JPH02169511A - Skin cosmetic - Google Patents

Abstract

PURPOSE:To obtain a skin cosmetic having excellent preventing effects on aging of skin and beautifying effects on skin, containing transglutaminase and a water-soluble polyhydric alcohol. CONSTITUTION:Transglutaminase, a protein modifying enzyme and a water-soluble polyhydric alcohol (e.g. propylene glycol) are blended with purified water and prepared to give the aimed substance. In order to enhance activity of transglutaminase, preferably a calcium salt is simultaneously blended. Additives such as oily substance, emulsifying agent, colorant, antiseptic, ultraviolet light absorber and antioxidant may be properly blended. The blend can be prepared into a form of lotion, milky lotion or cream. The amount of transglutaminase blended is 0.0001-0.1wt.%, the amount of the water-soluble polyhydric alcohol is 50-5,000 times as much as that of transglutaminase and the amount of the calcium salt is 0.001-0.5wt.%. Since color and fragrance of the cosmetic will not change with time, the cosmetic can be used without anxiety for a long period of time.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention provides skin anti-aging effects (improving rough skin effects, moisturizing effects, etc.) and skin beautifying effects, which are obtained by containing transglutaminase and water-soluble polyhydric alcohol. Regarding excellent skin cosmetics.

[Prior Art and Problems to be Solved by the Invention] The skin plays the role of protecting an individual from the external environment, that is, preventing the invasion of foreign substances and preventing loss of body fluids. Water in the skin has a gradient of water content that decreases as it goes from the dermis to the basal cell layer of the epidermis and then to the stratum corneum, and it constantly moves from the inside of the skin to the outer layer and transpires to the outside through the stratum corneum. ing.

This water evaporation is mainly controlled by the protective function of the dense cell tissue of the stratum corneum. It has been observed that in aged skin or very dry skin, the skin surface becomes dry and lacks smoothness, leading to exfoliation of corneocytes and rough skin. In such cases, it is recognized that the skin's ability to retain moisture is impaired. In other words, it is caused by the condition exceeding the normal control limit of the protective function of the stratum corneum, or by the weakening of the protective function.

As a way to solve these skin problems, if we can densify the structure of the stratum corneum and lamellar granules on the skin surface and activate their defense function, this will improve the moisture retention function of the skin and improve skin health. It is thought that the skin can be maintained in a healthy state and that dry skin can be improved or repaired. However, there are no skin cosmetics that actually improve the condition of the skin surface and truly improve aging skin and dry skin, but only hydrophilic skin moisturizers and oily skin softeners that provide appropriate moisture and oil content. is being incorporated into skin cosmetics. None of the ingredients exhibited skin aging prevention effects or skin beautification effects.

In addition, cosmetics using proteins improved by transglutaminase, which is involved in the construction of the stratum corneum of the skin (Japanese Unexamined Patent Publication No. 6
1-172807) has also been proposed, but it has not improved safety or oral stability.

An object of the present invention is to provide a skin cosmetic that has excellent effects on improving rough skin, preventing skin aging such as moisturizing effects, beautifying skin, safety, and stability over time.

[! [Means for Solving a Problem] Based on the above-mentioned idea, the present inventors conducted intensive research on the components of the biochemical production mechanism of the stratum corneum, the outermost layer of the skin, and found that protein-modifying enzymes When applied to aging or dry skin, skin cosmetics containing transglutaminase and water-soluble polyhydric alcohol act directly on the surface of the skin and the stratum corneum, the outermost layer of the skin. By making the structure more precise,
Improves dry skin, increases moisture retention, and improves skin image quality. Furthermore, they discovered that it has a beautifying effect that imparts flexibility and elasticity, leading to the completion of the present invention.

The present invention is a skin cosmetic containing transglutaminase and a water-soluble polyhydric alcohol.

Moreover, the skin cosmetic further contains a calcium salt.

Transglutaminase used in the present invention (EC2, 3,
2,13 (hereinafter abbreviated as TGase) is one of the enzymes that modify kumbaku substances, and it binds the γ-carboxylamide group of glutamine residues and the ε of lysine residues in proteins and peptides.
- catalyzes the reaction between ε-(T-glutamyl)lysine bonds and crosslink formation reaction through ε-(T-glutamyl)lysine bonds. T.G.
Ase exists in various tissues and blood cells of animals, and is particularly involved in the coagulation reaction of blood-derived fibrin protein and the keratinization reaction of epidermal cells and hair. Among these, TGase is an essential factor during epidermal keratinization, forms a corneocyte membrane, and builds up a very strong outermost layer of the skin. TGas
e is also known to catalyze cross-linking reactions of epidermal-derived proteins in vivo. Furthermore, it is also known that TGa se is activated by divalent calcium ions and catebusin D, which is a lysosomal enzyme.

TGase catalyzes the reaction between free glutamine residues and lysine residues mainly present in the outermost layer of the skin, resulting in ε-(T
- By forming crosslinks consisting of (glutamyl) lysine bonds, the surface structure is densified, improving dry skin, improving the image quality of the skin with a light path that retains moisture, and further increasing the flexibility and elasticity of the skin. It has a beautifying effect on the skin.

The TGa se used in the present invention is the liver of a mammal such as a guinea pig, rat, pig, cow, or sheep.

It can be extracted and purified by known methods from serum, platelets, 9 hair follicles, 1 epidermis, etc. and used. Moreover, those derived from microorganisms can also be used.

The water-soluble polyhydric alcohol used in the present invention together with the above-mentioned TGase acts to suppress the oral deterioration of the above-mentioned TGase in an aqueous solution, and includes, for example, ethylene glycol and propylene glycol.

Dipropylene glycol, 1,3-butylene glycol, 1,4-butylene glycol, dibutylene glycol, glycerin, diglycerin, glucose, maltose, maltitol, sucrose, fructose, xylitol, sorbitol, threitol, erythritol, etc. It will be done. These may be used alone or in combination of two or more.

Furthermore, in order to enhance the activity of TGase, it is preferable to use a calcium salt at the same time. For example, calcium chloride, calcium acetate, calcium lactate, calcium stearate, calcium gluconate, calcium carbonate, calcium oxide, and calcium hydroxide can be used. More preferably, *soluble calcium salts such as calcium chloride, calcium acetate, and calcium lactate are used to significantly enhance the activity.

Additionally, dextrins, cyclodextrins, and starches are traditionally used to stabilize enzymes.

Carboxymethylcellulose, hydroxymethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol
When a water-soluble polymer such as pectin, mannan, gum arabic gelatin, collagen, alginate, xanthan gum, etc. is combined with TGase and incorporated into a skin cosmetic, the stability is further improved.

Furthermore, TGase may be immobilized on polyalkylene glycol, dextran sulfate, etc. by a known method.
It is also possible to stabilize the protein by entrapping it with silk fibroin protein as described in Japanese Patent Application No. 23754.

The skin cosmetic of the present invention can be produced in the same manner as ordinary cosmetics using the above-mentioned TGa s e and a water-soluble polyhydric alcohol.
A skin low silane can be obtained by uniformly mixing a water-soluble polyhydric alcohol and a water-soluble polyhydric alcohol. Cosmetic emulsions and cosmetic creams can also be obtained by combining oily substances and emulsifiers. Of course, the above cosmetics include
Additives such as colorants, preservatives, ultraviolet absorbers, and antioxidants can be added as appropriate.

In the cosmetic composition of the present invention, the amount of transglutaminase blended is preferably set to 0.00001 wt% (hereinafter abbreviated as wt%) to 0.1 wt%. In other words, if it is less than 0.00001 wt%, the enzyme function is not sufficient, and even if it exceeds 0.1 wt%, there is no improvement in the effect commensurate with the increase, and there is a tendency for skin irritation to become stronger. It is. In addition, the blending amount of water-soluble polyhydric alcohol is 50 to 5000 of TGase.
It is preferable to set it so that it is twice as large (based on weight). If it is less than 50 times, the oral stability of the skin cosmetic will decrease and discoloration and odor will occur easily, and if it exceeds 5000 times, the skin cosmetic will feel sticky and undesirable.

Furthermore, the calcium salt used in the present invention contains zero
．． By incorporating OO1 to 0.5 wt%, the enzyme activity can be further enhanced.

〔Example〕

Next, the present invention will be explained based on examples.

The rough skin improvement effect test, keratin improvement effect test, practical test, and oral stability test shown in Examples were conducted as follows.

(Rough skin improvement effect test) The effect of continuous application for 4 weeks was investigated on the lower legs of 20 middle-aged and elderly subjects who complained of rough and dry skin.

Approximately 1 g of the sample was applied to the test site of the left lower leg of the subject once a day, and the condition of the skin before and after the test was judged according to the following criteria. No sample was applied to the right lower leg, which served as a control.

The criterion for skin dryness (for example, +→-1+10→±) was evaluated as valid, 1 level improvement was evaluated as slightly effective 9, and no change was evaluated as invalid. The test results are shown as the number of subjects who found the drug to be effective or somewhat effective.

(Test on the effect of improving keratin) A mending tape (manufactured by Chiban) was adhered to the subject's skin before and after the start of the above-mentioned rough skin improvement effect test, and when the tape was peeled off, the state of the keratinocytes attached to the tape was examined in detail using a scanning electron microscope. I looked into it. Then, the anti-exfoliation property of skin corneocytes was analyzed according to the following criteria, and the corneum improving effect was determined.

: Normal ± : Slight dryness, no flaking + : Mild dryness, flaking ++ : Moderate dryness, flaking +++ : Significant dryness, flaking Comparing the judgment results of the test site and control site before and after the test, the degree of skin dryness is 2 or more levels. In case of improvement, evaluation criteria for keratin improvement effect Evaluation points: 1 No two scales observed 2: Small scales scattered 3: Small to medium scales noticeable 4: Large scales noticeable evaluation is the evaluation score of the test area after 4 weeks of continuous application. A difference of 2 points or more from that of the control site was considered valid, a difference of 1 point was considered somewhat effective, and a difference of 0 points was considered invalid.

The judgment results were expressed as the number of subjects who answered that the test was effective or somewhat effective.

(Practical Test) Samples were applied to 20 female subjects (35 to 55 years old) who complained of rough skin, fine wrinkles, dry skin, etc. twice a day in the morning for three consecutive months, and the following items were evaluated immediately thereafter.

Evaluation items f'lA Moisture: Number of people who said their skin felt moisturized Smoothness: Number of people who said their skin became smooth Elasticity: Number of people who felt their skin became taut Irritation: Irritation caused by irritation when applied Number of people who answered that their skin got a rash2 Number of people who said that their skin got a rash during the period of use (daily stability test) Changes in color and odor after the sample was left in a thermostat at 45℃ for 3M under sealed and light-shielded conditions The presence or absence of was observed.

Example 1 J. Connellan et al. method (Journal of
Biological Chemistry Vol. 246, p. 1093,
TGase was prepared from guinea pig liver according to the method described in 1971) and Japanese Patent Application Laid-Open No. 175884. 0.25M in 500g of fresh guinea pig liver
A homogenate was prepared by adding a 1.5° C. sucrose solution using a Polytron (manufactured by Kinematica), and a crude fraction of TGase was obtained from the supernatant by centrifugation. This fraction was collected using DEAR cellulose column chroma I/graph (2 m
MEDTA, 5mM) Lis-HCl buffer PH7,5) and 10% agarose gel column chromatography (B
Purification was performed using iogel 1, 0.5M). Finally, TGase was obtained by ultrafiltration and lyophilization.

Using the TGase obtained by the above method, skin low silon was obtained by uniformly mixing these components into the following raw material composition.

Composition Amount (wt%) TGase
O,01 Calcium chloride 0.1 Glycerin 10. OO Purified water Remaining amount when the total amount is 100 A skin lotion was obtained by uniformly mixing these ingredients with the following raw material composition.

Composition Mixture ffi (wt%) TG
ase O,1 Calcium lactate 0.1 1.3-butylene glycol 10.0 Purified water
Remaining amount with a total amount of 100 Example 2 Using TGase obtained in the same manner as in Example 1, a skin lotion was obtained by uniformly mixing these ingredients with the following raw material composition.

Composition Formulation 1i (wt%) TGa
se O,0001 Calcium acetate
0.05 Propylene glycol 1. OOOO purified water
Remaining amount where the total amount is lOO Example 3 Using TGase obtained in the same manner as in Example 1, Example 4 Using TGase obtained in the same manner as in Example 1, the following raw material composition was prepared and these components were prepared. A skin lotion was obtained by uniformly mixing the ingredients.

Composition Formulation 1ii (wL%) TG
ase 0.1 Dipropylene glycol 5.0 Purified water Remaining amount when total amount is 100 Example 5 Using TC obtained in the same manner as in Example 1; An aqueous phase was prepared. Then, the water phase heated to <80°C was added to the oil phase heated to 80°C, and the mixture was rapidly cooled while stirring uniformly to obtain skin milk.

Composition Blend 1 (wt%) (oil phase) Liquid paraffin 20. OO Cetyl alcohol 5.00 Polyoxyethylene sorbitan monooleate 5.00 (Aqueous phase) Ga5e Calcium chloride diglycerin methylparaben Purified water 0.01 0.2 5.00 0, IO Remaining amount with total amount as 100 Example 6 Implementation Using TGase obtained in the same manner as in Example 1, an oil phase and an aqueous phase were prepared with the following raw material compositions. Then, an aqueous phase also heated to 80"C was added to the oil phase heated to 80"C, and the mixture was quickly cooled while stirring uniformly to obtain a skin cream.

Composition Blend It (wt%) (oil phase) Octyldodecyl myristate 35. OO Cetyl alcohol 5.00 Cetyl palmitate 2.00 Sorbitan sesquistearate 3.00 (aqueous phase) TGase O,01 Glycerin 15. OO booxyethylene sorbitan monooleate (20E, 0.) 5.00
Methylparaben 0.10 Purified water
Comparative Example 1 of Remaining Amount with Total Amount as 100 Using TGase obtained in the same manner as in Example 1, a skin lotion was obtained by uniformly mixing these ingredients with the following raw material composition.

Composition Amount (wt%) TGa
se 0.0
Calcium monochloride 0.1 Purified water
Comparative Example 2 of Remaining Amount with Total ■ as 100 A crosslinked polymeric protein was obtained by the method described in Example 1 of JP-A-61-172807. That is, milk αS1 casein 50
g to O,LM Tris-HCl buffer (pH 7,6) containing 5mM calcium chloride and 20mM dithiothreitol.
) 1ON, TC; aselg obtained in the same manner as in Example 1 of the present invention was added, and the mixture was incubated at 37°C for 30 minutes to react.

After the reaction was completed, the mixture was desalted by dialysis and freeze-dried to obtain a powdery white crosslinked polymer. A skin milk was obtained by blending 5% of this cross-linked polymer instead of TGase and stirring uniformly in the same manner as in Example 5, except that purified water was added thereto.

Comparative Example 3 A skin lotion was obtained by uniformly mixing the following raw material composition.

Composition Blend 1 (wt%) Glycerin 10.00 Purified water Remaining amount when the total amount is 100 For the 6 types of experimental products and 3MB comparative product obtained as above, each test was conducted according to the procedure described above. conducted and evaluated. The results are shown in Table 1. As can be seen from this table, all the implementation products are T
C; Comparative example 1 using ase alone. Comparative Example 2 containing a cross-linked protein obtained by the action of TGase and Comparative Example 3 using water-soluble polyhydric '2 alcohol alone showed superior effects in improving the skin.

Further, there is no irritation or rash to the skin as in Comparative Examples 1 and 2. In addition, the oral stability, which is a problem with comparative products, is not a problem at all with experimental products.

〔Effect of the invention〕

As mentioned above, the skin cosmetics of the present invention contain TGa
Since it contains se and a water-soluble polyhydric alcohol, using this product can prevent dryness and aging of the skin without causing rough skin. Furthermore, this cosmetic has the advantage that its color and odor do not change orally and can be used safely for a long period of time.

Claims (2)

[Claims] (1) A skin cosmetic containing transglutaminase and a water-soluble polyhydric alcohol. (2) The skin cosmetic according to claim 1, further comprising a calcium salt.