KR20030086849A - A composition for whitening containing crown daisy extract - Google Patents

Abstract

PURPOSE: Provided is a composition for skin whitening which contains a Chrysanthemum coronarium L. extract, as natural skin whitening material, selected from freeze-dried extract, hot water extract or ethanol extract. CONSTITUTION: A composition for skin whitening characteristically contains 0.01-10 wt.% of one or more kinds of Chrysanthemum coronarium L. extracts, as an active ingredient, selected from the group consisting of freeze-dried extract, hot water extract or ethanol extract. It inhibits tyrosinase activity to decrease the generation of melanin, and has extended storage.

Description

Whitening composition containing wormwood extract {A composition for whitening containing crown daisy extract}

The present invention relates to a whitening composition containing mugwort extract, and more particularly to a whitening composition containing a freeze-dried powder, hot water extract or ethanol extract of mugwort, which is a natural whitening material.

Wormwood chrysanthemum ( Chrysanthemum coronarium L.) spread from East of origin to East Asia and used as vegetables. Regions currently used as vegetables are India, Southeast Asia, China, Korea, Japan, etc. ˜3%, fat 0.3%, fiber 0.9%, ash 1.5%.

The content of vitamins in mugwort is 6,000 IU, vitamin B1 100r, vitamin B2 60r, and vitamin C 60mg per 100g of mugwort, and it also contains calcium as carotene and mineral which are contained in blush, etc. It can be said that it contains a large amount of lysine (lysine), an essential amino acid.

The skin color of the human body turns black when it is exposed to sunlight. This is because melanin (melanin), a kind of black pigment, is formed in the human body. It consists of black pigment and protein.

Melanin is tyrosine in melanocytes (tyrosine) as a substrate to act as a tyrosinase (tyrosinase) to produce a dopaquinone (dopaquinone) is synthesized through the oxidation and enzyme reaction.

Therefore, in order to inhibit the synthesis of melanin in the human skin, a substance having an effect of inhibiting the activity of tyrosinase or inhibiting some reactions during the production of melanin is required. As a tyrosinase inhibitor, Ascorbic acid, corticosteroids, retinoids, azelaic acid, hydroquinone, hydroxyanisole, kojic acid, arbutin, etc. However, these chemical synthetic materials have problems such as safety, stability and discoloration and deodorization during long-term storage.

In order to overcome the above problems, research has been continuously conducted to utilize the extracts of plants, which are natural substances, as a whitening ingredient. As a well-known technique, the whitening cosmetics using the ingredients of baekbaekpi (Korean Patent Publication No. 92-2109) , Japanese Patent Application No. 55-44375, Japanese Patent Application No. 64-26507, Japanese Patent Application No. 64-83009, Japanese Patent Application Publication No. 1-256587), Whitening cosmetics using ingredients of licorice ( Korean Patent Application Publication No. 92-2109, Japanese Patent Application Publication No. 60-214721, Japanese Patent Application Publication No. 63-23809, Japanese Patent Application Publication No. Hei 1-49706), and the like.

Also, whitening cosmetics using ingredients of peony (Japanese Patent Laid-Open No. 61-246109), whitening cosmetics using ingredients of cinnamon (Japanese Laid-Open Patent No. 63-30403), and whitening compositions using gardenia berry extract (Korean publication) Patent No. 2002-4896), anti-aging cosmetic composition containing the extract of Yu-geun-pi (Korea Patent Publication No. 2002-4896), whitening composition containing Cheonnyeonja extract (Korean Patent No. 2001-93861), pagoji extract A whitening composition (Korean Patent Publication No. 2001-93859) and a whitening composition (Korean Patent Publication No. 2001-93860) containing a sewage extract are disclosed.

It has been found that many of the above plant extracts act on tyrosinase to inhibit melanin synthesis, but such natural ingredients are insufficient to completely solve problems such as safety, stability, and discoloration potential while maintaining strong inhibitory activity. It is true.

Therefore, the present inventors conducted a study to solve the above problems by maintaining the stability using the wormwood extract, a natural material while inhibiting the activity of tyrosinase involved in the inhibition of melanocyte proliferation and the production of melanin to melanin The present invention was completed by finding out that a whitening composition capable of reducing the amount of produced was prepared.

Accordingly, it is an object of the present invention to provide a new whitening composition containing the mugwort extract as an active ingredient.

1 is a melanocyte proliferation inhibitory effect of mugwort extract according to the present invention,

2 is an effect of inhibiting the dopa oxidase activity of mugwort extract according to the present invention,

3 is a melanin production inhibitory effect of mugwort extract according to the present invention,

Figure 4 shows the morphology of B16 melanoma cells three days after treatment with 1000 ㎍ / ㎖ ethanol extract according to the present invention.

In the outposts of wormwood (crown daisy), many flavonoid derivatives have antioxidant properties, including caffeic acid, chlorogenic acid, and 3,5-dicafee. Oilquinic acid (3,5-dicaffeoylquinic acid), 4-reninyl-3,5-dicafeonylquinic acid (4-succinyl-3,5-dicaffeoylquinic acid), emodin, thiophene (N- Isobutyl-6 (2-thiophenyl) -2,4-hexadieneamide-1,8-dihydroxyandthroquinone (thiophene (N-isobutyl-6- (2-thiophenyl) -2,4-hexadienamide- 1,8-dihydroxyanthroquinone, quercetin and its glycoside derivatives, rutin and isoquercetin are known.

Vitamin C, carotinoids and other rich antioxidants in wormwood are known to prevent cardiovascular diseases, and chrysancorin has been reported to have antimicrobial activity (Takenaka M. et al., M Stability and bioavailability of antioxidants in garland (Chrysanthemum coronarium L.) Biosci Biotechnol Biochem . 2000 Dec; 64 (12): 2689-91; Gins VK et al., Oxyanthraquinones and flavonoids from garland chrysanthenum Prikl Biokhim Mikrobiol . 2000 May-Jun; 36 (3): 344-53; Wang H. et al., Purification of chrysancorin, a novel antifungal protein with mitogenic activity from garland chrysanthemum seeds Biol Chem . 2001 Jun; 382 (6): 947-51).

In the whitening composition according to the present invention, one or two or more wormwood extracts selected from the group consisting of lyophilized extract of mugwort, hot water extract of mugwort, and ethanol extract of mugwort are used.

The freeze-dried extracts of mugwort used in the present invention are added to a mixer or homogenizer, and then overnight in a deep freezer. Prepare by drying completely for 3 days.

The water soluble extracts of mugwort used in the present invention are added to distilled water to completely dried mugwort and extracted with hot water bath for 4-6 hours, after the filtered extract is overnight in a freezer, 2-3 in a freeze dryer. Prepared by drying for days.

In addition, ethanol soluble extracts of wormwood used in the present invention (ethanol soluble extracts) is added to the ethanol to completely dried mugwort and extracted for 4-6 hours at 45-55 ℃ over night the filtered extract in the freezer and then in a freeze dryer Prepare by drying completely for 2-3 days.

In the present invention, the effects of the above-mentioned mugwort extracts on melanocytes were verified through cell proliferation experiments, tyrosinase activity experiments, melanin amount measurement experiments, and the mugwort extracts used in the present invention were used for the proliferation of melanocytes. By inhibiting and inhibiting the activity of tyrosinase involved in the production of melanin as well as reducing the amount of melanin, it was confirmed that there is a whitening effect.

The whitening composition according to the present invention contains 0.01-10% by weight, preferably 0.01-5% by weight of the mugwort extract, but if it contains 0.01% by weight or less of mugwort extract, it does not have a whitening effect and contains 10% by weight or more. This is because there may be a problem in the safety or formulation formulation.

The whitening composition according to the present invention may be used in cosmetics such as cosmetic creams, lotions, cosmetic emulsions, beauty waters, liquid or solid soaps or syrups, ointments, creams, premises, tinctures, extracts, herbal preparations, acupuncture agents, premises It may be formulated as an oral or external preparation, and may contain a skin absorption promoting substance to enhance the whitening effect.

Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples. However, the technical scope of the present invention is not limited to these examples and test examples.

<Example 1> (Preparation of mugwort lyophilized extract)

1kg of wormwood freshly prepared by washing in water was homogenized evenly in a homogenizer, and then overnight overnight in a -70 ° C freezer, and then completely dried in a freeze dryer at -50 ° C under reduced pressure for 1 day 23g freeze-dried extract Was prepared.

<Example 2> (Preparation of mugwort hot water extract)

100 ml of distilled water was added to 5 g of the dried dried mugwort lyophilized extract prepared in Example 1, and extracted with hot water bath at 80 ° C. for 5 hours.

The extract obtained above was filtered through Whatman No. 2 filter paper overnight at -70 ° C freezer, and then completely dried in a freeze dryer at -50 ° C under reduced pressure for 3 days. 44.75 mg (8.951 g / wormwood 1 kg) extract was prepared.

<Example 3> (Preparation of mugwort ethanol extract)

100 g of 70% ethanol was added to 5 g of the fully dried mugwort lyophilized extract prepared in Example 1, and extracted at 50 ° C. for 5 hours.

The extract obtained by filtering the extract obtained above with a tree was overnight overnight in a -70 ° C freezer, and then completely dried in a freeze dryer at -50 ° C under reduced pressure for 3 days, and 10.78mg of ethanol extract (2.157g / wormwood) 1 kg) was prepared.

Example 4 (Preparation of Nutrients Containing Mugwort Extract)

5 parts by weight of mugwort hot water extract prepared in Example 2, 5 parts by weight of beeswax, 1 part by weight of polysorbate 60, 0.5 parts by weight of sorbitassesquiolade, 3 parts by weight of liquid paraffin, 3 parts by weight of squalane, capric triglyceride 3 parts by weight, glycerin 2.5 parts by weight, butylene glycol 2.5 parts by weight, propylene glycol 2.5 parts by weight, 0.1 parts by weight of carboxyvinyl polymer, 0.1 parts by weight of triethanol amine and a suitable amount of preservatives, pigments, flavors and purified water The nutritional cosmetics were prepared.

Example 5 Preparation of Cream Containing Mugwort Extract

10 parts by weight of wormwood ethanol extract prepared in Example 3, 8 parts by weight of stearyl alcohol, 2 parts by weight of stearic acid, 2 parts by weight of stearic acid cholesterol, 4 parts by weight of squalane, 6 parts by weight of 2-octyldodecyl alcohol, polyoxyethylene alcohol Cream was prepared by the conventional method using 3 weight part of esters, 2 weight part of glyceryl monostearic acid esters, 5 weight part of propylene glycol, and purified water.

<Test Example 1> (B16 melanoma cell culture)

B16 melanoma cells used for cell proliferation, tyrosinase activity, and melanin levels of wormwood extracts used in the whitening composition according to the present invention, 10% fetal bovine serum (fetal bovine serum) in RPMI1640 basic medium 1% penicillin-streptomycin was added to the culture medium at 37 ° C. and 5% CO _2. The culture medium was changed every three days.

Test Example 2 (Measurement of Cell Proliferation Inhibition Effect)

Mugwort freeze-dried extract (CE), mugwort hot water extract (WE) and mugwort hot water extract (WE) prepared in Examples 1 to 3 in order to determine the melanin cell growth inhibition effect of mugwort extract used in the whitening composition according to the present invention The results of measuring the proliferation rate of B16 melanoma cells treated with EE) are shown in Table 1 and FIG. 1.

Cell proliferation was measured by the following method. First, B16 melanoma cells incubated at a concentration of 2 × 10 ⁴ / dish in a 60 mm culture dish to confirm cell adhesion at a concentration of 10 μg / ml, 100 μg / ml and 1000 μg / ml, respectively. Cells were harvested by treatment with 0.25% Trypsin-EDTA solution on the third day.

500 μl of the harvested cell shaker was mixed and uniformly mixed with 10 ml of isotone solution and counted with a Coulter counter (ZM, Coulter Co., England). The calculation was as follows.

Total cell count = number of cells counted by the Coulter counter x 42 X (cell shake amount / ml)

TABLE 1 Cell Proliferation Rate

division Concentration 10 μg / ml 100 μg / ml 1000 μg / ml CE 98% 92% 59% WE 102% 100% 71% EE 99% 96% 69%

As shown in Table 1 and Figure 1, the wormwood extract 10 ㎍ / ㎖ and 100 ㎍ / ㎖ treated with a low melanocyte proliferation inhibition rate, 1000 ㎍ / ㎖ treated lyophilized extract (CE), mugwort hot water extract (WE) And wormwood ethanol extract (EE) both can be seen that the melanocyte proliferation inhibition rate is high.

<Test Example 3> (Measurement of Dopa Oxidase Activity Inhibition Effect)

Mugwort lyophilized extract (CE), mugwort hot water extract (WE) and mugwort ethanol extract (EE) prepared in Examples 1 to 3 to investigate the whitening effect of mugwort extract used in the whitening composition according to the present invention Table 2 and FIG. 2 show the results of measuring the activity inhibition rate of dopa-oxidase involved in the conversion of dopaquinone to dopaquinone during melanin production.

The activity inhibition rate of dopa oxidase was measured by the following method. First, B16 melanoma cells incubated at a concentration of 2 × 10 ⁴ / dish in a 60 mm culture dish to confirm cell adhesion at a concentration of 10 μg / ml, 100 μg / ml and 1000 μg / ml, respectively. Cells were harvested by treatment with 0.25% Trypsin-EDTA solution on the third day and counted with a Coulter counter.

Then, divide 2X10 ⁵ cells per treatment group into 1.7 ml tubes, add 100 μl of 1 mM L-dopa (Sigma Chemical Co., St, Louis, Missouri), and then dissolve the cell grinding solution at 37 ° C. for 1 hour. After the reaction, the absorbance was measured at 490 nm with a spectrophotometer.

TABLE 2 Dopa Oxidase Activity

division Concentration 10 μg / ml 100 μg / ml 1000 μg / ml CE 88% 75% 36% WE 97% 81% 39% EE 90% 78% 37%

As shown in Table 2 and FIG. 2, the activity inhibition rate of dopa oxidase was low in the 10 ㎍ / ml and 100 ㎍ / ml treatment of mugwort extract, but the freeze-dried extract (CE) and mugwort hot water extract ( Both WE) and wormwood ethanol extract (EE) showed high dopa oxidase activity inhibition rate.

<Test Example 4> (Melanin production amount measurement)

Mugwort lyophilized extract (CE), mugwort hot water extract (WE) and mugwort ethanol extract (EE) prepared in Examples 1 to 3 to investigate the whitening effect of mugwort extract used in the whitening composition according to the present invention The results of measuring the amount of melanin production by are shown in Table 3 and FIG.

Melanin production was measured by the following method. First, B16 melanoma cells incubated at a concentration of 2 × 10 ⁴ / dish in a 60 mm culture dish to confirm cell adhesion at a concentration of 10 μg / ml, 100 μg / ml and 1000 μg / ml, respectively. Cells were harvested by treatment with 0.25% Trypsin-EDTA solution on the third day and counted with a Coulter counter.

Then, 100 μl of 1N NaOH solution per 2 × 10 ⁵ cell number was added to each treatment group, and the cells were lysed at 37 ° C. for 12 hours, and then synthetic melanin (Sigma. Chemical Co., St, Louis, Misso) 10mg dissolved in 10ml of 1N NaOH to prepare 1mg / ml of stock solution, 700g / ml, 300g / ml, 100g / ml, 70g / ml, 30g / ml, 10g / A standard solution was prepared by diluting to ml, 7g / ml, 3g / ml, 1g / ml, 0.1g / ml and 0g / ml.

The sample and the standard solution were placed in a 96 well assy plate, and the absorbance was measured with a spectrophotometer at 490 nm, and then the measured absorbance value was applied to a standard straight line made of synthetic melanin, and converted into melanin amount.

Table 3 Melanin Production

division Concentration 10 μg / ml 100 μg / ml 1000 μg / ml CE 91% 78% 27% WE 99% 85% 35% EE 99% 82% 26%

As shown in Table 3 and FIG. 3, the melanin production inhibition rate was low in the 10 ㎍ / ㎖ and 100 ㎍ / ㎖ treatment mugwort extract, but lyophilized extract (CE), mugwort hot water extract (WE) and 1000 ㎍ / ㎖ treatment It can be seen that wormwood ethanol extract (EE) has a high melanin production inhibition rate.

<Test Example 5> (stability test)

In order to test the stability of the whitening composition containing wormwood extract according to the present invention, the nutrient lotion prepared in Example 4 and the cream prepared in Example 5 were used at 4 ° C, room temperature (25 ° C), 40 ° C and 50 ° C. After being left for months, the results of observing the stability of each composition are shown in Table 4.

TABLE 4 Stability Test Results

division Storage temperature (℃) 4 25 40 50 Example 4 stability stability stability stability Example 5 stability stability stability stability

As shown in Table 4, it can be seen that the whitening composition according to the present invention maintains stability even after a long time.

The whitening composition according to the present invention contains wormwood extract, which is a natural material, has a whitening effect by reducing the amount of melanin by inhibiting the activity of tyrosinase involved in the proliferation of melanocytes and the production of melanin. Stability can be maintained even during storage.

Claims (3)

Whitening composition containing mugwort extract as active ingredient The method of claim 1, The composition is a whitening composition characterized in that it contains 0.01-10% by weight of wormwood extract The method according to claim 1 or 2, The mugwort extract is a whitening composition characterized in that one or two or more selected from the group consisting of lyophilized extract of mugwort, hot water extract of mugwort, and ethanol extract of mugwort