KR102255580B1 - Cosmetic composition comprising Ilex paraguayensis extract and Tranexamic acid - Google Patents

Abstract

The present invention relates to a whitening, antioxidant and anti-wrinkle cosmetic composition containing a mixture of Ilex paraguayensis extracts and tranexamic acid as active ingredients. Since the mixture provided by the present invention has excellent melanocyte proliferation inhibitory activity, melanocyte cycle inhibitory activity, melanogenesis inhibitory activity, DPPH scavenging activity and elastase inhibitory activity, the mixture can be used as a whitening, antioxidant and anti-wrinkle cosmetic composition.

Description

Cosmetic composition comprising Ilex paraguayensis extract and Tranexamic acid as active ingredients

The present invention relates to a cosmetic composition comprising a mixture of mate extract and tranexamic acid as an active ingredient.

In general, it is known as melanin that is most closely associated with skin whitening. Human skin color is determined by the content and distribution of melanin inside the skin. In addition to genetic factors, it is affected by environmental or physiological conditions such as solar ultraviolet rays, fatigue, and stress. Melanin is synthesized from melanocytes in the epidermis of the skin, and an enzyme called tyrosinase acts on tyrosine, a kind of amino acid in the melanosome, an organelle within the melanocyte It is made through a non-enzymatic oxidation reaction after being converted to quinone. When such melanin synthesis occurs excessively in the skin, skin tone is darkened, and spots, freckles, senile blacks, and birthmarks are generated. Therefore, by inhibiting the synthesis of melanin pigment by inhibiting the tyrosinase activity in the skin, not only can the skin tone be brightened to realize skin whitening, but also skin hyperpigmentation such as spots and freckles caused by ultraviolet rays, hormones and genetic causes. It can improve calmness.

As the desire and interest in beauty has increased due to the recent development of technology and the improvement of the quality of life, it has become necessary to develop a whitening product that prevents excessive melanin production, and accordingly, research on the development of whitening agents and whitening cosmetics is actively underway. Several whitening substances are used today to treat melanogenesis disorders, but most of them are toxic to melanogenic cells and cause several side effects. For example, hydroquinone, which has excellent whitening effect, has cytotoxicity, and is known to have renal toxicity and carcinogenic potential. In addition, due to skin damage such as tissue blackness (Ochonosis), it is prohibited to use in whitening cosmetics in Europe. Kojic acid is known to have excellent whitening effect, but it is cytotoxic and can cause contact dermatitis or cancer.

Free radicals introduced from outside the body or generated in the body promote aging in the body or cause cancer. Therefore, research on antioxidants that inhibit oxidation by free radicals is being conducted, and antioxidants are known as phenolic compounds, flavonoids, tocopherols, vitamin C, selenium, and the like. However, when applied to the skin, antioxidant substances that exist in nature cannot be expected to have a substantial effect. On the other hand, synthetic antioxidants having excellent antioxidant power and low cost have a problem that their use is limited due to concerns about safety such as side effects on the human body.

The causes of wrinkle formation can be largely divided into wrinkle formation due to natural aging and wrinkle formation due to UV exposure (photoaging), and the mechanism of wrinkle formation due to the two causes has not yet been accurately identified. Common phenomena found in relation to skin changes caused by natural aging and photoaging are the reduction and transformation of collagen and elastin, the main constituent proteins in the dermis, and reduction of skin elasticity due to the reduction of hyaluronic acid, the matrix of the dermis. In particular, according to the results of the studies so far, it is known that the reduction and transformation of collagen and elastin, the main proteins in the dermis, are directly involved in the formation of wrinkles and the reduction of elasticity of the skin. Collagen is a major matrix protein produced by fibroblasts of the skin and is present in the extracellular interstitial. Its important functions include skin mechanical firmness, connective tissue resistance and tissue bonding, support for cell adhesion, cell division and differentiation (organic Growth or induction of wound healing) is known. This collagen is reduced by age and photoaging caused by UV irradiation, which is known to be closely related to the formation of wrinkles in the skin. In addition, as the results of extensive research on skin aging have recently been reported, an important function of collagen in the skin has been revealed. Active ingredients that promote collagen synthesis to improve wrinkles are known. For example, retinoic acid, TGF (transforming growth factor), animal placenta-derived proteins, betulinic acid, and chlorella extract are known as collagen synthesis promoting substances.

As described above, research and development having whitening, antioxidant and anti-wrinkle effects are being actively progressed in various fields, and in particular, research using natural substances has been continued in order not to cause toxicity or irritation to the skin.

As for the cosmetic composition technology using plant extracts, in Korea, Korean Patent Publication No. 10-1789516 ``Antioxidant composition containing mate, Eoseongcho, octagonal and seaweed complex fermented extracts as active ingredients and a method for manufacturing the same'', registered patents in the Republic of Korea. Publication No. 10-1077229 “Method for preparing a megastigman derivative isolated from mate tea and a composition for improving wrinkles containing the same as an active ingredient”, Korean Patent Publication No. 10-1848887 “Tranexamic acid having skin whitening activity- Peptides and their uses”, Korean Patent Publication No. 10-1511252, “a composition for skin whitening containing white extract and tranexamic acid as active ingredients”, and the like are disclosed.

Accordingly, the present inventors have found that a cosmetic composition containing a mixture of mate (Ilex paraguayensis ) extract and tranexamic acid as an active ingredient has excellent melanocyte proliferation inhibitory activity, melanocyte cycle inhibitory activity, melanin production inhibitory activity, and DPPH scavenging. The present invention was completed by discovering the fact that it can simultaneously provide whitening, antioxidant and anti-wrinkle effects by simultaneously exhibiting the activity and the elastase inhibitory activity.

Korean Registered Patent Publication No. 10-1789516 Republic of Korea Patent Publication No. 10-1077229 Korean Patent Publication No. 10-1848887 Republic of Korea Patent Publication No. 10-1511252

An object of the present invention is to provide a cosmetic composition that can simultaneously impart whitening, antioxidant and anti-wrinkle effects.

Another object of the present invention is to provide a cosmetic product comprising the composition.

In order to achieve the above object, the present invention provides a whitening, antioxidant and anti-wrinkle cosmetic composition comprising a mixture of mate (Ilex paraguayensis) extract and tranexamic acid as an active ingredient.

In the present invention, the mate extract is characterized in that it is extracted with a solvent selected from the group consisting of water, an organic solvent, and a mixed solvent thereof.

In the present invention, the mixture of the mate extract and tranexamic acid is characterized in that the mate extract and tranexamic acid are mixed in a weight ratio of 1:1 to 20.

In the present invention, the mixture of the mate extract and tranexamic acid is characterized in that it is contained in an amount of 0.01 to 30% by weight based on the total weight of the composition.

In the present invention, the mixture of the mate extract and tranexamic acid is characterized in that it has a melanocyte proliferation inhibitory activity, a melanocyte cycle inhibitory activity, a melanogenesis inhibitory activity, a DPPH scavenging activity, and an elastase inhibitory activity.

In addition, the present invention provides a cosmetic comprising the composition.

The mixture provided from the present invention has excellent melanocyte proliferation inhibitory activity, melanocyte cycle inhibitory activity, melanin production inhibitory activity, DPPH scavenging activity and elastase inhibitory effect, so it can be used as a cosmetic composition for whitening, antioxidant and anti-wrinkle. .

1 is a graph showing the cell proliferation inhibitory activity of the mouse melanoma cell line (B16F10 cell) of Example 1. FIG.
Figure 2 is a graph showing the cell cycle inhibitory activity of the mouse melanoma cell line (B16F10 cell) of Example 1.
3 is a graph showing the activity of inhibiting the synthesis of extracellular melanin in Example 1. FIG.
4 is a graph showing the evaluation of free radical scavenging activity of Example 1. FIG.
5 is a graph showing the elastase inhibitory activity of Example 1.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by an expert skilled in the art to which the present invention belongs. In general, the nomenclature used in this specification is well known and commonly used in the art.

In the entire specification of the present application, when a part "includes" a certain component, it means that other components may be further included rather than excluding other components unless otherwise specified.

According to one embodiment of the present invention, the present invention relates to a cosmetic composition for whitening, antioxidant and anti-wrinkle comprising a mixture of mate (Ilex paraguayensis) extract and tranexamic acid as an active ingredient.

The mate extract according to the present invention may be obtained through various extraction methods known in the field to which the present invention belongs, and specifically, may be extracted with a solvent selected from the group consisting of water, an organic solvent, and a mixed solvent thereof. . Among them, extraction with ethanol is more preferable in terms of stably extracting the active ingredient.

The water may be alkaline water, and the organic solvent may be an anhydrous or aqueous lower alcohol having 1 to 4 carbon atoms, such as ethanol, propanol, methanol, n-butanol, acetone, 1,3-butylene glycol, etc. Organic solvent; Non-polar organic solvents such as ether, hexane, benzene, chloroform, and ethyl acetate; Vegetable organic solvents such as soybean oil and sesame oil; And one or more of these mixtures. The organic solvent may be used alone or in combination of two or more.

As a solvent used in the extraction, it may be used as a mixed solvent including one or more organic solvents of ethanol, methanol, n-butanol, and propanol, and 1,3-butylene glycol or water in a solvent thereof. When the mixed solvent is used as the extraction solvent, the extraction efficiency of the active ingredient may be further improved. At this time, when using the mixed solvent, it may be appropriately mixed and used according to an arbitrary purpose.

Specifically, the content of the extraction solvent may be 1 to 500 times the weight of the mate solid content, more preferably 5 to 200 times, most preferably 10 to 100 times.

In addition, the extract may be extracted by heating or at room temperature under conditions in which the active ingredient of the extract is not destroyed or the destruction is minimized. The extraction method is not particularly limited, and according to the type of the extract and the method of extraction, for example, boiling water extraction, reflux cooling extraction, ultrasonic extraction, cold sedimentation extraction, and percolation extraction method may be used.

In the extracting step, the number of extractions may be specifically repeated 1 to 5 times. In this case, the extraction efficiency of the active ingredient may be further improved.

For example, the step of extracting may include adding an extraction material and an extraction solvent to an extractor equipped with a reflux cooler, and then performing heating at 50 to 100° C. for 4 to 20 hours with reflux cooling and extracting. At this time, the heating-reflux cooling extraction method is a method of extracting by refluxing while heating, and then cooling to extract. Specifically, since the evaporation phenomenon may occur due to heating and solvent loss may occur, the cooling process is performed together to minimize the loss of solvent. Means the extraction method. Alternatively, in the step of extracting, the extraction solvent may be added to the extraction material, followed by immersion at 5 to 37°C for 1 to 15 days, followed by cold precipitation.

In the step of extracting as another method, after the extraction solvent is added to the extraction material, percolation may be performed for 1 to 200 hours.

In another method, the extraction may be performed by adding an extraction solvent to the extraction material, followed by boiling at 50 to 100° C. for 0.5 to 72 hours.

In the step of extracting as another method, ethanol of 10 to 100 times the weight of the extracted material is added based on the dry solids of Mate, extracted for 1 to 72 hours at 20 to 80°C, filtered and concentrated to form a concentrate. The extract of can be obtained.

In addition, the extraction material of the present invention includes not only the extraction material extracted by the above-described extraction solvent, but also the extraction material extracted through a conventional purification process. For example, the active fraction obtained through various purification methods additionally carried out, such as separation using an ultrafiltration membrane having a certain molecular weight cut-off value, is also included in the extract of the present invention.

Filtration is a process of removing suspended solid particles from the extract, and filtering particles using cotton, nylon, paper, etc., or ultrafiltration, cryofiltration, centrifugation, etc. may be used, but is not limited thereto.

The step of concentrating and then drying the filtrate includes, but is not limited to, freeze drying, vacuum drying, hot air drying, spray drying, vacuum drying, foam drying, high frequency drying, infrared drying, and the like. In some cases, a process of pulverizing the final dried extract may be added.

According to one embodiment of the present invention, the mate extract may be processed and used in various forms, such as an extract, a powder obtained by drying the extract, and a concentrate obtained by concentrating and filtering the extract. In addition, the extract is not particularly limited, but may be used in the form of, for example, tink, concentrate, extract, flow extract, etc. containing hydrous alcohol as a leaching solvent.

According to an embodiment of the present invention, the mixture of mate extract and tranexamic acid is preferably made by mixing the mate extract and tranexamic acid in a weight ratio of 1:1 to 20, and mixing in a weight ratio of 1:1 to 10 What is made is most preferable in terms of improving all of the whitening, antioxidant and anti-wrinkle effects. If it is out of the above range, it is not preferable because the skin cosmetic effect may be reduced.

According to an embodiment of the present invention, the mixture of mate extract and tranexamic acid obtained from the above method may be included in an amount of 0.01 to 30% by weight, more preferably 0.1 to 10% by weight, based on the total weight of the composition. have. If the extract is less than 0.01% by weight based on the total weight of the composition, whitening, antioxidant and anti-wrinkle effects cannot be sufficiently obtained, so the function as a composition decreases, and if it exceeds 30% by weight, sufficient efficacy even if it does not contain more Not only is it uneconomical because it can represent, but also it is not preferable because it is difficult to use as a cosmetic due to deterioration of the stability and spreadability of the formulation.

Since the mixture of the mate extract and tranexamic acid can simultaneously impart whitening, antioxidant and anti-wrinkle effects, there is an effect that can realize not only skin beauty but also fundamental skin health improvement.

As described above, since the mixture of mate extract and tranexamic acid has excellent melanocyte proliferation inhibitory activity, melanocyte cycle inhibitory activity, melanogenesis inhibitory activity, DPPH scavenging activity and elastase inhibitory activity, whitening, antioxidant and wrinkle It can be used as a cosmetic composition for preventing.

According to one embodiment of the present invention, the cosmetic composition is purified water (water), glycerin (glycerin), myristic acid (myristic acid), butylene glycol (butylene glycol), potassium hydroxide (potassium hydroxide), polyethylene Glycol (polyethylene glycol), stearate, stearic acid, lauric acid, lauramide DEA, glyceryl stearate, baehenyl alcohol ( behenyl alcohol), microcrystalline wax, phenoxyethanol, ethylhexylglycerin, tocopheryl acetate, 1,2-hexanediol, car Caprylyl glycol, hydroxyethylene cellulose, xanthan gum, collagen, sodium citrate, citric acid, magnesium ascorbate And it may include one or more selected from the group consisting of fragrance (fragrance).

In addition, the cosmetic composition of the present invention may be blended with other ingredients that are usually blended in the cosmetic, if necessary. For example, oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation accelerators, coolants , A limiting agent or purified water is preferred, but is not limited thereto.

According to one embodiment of the present invention, when used as the cosmetic composition, skin ointment, cream, softening lotion, astringent lotion, nutrient lotion, pack, mask sheet, essence, hair tonic, shampoo, conditioner, hair conditioner, hair treatment Ment, spray, gel, gel ointment, patch, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, cream, massage cream, nourishing cream, eye cream, moisture cream, hand cream, Foundation, nutritional essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream, cleansing water, powder, body lotion, and may have any one formulation selected from the group consisting of a body cleanser, but is not limited thereto. The composition of each of these formulations may contain various bases and additives necessary and appropriate for formulation of the formulation, and the types and amounts of these components can be easily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, titanium oxide, etc. may be used as carrier components. I can.

When the formulation of the present invention is a body gel, in addition to the above-described active ingredients, ingredients generally used in the manufacture of body gels, such as carbomer, triethanolamine, menthol, ethanol, glycerin, disodium EDTA, methylparaben, fragrance, etc. Can be used. Here, carbomer and triethanolamine act as a thickening agent, menthol acts as a fragrance ingredient, ethanol functions as a refreshing sensation, glycerin acts as a moisturizer, and disodium EDTA functions as a metal chelate. And, methylparaben is added to function as a preservative. In addition, in addition to menthol fragrance, various fragrances such as herbal fragrance may be added to the fragrance.

In addition to the above-described specific ingredients, it is also within the scope of the present invention to apply known substances widely used in the present technical field to achieve the functions of a thickener, a refreshing sensation, a fragrance ingredient, a moisturizer, and the like described above. Can be included.

When the formulation of the present invention is a powder or spray (atomizing agent), lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be added as a carrier component. Propellants such as carbon, propane/butane or dimethyl ether may be included.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is added as a carrier component, examples of which include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol , 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, and the like.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, crystallites Sex cellulose, aluminum metahydroxide, bentonite, agar or tracanth, and the like can be used.

When the formulation of the present invention is a surfactant containing cleansing, as a carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether Sulfate, alkylamidobetaine, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters may be used.

According to one embodiment of the present invention, the cosmetic composition for whitening, antioxidant and anti-wrinkle may be prepared as a composition such as body gel, body lotion, body cream, body oil, etc. for promoting fat decomposition by adding conventional additives, and It can also be manufactured in types. In this case, the cosmetic composition is preferably used as a method of transdermal administration by directly applying or spraying on the skin.

According to an embodiment of the present invention, the amount of use of the cosmetic composition for whitening, antioxidant and anti-wrinkle can be appropriately adjusted according to individual differences, formulations, and shapes such as age and skin fat condition, and has whitening, antioxidant and anti-wrinkle effects. It can be usefully used as a composition for.

The composition obtained from the above method exhibits melanocyte proliferation inhibitory activity, melanocyte cycle inhibitory activity, melanin production inhibitory activity, DPPH scavenging activity and elastase inhibitory activity to provide excellent whitening, antioxidant and anti-wrinkle cosmetics. .

Hereinafter, the present invention will be described in more detail through examples. These examples are only for describing the present invention in more detail, and that the scope of the present invention is not limited by these examples according to the gist of the present invention, to those of ordinary skill in the technical field to which the present invention pertains. It will be self-evident.

<Examples 1 to 4> Preparation of a mixture of mate extract and tranexamic acid

10 g of dried mate (Mate, Jecheon herbal medicinal herbs) and 70% ethanol were added at 10 times the weight of the dried mate, and extracted by heating and refluxing cooling extraction at 80° C. for 2 hours. After extraction, the filtrate obtained by filtration through a 0.45 μm filter was concentrated under reduced pressure and freeze-dried to prepare a powdery mate extract. The prepared powdered mate extract and powdered tranexamic acid were mixed in the amount (g) shown in Table 1 below, and then 100 mL of purified water was added and filtered through a 0.45㎛ filter to obtain a mixture of the final mate extract and tranexamic acid. Was prepared.

Example One 2 3 4 Mate extract 0.2 0.2 0.2 0.2 Tranexamic acid 2 One 0.2 4

<Comparative Examples 1 to 4> Preparation of a mixture of mate extract and tranexamic acid

A mixture of mate extract and tranexamic acid was prepared by mixing the mate extract and tranexamic acid in powder form prepared in the same manner as in Example 1 in the amount (g) shown in Table 2 below.

Comparative example One 2 3 4 Mate extract 0.2 - 0.2 0.2 Tranexamic acid - 2 0.1 0.04

<Comparative Example 5> Preparation of Baekchul extract

10 g of dried Baekchul (Baekchul, Jecheon Herbal Herbs) and 70% ethanol were added at 10 times the weight of the dried Baekchul, followed by heating at 80° C. for 2 hours, followed by cooling extraction with reflux. After extraction, the filtrate obtained by filtration through a 0.45 μm filter was concentrated under reduced pressure and freeze-dried to prepare a powdery extract of Baekchul. 100 mL of purified water was added to 0.2 g of the prepared powdered Baekchul extract and filtered through a 0.45 μm filter to prepare a final Baekchul extract.

<Experimental Example 1> Cytotoxicity test: WST (Water-soluble tetrazolium salt) assay

Cytotoxicity experiments were conducted using HaCaT cells and B16F10 melanoma cells, which are human keratin cell lines, using Examples 1 to 4 and Comparative Examples 1 to 5 to determine whether they have toxicity to skin cells.

In order to confirm whether it has toxicity to human keratinocyte line, HaCaT cells and mouse melanoma cell line, B16F10 cells, using DMEM (Dulbecco's Modified Eagle Medium) medium, 5X10 ³ cells/well and 4X10 ³ cells/well After dispensing each of the cells at the concentration, the cells were cultured for 24 hours at _{37°C and 5% CO 2.} In the cells cultured for 24 hours, Examples 1 to 4 and Comparative Examples 1 to 5 were mixed with a medium to a final concentration of 4%, treated in each well, and cultured for 24 hours at _{37°C and 5% CO 2} , After removing the culture medium and treating each well with WST-1 assay solution (ez-cytox) for 2 hours, the absorbance was measured at 450 nm with an ELISA reader, and the cell viability was calculated as shown in Equation 1 below. .

[Equation 1]

Cell viability (%) = [(absorbance of sample treated group/absorbance of untreated group) X 100]

HaCaT cell viability (%) B16F10 cell viability (%) Example 1 105.00±2.91 98.67±4.76 Example 2 103.11±1.59 106.55±6.38 Example 3 98.29±4.63 96.41±4.63 Example 4 101.73±3.33 108.39±4.80 Comparative Example 1 107.99±2.89 96.05±2.22 Comparative Example 2 107.27±2.64 99.47±4.77 Comparative Example 3 103.00±3.08 101.55±4.86 Comparative Example 4 96.50±2.54 93.01±2.99 Comparative Example 5 98.76±4.05 95.88±2.67

As a result, as shown in Table 3 above, it was confirmed that cytotoxicity was not observed at the 4% treatment concentration of Examples 1 to 4, so that it could be used as a safe material for cosmetics.

<Experimental Example 2> Cell proliferation test: WST (Water-soluble tetrazolium salt) assay

Cell proliferation experiments were performed using B16F10 melanoma cells in order to confirm whether it has a proliferation inhibitory effect on melanocytes using Examples 1 to 4 and Comparative Examples 1 to 5.

To determine whether it affects the proliferation of B16F10 cells, which is a melanoma cell line in mice, cells were dispensed at a concentration ^{of 3X10 3 cells/well in a 96-well plate using DMEM (Dulbecco's Modified Eagle Medium) medium, and then at 37°C and} Incubated for 24 hours in 5% CO _{2 conditions.} Examples 1 to 4 and Comparative Examples 1 to 5 were mixed with a medium to a final concentration of 4% in cells cultured for 24 hours, treated in each well, and cultured for 72 hours at _{37°C and 5% CO 2} , After removing the culture medium and treating each well with WST-1 assay solution (ez-cytox) for 2 hours, the absorbance was measured at 450 nm with an ELISA reader, and the cell proliferation rate was calculated as shown in Equation 2 below. .

[Equation 2]

Cell proliferation rate (%) = [(absorbance of sample treated group/absorbance of untreated group) X 100]

B16F10 cell proliferation rate (%) Example 1 19.48±3.49 Example 2 37.55±3.04 Example 3 42.38±2.64 Example 4 23.80±3.08 Comparative Example 1 47.69±2.22 Comparative Example 2 68.06±2.99 Comparative Example 3 45.86±2.94 Comparative Example 4 46.97±3.14 Comparative Example 5 57.83±3.48

As a result, as shown in Table 4, the cell proliferation inhibitory activity shown in the 4% treatment concentration of Examples 1 to 4 compared to the cell proliferation inhibitory activity of B16F10 melanoma cells shown at the 4% treatment concentration of Comparative Examples 1 to 5 was remarkably. It was confirmed that it was excellent, and among them, it was confirmed that the B16F10 melanoma cell proliferation inhibitory effect of Example 1 was the most excellent.

In addition, as shown in FIG. 1, when the mixture of the mate extract and tranexamic acid of Example 1 was treated by concentration, a concentration-dependent cell proliferation inhibitory effect was observed in B16F10 melanoma cells, and it was confirmed that there is an effect on whitening. .

<Experimental Example 3> Cell cycle analysis test: cell cycle analysis

The cell cycle refers to a complex process that regulates the growth and proliferation of cells, repair of damaged DNA, etc. If the cell cycle is interrupted by a specific factor, cells die because cell proliferation does not occur. Therefore, when the mixture of mate extract and tranexamic acid inhibits the cell cycle of melanocytes, it may exhibit an effect of ultimately blocking the production of melanin by inhibiting the proliferation of melanocytes. Accordingly, a cell cycle analysis test was performed using B16F10 melanoma cells in order to confirm whether the mixture of the mate extract and tranexamic acid of Example 1 had a cell cycle inhibitory effect on melanocytes.

To determine whether it affects the cell cycle of B16F10 cells, the melanoma cell line of the mouse, cells were dispensed at a concentration ^{of 5X10 5 cells/plate in a 60 Ø plate using DMEM (Dulbecco's Modified Eagle Medium) medium, and then} Incubated for 24 hours in 5% CO _{2 conditions.} In cells cultured for 24 hours, the mixture of the mate extract and tranexamic acid of Example 1 was mixed with a medium so that the final concentration was 0% and 4%, respectively, and treated on a plate, followed by treatment at 37°C and 5% CO ₂ for 24 hours. While incubating. After 24 hours, the reacted cells were collected for each sample and centrifuged at 250Xg for 5 minutes at 4℃ to obtain a cell pellet, washed twice with DPBS, and fixed at -20℃ with 70% ethanol. The fixed cells were washed twice with DPBS, suspended in 0.5 mL of FxCycle™ PI/RNase Staining Solution, reacted for 20 minutes at dark room temperature, and analyzed the cell cycle using flow cytometry.

As a result, as can be seen in FIG. 2, when the mixture of the mate extract and tranexamic acid of Example 1 was treated to confirm the cell cycle analysis results of B16F10 melanoma cells, the example 1 compared to the untreated group without any treatment A remarkable increase in the cell cycle inhibitory activity of the G1 phase was observed at the 4% treatment concentration, confirming that the melanin cell cycle inhibitory effect was excellent.

<Experimental Example 4> Whitening efficacy test: Confirmation of extracellular melanin production

In order to examine the whitening effect using Examples 1 to 4 and Comparative Examples 1 to 5, an experiment to confirm the amount of extracellular melanin production was conducted using B16F10 melanoma cells.

^{To check the amount of extracellular melanin produced by B16F10 melanoma cells, the cells were dispensed at a concentration of 3X10 4} cells/well in a 24-well plate using DMEM (Dulbecco's Modified Eagle Medium) medium, and then 37°C and 5% CO ₂ conditions. Incubated for 24 hours at. Examples 1 to 4 and Comparative Examples 1 to 5 together with α-MSH 50nM for inducing melanin production in cells cultured for 24 hours were mixed with a medium so that the final concentration was 4%, and treated in each well, and then at 37°C and After incubation for 72 hours in 5% CO ₂ conditions, the culture solution was recovered. The culture medium reacted for 72 hours was taken 1 mL to measure the amount of extracellular melanin produced, and the absorbance was measured at 450 nm using an ELISA plate reader. The melanin production inhibition rate was calculated based on the absorbance of the blank using DPBS instead of the sample solution, and arbutin (β-arbutin) was used as a positive control.

Extracellular melanin production (%) Example 1 29.99±1.38 Example 2 35.97±1.02 Example 3 41.44±2.43 Example 4 33.33±1.42 Comparative Example 1 63.06±2.99 Comparative Example 2 76.04±1.90 Comparative Example 3 58.36±0.58 Comparative Example 4 60.40±2.35 Comparative Example 5 67.44±2.47 β-arbutin 40.27±1.84

As a result, as shown in Table 5, it was confirmed that the reduction in the amount of extracellular melanin produced at the 4% treatment concentration of Examples 1 to 4 compared to the amount of extracellular melanin produced at the 4% treatment concentration of Comparative Examples 1 to 5 was remarkably excellent. Among them, it was confirmed that the reduction in the amount of extracellular melanin production in Example 1 was the most excellent.

In addition, as shown in Figure 3, when the mixture of the mate extract and tranexamic acid of Example 1 was treated by concentration, a concentration-dependent decrease in the amount of extracellular melanin production was observed in B16F10 melanoma cells, confirming that the whitening activity was excellent. I did.

<Experimental Example 4> Whitening efficacy test: Confirmation of the amount of melanin produced in cells

In order to examine the whitening effect using Examples 1 to 4 and Comparative Examples 1 to 5, an experiment to confirm the amount of melanin production in cells was performed using B16F10 melanoma cells.

^{To check the intracellular melanin production of B16F10 melanoma cells, cells were dispensed at a concentration of 3X10 4} cells/well in a 24-well plate using DMEM (Dulbecco's Modified Eagle Medium) medium, and then 37°C and 5% CO ₂ conditions. Incubated for 24 hours at. Examples 1 to 4 and Comparative Examples 1 to 5 together with α-MSH 50nM for inducing melanin production in cells cultured for 24 hours were mixed with a medium so that the final concentration was 4%, and treated in each well, 37°C and Incubated for 72 hours in 5% CO _{2 conditions.} After incubation, washed twice with DPBS and treated with trypsin-EDTA to recover the cells, and then collected the cells with a centrifuge to remove the supernatant. Cells were treated with 1N NaOH solution to which 10% dimethyl sulfoxide (DMSO) was added and reacted at 65° C. for 1 hour, and then absorbance was measured at 410 nm using an ELISA plate reader to measure the amount of melanin production. The melanin production inhibition rate was calculated based on the absorbance of the blank using DPBS instead of the sample solution, and arbutin (β-arbutin) was used as a positive control.

Melanin production in cells (%) Example 1 35.97±1.02 Example 2 38.09±2.49 Example 3 42.94±3.68 Example 4 39.52±2.67 Comparative Example 1 53.73±3.04 Comparative Example 2 82.20±1.90 Comparative Example 3 50.41±2.44 Comparative Example 4 53.03±1.47 Comparative Example 5 64.94±1.15 β-arbutin 60.65±2.91

As a result, as shown in Table 6, it was confirmed that the decrease in the amount of melanin production in the cells exhibited at the 4% treatment concentration of Examples 1 to 4 compared to the amount of melanin produced in the cells at the 4% treatment concentration of Comparative Examples 1 to 5 was remarkably excellent. Among them, it was confirmed that the decrease in the amount of melanin production in the cells of Example 1 was the most excellent.

<Experimental Example 5> Antioxidant test: DPPH scavenging activity

In order to examine the antioxidant effect using Examples 1 to 4 and Comparative Examples 1 to 5, DPPH (2,2-diphenyl-1-picryl hydrezyl) radical scavenging activity was evaluated.

Examples 1 to 4 and Comparative Examples 1 to 5 were mixed with 450 μl of 55 μM DPPH solution to a final concentration of 4% and reacted at 4° C. for 30 minutes. After the reaction, each sample was placed in a 96-well plate and absorbance was measured at 520 nm using an ELISA reader. At this time, a sample without a sample was used as a control, and the free radical scavenging activity (%) of the sample was calculated as shown in Equation 3 below. At this time, ascorbic acid was used as a positive control for scavenging radicals, and when the free radical scavenging ability was 50%, the concentration of ascorbic acid was 1.92 μg/mL.

[Equation 3]

Free radical scavenging rate (%) = [(1-absorbance of the sample treatment group / absorbance of the untreated group) X 100]

DPPH radical scavenging rate (%) Example 1 89.42±0.39 Example 2 81.95±1.57 Example 3 73.88±0.45 Example 4 77.46±0.45 Comparative Example 1 63.62±0.52 Comparative Example 2 9.89±1.50 Comparative Example 3 64.16±0.98 Comparative Example 4 63.90±1.12 Comparative Example 5 58.53±0.92 Ascorbic acid (IC ₅₀ ) 1.92㎍/mL

IC ₅₀ means the sample concentration when the free radical scavenging ability is 50% by the added sample. As a result, as shown in Table 7, it was confirmed that the DPPH radical scavenging activity exhibited at the 4% treatment concentration of Examples 1 to 4 compared to the DPPH radical scavenging activity result shown at the 4% treatment concentration of Comparative Examples 1 to 5 was remarkably excellent. Among them, it was confirmed that the DPPH radical scavenging activity of Example 1 was the most excellent.

In addition, as shown in FIG. 4, when the mixture of the mate extract and tranexamic acid of Example 1 was treated by concentration, a concentration-dependent increase in DPPH radical scavenging activity was observed, confirming that the antioxidant activity was excellent.

<Experimental Example 6> Anti-wrinkle efficacy test: Elastase inhibition activity

In order to examine the anti-wrinkle effect using Examples 1 to 4 and Comparative Examples 1 to 5, the Elastase inhibitory activity was confirmed.

Examples 1 to 4 and Comparative Examples 1 to 5 were dissolved in 10 mM Tris-HCL buffer (pH 8.0) to a final concentration of 4%, and a substrate having a concentration of 1.6 mM N-succinyl-(Ala)3-p-nitroanilide and Elastase from procine pancreas Type IV 0.4 U/mL was mixed and reacted at 25° C. for 5 minutes, and the absorbance was measured at 410 nm by placing in a 96-well plate. At this time, the sample without the sample was used as a control group, and the elasticity inhibition activity (%) of the sample was calculated as shown in Equation 4 below.

At this time, oleanolic acid was used as a positive control, and when the elastase inhibitory activity was 50%, the concentration of oleanolic acid was 7.49 μg/mL.

[Equation 4]

Elastase inhibitory activity (%) = [(1-absorbance of the sample treated group / absorbance of the untreated group) X 100]

Elastase inhibitory activity (%) Example 1 39.68±1.26 Example 2 31.19±0.43 Example 3 25.05±1.27 Example 4 29.08±0.60 Comparative Example 1 12.09±0.20 Comparative Example 2 8.98±1.03 Comparative Example 3 14.05±0.23 Comparative Example 4 12.97±0.35 Comparative Example 5 4.95±0.96 Oleanolic acid (IC ₅₀ ) 7.49㎍/mL

As a result, as shown in Table 8, the elastase inhibitory effect exhibited at the 4% treatment concentration of Examples 1 to 4 compared to the result of the elastase inhibitory activity shown at the 4% treatment concentration of Comparative Examples 1 to 5 was remarkably excellent. It could be confirmed, and among them, it was confirmed that the elastase inhibitory effect of Example 1 was the most excellent.

In addition, as shown in FIG. 5, when the mixture of the mate extract and tranexamic acid of Example 1 was treated by concentration, a concentration-dependent increase in elastatase inhibitory activity was observed, confirming that the anti-wrinkle effect was excellent.

<Production Example 1> Preparation of lotion

A lotion was prepared by a conventional method using the ingredients shown in Table 9 below.

ingredient Content (% by weight) Mixture of Example 1 1.00 Hydroxyethylene cellulose
(2% aqueous solution) 12.00 Xanthan gum (2% aqueous solution) 2.00 1,3-butylene glycol 6.00 glycerin 4.00 Sodium hyaluronate (1% aqueous solution) 5.00 Purified water Balance system 100.0

<Production Example 2> lotion production

A lotion was prepared by a conventional method using the ingredients shown in Table 10 below.

ingredient Content (% by weight) Mixture of Example 1 3.00 Ascorbic acid-2-magnesium phosphate salt 1.00 Water-soluble collagen (1% aqueous solution) 1.00 Sodium citrate 0.01 Citric acid 0.05 1,3-butylene glycol 3.00 Purified water Balance system 100.0

<Production Example 3> Cream preparation

A cream was prepared by a conventional method using the ingredients shown in Table 11 below.

ingredient Content (% by weight) Mixture of Example 1 3.00 Polyethylene glycol monostearate 2.00 Self-emulsifying glycerin monostearate 5.00 Cetyl alcohol 4.00 Squalene 6.00 Tri2-ethylhexane glyceryl 6.00 Sphingoglycolipid 1.00 1,3-butylene glycol 7.00 Purified water Balance system 100.0

<Production Example 4> Preparation of ointment for external use

An ointment for external use was prepared by a conventional method using the components shown in Table 12 below.

ingredient Content (% by weight) Mixture of Example 1 4.00 Stearic acid 15.00 Cetearic Alcohol 1.00 Cetyl alcohol 4.00 Potassium hydroxide 0.70 glycerin 5.00 1,3-butylene glycol 3.00 antiseptic Appropriate amount Purified water Balance system 100.0

As described above, specific parts of the present invention have been described in detail, and it will be apparent to those of ordinary skill in the art that these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereby. will be. Accordingly, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (6)

Mate ( Ilex paraguayensis ) extract and tranexamic acid (Tranexamic acid) containing a mixture in a weight ratio of 1:10 as an active ingredient,
The mate ( Ilex paraguayensis ) extract is whitening, antioxidant, characterized in that obtained by adding mate and ethanol solvent to an extractor equipped with a reflux cooler and then extracting by heating and reflux cooling extraction method at 50 to 100°C for 4 to 20 hours. And anti-wrinkle cosmetic composition.


delete delete The cosmetic composition for whitening, antioxidant and anti-wrinkle according to claim 1, wherein the mixture of the mate extract and tranexamic acid is contained in an amount of 0.01 to 30% by weight based on the total weight of the composition.
The method of claim 1, wherein the mixture of the mate extract and tranexamic acid has a melanocyte proliferation inhibitory activity, A cosmetic composition for whitening, antioxidant and anti-wrinkle, characterized in that it has melanin cell cycle inhibitory activity, melanin production inhibitory activity, DPPH scavenging activity, and elastase inhibitory activity.
A cosmetic comprising the composition of any one of claims 1, 4 and 5.

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