KR20030025716A - Melanin synthesis inhibition compound and its composition contaning Dioscorea rhizoma extract - Google Patents

Abstract

PURPOSE: A skin whitening agent containing an extract of Dioscoreae Rhizoma is provided which exhibits a better skin whitening effect than albutin and hydroxyanisole and has no skin irritation even when used for a long period of time. CONSTITUTION: Dioscoreae Rhizoma is ground, added 5 times with 95% alcohol and extracted at 4 to 40deg.C for 48hr, the extraction process is repeated two times or more, the filtrate is dried in a rotary vacuum drier at 60deg.C. A skin whitening cosmetic contains 0.00001 to 10% by weight of the Dioscoreae Rhizoma extract, based on the dried weight of the Dioscoreae Rhizoma extract.

Description

Whitening composition containing acid extract {Melanin synthesis inhibition compound and its composition contaning Dioscorea rhizoma extract}

The present invention relates to a whitening cosmetics containing the extract of the herb, which is an herbal medicine that is dried and steamed as it is by removing the bark of the plant root stem belonging to the hemp and yam family.

Melanin (melanin) is a brown or black polymer pigment produced by the oxidation of phenols widely distributed in nature is a biopolymer material strongly bound to protein. In our surroundings, melanin synthesis is observed in browning, which occurs when the cut surfaces of apples, potatoes, and bananas are exposed to pneumoconiosis, or in animal skin, feathers, skin, hair, and eyes. Overproduction of melanin is known to form blemishes, freckles, promote skin aging, and induce skin cancer in the human body. In foods, it also degrades the quality of vegetables, fruits, and fish.

The biosynthesis of melanin is reported to be mainly caused by the action of tyrosianse. Animal melanin catalyzed by tyrosinase is divided into yellow-red-brown pheomelanin and brown-black eumelanin. By the 1980s, melanin was thought to be biosynthesized via the Raper-Mason pathway from L-tyrosine to dopa (L-3,4-dihydroxy-phenylalanine). However, Kameyama et al. Noted that melanin production in mammals is not related to tyrosine, but also to tyrosine regulatory proteins (TRP1), dopachrome tautomerase, and melanogenic inhibitors that are present in cells. It has been reported that it is regulated by action and melanin stimulating hormone (MSH).

Hydroxyanisole (4-hydorxyanisol), 5-hydoroxyindole, and hydroquinone (hydorquinone), known as tyrosinase inhibitors, show potent melanin biosynthesis inhibitory activity but cause pigment cell degeneration or lethality. It has side effects that impair its original function. Kojic acid (5-hydorxy-2- (hydroxymethyl-v-pyrone)) and arbutin (hydroquinone-β-D-gluco pyranoside) are already commercialized and used as whitening agents in cosmetics and foods. Although it is used, problems such as low inhibitory activity, discoloration during use, and instability of the substance itself have been raised, and most substances have been reported to cause cancer, mutations, or have strong toxicity. Therefore, the development of inhibitors with low cytotoxicity and high melanin biosynthesis inhibitory activity is required.

The two most widely used methods of searching for whitening agents are two types of screening methods: a search for tyrosinase inhibitors using mushroom-derived tyrosinase and a method for evaluation at the cellular level using melanin synthetic cells in animals. The former has the advantage of being simpler and less time-consuming and costly, but the disadvantage is that it is often inconsistent with actual clinical effects. The latter can be more comprehensively selected for whitening agents, but the cost, time and effort are much higher. The disadvantage is that it costs.

In order to compensate for this drawback, the present inventors have introduced a method of searching for substances that selectively inhibit the melanin synthesis of fungal spores using fungi synthesizing melanin and selectively acting on the melanin production site of cells.

1 is a diagram comparing the degree of inhibition of melanogenesis in B-16 melanoma cells derived from rats by treatment of the extract and arbutin, a conventional whitening substance.

The present invention relates to a whitening cosmetic containing an acid extract, and more particularly, to remove the bark of the plant root stem belonging to the hemp and yam family, pulverized and dried as it is, to add an organic solvent extract to the conventional basic cosmetics It is about a whitening cosmetics which makes an effect.

Whitening of various plant extracts by measuring melanin synthesis in melanin-synthesizing melanoma to detect melanogenesis inhibitors effective in humans, and B-16 melanoma cells derived from rats similar to human melanocytes The effect was searched.

As a result, the drug extract was found to have no irritation as a result of applying it to human skin as a very good melanin inhibitor even at low concentrations, thus completing the present invention.

Example 1.

The preparation method of the acid extract contained in the whitening cosmetics of the present invention is as follows. After diluting 1 kg of Dioscorea Rhizoma, 95% functional alcohol, which is 5 times the amount of distillate, was added and extracted two times or more for 48 hours at 4-40 ° C., followed by filtration through Whatman 42 filter paper. The filtrate was dried at 65 ° C. with a rotary vacuum evaporator to obtain a dry weight increase of 56 g.

Experimental Example 1: Inhibitory effect of medicinal herb extract on melanin synthesis in melanin synthesis fungi Asp. Put the PDA medium containing the spore suspension of the miser and solidify. The extract of Example 1 was dissolved in 50 µl ethanol and diluted sequentially from the concentration of 500 µg / ml, and directly added to the surface of the medium. Synthesis was visually observed. At this time, hydroxyanisole was used as a control. The experimental results are shown in Table 1.

TABLE 1

Inhibition of melanin synthesis according to concentration showed stronger effect than hydroxy anisole used as a control, and there was no lethality of strain at high concentration. In the case of hydroxyanisole, it shows a strong whitening effect, but has side effects such as pigment cell degeneration and lethality.

Experimental Example 2 Effects of Pesticide Extracts on Melanin Production in Melanocytes

Rat B16-melanoma cells were suspended in DMEM (Dulbescco's Modified Eagle's Medium) medium containing 10% FCS (fetal calf serum) at a concentration of 5 × 10 ³ cells / ml, followed by suspension of 5 ml of tissuue culture. Put it in a flask (Falcon, Becton dickinson) and add the assay sample by concentration and incubate at 37 ℃ with 5% CO ₂ -95% air condition. After 4 days of incubation, the cells adsorbed on the bottom of the flask were washed with PBS (phosphate buffered saline), separated with 0.05% trypsin and 0.53 mM EDTA solution, the cells were centrifuged for 10 minutes, collected in a tube, and the number of cells was measured. It was. After constant adjustment of the number of cells in each tube, 5% trichloroacetate (TCA) was treated and stirred, and centrifuged to wash the precipitated melanin with PBS. 1N NaOH was dissolved in the melanin precipitated, and the absorbance was measured at 475 nm. Melanin concentration was determined from the standard concentration curve of synthetic melanin (sigma) and arbutin was used as a control.

The experimental results are shown in Table 2.

TABLE 2

Experimental results show that the extract is a powerful whitening agent that inhibits melanin synthesis in melanocytes even at concentrations four times lower than arbutin.

Example 2

Prescription examples of cosmetic thickening cosmetics (skin) containing acid extracts are shown in Table 3.

TABLE 3

Experimental Example 3: Safety experiment on the human body of the formulation containing the acid extract

In order to determine the degree of skin irritation with respect to the composition according to the present invention, a skin patch test on the whitening cosmetic according to Example 2 was performed on 20 subjects. 20 μl of the softening water of Example 2 was added to a fin chamber, and the patch was applied to the human upper arm for 24 hours. The skin reaction was examined within 1 hour after the patch was removed, and then examined again the following day (after 48 hours). Skin response was determined by the test criteria in Table 4, and the average response rate was calculated according to the following formula.

TABLE 4

The results are shown in Table 5.

TABLE 5

※ Criteria for average response rate (%) of human patch experiment

Less than 1: no stimulation, less than 1-3: slight stimulation less than 3-5: weak stimulation

5 or more: strong stimulus

As can be seen from Table 5, the whitening cosmetic composition according to the present invention can be said to be safe for the human body because it belongs to the non-irritating range.

The whitening effect of the extract of the present invention is the most effective whitening effect of the extract of the present invention compared with the arbutin and hydroxyanisole developed as a conventional whitening material. No irritation to

Claims (3)

After extracting pulverized Dioscorea rhizoma, 95% functional alcohol, which is 5 times higher, was added and extracted twice or more for 48 hours at 4-40 ℃, and then filtered and the filtrate was dried by rotary evaporator at 65 ℃. Whitening cosmetics containing The whitening cosmetic according to claim 1, wherein the acid medicine is a Chinese herb which is dried and dried as it is removed from the bark of the plant root stem belonging to the hemp and the yam family. The whitening cosmetic according to claim 1, wherein the acid extract is contained in an amount of 0.00001 to 10% by weight based on the dry weight of the cosmetic.