KR102336538B1 - Composition for skin whitening comprising natural complex extracts with the effect of skin whitening - Google Patents

Abstract

The present invention relates to a skin whitening composition containing natural complex extracts having a skin whitening effect. More specifically, the present invention provides a skin whitening composition comprising Sanguisorba officinalis extract and seaweed extract, wherein the seaweed extract contains at least one selected from the group consisting of Undaria pinnatifida, Laminariales and Porphyra. In addition, the skin whitening composition further comprises Camellia sinensis.

Description

Composition for skin whitening comprising natural complex extracts with the effect of skin whitening

The present invention relates to a skin whitening composition containing a natural complex extract having a skin whitening effect, and more particularly, to a skin whitening composition containing a cucumber extract and seaweed extract.

Recently, interest in skin beauty is increasing, and pigmentation such as spots and freckles in the skin is caused by melanin production or proliferation of melanocytes due to ultraviolet rays.

Melanin is a black-brown biopolymer, produced in melanocytes, secreted from the cells in the form of melanosomes, and moved to the keratinocytes of the epidermis. The melanin biosynthetic cycle is a function of tyrosinase, TRP-1 (tyrosinase-related protein 1, DHICA oxidase), TRP-2 (tyrosinase-related protein 2, dopachrome tautomerase) enzymes specifically expressed in melanocytes. It is caused by complex physiological processes regulated by

It is known in the art that tyrosinase activity inhibitors such as arbutin, kojic acid, and linoleic acid have an effect of inhibiting melanin pigmentation. However, synthetic whitening agents recently used in skin whitening cosmetics are toxic to melanocytes and cause various side effects, so a new whitening agent is required.

Therefore, in order to solve the problems of raw materials having a skin whitening effect, and to develop a natural whitening agent with relatively low toxicity, it is possible to suppress abnormal melanin production from natural substances that have skin whitening action while ensuring safety. Efforts have been made to find extracts with

Cucumber grass (Sanguisorba officinalis L.) is a perennial grass of the Rosaceae family. The leaves of this plant are similar to those of elm trees and the freshly sprouted root leaves crawl on the ground, so it was called Jiyu. Roots and rhizomes contain 12-20% mixed tannins, gallic acid, ellagic acid, saponins, and poterin.

Seaweed (Undaria pinnatifida) is a seaweed with wide, flat leaves and spread in the shape of wings. The color is mainly black brown or yellowish brown, and the length is about 1~2 meters and the width is about 60cm. It usually grows from autumn to winter and propagates by spores in late spring or first summer. It has been widely used for food since ancient times, and it is very good for nutrition for children and pregnant women who are developing due to its high content of iodine and calcium.

Kelp refers to the brown algae kelp family, and there are about 20 species on the Pacific coast. It is 1.5-3.5 m long and 25-40 cm wide. It is a large sea horse, and the 2-4 year-old thallus (葉体) is a spore generation (胞子 generations), and the division of stem, leaf, and root is clear from the outside. The leaves are long and band-shaped, and slightly below the middle part is the widest. It has been used for food in Japan and China as well as Korea since ancient times and is a raw material for iodine.

Seaweed (Porphyra tenera) is a seaweed belonging to the family Porphyra, and grows by attaching to the reef of the sea like moss. It is 14-25 cm long and 5-12 cm wide. The body is long elliptical or egg-shaped like a string, with folds at the edges. The upper part of the body is reddish-brown, and the lower part is bluish green. Since it contains a lot of protein and vitamins, it has been used as a nutrient-rich food since ancient times.

Green tea (綠茶, Thea sinensis L.) is a tea made from unfermented tea leaves. Green tea contains a large amount of vitamins and minerals, which help enzyme activity to facilitate metabolism.

Therefore, while the present inventors were searching for natural substances as new materials with excellent skin whitening effect without side effects in the living body, they focused on cucumber grass and seaweed extracts and tested their functions. However, it was confirmed that it was not used for whitening purposes.

Unexamined Patent Publication No. 10-2019-0003011 (2019.01.09. Announcement) Laid-open Patent Publication No. 10-2012-0061733 (Announcement on June 13, 2012)

An object of the present invention is to provide a skin whitening composition comprising, as an active ingredient, an extract of cucumber grass and seaweed having excellent safety while having a skin whitening action in order to develop a natural whitening agent with relatively low toxicity.

In order to solve the above technical problem, the present invention provides a composition for skin whitening comprising an extract of cucumber grass and seaweed. The seaweed provides a composition for skin whitening that is at least one selected from the group consisting of seaweed, kelp and laver. The composition for skin whitening may further include green tea. It provides a cosmetic composition for skin whitening, a pharmaceutical composition, a quasi-drug, and a health functional food comprising the composition for skin whitening.

Since the composition for skin whitening according to the present invention contains cucumber grass and seaweed extract as active ingredients, it does not show toxicity to cells and has an effect of inhibiting melanin production, so skin whitening cosmetic compositions, pharmaceutical compositions, quasi-drugs and health functional foods can be usefully used as

1 is a graph confirming the cytotoxicity by each extract treatment.
Figure 2 is a graph confirming the melanin synthesis inhibitory effect by each extract treatment.
Figure 3 is a graph confirming the melanin synthesis inhibitory effect through the combination of cucumber extract and kelp, seaweed, laver or green tea extract.
Figure 4 is a graph confirming the melanin synthesis inhibitory effect through the combination of the two extracts in the group consisting of cucumber extract and kelp, seaweed, and green tea.
5 is a graph confirming the effect of inhibiting melanin synthesis through the combination of cucumber grass, kelp, laver and green tea extract and the combination of cucumber grass, kelp, seaweed, laver and green tea extract.

Hereinafter, the present invention will be described in detail through specific examples. However, the following examples are only illustrative of embodiments of the present invention, and the scope of the present invention is not limited to the following examples, and does not depart from the technical spirit of the present invention described in the claims. It will be apparent to those of ordinary skill in the art to which the present invention pertains that various modifications are possible within.

Hereinafter, embodiments and technical features of the present invention according to each step will be described in detail.

As used herein, the term "skin whitening" refers to the overall whitening of the basic skin tone, age spots, melasma, melasma, freckles, post-inflammatory hyperpigmentation, or hyperpigmentation disorders including sun-induced pigmentation defects. It refers to anything that reduces or improves melanin in the skin, including whitening.

The present invention provides a composition for skin whitening comprising an extract of cucumber grass and seaweed as an active ingredient in order to solve the above-described technical problem. Cucumber grass and seaweed may be a mixed extract in a weight ratio of 1-100: 1-100. Cucumber grass and seaweed may be a mixed extract in a weight ratio of 1-50: 1-50. Cucumber grass and seaweed may be a mixed extract in a weight ratio of 1 to 10: 1 to 10. Cucumber grass and seaweed may be a mixed extract in a weight ratio of 1-5: 1-5.

The seaweed is characterized in that at least one selected from the group consisting of seaweed, kelp and laver.

The composition for skin whitening may further include green tea. Cucumber grass, seaweed and green tea may be extracts mixed in a weight ratio of 1-100: 1-100: 1-100. Cucumber grass, seaweed and green tea may be extracts mixed in a weight ratio of 1-50: 1-50: 1-50. Cucumber grass, seaweed and green tea may be extracts mixed in a weight ratio of 1 to 10: 1 to 10: 1 to 10. Cucumber grass, seaweed and green tea may be extracts mixed in a weight ratio of 1-5: 1-5: 1-5.

The skin whitening composition provides a skin whitening composition comprising cucumber grass, kelp, seaweed and green tea extract. Cucumber grass, kelp, seaweed and green tea may be extracts mixed in a weight ratio of 1-100: 1-100: 1-100: 1-100. Cucumber grass, kelp, seaweed and green tea may be extracts mixed in a weight ratio of 1-50: 1-50: 1-50: 1-50. Cucumber grass, kelp, laver and green tea may be extracts mixed in a weight ratio of 1 to 10: 1 to 10: 1 to 10: 1 to 10. Cucumber grass, kelp, laver and green tea may be extracts mixed in a weight ratio of 1-5: 1-5: 1-5: 1-5.

The skin whitening composition provides a skin whitening composition comprising cucumber grass, kelp, seaweed, seaweed and green tea extract. Cucumber grass, kelp, seaweed, laver and green tea may be extracts mixed in a weight ratio of 1-100: 1-100: 1-100: 1-100: 1-100. Cucumber grass, kelp, seaweed, laver and green tea may be extracts mixed in a weight ratio of 1-50: 1-50: 1-50: 1-50: 1-50. Cucumber grass, kelp, seaweed, laver and green tea may be extracts mixed in a weight ratio of 1-10: 1-10: 1-10: 1-10: 1-10. Cucumber grass, kelp, seaweed, laver and green tea may be extracts mixed in a weight ratio of 1-5: 1-5: 1-5: 1-5: 1-5.

The cucumber plant may use one or more parts selected from the group including flowers, leaves, stems, fruits, seeds, roots and outposts. Specifically, the Cucumber extract of the present invention is most preferably prepared using the root of Cucumber.

As the extraction method of the extract, a conventional method in the art, such as hot water extraction, cold extraction, hot extraction, pressure extraction, reflux extraction, or ultrasonic extraction, may be used.

The extract is an extract soluble in a solvent selected from water, an organic solvent, or a mixture thereof, and preferably, the organic solvent is soluble in alcohol, hexane, acetone, ethyl acetate, butylene glycol or propylene glycol having 1 to 10 carbon atoms. one extract. Preferably, the alcohol having 1 to 10 carbon atoms is an alcohol having 1 to 5 carbon atoms. Preferably, the alcohol having 1 to 5 carbon atoms is methanol, ethanol, or propanol.

The skin whitening composition of the present invention may be used as a cosmetic composition, a pharmaceutical composition, a quasi-drug or a health functional food.

The cosmetic composition containing the composition of the present invention is not particularly limited in its formulation, and for example, lotion, cream, lotion, essence, pack, powder, lipstick, makeup base, foundation, mascara, ointment, gel, patch, and cosmetic liquid. , cleansing foam, cleansing cream, cleansing water, soap, shampoo, rinse, or spray, but is not limited thereto. A person skilled in the art can easily adopt and use a suitable cosmetically acceptable carrier according to the type of formulation.

Pharmaceutical compositions comprising the composition of the present invention include ointments, patches, gels, creams or sprays, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, tablets, granules, powders, capsules, liquid solutions, formulations such as pills, but are not limited thereto. A person skilled in the art can easily adopt and use a suitable pharmaceutically acceptable carrier depending on the type of formulation.

<Example 1> Preparation of cucumber grass, green tea and seaweed extract

1) Preparation of Cucumber Extract

To prepare a cucumbergrass extract, the oil, which is the root of cucumbergrass, was washed with primary distilled water, dried at 40°C for 24 hours using a dry oven to completely remove moisture, and then finely pulverized with a grinder and used as a sample. 500 ml of hot water made with primary distilled water was added to 100 g of a pulverized sample of the dried fat oil, stirred at 100° C. for 1 hour, and then extracted. The extracted solution was filtered twice with Whatman paper filter paper (Whatman no. 2), and the filtrate was filtered through a 0.45 μM filtration system (Corning, USA) to obtain an extract. The extracted solution was frozen in a cryogenic freezer and then freeze-dried using a freeze dryer to use the sample as the final extract. Each obtained sample was stored frozen and used as a sample for this experiment.

2) Preparation of green tea extract

To prepare a green tea extract, 500 ml of hot water made with primary distilled water was added to 100 g of dried green tea leaves and extracted for 1 hour. The extracted solution was filtered with filter paper, frozen in a cryogenic freezer, and then freeze-dried using a freeze dryer to use the sample as the final extract. Each obtained sample was stored frozen and used as a sample for this experiment.

3) Preparation of seaweed extract

To prepare a seaweed extract, kelp, seaweed, and seaweed collected from Shinan-gun were washed with distilled water to remove moisture, and then kelp and seaweed were finely ground and used as samples. To 100 g of each pulverized sample, 500 ml of 80% (v/v) ethanol was added, stored refrigerated for 7 days, and extracted by heating at 100° C. for 4 hours. The extract was filtered three times with Whatman paper filter paper (Whatman no. 2) to obtain an extract. The extract was frozen in a cryogenic freezer and then freeze-dried using a freeze dryer to use the sample as the final extract. Each obtained sample was stored frozen and used as a sample for this experiment.

<Example 2> Confirmation of cytotoxicity of cucumber grass, green tea and seaweed extracts

Cucumber extract, green tea extract, kelp extract, seaweed extract, and seaweed extract prepared in Example 1 were prepared at a concentration of 10-50 μg/mL, and then, in order to confirm whether they exhibit safety against skin cells, cytotoxicity was checked. Mice-derived melanocytes, Mel-ab cells (a spontaneously immortalized murine melanocyte cell line) were cultured using DMEM medium (WelGENE, South Korea). 10% (v/v) fetal bovine serum (FBS, Gibco, USA) and 50 μg/mL antibiotic (penicillin-streptomycin) were added to the culture medium, and 100 nM 12-Otetradecanoylphorbol-13 was added to induce melanin biosynthesis. -acetate (TPA) and 1 nM cholera toxin (CT) were added and cultured. During the experiment, cell lines were cultured in a 5% CO2 incubator.

The cultured Mel-ab cells were each dispensed in a 24-well plate at 5 x 104 cells/well and cultured for 24 hours, each extract was diluted according to the concentration and added, and then cultured again in the same environment for 24 hours. To use as a negative control, Mel-ab cells were treated with PBS or ethanol solution and cultured under the same conditions. After completion of the culture, the cells were stained with 0.1% (w/v) crystal violet solution for 5 minutes, and then treated with 95% (v/v) ethanol for 15 minutes, and absorbance was measured at absorbance at 590 nm. The density was checked. All treatment experiments were repeated three times. The cytotoxicity (%) of each extract was calculated using the measured absorbance value.

As a result of the experiment, as shown in FIG. 1 , as a result of confirming the cytotoxicity of the cucumber extract against Mel-ab cells, it was determined that there was no cytotoxicity up to a concentration of 10-50 μg/mL. In addition, green tea extract, kelp extract, seaweed extract and seaweed extract were also measured to have no cytotoxicity at a treatment concentration of 10 μg/mL.

As a result of co-treatment with each extract, cells were not only co-treated with each extract at a treatment concentration of 10 μg/mL, but also when co-treated with a treatment concentration of 50 μg/mL of cucumber extract and 10 μg/mL of the remaining extracts. It was confirmed that there was no toxicity.

<Example 3> Melanin synthesis inhibitory effect of cucumber grass, green tea and seaweed extracts

In order to confirm the pigmentation inhibitory effect of the combined treatment of cucumber extract and other extracts, cucumber extract, seaweed extract, kelp extract, seaweed extract, and green tea extract are treated in cells that cause melanin pigment accumulation to be produced in each treatment group. Melanin pigment was quantified.

After dispensing the cultured Mel-ab cells at 2 x 105 cells/well each in a 6-well plate and culturing for 24 hours, hot water cucumber extract (10-50 μg/mL), green tea extract (10 μg/mL), kelp extract ( 10μg/mL), seaweed extract (10μg/mL) and seaweed extract (10μg/mL) were diluted according to each concentration and added to the experimental medium, and then the cells were cultured. A negative control group was treated with PBS, and 500 μM arbutin as a positive control group was treated with Mel-ab cells under the same conditions. Samples were treated in each control group and test group, and the cells were allowed to react for 5 days. After removing the medium on the 5th day after the extract treatment, 550 μl of 1 N NaOH was treated and the reaction was performed in an incubator at 37° C. for 10 minutes, followed by reaction in a water bath at 100° C. for 30 minutes to dissolve melanin. After dissolution, the absorbance of the medium was measured at 400 nm to confirm the degree of melanin pigment production. And in order to check the cytotoxicity caused by the sample treatment for 5 days, the protein of each treatment group was quantified using the dissolved medium using the Bradford method. Melanin content by each sample was measured by calculating the melanin content per unit protein using the quantified protein value. All sampling experiments were repeated three times.

As a result, as shown in FIG. 2, the melanin content in the Mel-ab cells was increased in the PBS-treated group (100±1.55%), and the melanin content was decreased in 500 μM arbutin and each extract as a positive control. As a result of comparing the melanin content in each extract, arbutin (80.71±3.02%), cucumber extract (10μg/mL, 82.19±1.29%), cucumber grass extract (50μg/mL, 70.55±2.46%), kelp extract (10μg/mL) , 93.8±6.69%), seaweed extract (10μg/mL, 90.91±6.52%), laver extract (10μg/mL, 97.88±1.22%), green tea extract (10μg/mL, 85.69±1.64%) Melanin content was measured.

<Example 4> Melanin synthesis inhibitory effect through the combination of cucumber grass extract and kelp, seaweed, laver or green tea extract

The degree of melanin production by the combination of cucumber grass extract, kelp extract, seaweed extract, laver extract, and green tea extract was confirmed. Cucumber and seaweed extracts, cucumber grass and kelp extracts, cucumber grass and laver extracts, or cucumber grass and green tea extracts were added in the same weight ratio to the experimental medium so that the total concentration was 10 μg/mL, and then the cells were cultured.

As a result, as shown in FIG. 3(a), as a result of the combined treatment of the two extracts in Mel-ab cells, cucumber grass and seaweed extract, cucumber grass and kelp extract, cucumber grass and seaweed extract, cucumber grass and green tea extract were combined, respectively, 79.27 The melanin content of ±3.12%, 90.27±4.68%, 86.68±0.89%, and 78.72±0.47% was measured.

In particular, when the melanin biosynthesis inhibitory effect is calculated, as shown in FIG. 3(b), the melanin biosynthesis inhibitory effect of 20.73% and 21.28% of the melanin biosynthesis inhibitory effect was confirmed in the combination of cucumber grass and seaweed extract, cucumber grass and green tea extract, respectively, when the two extracts were treated together. The compounding effect by simple sum expected from the melanin biosynthesis inhibitory effect of the single extract confirmed in 3 (1/2 of the sum of the melanin biosynthesis inhibitory effect of the two single extracts of Example 3), which is 13.45%, 16.06% more effective than A synergistic effect was confirmed, and among them, the most outstanding synergistic effect was confirmed when the cucumber grass and seaweed extracts were combined.

<Example 5> Melanin synthesis inhibitory effect through the combination of two extracts from the group consisting of cucumber grass extract and kelp, seaweed, and green tea

In order to find a combination that shows a synergistic effect other than seaweed when combined with cucumber extract, two extracts from the group consisting of cucumber grass, kelp, seaweed, and green tea were added in the same weight ratio to the experimental medium so that the total concentration of the mixture was 10 μg/mL. Then, the cells were cultured.

As a result of the combined treatment, as shown in Fig. 4(a), 91.93±3.02%, 83.33±1.27, respectively, in the formulation of cucumber grass, kelp and laver extract, the combination of cucumber grass, kelp and green tea extract, and the combination of cucumber grass, laver and green tea extract %, the melanin content of 78.62±2.03% was measured.

In particular, when calculating the melanin biosynthesis inhibitory effect, as shown in Fig. 4(b), when the three extracts were treated together, the melanin biosynthesis inhibitory effect of 21.38% was confirmed in the combination of cucumber grass, seaweed and green tea extracts. A synergistic effect superior to that of 11.41%, which is the compounding effect by simple sum expected from the melanin biosynthesis inhibitory effect (1/3 of the sum of the melanin biosynthesis inhibitory effect of the three single extracts of Example 3), was confirmed.

<Example 6> Melanin synthesis inhibitory effect through a combination of cucumber grass, kelp, seaweed and green tea extract and a combination of cucumber grass, kelp, seaweed, laver and green tea extract

A combination of cucumber grass, kelp, seaweed and green tea extract and a combination of cucumber grass, kelp, seaweed, laver and green tea extract were added to the test medium so that the total concentration of the mixture was 10 μg/m in the same weight ratio, respectively, and then the cells were cultured. .

As a result of the treatment, as shown in FIG. 5(a), melanin content of 72.52±0.97% and 70.45±0.92%, respectively, was measured.

In particular, when calculating the melanin biosynthesis inhibitory effect, as shown in FIG. 5(b), 27.48% and 29.55% of melanin in the combination of cucumber grass, kelp, laver and green tea extract and the combination of cucumber grass, kelp, seaweed, laver and green tea extract, respectively The biosynthesis inhibitory effect was confirmed and the compounding effect by simple summation expected from the melanin biosynthesis inhibitory effect of the single extract confirmed in Example 3 (four combinations are 1/4 of the sum of the melanin biosynthesis inhibitory effect of the four single extracts of Example 3) , five formulations showed a synergistic effect superior to that of 10.11% and 9.9%, which is 1/5 of the sum of the sum of the melanin biosynthesis inhibitory effects of the five single extracts of Example 3), and 500 μM arbutin (19.29%) as a comparison group It was confirmed that the effect was superior to that of the treated group.

Cucumber, kelp, seaweed, seaweed and green tea extract were added to the test medium in a weight ratio of 5:1:1:1:1 so that the total concentration of the mixture was 10μg/m, and then the cells were cultured. A melanin content of 64.1±1.5% was determined.

When the melanin biosynthesis inhibitory effect is calculated, the melanin biosynthesis inhibitory effect of 35.9% was confirmed, and the compounding effect by simple summation expected from the melanin biosynthesis inhibitory effect of the single extract confirmed in Example 3 (the biosynthesis inhibitory effect of the cucumber extract of Example 3) 5/9 and 1/9 of the biosynthesis inhibitory effect of the remaining extracts), a synergistic effect superior to that of 15.92% was confirmed.

Claims (10)

delete including cucumbergrass and seaweed extracts,
The composition for skin whitening wherein the seaweed is at least one selected from the group consisting of seaweed, kelp and laver.
The composition for skin whitening according to claim 2, wherein the seaweed is seaweed.
The composition according to claim 2, wherein the seaweed is seaweed, and the composition further comprises green tea extract.
The composition for skin whitening according to claim 2, wherein the seaweed is kelp and seaweed, and further comprises green tea extract.
The composition for skin whitening according to claim 2, wherein the seaweed is kelp, seaweed and seaweed, and further comprises a green tea extract.
The cosmetic composition for skin whitening according to any one of claims 2 to 6, comprising the composition for skin whitening.
The pharmaceutical composition for skin whitening according to any one of claims 2 to 6, comprising the composition for skin whitening.
The quasi-drug for skin whitening according to any one of claims 2 to 6, comprising the composition for skin whitening.
The health functional food for skin whitening according to any one of claims 2 to 6, comprising the composition for skin whitening.

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