JP3236347B2 - Whitening cosmetics - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a whitening cosmetic having a skin whitening effect, and more particularly to a whitening cosmetic in which a crude drug extract having a whitening effect based on a melanin production inhibitory action is blended as an active ingredient.

[0002]

2. Description of the Related Art In general, it is considered that darkening, spots, freckles, and the like due to sunburn are caused by the production of melanin, which is a black-brown amorphous pigment, and this melanin causes external stimuli such as ultraviolet rays on the skin. When it is received, tyrosinase (tyrosine oxidase) present in the melanocytes of the skin is activated, and tyrosine, one of the amino acids constituting protein, is oxidized and produced. Therefore, since an effect of whitening skin is expected by suppressing the activity of tyrosinase involved in melanin production, it has been proposed to add a tyrosinase activity inhibitory component to cosmetics.

Conventionally, as a whitening cosmetic having a whitening effect, as disclosed in JP-B-55-43443, "Whitening cosmetic" and JP-B-54-974, "Composition of crude drug extract", ascorbic acid or What blended the derivative is known. In addition, skin external preparations containing arbutin (JP-A-60-16906), bleaching cosmetics containing kojic acid (Japanese Patent Publication No. 32-8100), plant components (JP-A-63-2913, etc.) ) Or animal components (JP-A-63-8312, etc.) are known as having a whitening effect.

[0004] However, many of the above-mentioned conventional cosmetics do not show a sufficient whitening effect, and some of them have problems in safety to the skin such as insufficient storage stability and irritation. There were many.

[0005]

SUMMARY OF THE INVENTION The present invention solves the above-mentioned problems of the prior art, and provides a whitening cosmetic having an excellent skin whitening effect, sufficient storage stability and high safety. The purpose is to do.

[0006]

Means for Solving the Problems The present inventors have made intensive studies to solve the above problems, and as a result, it has been found that an extract of dried black beans has an inhibitory effect on tyrosinase enzyme activity and an inhibitory effect on melanin production in melanoma cells. It has been found that the present invention can be provided.

That is, the present invention provides a skin-whitening cosmetic comprising an extract of a dried black bean.

[0008]

The cosmetic of the present invention can be manufactured by the following method. First, a dried product of black beans or a crushed product of the dried product is extracted by heating using an extraction solvent. In addition, as the extraction solvent, alcohol (methanol, ethanol, propanol, isopropyl alcohol, or the like), water, or the like can be used, and a mixed solution thereof can also be used. For example, if the alcohol concentration is 20-70
Extraction at 50 ° C. for 1 hour using a hydroalcoholic solution at a high concentration provides good extraction efficiency.

After extraction, the extract is filtered to obtain an extract. The obtained extract is further concentrated under reduced pressure while heating at a temperature of 60 ° C. or lower to dryness, and the dried extract is collected and blended into cosmetics. In addition, you may mix the said extract with cosmetics as it is.

[0010] The present invention shows that the extract of the herbal medicine (dried black bean) thus obtained exhibits a superior melanin production inhibitory action at a lower concentration than the conventionally used ascorbic acid. It has been confirmed by those and others that a whitening cosmetic having a whitening effect can be obtained by adding 0.01 to 5.0% of this extract as an active ingredient.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the scope of the present invention is not limited by the following examples.

[0012]

Embodiment 1 In this embodiment, an example of a method for extracting crude drugs will be described. First, about 100 g of dried mulberry straw, which is a crude drug, was pulverized with a mixer, and the pulverized substance and 500 ml of 50% ethyl alcohol were placed in a flask, and refluxed at 50 ° C. for 1 hour with stirring. After the extraction, this solution was subjected to suction filtration, and the obtained filtrate was concentrated under reduced pressure at 50 ° C. using an evaporator. Next, the obtained concentrated liquid was dried under reduced pressure to obtain 9.2 g of brown crystals (extract).

In the same manner as described above, 15.6 g, 8.8 g and 11.9 g of extracts were obtained from about 100 g of the dried herbs of black beans, soybeans and chicken blood wisteria, respectively.

[0014]

Example 2 In this example, the tyrosinase activity-inhibiting activity of the extracts of Mulberry, Black bean, Soot and Chicken wisteria obtained in Example 1 was measured. The tyrosinase activity inhibitory effect was measured by a method of quantifying dopachrome produced from dopa by tyrosinase by measuring absorbance at 475 nm. In measuring the tyrosinase activity inhibitory action, the following reaction reagents were used. (A) Sodium succinate buffer (100 mM, pH 5.
5) (b) Mushroom tyrosinase (manufactured by SIGMA) solution (prepared at 270 units / ml with the buffer of (a)) (c) L-DOPA (manufactured by Wako Pure Chemical Industries, Ltd.) solution (6 mM First, prepare 1.8 ml of buffer (a) and (b) in a test tube.
0.1 ml of tyrosinase solution, and add 2% (w
/ v) concentration of sample solution (aqueous solution of extract obtained in Example 1)
0.1 ml was added, and the mixture was incubated in a constant temperature water bath at 30 ° C. for 15 minutes. Next, 1 ml of the L-DOPA solution of (c) was added to the test tube.
In addition, after stirring, the test tube was set in a reciprocating shaker at about 45 ° in a thermostat chamber at 30 ° C., and shaken for 40 minutes (reciprocation frequency 150 / min). After shaking, use a spectrophotometer at 475 nm
Was measured, and the measured value was designated as A.

On the other hand, the absorbance at 475 nm was measured in the same manner as above except that the buffer (a) was added instead of the sample solution as a control. In addition, instead of the L-DOPA solution as a blank,
475 in the same manner as above except that buffer was added.
The absorbance at nm was measured, and the measured value was designated as C.

The tyrosinase activity inhibition rate of the sample solution was calculated from the measured value of the absorbance at 475 nm. The calculation of the tyrosinase activity inhibition rate is performed using the following formula,
The results are shown in Table 1.

Tyrosinase activity inhibition rate (%) = ｛1-
(AC) / B｝ × 100

[0018]

[Table 1]

As can be seen from Table 1, the extract of mulberry straw, black soybean, soybean and chicken blood wister can strongly inhibit the activity of tyrosinase even at a low concentration of 2% (w / v) aqueous solution. It was confirmed that the compound had an excellent tyrosinase activity inhibitory action.

[0020]

Example 3 In this example, the melanin production inhibitory effect of each of the extracts of Mulberry, Black Bean, Soybean, and Chicken Wisteria obtained in Example 1 was measured.

First, B16 melanoma cells (B16-F0, ATCC), which are malignant melanoma cells derived from mice that produce melanin,
No. CRL-6322) was cultured in Eagle's MEM medium supplemented with fetal bovine serum to a final concentration of 10%, and the cells were added to each well of a 6-well plate (FALCON) at 3 × 10 5
⁶ ml of the above medium containing ³ cells / ml was added and cultured in a CO ₂ incubator (5% CO ₂ , 37 ° C.) for 5 days.

Next, the medium was treated with 0.03% theophylline.
New Eagle MEM medium (6 ml) containing (SIGMA)
After adding an appropriate amount of a sample solution (aqueous solution of the extract obtained in Example 1) to each well, the cells were further cultured for 3 days. After completion of the culture, the medium was discarded, 1 ml of physiological saline was added to each well, and the cells adhered to the bottom of the well were suspended using a scraper to scrape the cells. Next, the cell suspension is transferred to a microcentrifuge tube using a pipette.
(1.5 ml volume, manufactured by Eppendorf) and centrifuged (1,000 × g, 15 minutes).

On the other hand, as a control, the same test as above was conducted by adding sterile water instead of the sample solution. In addition, as an experimental group for comparing the whitening of the cells, a 2% L-ascorbic acid aqueous solution was used instead of the sample solution (a) 60 μl, (b) 150 μl.
μl and (c) 300 μl were added, and the same test was performed.

Next, the degree of whitening of the pelleted cells was visually compared to determine the melanin production inhibitory effect. At that time, the degree of whiteness of the cells in the control experiment (sterilized water-added) was "-", and the degree of whiteness of the cells in the comparative experiment to which L-ascorbic acid was added was (a): "+", ( b)
: “++”, (c): As “++++”, the degree of whiteness of the cells when the sample solution was added was −, +, ++, +++
It was visually determined which one of the two corresponded, and the strength of the melanin production inhibitory effect of the sample solution was determined in four steps. The results are shown in Table 2.

[0025]

[Table 2]

As can be seen from Table 2, each of the extracts of Mulberry, Aoko and Chicken Wisteria showed an inhibitory effect on melanin production at a concentration of 50 μg / ml equivalent to 200 μg / ml of L-ascorbic acid. It was confirmed that melanin production was suppressed at a lower concentration than ascorbic acid. Also, the black bean extract is 200
At the concentration of μg / ml, the same melanin production inhibitory action as L-ascorbic acid was shown. In addition, each extract contains 800 μg /
There was no toxicity to the cells even at the high concentration of ml.

[0027]

Example 4 In this example, an example of blending the crude drug Mulberry extract obtained in Example 1 with a whitening cosmetic will be described.

(Weight%) Mulberry extract 1.0 Glycerin 5.0 Polyoxyethylene sorbitan monolaurate 1.5 Ethanol 10.0 Appropriate amount Preservative, antioxidant Appropriate amount of pigment Appropriate amount of purified water

[0029]

EFFECT OF THE INVENTION The extract of dried black beans, which is a crude drug,
It has an excellent tyrosinase activity inhibitory action and an inhibitory action on melanin production in melanoma cells, and the whitening cosmetic of the present invention containing this extract exhibits an excellent skin whitening effect. In addition, the extract of dried black beans blended in the whitening cosmetic composition of the present invention exhibits an excellent skin whitening effect in a small amount and has low toxicity to cells, and is therefore highly safe.

────────────────────────────────────────────────── ─── Continuation of the front page (72) Masao Takahashi, Inventor 4-11, Kobunacho, Nihonbashi, Chuo-ku, Tokyo Inside Junsei Sangyo Co., Ltd. (56) References JP-A-4-356416 (JP, A) JP-A-Hei 5-97649 (JP, A) JP-A-4-342519 (JP, A) JP-A-56-113284 (JP, A) JP-A-3-223217 (JP, A) JP-A-3-501843 (JP, A) A) (58) Field surveyed (Int. Cl. ⁷ , DB name) A61K 7/00 A61K 7/48 A61K 35/78

Claims (1)

(57) [Claims] 1. A skin whitening cosmetic comprising an extract of dried black beans .