KR20230156285A - Cosmetic composition comprising enzyme treated fraxinus rhynchophylla - Google Patents

Abstract

According to an embodiment of the present invention, a cosmetic composition is provided. The cosmetic composition may include an enzyme-treated product of Fraxinus rhynchophylla as an active ingredient.

Description

Cosmetic composition containing ash tree enzyme treatment {COSMETIC COMPOSITION COMPRISING ENZYME TREATED FRAXINUS RHYNCOPHYLLA}

The present invention relates to a cosmetic composition containing an ash tree enzyme-treated product. Specifically, a cosmetic composition containing an ash tree enzyme-treated product obtained by treating an ash tree extract with an enzyme as an active ingredient, thereby providing excellent anti-inflammatory and antioxidant effects. It relates to composition.

The skin of modern people is affected by external stimuli such as chemical irritants, environmental pollution, and ultraviolet rays, which cause inflammatory allergic diseases such as acne and atopic dermatitis. Due to the decline in skin function due to aging, active harmful oxygen increases and melanin is deposited in cells. This causes spots, wrinkles, etc.

To solve this problem, functional cosmetics with anti-inflammatory and antioxidant effects are being developed. In particular, as the demand for cosmetics using natural water that does not irritate the skin and cause trouble increases, much research is being conducted to discover natural ingredients and increase their efficacy.

However, there are limitations in extracting active ingredients with specific efficacy from natural products, and even if extracted, the yield is not high, so there is a problem in that the anti-inflammatory and antioxidant effects are not substantially realized.

KR 10-2019-0089309 A

The present invention is intended to solve the above problems, and its purpose is to provide a cosmetic composition with excellent anti-inflammatory and antioxidant effects by containing as an active ingredient an enzyme-treated ash tree extract obtained by treating an ash tree extract with an enzyme. do.

The technical problems of the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned will be clearly understood by those skilled in the art from the description below.

According to an embodiment of the present invention, a cosmetic composition is provided. The cosmetic composition may include an enzyme-treated product of Fraxinus rhynchophylla as an active ingredient.

Additionally, the ash tree enzyme-treated product may include a reaction product produced by treating an ash tree extract with an enzyme.

Additionally, the ash tree extract can be extracted from ash tree with ethanol.

Additionally, the enzyme may be a glycolytic enzyme.

Additionally, the enzyme may be one or more selected from the group consisting of Celluclast and Viscozyme.

Additionally, the enzyme may be Viscozyme.

Additionally, the ash tree enzyme-treated product may be a supernatant of the reaction products produced by treating ash tree extract with enzymes.

Additionally, the ash tree enzyme-treated product may be in the form of powder obtained by freeze-drying the supernatant.

In addition, the ash tree enzyme-treated product may contain a higher content of esculetin compared to the enzyme-untreated ash tree extract.

Additionally, the ash tree enzyme-treated product may contain esculetin converted from esculin.

Additionally, the cosmetic composition may have excellent anti-inflammatory and antioxidant effects.

According to an embodiment of the present invention, a method for producing an ash tree enzyme-treated product is provided. The method includes extracting an ash tree with ethanol to obtain an ash tree extract; Proceeding with an enzyme reaction by treating the ash tree extract with an enzyme in a slightly acidic buffer solution; It may include obtaining a supernatant from the reaction product produced by the enzyme reaction.

Additionally, in the above method, the enzyme may be Viscozyme.

According to the present invention, the content of esculetin can be significantly increased by treating ash tree extract with enzymes. The cosmetic composition according to the present invention has excellent anti-inflammatory or antioxidant effects by containing the enzyme-treated ash extract as an active ingredient.

Additionally, because the present invention uses a biotechnological method such as an enzyme reaction, the content of esculetin can be increased under skin-friendly conditions that do not cause skin toxicity or irritation. In addition, since no toxic ingredients are used as raw materials for enzyme reaction, no special purification process is required, and active ingredients can be obtained from ash tree extract in an environmentally friendly way without generating wastewater.

In order to more fully understand the drawings cited in the detailed description of the present invention, a brief description of each drawing is provided.
Figures 1 to 3 are diagrams related to experiments confirming the esculetin content of compositions for Examples and Comparative Examples of the present invention.

Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by a person skilled in the art to which the present invention pertains. In general, the nomenclature used herein is well known and commonly used in the art. Additionally, when describing embodiments of the present invention, if detailed descriptions of related known configurations or functions are judged to impede understanding of the embodiments of the present invention, the detailed descriptions will be omitted. In addition, embodiments of the present invention will be described below, but the technical idea of the present invention is not limited or limited thereto and may be modified and implemented in various ways by those skilled in the art.

The cosmetic composition of the present invention may include an enzyme-treated ash tree. Specifically, the cosmetic composition of the present invention may include a reaction product produced by treating an ash tree extract with an enzyme.

Ash tree ( Fraxinus rhynchophylla ) is a deciduous broad-leaved tree of the Ash tree family of the Ash tree family, and is known to have anti-inflammatory and antioxidant effects. Ash tree has been mainly used as an herbal medicine material, but has not received much attention as a raw material for cosmetic compositions. This is because it is difficult to extract the active ingredients from the ash tree, and the skin-improving activity of the active ingredients is unknown. The present inventor confirmed that active ingredients can be obtained in high yield by treating ash tree extract with enzymes, and completed the present invention by applying this to a cosmetic composition.

In the present invention, 'ash tree extract' means extraction of ash tree using a solvent. Depending on the embodiment, the ash tree extract may be obtained by extracting ash tree using ethanol as a solvent. However, it is not limited to this, and various solvents applicable in the relevant technical field can be used alone or in combination. For example, water (or distilled water) as an extraction solvent; lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol, and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol, and propylene glycol; and hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane; Or a mixture thereof can be used.

The ash tree used in ash tree extract may include all parts of the ash tree, including leaves, branches, stems, flowers, fruits, roots, and root bark.

Depending on the embodiment, the enzyme used in the present invention may be a glycolytic enzyme. By treating ash tree extract with glycolytic enzymes, the bioavailability of the active ingredient can be increased by converting glycosides into non-glycosides. In particular, the enzyme used in the present invention may be one or more selected from the group consisting of Celluclast and Viscozyme, and preferably may be Viscozyme.

'Celluclast' is an enzyme isolated after submerged fermentation of the fungus Trichoderma resei and is a type of cellulase. In addition, 'Viscozyme' is an enzyme derived from Aspergillus aculeatus and contains arabinase, carbohydrase, cellulase, and beta-glucanase. It is a complex enzyme containing Glucanase, Hemicellulase, and Xylanase.

When an enzyme selected from the group consisting of celluclast and viscozyme, especially viscozyme, is treated with ash tree extract, an enzyme reaction occurs and the content of esculetin can be greatly increased. This is because the sugar in the esculin glycoside contained in the ash tree extract is decomposed by the enzyme, thereby greatly increasing the content of esculetin.

Esculin is a glycoside of esculetin with glucopyranoside bound to esculetin. When the sugar is separated from esculetin, it can be converted to esculetin, a non-glycoside. . Esculetin is a coumarin derivative and is known to have anti-inflammatory and antioxidant effects, including suppressing the expression of histamine, an inflammation mediator, and inflammatory cytokines related to atopic dermatitis, and scavenging hydroxyl radicals and reactive oxygen species. In addition, compared to esculin, which is a glycoside, esculetin, which is a non-glycolyte, is very advantageous in terms of bioavailability.

Accordingly, the enzyme-treated ash tree material of the present invention contains a high content of esculetin converted from esculin, and has the characteristic of being very excellent in at least one of anti-inflammatory and antioxidant effects. Since the ash tree enzyme-treated product of the present invention contains a higher content of esculetin compared to the ash tree extract itself (untreated with enzymes) and the case of fermenting ash tree using yeast, it has anti-inflammatory or antioxidant effects. Even better.

The ash tree enzyme-treated product of the present invention can be produced through the following method. First, ash tree extract can be obtained by extracting ash tree with ethanol. For example, ash tree extract can be extracted using extraction methods commonly used in the art, such as reflux extraction, supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction, or ultrasonic extraction. Here, 'ash tree' may include all parts of the ash tree, including leaves, branches, stems, flowers, fruits, roots, and root bark, and may include the ash tree itself or the dried product of the ash tree.

After obtaining the ash tree extract, the enzyme reaction can be performed by treating the ash tree extract with an enzyme in a slightly acidic buffer solution. At this time, the buffer solution may have a pH of about 4 to 6, and preferably may have a pH of about 5, but is not limited thereto.

Depending on the embodiment, the enzyme treated with the ash tree extract may be one or more selected from the group consisting of celluclast and viscozyme, and preferably may be viscozyme. When an ash tree extract is treated with an enzyme selected from the group consisting of Celluclast and Viscozyme to cause an enzymatic reaction, especially when Viscozyme is used, the content of esculetin can be very high.

In this way, after treating the ash tree extract with an enzyme, the reaction product produced by this enzyme reaction can be used as the ash tree enzyme-treated product of the present invention, and the filtrate obtained after removing the enzyme from the reaction product can also be used. Additionally, the supernatant located at the top can be separated from the reaction product produced by the enzyme reaction and used as an enzyme-treated ash tree of the present invention. That is, the ash tree enzyme-treated product of the present invention may include a reaction product produced by treating an ash tree extract with an enzyme.

Depending on the embodiment, the ash tree enzyme-treated product may be a liquid formulation or a powder form obtained by freeze-drying the reaction product or supernatant. At this time, the drying method may be applied not only by freeze drying, but also by various methods commonly used in the technical field, such as vacuum drying, hot air drying, and spray drying. In addition, the reaction product of ash tree extract treated with enzymes can be manufactured or processed in various ways, such as distilling under reduced pressure or concentrating the reaction product.

As such, the present invention can increase the content of esculetin under skin-friendly conditions because it uses a biotechnological method of treating ash tree extract with enzymes. Specifically, the enzyme reaction used in the present invention can be friendly to the skin because the enzyme reaction proceeds in a non-toxic solvent, low temperature, and slightly acidic buffer solution. In addition, the enzyme reaction used in the present invention does not require a special purification process because no toxic components are used among the enzyme reaction raw materials, and the active ingredient can be obtained from ash tree extract in an environmentally friendly manner without generating waste water. . Therefore, compared to the method of extracting the active ingredient through a chemical reaction such as an acid catalytic reaction, there is an advantage in that the active ingredient of the ash tree extract can be obtained in a skin-friendly and environmentally friendly method.

Depending on the embodiment, the present invention can provide a cosmetic composition containing enzyme-treated ash tree as an active ingredient. For example, it is manufactured as basic cosmetics such as lotion, nutritional lotion, nutritional cream, massage cream, essence, soap, detergent, cleansing cream, cleansing water, lipstick, lip balm, mascara, blusher, shading, highlighter, makeup base, It can be manufactured into color cosmetics such as foundation, pact, concealer, skin cover, etc. It can also be manufactured into hair cleaning products such as shampoo, rinse, hair conditioner, hair gel, etc. Additionally, for example, it may be manufactured as a product that is attached to the skin, such as a pack, hydrogel slimming patch, etc., or may be manufactured as a product that is sprayed on the skin, such as a mist or aerosol.

Depending on the embodiment, the cosmetic composition of the present invention may be provided in a formulation commonly prepared in the art. For example, it may be provided in the form of a solution, an emulsion obtained by dispersing the oil phase in the water phase, an emulsion obtained by dispersing the water phase in the oil phase, suspension, emulsion, solid, gel, oil, powder, paste, or foam aerosol. You can.

Depending on the embodiment, the present invention can provide not only a cosmetic composition but also a pharmaceutical composition containing an enzyme-treated ash tree as an active ingredient. For example, it can be manufactured as a pharmaceutical composition for various purposes such as anti-inflammatory to suppress inflammatory response, antioxidant to inhibit cell aging or oxidation, and skin whitening to improve blemishes, freckles, moles, and skin pigmentation. You can. More specifically, it can also be manufactured into medicinal cosmetics such as medicinal lotion, medicinal cream, medicinal emulsion, and medicinal foundation.

Depending on the embodiment, other ingredients commonly used in the art, such as thickeners, pH adjusters, isotonic agents, surfactants, stabilizers, preservatives, preservatives, ultraviolet rays, etc., to the extent that they do not impair the effect of the present invention. It may further include absorbents, disinfectants, moisturizers, blockers, antioxidants, organic pigments, inorganic pigments, fragrances, vitamins, etc. The mixing amount of the above components can be easily selected by a person skilled in the art within a range that does not impair the purpose and effect of the present invention.

Below, examples are shown to further explain the present invention in more detail, but the present invention is not limited thereto.

Manufacturing example

(1) Preparation of Examples 1 to 3 and Comparative Examples 1 to 4

Ash tree extract was obtained by adding 70% ethanol to the ash tree bark at a ratio of 1:4. Afterwards, 1% of ash tree extract and 1% of the enzyme Viscozyme were treated in a buffer solution of pH 5 containing disodium phosphate and citric acid, and then incubated in an incubator at 35°C and 150rpm. The enzyme reaction was performed for 18 hours under these conditions. After completion of the enzyme reaction, the enzyme was precipitated at 90°C for 10 minutes and centrifuged to obtain the supernatant, thereby preparing the ash tree enzyme-treated product of Example 1. In addition, the supernatant was freeze-dried for 3 days to prepare the ash tree enzyme-treated product of Example 2 in powder form.

In order to compare the effects of the presence or absence of enzyme treatment, enzyme-untreated ash tree extracts of Comparative Examples 1 and 2 were prepared by excluding the enzyme treatment process from the preparation methods of Examples 1 and 2, respectively. In addition, in order to compare the effects depending on the type of enzyme, the enzymes listed in Table 1 below were used in the same manner as in Example 1, and Example 3 and Comparative Example were prepared under pH and incubator temperature conditions suitable for each enzyme. Ash tree enzyme-treated products 3 and 4 were prepared.

(2) Preparation of Comparative Examples 5 and 6

Ash tree extract was obtained by adding 70% ethanol to the ash tree bark at a ratio of 1:4. Afterwards, 1% ash tree extract, 2% sugar, 5% yeast, and 92% distilled water were mixed and fermented at room temperature for 3 days. After completion of the fermentation reaction, the yeast was removed, sterilized at 90°C for 30 minutes, and then centrifuged to obtain the supernatant, thereby preparing the fermented ash extract of Comparative Example 5.

Additionally, 10% ash bark, 2% sugar, 5% yeast, and 83% distilled water were mixed and fermented at room temperature for 3 days. After completion of the fermentation reaction, the ash tree bark and yeast were removed, sterilized at 90°C for 30 minutes, and then centrifuged to obtain a supernatant, thereby preparing a fermented ash tree raw material of Comparative Example 6.

Experiment example

(1) Check the content of esculetin

In order to determine the content of esculetin in Examples 1 to 3 and Comparative Examples 1 to 6, they were analyzed by HPLC (High Performance Liquid Chromatography), and the results are shown in Table 2 and Figures 1 to 3.

As a result of the experiment, in the case of the liquid formulation, it was confirmed that Examples 1 and 3 each had a very high content of esculetin, totaling more than 1300 ppm. In particular, in the case of Example 1, which used viscozyme as an enzyme, the content of esculetin The content was confirmed to be very high, exceeding 1600ppm. On the other hand, in the case of Comparative Example 1 and Comparative Examples 3 to 6, the content of esculetin was confirmed to be lower than that of Examples 1 and 3. That is, in the case of a liquid formulation, when the ash tree extract was not treated with enzymes (Comparative Example 1), or when other types of enzymes such as BAN and Pluszyme were used (Comparative Examples 3 and 4), fermentation was performed using yeast. Compared to one case (Comparative Examples 5 and 6), when Celluclast and Viscozyme were used as enzymes, it was found that a higher content of esculetin was contained.

Additionally, in the case of the powder formulation, Example 2 was confirmed to have a very high esculetin content of 33954.61 ppm. On the other hand, in the case of Comparative Example 2, the esculetin content was confirmed to be very low at 10822.09ppm. That is, even in the case of a powder formulation, it was found that the ash tree extract treated with enzyme contained a content of esculetin that was more than 3.14 times higher than when the ash tree extract was not treated with the enzyme.

(2) Evaluation of anti-inflammatory efficacy

1) Cytotoxicity evaluation

For Example 2 and Comparative Example 2, a cytotoxicity test was performed according to the MTT assay, and the results are shown in Table 3.

The specific experimental method is as follows.

100 μL of the RAW 264.7 cell line was dispensed at an appropriate concentration per well in a 96 well plate and cultured for 24 hours under 37°C and 5% CO ₂ incubator conditions. Samples were treated with the cultured cells at different concentrations and then cultured for 24 hours at 37°C in a 5% CO ₂ incubator. 3 hours before the completion of incubation, 10 μL of 5 mg/mL MTT was treated and cultured under the same conditions for 3 hours. After completion of incubation, the culture medium was discarded, 100 μL of DMSO was dispensed, and the mixture was stirred at room temperature for 10 minutes. Absorbance was measured at 570 nm using ELISA (Bio-Rad, USA). Cell viability was derived using the formula (absorbance of experimental group/absorbance of control group) × 100.

As a result of the experiment, it was found that Example 2 and Comparative Example 2 did not exhibit cytotoxicity at a concentration of 1000 μg/mL or less.

2) Anti-inflammatory evaluation

A NO assay was performed to evaluate the ability to inhibit NO production for Example 2 and Comparative Example 2, and the results are shown in Table 4.

The specific experimental method is as follows.

100 μL of the RAW264.7 cell line was dispensed at an appropriate concentration per well in a 96 well plate and cultured for 24 hours under 37°C and 5% CO ₂ incubator conditions. In order to induce the production of NO in cultured cells, the medium was exchanged with 1㎍/mL of lipopolysaccharide (LPS), and then the samples were diluted according to concentration using cell medium, treated with cells, and re-cultured for 24 hours. . The amount of NO produced was determined by mixing 100 μL of cell culture supernatant and 100 μL of Griess reagent and reacting for 10 minutes in a 96-well plate. Absorbance was measured at 540 nm using ELISA (Bio-Rad, USA). The amount of NO was calculated by comparing it with the standard curve for each concentration of sodium nitrate (NaNO ₂ ), and the anti-inflammatory effect was 100 - [{NO conc. (μM) _{comparison group 2 or experimental group} }/{NO conc. (μM) _{comparison group 1} }×100] was derived from the formula. Since nitric oxide is an inflammation-causing factor, the higher the inhibitory activity of nitric oxide is measured, the higher the anti-inflammatory effect can be evaluated.

As a result of the experiment, it was confirmed that Example 2 had an excellent anti-inflammatory effect as the NO inhibitory activity was measured to be higher than that of Comparative Example 2 at the concentration of each sample. In particular, at a concentration of 100 ㎍/mL, it was found that the anti-inflammatory effect of the enzyme-treated ash tree extract was more than 1.6 times better than that of the untreated ash tree extract.

(3) Evaluation of antioxidant efficacy

The antioxidant efficacy of Example 2 and Comparative Example 2 was evaluated, and the results are shown in Table 5.

The specific experimental method is as follows.

The 0.5mM DPPH solution and the sample were mixed in a 1:1 ratio, blocked from light and mixed for 30 minutes at room temperature, and then the absorbance at 540 nm was measured using ELISA (Bio-Rad, USA). The antioxidant effect was derived by the formula 100 - [{by sample (OD _DPPH -OD _ethanol )}/{dH ₂ O (OD _DPPH -OD _ethanol )}×100], and DPPH, which contains active oxygen, is scavenged by antioxidant substances. Therefore, the higher the scavenging ability is measured, the higher the antioxidant efficacy can be evaluated.

As a result of the experiment, it was confirmed that Example 2 had an excellent antioxidant effect as the DPPH free radical scavenging ability was measured to be higher than that of Comparative Example 2 at the concentration of each sample. Specifically, at a concentration of 100 ㎍/mL, it was found that the antioxidant effect of the enzyme-treated ash tree extract was more than 1.75 times better than that of the enzyme-treated ash tree extract.

As the specific parts of the present invention have been described in detail above, those skilled in the art will understand that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (5)

In the method of producing an ash tree enzyme-treated product,
Extracting ash tree with ethanol to obtain ash tree extract;
Performing an enzyme reaction by treating the ash tree extract with an enzyme in a buffer solution of pH 4 to 6 containing disodium phosphate and citric acid; and
Comprising the step of obtaining a supernatant from the reaction product produced by the enzyme reaction,
The enzyme is at least one selected from the group consisting of Celluclast and Viscozyme,
The ash tree enzyme-treated product is in the form of powder obtained by freeze-drying the supernatant,
A method wherein the ash tree enzyme-treated product contains esculetin converted from esculin. According to claim 1,
The ash tree enzyme-treated product is characterized in that it contains a higher content of esculetin compared to the enzyme-untreated ash tree extract. According to claim 1,
A method characterized in that at least one of anti-inflammatory and antioxidant effects is excellent. An enzyme-treated ash tree product prepared by the method according to any one of claims 1 to 3. A cosmetic composition comprising the enzyme-treated ash tree according to claim 4 as an active ingredient.