of alkyl esters such as ethyl acetate; hydrocarbons such as ethyl acetate; hydrocarbons such as benzene and hexane; ethers such as diethyl ether; halogenated alkanes such as dichloromethane and chloroform; It can be used after purification. Furthermore, the above compound can be prepared by chemical synthesis.

In addition, grabrizine, glabrene, liquiritin, isoliquiritin and licorice extract, placenta extract, carotenoids and animal and plant extracts, irasen extract, sunflower seed extract, ibukitorano extract, ages extract containing these , Ogon extract, Ononis extract, Seaweed extract, Kujin extract, Sesame extract, Plant extract containing linoleic acid, Plant extract containing linolenic acid, Saicin extract, Hawthorn extract, Shirayuri extract , Peony extract, Sempukuka extract, Sakuhakuhi extract, soybean extract, tea extract, Toki extract, sandalwood extract, beech extract, grape seed extract, hop extract, mica extract, yukinoshita extract, The preparation of Yokuinin extract is not particularly limited. It is extracted under heating from a low temperature with an equivalent organic solvent. Examples of the extraction solvent include water; lower monohydric alcohols such as methyl alcohol and ethyl alcohol; liquid polyhydric alcohols such as glycerin, propylene glycol, and 1,3-butylene glycol; ketones such as acetone and methyl ethyl ketone; and ethyl acetate. One or more of alkyl esters; hydrocarbons such as benzene and hexane; ethers such as diethyl ether; and halogenated alkanes such as dichloromethane and chloroform can be used. In particular, one, two or more mixed solvents of water, ethyl alcohol, and 1,3-butylene glycol are particularly suitable.

The amount of Thr-Arg, Leu-Leu-Arg, Thr-Lys-Pro-Arg, Tyr-Ile-Gly-Ser-Arg, Arg ethyl ester showing serine protease inhibitory action used in the present invention is 0.0001 to 10.0% by weight is preferable, and the range of 0.01 to 1.0% by weight is particularly optimal.

Ellagic acid and its derivatives and their salts, endothelin antagonists, ascorbic acid and its derivatives and their salts, glutathione and their derivatives and their salts, cysteine and its derivatives and their salts, resorcin and their use as melanin production inhibitors Derivatives thereof and salts thereof, hydroquinone and derivatives thereof, and salts thereof, glabrizine, glabrene, liquiritin, isoliquiritin and licorice extract containing them, placenta extract, carotenoids and animal and plant extracts containing them, irasen extract , Sunflower seed extract, Ibukitorano extract, Ages extract, Ogon extract, Ononis extract, Seaweed extract, Kujin extract, Gokahi extract, Plant extract containing linoleic acid, Plant containing linolenic acid Extract, Saishin extract, Hawthorn extract, Shirayuri extract, Peonies extract, Sempukuka extract, Sakuhakuhi extract, Soybean extract, Tea extract, Toki extract, Sandalwood extract, Beech extract, Grape seed extract Products, hop extract, mica squid extract, yukinoshita extract, yokoinin extract, dried by drying each material, then finely pulverized 1 to 1000 times, especially 10 to 100 times the solvent In the case of normal temperature extraction, it is preferably performed at 0 ° C. or higher, particularly 20 ° C. to 40 ° C. for 1 hour or longer, particularly 3 to 7 days. Moreover, you may heat-extract at 60-100 degreeC for 1 hour.

Each of the above-mentioned extracts obtained under the above conditions may be used as an extracted solution. However, if necessary, it is necessary to appropriately use a concentrated and powdered product after filtration or the like. I can do it.

The blending amount of the melanin production inhibitor in the cosmetic of the present invention is preferably 0.0001 to 20.0% by weight, particularly 0.01 to 10.0% by weight in terms of the evaporated and dried content. .

In addition to the above essential components, the cosmetics of the present invention are aqueous components, oily components, plant extracts, animal extracts, powders, excipients, surfactants, oils, alcohols, pH adjusters, preservatives, antioxidants. It is prepared by mixing agents and thickeners, sweeteners, pigments, fragrances, and the like as necessary and blending appropriately. The dosage form of the cosmetic of the present invention is not particularly limited, and can be various dosage forms such as lotion, milky lotion, cream, pack, powder, spray, ointment, dispersion, and cleaning agent.

Hereinafter, examples relating to the whitening effect by the serine protease inhibitor and the melanin production inhibitor according to the present invention will be shown and application examples of cosmetics using the raw materials will be described, but the examples are not limited thereto. Needless to say.

Commercially available reagents were used for Thr-Arg, Leu-Leu-Arg, Thr-Lys-Pro-Arg, Tyr-Ile-Gly-Ser-Arg, and Arg ethyl ester showing serine protease inhibitory action. Thr-Arg is Thr-Arg from Sigma, Leu-Leu-Arg is Leupeputin from Wako Pure Chemical Industries, Thr-Lys-Pro-Arg is Tuftsin from Wako Pure Chemical Industries, Tyr-Ile-Gly- Ser-Arg used Laminin Pentapeptide from Wako Pure Chemical Industries, and Arg ethyl ester used Sigma L-arginine ethyl ester dihydrochloride.

(Measurement of trypsin inhibitory activity)
(Reagent)
・ Phosphate buffer solution: Mix 0.1M-NaH ₂ PO ₄ and 0.1M-Na ₂ HPO ₄ at 5:25 to prepare PH7.4.
-Substrate solution: Add 0.6 g of Hammerstein Cazein to 80 ml of 0.05M-Na ₂ HPO ₄ and dissolve by heating in a boiling water bath. After cooling, adjust PH to 7.4.
Trypsin solution: 1: 250 trypsin (DIFCO) is dissolved in a phosphate buffer so as to be 10 μg / ml.
-Precipitation reagent: Prepare solutions of trichloroacetic acid, sodium acetate, and acetic acid solutions to concentrations of 0.11M, 0.22M, and 0.33M, respectively.
Sample solution: Prepare by adding water or ethanol to a predetermined concentration.
(Measurement)
Add 0.1 ml of sample solution to 0.9 ml of casein substrate solution and mix. Further, add 2 ml of trypsin solution, stir and leave at 37 ° C. for 10 minutes. Add 3 ml of precipitation reagent, stir well and let stand at room temperature for 15 minutes. Thereafter, the mixture is centrifuged at 3.000 rpm for 15 minutes, and the supernatant is taken and the absorbance at 280 nm is measured.
(Measurement of inhibition rate)
• For control (C), trypsin solution is added after the precipitation reagent.
-The control (T) uses distilled water instead of the sample.
・ The test section (S) was obtained by performing the above operation using each sample.

Table 1 shows the trypsin inhibition rates of various samples. Benzamidine is a substance used in various applications as a trypsin inhibitor, and showed an inhibition rate of 10.3% at a concentration of 0.1%. Arg-Ala and Arg-glutamate were tested as dipeptides with Arg at the N-terminus, and showed only 1.7% and -3.3% inhibitory activity at 0.1% concentration. Further, Gly-Gly was tested as a dipeptide not containing Arg, but at a concentration of 0.1%, it was -4.5%, indicating no inhibitory activity. In contrast, Thr-Arg, a dipeptide having an Arg at the C-terminal, showed 14.6% inhibitory activity at 0.1% concentration. Arg ethyl ester exhibited 11.1% inhibitory activity at 0.1% concentration. Furthermore, C-terminal arginine tripeptide, tetrapeptide, and pentapeptide Leu-Leu-Arg, Thr-Lys-Pro-Arg, and Tyr-Ile-Gly-Ser-Arg were tested. 65.0%, 29.4% and 52.2% showed high inhibitory activity. In addition, Gly-Tyr-Ala, Tyr-Gly-Gly, and Val-Tyr-Val were tested as tripeptides that do not contain arginine, but at 0.1% concentration, the inhibitory activity was 2.5%, 3.6%, and 0.3%, respectively. There was no inhibitory activity. From the above results, it was found that the peptide having arginine at the C-terminus has high trypsin inhibitory activity.

Next, although the example of the formulation example of the cosmetics which mix | blended various components of this invention is shown, this invention is not limited to this.
Examples of cosmetic formulations

(1) Cosmetic cream
(weight%)
a) Beeswax
2.0
b) Stearyl alcohol
5.0
c) Stearic acid
8.0
d) Squalane
10.0
e) Self-emulsifying glyceryl monostearate
3.0
f) Polyoxyethylene cetyl ether (20E.O.)
1.0
g) Jojoba oil
10.0
h)
Tyr-Ile-Gly-Ser-Arg
0.5
i) Ascorbic acid glucoside 2.0
j) 1,3-butylene glycol
5.0
k) Potassium hydroxide
0.3
l) Preservatives and antioxidants
Appropriate amount
m) Purified water
The remaining preparation methods a) to f) are dissolved by heating and kept at 80 ° C. Heat up to H) to m), keep at 80 ° C., add to a) to f), emulsify, and cool to 40 ° C. with stirring. Then g) is added and stirred to dissolve uniformly.

(2) Latex
(weight%)
a) Beeswax
0.5
b) Petrolatum
2.0
c) Squalane 8.0
d) Sorbitan sesquioleate
0.8
e) Polyoxyethylene oleyl ether (20E.O.) 1.2
f) Thr-Arg
1.0
g) Arbutin
2.0
h) 1,3-butylene glycol
7.0
i) Carboxyvinyl polymer
0.2
j) Potassium hydroxide
0.1
k) Purified water
The rest
l) Preservatives and antioxidants
Appropriate amount
m) ethanol
7.0
The process up to production methods a) to e) are dissolved by heating and kept at 80 ° C. Heat up to h) to l), keep at 80 ° C., add to a) to e), emulsify, and cool to 50 ° C. with stirring. Add f), g), m) at 50 ° C., stir to 40 ° C. and cool.

(3) Lotion
(weight%)
a) Leu-Leu-Arg
0.1
b) Magnesium ascorbate phosphate 1.0
c) Glycerin
5.0
d) Polyoxyethylene sorbitan monolaurate (20E.O.) 1.0
e) ethanol
6.0
f) Fragrance
Appropriate amount
g) Preservatives and antioxidants
h) Purified water
The remaining preparation methods a) and b) are mixed uniformly. c) to h) are mixed and dissolved uniformly. Mix two agents when using.

(4) Lotion
(weight%)
a)
Thr-Lys-Pro-Arg 0.05
b) Glutathione 0.1
c) Glycerin
5.0
d) Polyoxyethylene sorbitan monolaurate (20E.O.) 1.0
e) ethanol
6.0
f) Fragrance
Appropriate amount
g) Preservatives and antioxidants
h) Purified water
Mix the remaining preparation methods a) to b). c) to h) are mixed and dissolved uniformly. Mix two agents when using.

(5) Face wash
(weight%)
a) Thr-Arg
0.5
b) Ascorbic acid glucoside
2.0
c) Peonies extract 0.5
d) Talc
The rest
e) Cellulose
20.0
f) Potassium myristate
30.0
g) Sodium lauryl phosphate
10.0
h) Fragrance
Appropriate amount
i) Preservative
Appropriate amount of production method a) to i) are mixed and stirred and dispersed well to make uniform.

[Effect confirmation test]
(1) As a test subject for human effect by application, 20 females aged 25 to 55 years were allowed to use each of the products of the present invention and the comparative product twice a day (morning and night) for 2 consecutive months. The state of the part was compared before and after the test, and the improvement effect was investigated. In this test, the cosmetics indicated by 0025 as Example 1 were used. In Comparative Example 1, cosmetics obtained by removing Tyr-Ile-Gly-Ser-Arg from the cosmetics shown in 0025 were used. A cosmetic was prepared by removing ascorbic acid glucoside from the cosmetic shown in 0025, and the effect of its use was investigated. While using cosmetics containing the active ingredient of the present invention daily, skin smears and whitening effects were tabulated before and after application for 2 months to confirm the effects. The results and evaluation criteria are shown in Tables 2 and 3.

As is clear from Table 2, Comparative Example 1 consisting only of a conventional melanin production inhibitor
Compared with Comparative Example 2 consisting only of a serine protease inhibitor, Example 1 consisting of a melanin production inhibitor and a serine protease inhibitor gave a total score of 73 points. On the other hand, Comparative Example 1 had 51 points and Comparative Example 2 had a total score of 49 points, indicating a high whitening effect of the product of the present invention. In addition, 20 subjects who used Example 1 did not generate any panelists who felt abnormal skin during the test period.

Cosmetics containing the melanin production inhibitor and serine protease inhibitor of the present invention have excellent whitening effect and high safety, and therefore can be applied to various whitening cosmetics.

Claims (3)

Thr-Arg, Leu-Leu- Arg, 1 or more kinds of serine protease activity inhibitors comprising a compound selected from the group consisting of Thr-Lys-Pro-Arg. A whitening cosmetic comprising the serine protease activity inhibitor according to claim 1. The serine protease activity inhibitor according to claim 1, ellagic acid and derivatives thereof, and salts thereof, endothelin antagonists, ascorbic acid and derivatives thereof, and salts thereof, glutathione and derivatives thereof, salts thereof, cysteine and derivatives thereof, and Salts thereof, resorcin and derivatives thereof, and salts thereof, hydroquinone and derivatives thereof, and salts thereof, glabrizine, glabrene, liquiritin, isoliquiritin and licorice extract, placenta extract, carotenoids containing these, and animals and plants containing these Extract, Ireisen extract, Sunflower seed extract, Ibukitorano extract, Agetsu extract, Ogon extract, Ononis extract, Seaweed extract, Kudin extract, Sesame extract, Oil and fat containing linoleic acid, Linolenic acid Including Oil, Saishin extract, Hawthorn extract, Shirayuri extract, Peonies extract, Sempukuka extract, Sakuhakuhi extract, Soybean extract, Tea extract, Toki extract, Juniper extract, Beech extract, Grape seed extract things, hop extract, Maikaika extract, saxifrage extract, whitening cosmetics, characterized in that contain one or more melanin production inhibitors selected from Coix extract.