JP2008110927A - Cosmetic containing royal jelly protein hydrolyzate - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide cosmetics intended for the applications of a royal jelly protein hydrolyzate as an ingredient thereof, particularly skin-brightening cosmetics, skin care cosmetics, antiaging cosmetics, cosmetics for lustrous skin and cosmetics for ameliorating/preventing wrinkles, and to provide a hyaluronic acid production promoter. <P>SOLUTION: The hyaluronic acid production promoter containing a protein hydrolyzate as a royal jelly material is provided. Skin-brightening cosmetics containing the hyaluronic acid production promoter are also provided. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a cosmetic containing a hyaluronic acid production promoter, and more particularly to uses such as whitening, skin care, anti-aging, beautiful skin, wrinkle improvement / wrinkle prevention, and hyaluronic acid production promotion.

  Currently, royal jelly is widely used around the world, such as health foods, pharmaceuticals, and cosmetics, and various pharmacological and nutritional physiological effects such as vasodilation, blood pressure lowering, and antibacterial effects have been reported. Royal jelly is a milky substance synthesized in the salivary glands of bees and secreted from the hypopharyngeal gland and the large vagina in the head. It is something to give to. Royal jelly contains protein, vitamins, and minerals in a well-balanced manner, and the queen bee that continues to eat it for a lifetime has a life 40 times longer than that of a worker bee and the ability to lay 2000 eggs a day. Research on the active ingredients of this royal jelly has been focused on low molecular weight compounds such as fatty acids, amino acids, vitamins such as 10-hydroxy-2-decenoic acid.

  On the other hand, royal jelly usually contains 10% or more protein, but research on the molecular structure and physiological function of this protein is delayed. Moreover, alcohol is extracted during the production of a drink containing a royal jelly, and the protein is separated as a precipitate. However, at present, the protein is not effectively used.

Regarding the processed protease of royal jelly protein, calcium absorption promoting action (Non-patent document 1), blood glucose level lowering action (Non-patent document 2) are known, antioxidant action (Non-patent document 3), collagen production promoting action (patent) Reference 1) and antihypertensive action (Patent Document 2) have also been reported.
JP2003-171258 JP 2005-255670 A Teruko Nakasa, Food and Development, 34.5 (1999) Shuichiro Ueda, Food Style 21, 6, 5 (2002) Japan Royal Jelly Fair Trade Council "Study on Royal Jelly" 2001 Research Report

  An object of the present invention is to provide a cosmetic application of a royal jelly protein hydrolyzate, in particular, a whitening cosmetic, a skin care cosmetic, an anti-aging cosmetic, a beautiful skin cosmetic, and a wrinkle improving / wrinkle preventing cosmetic.

  Another object of the present invention is to provide a hyaluronic acid production promoter.

  As a result of repeated studies in view of the above problems, the present inventor has found that the protein hydrolyzate of royal jelly material has a strong hyaluronic acid production promoting action, and based on this hyaluronic acid promoting action, skin care, whitening, anti-aging (anti-aging) ), Cosmetics having various effects such as beautiful skin, wrinkle improvement / wrinkle prevention, etc. were found.

The present invention provides the following hyaluronic acid production promoter, whitening cosmetics, skin care cosmetics, anti-aging cosmetics, skin beautifying cosmetics, wrinkle improving / wrinkle preventing cosmetics.
1. Hyaluronic acid production promoter containing protein hydrolyzate of royal jelly material.
2. Item 2. A whitening cosmetic comprising the hyaluronic acid production promoter according to Item 1.
3. Item 11. A skin care cosmetic comprising the hyaluronic acid production promoter according to Item 1.
4). Item 6. An anti-aging cosmetic comprising the hyaluronic acid production promoter according to Item 1.
5. Item 1. A skin beautifying cosmetic comprising the hyaluronic acid production promoter according to Item 1.
6). A cosmetic for improving or preventing wrinkles comprising the hyaluronic acid production promoter according to Item 1.

  ADVANTAGE OF THE INVENTION According to this invention, the trouble regarding skin, such as a spot, dullness, rough skin, a spot, sagging, can be solved by applying the cosmetics containing a royal jelly protein hydrolyzate to skin.

  In the present invention, the royal jelly material used as a protein raw material means a material containing the protein of royal jelly, and is not particularly limited as long as this requirement is satisfied. For example, raw royal jelly, dried royal jelly, royal jelly is ethanol precipitated. Protein fraction obtained by protein denaturation treatment such as ammonium sulfate treatment and heat treatment (including ethanol precipitate) or royal jelly is extracted, filtered, centrifuged, chromatographed, etc. Means a material containing protein.

The royal jelly may be anything, and widely used by bees secreted to grow queen bees. Specifically, the royal jelly is a milky white jelly-like substance produced by mixing secreted products of the bees from the hypopharyngeal gland and the greater vagina.
As the protein fraction derived from royal jelly, a protein fraction obtained by alcohol precipitation of royal jelly can be preferably used. In particular, it is preferable to use an ethanol-denatured protein of royal jelly that is by-produced as a water-insoluble protein during the production of a royal jelly beverage. The ethanol-denatured protein can be obtained as a protein having a low solubility in water of a residue obtained by extracting a physiologically active substance for beverages such as decenoic acid from royal jelly using about 50 to 100% by volume of ethanol.

  The hydrolyzate of the royal jelly material of the present invention can be obtained by performing hydrolysis with a proteolytic enzyme. Proteolytic enzymes include endopeptidases and exopeptidases, and one or more of these can be used.

  The proteolytic enzyme used in the present invention is derived from animals (for example, trypsin, chymotrypsin, pepsin, pancreatin, etc.), derived from plants (for example, papain, etc.), or derived from microorganisms (for example, lactic acid bacteria, yeast, mold, Bacillus subtilis). And exopeptidases derived from microorganisms (eg, lactic acid bacteria, Aspergillus, Rhizopus, etc.).

  Preferable examples of the proteolytic enzyme include, for example, Streptomyces griseus-produced peptidase (trade name: actinase AS), Aspergillus oryzae-produced peptidase (trade name: protease A, flavorzyme), Aspergillus・ Production of peptidase from Aspergillus melleus (trade name: Protease P), production of peptidase from Aspergillus oryzae (trade name: Ummamizyme G, Promod 192P, Promod 194P, Sumiteam FLAP), production of Aspergillus sojae Peptidase (trade name: Sternzyme B15024), Aspergillus genus peptidase (trade name: cochlase P), Rhizopus oryzae peptidase (trade name: peptidase R), Bacillus subtilis (Baci) llus subtilis-produced peptidase (trade name: orientase 22BF, nucleisin), Bacillus licheniformis-produced peptidase (trade name: Alcalase), Bacillus stearothermophilus-produced peptidase (product) Name: Protease S), Bacillus amyloliquefaciens production peptidase (trade name: Neutase), Bacillus genus peptidase (trade name: Protamex).

  Furthermore, a mutant enzyme in which one or more amino acids are substituted, added, deleted, inserted, or the like may be used for the proteolytic enzyme. Such a mutant proteolytic enzyme is also used for enzymatic degradation of the royal jelly material of the present invention.

  The conditions for treating the royal jelly material with a proteolytic enzyme include, for example, about 15 ° C. to about 60 ° C., preferably about 20 ° C. to about 50 ° C., particularly around 37 ° C. in an aqueous solvent, preferably about 1 to 72 hours, preferably It can implement preferably by processing for about 3 to 48 hours. The amount of proteolytic enzyme used is, for example, about 0.01 to 3 g, preferably about 0.05 to 1 g, per 100 g of protein. As the proteolytic enzyme, a free enzyme may be used, or an enzyme immobilized on a carrier may be used.

  Water can be suitably used for the reaction solution, but it may be carried out in an aqueous solvent such as hydrous ethanol containing a small amount of a water-miscible organic solvent such as ethanol.

  The reaction solution of the enzymatic degradation product is subjected to heat treatment, and if necessary, post-treatment such as centrifugation, filtration, extraction, desalting, freeze drying or spray drying (or spray drying) to obtain a protein degradation product. Can do. For example, when raw royal jelly or dried royal jelly is used as the royal jelly material, components such as saccharides derived from royal jelly can be removed.

  In one preferred embodiment of the present invention, the royal jelly material proteolysate of the present invention has an average molecular weight of about 300 to 7000, particularly about 500 to 6000, as measured by a gel filtration column.

  The proteolysate of the present invention may be produced by desalting as necessary after enzyme treatment, and it is also possible to collect fractions having a stronger hyaluronic acid production promoting effect by treatment with a column or the like. .

  The compounding quantity of the royal jelly protein hydrolyzate in this invention is 0.0005 to 20.0 weight% as a dry substance in the cosmetics whole quantity, Preferably it is 0.001 to 10.0 weight%. If it is less than 0.0005% by weight, the effect of the present invention is not sufficiently exhibited, and if it exceeds 20.0% by weight, it is difficult to make a preparation. Moreover, even if it mix | blends 10.0 weight% or more, the improvement of the big effect is not seen. Furthermore, the royal jelly protein hydrolyzate is particularly preferably applied to the skin at a concentration of 0.01 to 0.1% by weight.

  Hyaluronic acid is an acidic mucopolysaccharide synthesized by hyaluronic acid synthase (HAS) present in the membrane of dermal fibroblasts (Y. Sugiyama et al., J. Invest. Dermatol., 110, 116 (1998)). Hyaluronic acid contributes to retention of water in the cell gap, retention of cells based on the formation of a jelly-like matrix in the tissue, and maintenance of tissue lubricity and flexibility (BIOINDUSTRY Vol8, 346 (1991) ). Hyaluronic acid has been reported to decrease with age (K. Matsuoka et al. Aging, 1, 47 (1989)) or to denature (LAMMeyer et al. J. Invest. Dermatol., 102, 385 (1994)) Such changes are said to contribute to dry skin, wrinkles, sagging and the like.

  Since the royal jelly protein hydrolyzate used in the present invention has an excellent hyaluronic acid production promoting action on human skin, the cosmetics containing the hydrolyzate give skin elasticity and moisture. , Prevents skin aging, eliminates or reduces wrinkles, makes skin fresh, makes spots and dullness unnoticeable, has a beautifying effect, and maintains healthy skin, whitening cosmetics, anti-aging cosmetics, beautiful skin It is useful as cosmetics, skin care cosmetics, wrinkle improving / preventing cosmetics, and the like.

  In addition to the royal jelly protein hydrolyzate, the cosmetics of the present invention include components usually used in cosmetics, such as whitening agents, moisturizers, antioxidants, oily components, ultraviolet absorbers, surfactants, thickeners, Alcohols, powder components, color materials, aqueous components, water, various skin nutrients, and the like can be appropriately blended as necessary.

  The cosmetic dosage form of the present invention includes an aqueous solution system, a solubilization system, an emulsification system, a powder system, an oil liquid system, a gel system, an ointment system, an aerosol system, a water-oil two-layer system, and a water-oil-powder three-layer system. A wide range of dosage forms can be adopted. For example, for basic cosmetics, face wash, lotion, milky lotion, cream, gel, essence, beauty liquid, pack, mask, etc., for makeup cosmetics, foundation, lipstick, blusher, eye shadow, eyeliner , Mascara, etc., facial cleanser, massage agent, cleansing agent, after shave lotion, pre-shave lotion, shaving cream, body soap, soap, shampoo, rinse, hair treatment, hair conditioner, hair art agent, hair restorer Antiperspirants, hygiene products and the like.

EXAMPLES Hereinafter, although an Example demonstrates this invention, this invention is not limited by this Example.
Production Example 1
250 g of fresh royal jelly was weighed into a 500 mL beaker, 20 mL of ion exchange water was added, and the mixture was stirred until uniform. 47 mL of 2N NaOH was added and adjusted to pH 8.7. 2.5 g of enzyme BP (SternzymeBP14202; bacilillolysin derived from Bacillus subtilis manufactured by SternEnzyme) was added, and 33 mL of ion-exchanged water was further added and stirred until uniform. The beaker was immersed in a constant temperature water bath and reacted at 45 ° C. for 15 hours while stirring the royal jelly solution with a three-one motor. The temperature of the thermostatic bath was raised and the royal jelly solution was heated while stirring with a three-one motor. After reaching 80 ° C., the mixture was immediately cooled with water and freeze-dried to obtain a royal jelly protein hydrolyzate as a lyophilized product.

Example 1: Promotion of hyaluronic acid production (in vitro)
Normal human fibroblast cell line WI-38 cells were adjusted to 1 × 10 ⁵ cells / ml with 10% FCS (fetal bovine serum) -containing RPMI-1640 medium (cell suspension), and cell suspension per well 100 μl of the solution was added. Then, this plate was incubated overnight at 37 ° C. in 5% CO ₂ -95% air, and the medium was changed with a mixture of 150 μl of RPMI-1640 medium and 40 μl of PBS (−), and the royal jelly protein hydrolyzed protein obtained in Production Example 1 was replaced. The degradation product was added to each well to a concentration of 100 μg / mL (final concentration), and incubated at 37 ° C. for 48 hours in 5% CO ₂ -95% air. The culture was centrifuged at 1500 rpm for 15 minutes to obtain a supernatant. The amount of hyaluronic acid in the obtained supernatant was measured using a commercially available hyaluronic acid measurement kit (Seikagaku Corporation).

  In addition, the hyaluronic acid concentration in the supernatant was measured in the same manner for the untreated raw royal jelly lyophilizate instead of the royal jelly protein hydrolyzate obtained in Production Example 1 or for these control groups. The hyaluronic acid production promotion rate was calculated with the amount of hyaluronic acid when 0.01 ml of water was added instead of the sample being 100. The results are shown in Table 1.

  From the results in Table 1, it was revealed that the royal jelly protein hydrolyzate of the present invention has an excellent hyaluronic acid production promoting action for human cells.

Example 2: Promotion of hyaluronic acid production (in vivo)
An aqueous solution of a royal jelly protein hydrolyzate and a base (water, control) were applied to the skin of a hairless mouse (Hos: hr-1) for 2 weeks at 100 μl / 5 cm ² / animal / day, and the skin was applied 24 hours after the final application. Collected. The collected skin of hairless mice was treated by the following method to determine the amount of hyaluronic acid in the skin.

  Hyaluronic acid was quantified as follows.

  Mouse skin degreased with acetone is cut into small pieces, then subjected to alkali treatment, neutralization, pronase E (Sigma) treatment, trichloroacetic acid treatment and dialysis (Spectra / POR1, fractional molecular weight 6000 to 8000, manufactured by Spectrum). A sample for quantitative determination of hyaluronic acid was prepared. Hyaluronic acid was quantified by ELISA (AJ Fosang et al., Matrix, 10, 306-313, 1990) using biotin-labeled hyaluronic acid binding protein (Seikagaku Corporation). The hyaluronic acid content (relative value%) per dry skin weight relative to the control was used as an index of the skin hyaluronic acid content.

As a result of the above test, it was found that the royal jelly protein hydrolyzate promotes the production of hyaluronic acid in the skin, and a high effect can be obtained particularly at a concentration of 10 to 100 μg / ml.
Formulation Example 1: Lotion Toner lotion was prepared by a conventional method based on the following formulation. In addition, what was prepared in said manufacture example 1 was used as a royal jelly protein hydrolyzate.
Toner lotion composition Composition amount (%)
Ethanol 10.00
Sorbit solution 5.00
Methylparaben 0.05
Tripolyoxyethylene ether phosphate 0.40
Royal jelly protein hydrolyzate 0.01
Citric acid 0.02
Sodium citrate 0.05
Purified water remaining
(Total) 100.00

Formulation Example 2: Cosmetic liquid A cosmetic liquid was prepared by a conventional method based on the following formulation. In addition, what was prepared in said manufacture example 1 was used as a royal jelly protein hydrolyzate.
Cosmetic liquid prescription composition
Ethanol 8.00
1,3 Butylene glycol 5.00
Methylparaben 0.10
Polyoxyethylene cetyl ether 0.20
Royal jelly protein hydrolyzate 0.05
Xanthan gum 0.05
Hydroxyethyl cellulose 0.10
Edetic acid disodium 0.01
Purified water remaining
(Total) 100.00

Formulation Example 3: Shampoo Shampoo was prepared by a conventional method based on the following formulation. In addition, what was prepared in said manufacture example 1 was used as a royal jelly protein hydrolyzate.
Composition Blending amount (%)
Lauryl sulfate triethanolamine 20.00
Sodium polyoxyethylene lauryl ether sulfate 10.00
Sodium alkyl sulfate solution 10.00
Palm oil fatty acid amidopropyl betaine solution 10.00
Polyoxyethylene lauryl ether 1.00
Coconut oil fatty acid diethanolamide 3.00
Palm oil fatty acid monoethanolamide 0.45
Ethylene glycol distearate 0.50
Methylparaben 0.10
Glycerin 5.00
O- [2-hydroxy-3- (trimethylammonio) chloride
Propyl] hydroxyethyl cellulose 0.20
Polyoxyethylene / methylpolysiloxane copolymer 0.10
Royal jelly protein hydrolyzate 0.05
Purified water remaining
(Total) 100.00

Formulation Example 4: Emulsion An emulsion was prepared by a conventional method based on the following formulation. In addition, what was prepared in said manufacture example 1 was used as a royal jelly protein hydrolyzate.
Composition Blending amount (%)
Soft lanolin fatty acid cholesteryl 10.00
POE (20) sorbitan monostearate 1.80
Glycerol monostearate 1.20
Royal Jelly Protein Hydrolyzate 0.10
Glycerin 3.00
Methylparaben 0.10
Ethylparaben 0.10
Perfume appropriate amount
Purified water balance
(Total) 100.00

Formulation Example 5: Cream A cream was prepared by a conventional method based on the following formulation. In addition, what was prepared in said manufacture example 1 was used as a royal jelly protein hydrolyzate.
Composition Blending amount (%)
Macadamia nut oil fatty acid cholesteryl 10.00
Stearic acid 7.50
Cetostearyl alcohol 1.50
Triisooctanoic acid glycerin 12.00
Lanolin 3.00
Cetyl palmitate 4.00
PEG monostearate (40) 2.00
Glycerol monostearate 5.00
Royal Jelly Protein Hydrolyzate 0.10
Preservative Appropriate amount Antioxidant Appropriate amount Fragrance Appropriate amount
Purified water remaining
(Total) 100.00

Test Example About the lotion obtained in Formulation Example 1 (invention) and the lotion having the same composition except that it does not contain the royal jelly protein hydrolyzate (control), 10 panelists gave moisturizing action (moist feeling), Four items of whitening action (improvement of dullness), anti-aging (improvement of wrinkles), and beautification action (skin tension, gloss) were tested.

  As a result, the lotion of the present invention containing the royal jelly protein hydrolyzate was highly evaluated for all items as compared with the control lotion containing no hydrolyzate.

  Moreover, although data are not shown, it became clear that the improvement of the evaluation of the above four items is related to the increase in the amount of hyaluronic acid in the panelists, and the improvement of these evaluations is based on the hyaluronic acid production promoting action.

Claims (6)

Hyaluronic acid production promoter containing protein hydrolyzate of royal jelly material. A whitening cosmetic comprising the hyaluronic acid production promoter according to claim 1. A skin care cosmetic comprising the hyaluronic acid production promoter according to claim 1. An anti-aging cosmetic comprising the hyaluronic acid production promoter according to claim 1. A skin care cosmetic comprising the hyaluronic acid production promoter according to claim 1. A cosmetic for improving or preventing wrinkles comprising the hyaluronic acid production promoter according to claim 1.