JP2003144095A - Skin care preparation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a skin care preparation, a food and drink for skin care and cosmetic useful for prevention of chapping, wrinkles, reduction of elasticity of the skin or the like. SOLUTION: The skin care preparation, the food and drink for skin care and the cosmetic includes a basic protein fractionation originating from milk or a basic peptide fraction obtained from hydrolysis of the protein with a protease such as pepsin, pancreatin, or the like. The basic protein fraction and the basic peptide fraction have a function of increasing amount of collagen in the skin.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a skin beautifying agent, a skin beautifying food and drink, and a cosmetic which are useful for preventing rough skin, wrinkles, elasticity deterioration and the like.

[0002]

2. Description of the Related Art In recent years, research on skin aging has been advanced, and in terms of macroscopic causes of skin aging, in addition to the decay of metabolism due to aging, the effects of sunlight (ultraviolet rays), drying, oxidation, etc. It has been confirmed that they are involved in a complicated way. It has become clear that collagen fibers, which are the most major matrix component of the dermis, are prominently reduced by the action of these factors. When the tension and tension retention mechanism such as elasticity held by collagen fibers is destroyed by the action of ultraviolet rays or the like, the skin becomes wrinkled or sagging and becomes aged.
Collagen can also retain water in its molecule, which helps to keep the skin moist, so when external factors destroy the collagen, the skin becomes dry. , Will be in a rough state. From the above, there has been a demand for a skin beautifying agent which can prevent the aging of the skin by promoting the biosynthesis of collagen, which is one of the important components of the dermis layer, and has no problem in safety.

[0003]

In view of these problems, the present inventors have been earnestly searching for a substance showing a collagen production promoting action which is widely contained in food materials. It was found that the basic protein fraction of or the basic peptide fraction obtained by degrading the basic protein fraction with a proteolytic enzyme such as pepsin or pancreatin can increase the amount of skin collagen. Then, they have found that this basic protein fraction or basic peptide fraction can be used as an active ingredient of a skin beautifying agent, a skin beautifying food and drink, and a cosmetic, and have completed the present invention. Therefore, an object of the present invention is to provide a skin beautifying agent containing a milk-derived basic protein fraction or basic peptide fraction having this skin collagen production promoting activity as an active ingredient. Moreover, this invention makes it a subject to provide the food / beverage products and cosmetics for beautiful skin which mix | blended these fractions.

[0004]

Means for Solving the Problems The feature of the skin-care agent of the present invention is that a basic protein fraction derived from milk or a basic peptide fraction obtained by degrading the basic protein fraction with a proteolytic enzyme is effective. It is to make it an ingredient. The milk-derived basic protein fraction is obtained from milk of mammals such as milk, human milk, goat milk, and sheep milk, and the basic peptide fraction is milk-derived basic protein fraction. It can be obtained by reacting a proteolytic enzyme with.

The basic protein fraction derived from milk has the following properties as shown in Test Examples 1 to 4 described later. 1) According to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the molecular weight was 3,000 ~ 8
It consists of several proteins in the range of 0,000. 2) 95% by weight or more is protein and contains a small amount of fat and ash. 3) The protein mainly consists of lactoferrin and lactoperoxidase. 4) The amino acid composition of the protein contains 15% by weight or more of basic amino acids such as lysine, histidine, and arginine.

Such a basic protein fraction is obtained by, for example, contacting a dairy raw material such as skim milk or whey with a cation exchange resin to adsorb the basic protein, and adsorbing the basic protein fraction onto the resin. Eluate with 0.1M to 1M salt concentration eluate, collect this eluate fraction, desalt and concentrate by reverse osmosis (RO) membrane or electrodialysis (ED) method, and dry if necessary. Can be obtained by

As a method for obtaining a milk-derived basic protein fraction, milk or a milk-derived raw material is brought into contact with a cation exchanger to adsorb the basic protein, and then the cation exchanger is adsorbed. PH of the basic protein fraction
Method obtained by eluting with an eluent having an ionic strength of more than 5 and an ionic strength of more than 0.5 (JP-A-5-202098), method using an alginic acid gel (JP-A-61-246198), inorganic porous particles A method for obtaining whey from whey (JP-A-1-86839), a method for obtaining a milk using a sulfated ester compound (JP-A-63-255300), and the like are known. In the present invention, The basic protein fraction obtained by such a method can be used.

The milk-derived basic peptide fraction has the same amino acid composition as the basic protein fraction. For example, the milk-derived basic protein fraction obtained by the above-mentioned method is treated with pepsin. A peptide composition having an average molecular weight of 4,000 or less can be obtained by acting a proteolytic enzyme such as trypsin, chymotrypsin, etc. and, if necessary, a proteolytic enzyme such as pancreatin.

The skin beautifying agent of the present invention exhibits a skin beautifying effect upon oral administration or application. When orally administering the skin-care agent of the present invention, the milk-derived basic protein fraction or basic peptide fraction of the active ingredient can be used as it is, but according to a conventional method, a powder, a granule, It can also be used by formulating into tablets, capsules, drinks and the like. Furthermore, it is also possible to mix these basic protein fractions and basic peptide fractions as they are or after formulating them into nutritional supplements, foods and drinks, etc. Further, by adding a basic protein fraction or a basic peptide fraction together with a component such as vitamin C, which has been considered to have an effective action on collagen production, further skin-beautifying action can be expected. Since the milk-derived basic protein fraction or the basic peptide fraction is relatively stable to heat, it is likely that a raw material containing the milk-derived basic protein fraction or the basic peptide fraction is usually used. It is also possible to perform heat sterilization under the conditions.

When applying the skin beautifying agent of the present invention, it should be prepared into various dosage forms such as liquid preparations, solid preparations and semi-solid preparations by blending it with known components which are usually used depending on the purpose of use. The preferred compositions include ointments, gels, creams, sprays, patches, lotions, powders, granules, tablets and the like. For example, the skin collagen synthesis promoter of the present invention, hydrocarbons such as petrolatum, stearyl alcohol, higher fatty acid lower alkyl esters such as isopropyl myristate, animal fats and oils such as lanolin, polyhydric alcohols such as glycerin, glycerin fatty acid ester, mono Cosmetics and pharmaceuticals can be produced by mixing with polyethylene glycol stearate surfactants, inorganic salts, waxes, resins, water and, if necessary, preservatives such as methyl paraoxybenzoate and butyl paraoxybenzoate.

The effective dose of the skin-care agent of the present invention by oral administration varies depending on age, therapeutic effect, pathological condition, etc., but according to the results of animal experiments using rats, it is basic to show skin collagen production promoting action. It was found that the protein fraction or basic peptide fraction should be ingested in an amount of 20 mg or more per 1 kg of rat body weight. Therefore, usually
Since an effect can be expected by ingesting 20 mg or more of a basic protein fraction or a basic peptide fraction per adult per day, it may be added to foods and drinks or administered as a medicine so as to secure this required amount.

The effective amount of the skin beautifying agent of the present invention applied varies depending on the dosage form, but is preferably 0.001% to 2% by weight based on the total amount of the composition to be applied, so that the basic protein fraction or the base is contained. The active peptide fraction may be blended. However, a compound such as a bath agent that is diluted at the time of use can be further increased in the compounding amount. Next, the present invention will be described in detail by showing Examples and Test Examples.

[0013]

Example 1 A column (diameter 5) filled with 400 g of cation exchange resin sulfonated chitopearl (manufactured by Fuji Spinning Co., Ltd.)
(cm x height 30 cm) is thoroughly washed with deionized water, and then 40 liters of unsterilized skim milk (pH 6.7) is applied to this column at a flow rate of 25 ml / min.
It was passed at. After passing through the column, thoroughly wash this column with deionized water, and then add 0.08 M sodium chloride in 0.02 M carbonate buffer (p
The basic protein fraction adsorbed on the resin was eluted with H 7.0). Then, this eluate was desalted with a reverse osmosis (RO) membrane, concentrated, and then freeze-dried to obtain 21 g of a powdery basic protein fraction.

[0014]

[Test Example 1] The basic protein fraction obtained in Example 1 was measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the molecular weight was distributed in the range of 3,000 to 80,000. It was

[0015]

Test Example 2 The basic composition of the basic protein fraction obtained in Example 1 was analyzed. The results are shown in Table 1. As shown in this table, most of this fraction is protein.

[0016]

[Table 1]

[0017]

[Test Example 3] The basic protein fraction obtained in Example 1 was hydrolyzed with 6N hydrochloric acid at 110 ° C for 24 hours, and then its amino acid composition was measured with an amino acid analyzer (L-8500 type, manufactured by Hitachi, Ltd.). analyzed. The results are shown in Table 2. This basic protein fraction contains 15% by weight or more of basic amino acids in the amino acid composition.

[0018]

[Table 2]

[0019]

Example 2 A column (diameter 100) filled with 30 kg of cation exchange resin SP Toyopearl (manufactured by Tosoh Corporation)
cm x 10 cm in height) was thoroughly washed with deionized water, and this column was heat-sterilized at 121 ° C for 30 seconds.
6.2) was passed through at a flow rate of 10 l / min. After the passage, the column was thoroughly washed with deionized water, and the basic protein fraction adsorbed on the resin was eluted with 0.1 M citrate buffer (pH 5.7) containing 0.9 M sodium chloride. Then, the eluate was desalted by electrodialysis (ED), concentrated, and then freeze-dried to obtain 183 g of a powdery basic protein fraction.

[0020]

Example 3 Basic protein fraction obtained in Example 1
After 50 g was dissolved in 10 liters of distilled water, 1% pancreatin (manufactured by Sigma) was added and reacted at 37 ° C for 2 hours. After the reaction, the enzyme was inactivated by heating at 80 ° C. for 10 minutes to obtain 48.3 g of a basic peptide fraction.

[0021]

TEST EXAMPLE 4 With respect to the basic protein fraction obtained in Example 1 and the basic peptide fraction obtained in Example 3, the collagen production promoting action was examined by animal experiments using rats. 7-week-old Wistar male rats were administered with saline (group A), the basic protein fraction obtained in Example 1 was administered at 20 mg per 1 kg of rat body weight (Group B), and in Example 1. The obtained basic protein fraction was used for rat weighing 1 kg.
200 mg per group (C group), basic peptide fraction obtained in Example 3 at 20 mg per 1 kg of rat body weight (D group), basic peptide fraction obtained in Example 3 at rat Divided into 5 test groups (n = 6) of 200 mg / kg body weight (group E), each of which was administered once daily with a sonde 10
Raised for a week.

Regarding the amount of collagen in the skin,
After treatment according to the method of Mni et al. (see Arch. Biochem. Biophys, page 292, 1967), the amount of hydroxyproline contained in the soluble fraction was measured. Hydroxyproline is a special amino acid contained only in collagen and accounts for about 10% of all the amino acids that make up collagen, so the amount of collagen can be estimated. (Ryuji Asano et al. Bio Industor
y, p. 12, 2001) The results are shown in Table 3.

[0023]

[Table 3]

According to this, the amount of hydroxyproline in the soluble fraction after 10 weeks was significantly higher in group B, group C, group D and group E than in group A. From this, it was revealed that the basic protein fraction and the basic peptide fraction have an action of promoting skin collagen production, and it was suggested that they are useful as a skin beautifying agent. Further, it was revealed that this skin collagen production promoting action was observed when a basic protein fraction or a basic peptide fraction was administered at a minimum of 20 mg per 1 kg of rat body weight.

[0025]

[0026]

[Example 4] A skin beautifying beverage having the composition shown in Table 4 was produced.
The produced beverage had a good flavor, the flavor did not deteriorate even after storage at room temperature for 1 year, and there was no problem such as precipitation.

[0027]

[Table 4]

[0028]

Example 5 A dough having the composition shown in Table 5 was prepared, molded, and then roasted to manufacture a biscuit for beautiful skin.

[0029]

[Table 5]

[0030]

Example 6 A skin beautifying agent having the composition shown in Table 6 was produced.

[0031]

[Table 6]

[0032]

Test Example 5 With respect to the basic protein fraction obtained in Example 1 and the basic peptide fraction obtained in Example 3, a normal human fibroblast cell line [CCD45SK (ATCCRL collected from the skin of a Caucasian woman was used. 1506)] was used to examine the collagen production promoting action. 10% fetal bovine serum
(Hereinafter abbreviated as FBS) containing MEM medium (Dainippon Pharmaceutical Co., Ltd., 10-10
1), normal human fibroblast cell line 4 × 10 ⁴ cells / well / 0.4
The cells were seeded on a 24-well plate so that the amount became ml, and cultured at 37 ° C. for 24 hours under 5% carbon dioxide and saturated steam, and then replaced with a MEM medium containing 0.6% by volume FBS. Then, the basic protein fraction obtained in Example 1 and the basic peptide fraction obtained in Example 3 were added to each well at 0.1% by volume and cultured for 24 hours, followed by β -Aminopropionitrile 50 μg / mL, tritium-L-proline 1 μCi / ml
The resulting mixture was added as described above and further cultured for 24 hours to obtain a culture solution. From the culture broth thus obtained, the method of Webster et al. (Analytical Biochemistry, page 220, 1979)
According to the procedure described above, the collagen fraction was fractionated, and the radioactivity incorporated in the collagen fraction was measured. As a control, the same test was conducted without adding the basic protein fraction and the basic peptide fraction. The results are shown in Table 7.

[0033]

[Table 7]

According to this, the group to which the basic protein fraction and the basic peptide fraction were added produced about twice as much collagen as the group to which the basic protein fraction and the basic peptide fraction were not added. Exhibited ability. From this, it was clarified that the basic protein fraction and the basic peptide fraction act on skin fibroblasts to promote collagen production, and it was suggested that they are useful as a skin beautifying agent.

[0035]

Example 7 A lotion having the composition shown in Table 8 was produced by a conventional method.

[0036]

[Table 8]

[0037]

Example 8 A cream having the composition shown in Table 9 was produced by a conventional method.

[0038]

[Table 9]

[0039]

Test Example 6 Using the lotion obtained in Example 7 and the cream obtained in Example 8, an actual use test was conducted. As a comparative product, the one obtained by removing the basic protein fraction in the formulation of Examples 7 and 8 was used. 40 adult women with dry skin with facial lumps and wrinkles
Each person is randomly divided into 4 groups (A to D groups), the lotion of the present invention is applied to the face of group A, the comparative lotion is applied to the face of group B, and C
The fingers of the group were applied with the cream of the present invention and the fingers of the group D were applied with the cream of the comparative product twice a day, respectively, in the same manner as in the normal use state. As a result, it was proved that the lotion of the product of the present invention was more excellent in dry feeling and rough skin improvement than the lotion of the comparative product, and was excellent in the skin beautifying effect. Further, the cream of the present invention also showed a remarkable improvement as compared with the cream of the comparative product, and it was revealed that it has an effect of suppressing natural aggravation such as rough skin.

[0040]

EFFECTS OF THE INVENTION The skin beautifying agent, food and drink for skin beautification and lotion of the present invention have an action of promoting collagen production in the skin,
It is useful for the prevention and treatment of skin aging.

Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 7/48 A61P 17/16 38/00 43/00 105 A61P 17/16 A61K 37/02 43/00 105 A23L 2 / 00 F (72) Inventor Hiroshi Kawakami 204-5 Fujima, Kawagoe City, Saitama Prefecture F-reference (reference) 4B017 LC03 LK15 LK18 LK23 LP06 LP08 4B018 LB01 LB08 LE05 MD20 MD22 MD71 MD90 ME14 MF01 MF12 4C083 AA082 AC022 AC072 AC122 AC442 AC482 AC302 AC302 AC482 AC482 AC302 AD202 AD222 AD411 AD412 AD512 CC04 CC05 EE12 4C084 AA02 AA03 BA03 BA31 CA38 NA14 ZA892 ZB212

Claims (7)

[Claims] 1. A skin beautifying agent containing a basic protein fraction derived from milk as an active ingredient. 2. The skin beautifying agent according to claim 1, wherein the milk-derived basic protein fraction is a fraction containing 15% by weight or more of basic amino acids in its amino acid composition. 3. The milk-derived basic protein fraction is prepared by contacting milk or a milk-derived raw material with a cation exchange resin to adsorb the basic protein, and the fraction adsorbed to the resin is added with a salt concentration of 0.1 M to The skin beautifying agent according to claim 1, which is a fraction obtained by elution with a 1 M eluate. 4. A skin beautifying agent comprising a basic peptide fraction obtained by degrading the milk-derived basic protein fraction according to any one of claims 1 to 3 with a proteolytic enzyme as an active ingredient. 5. The skin-care agent according to claim 4, wherein the proteolytic enzyme is at least one proteolytic enzyme selected from the group consisting of pepsin, trypsin, chymotrypsin and pancreatin. 6. A food or drink for beautiful skin containing the milk-derived basic protein fraction or basic peptide fraction according to any one of claims 1 to 5. 7. A cosmetic comprising the milk-derived basic protein fraction or basic peptide fraction according to any one of claims 1 to 5.