KR20170058562A

KR20170058562A - METHOD FOR PREPARING AN LICORICE EXTRACT OF Nrf-2 ACTIVATOR, AND COSMETIC COMPOSITION COMPRISING THE SAME

Info

Publication number: KR20170058562A
Application number: KR1020150162285A
Authority: KR
Inventors: 박정옥; 주철규; 박세정; 박진오; 박수남
Original assignee: 대봉엘에스 주식회사
Priority date: 2015-11-19
Filing date: 2015-11-19
Publication date: 2017-05-29
Also published as: KR101866742B1

Abstract

The present invention relates to use of a licorice extract for enhancing activity of Nrf-2, a method for preparing the same, and a cosmetic composition comprising the same. More specifically, the method for preparing the composition for enhancing activity of Nrf2 comprises the following steps: extracting the Nrf2 activating composition, comprising a licorice extract as an active ingredient, and licorice with an alcohol solvent having 1-4 carbon atoms; obtaining powder by freezing and drying the extract; obtaining a fraction from the powder by using water and ethyl acetate; and obtaining a water fraction from the fraction. According to the present invention, the cosmetic composition has an effect of enhancing ARE activity related to Nrf-2. When the cosmetic composition is applied to skin, homeostasis of the skin, exposed to the outside all the time and suffering from stress, is maintained. The cosmetic composition has an excellent effect of preventing skin aging caused by active oxygen. Therefore, a hypoallergic, skin-friendly, and antiaging cosmetic product in an advanced concept can be developed.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for producing a licorice extract and a cosmetic composition containing the same,

The present invention is the result of the project "Establishment of standards and extraction process of domestic raw materials for functional materials" in R & D Agency's research project "Standardization of Domestic Natural Resources Quality to Replace Imports of Functional Beauty Materials" (Project No. PJ008489).

TECHNICAL FIELD The present invention relates to a method for producing a licorice extract promoting Nrf2 activity and a cosmetic composition containing the same.

In the cell, active oxygen occurs naturally in the energy combustion process, and the antioxidative reaction which can neutralize these active oxygen occurs at the same time. When the active oxygen production reaction and the antioxidation reaction occur in a balanced manner, the human body is normally grown and metabolized. However, when the generated active oxygen exceeds the capacity of the antioxidant that can be treated in cells, the cell falls into oxidative stress and causes various diseases such as cancer, diabetes, heart disease and aging.

To prevent this, cells develop complex signaling mechanisms to detoxify harmful substances and maintain cellular redox homeostasis. One of these signaling mechanisms protects cells and induces the expression of detoxifying enzymes. In the first step, hydroxylation of the xenobiotic, which promotes the production of reactive oxygen species by cytochrome P450 enzyme, It is known to produce water. Afterwards, phase II system enzymatic reaction converts to hydrophilic xenobiotics. Finally, the aqueous external substrate produced through Phase I / II reaction is eventually excreted through the phase III reaction.

NF-E2 related factor 2 (Nrf-2) is known to be a major regulator of a series of proteins called phase II enzymes that protect the body from oxidative stress and serve as a detoxifying agent for xenobiotic. In the absence of oxidative stress, cytoplasmic Nrf2 is bound to Keap1 protein and is constantly ubiquitinated and degraded, thus its activity is inhibited. On the other hand, in the presence of excessive oxidative stress, Nrf2 is liberated from Keap1 protein and transported into the nucleus, which binds to the antioxidant response element (ARE) present in the antioxidant enzyme gene promoter and expresses the antioxidant enzyme, . Through these roles, it is known that Nrf2 plays an important role in cell homeostasis.

Recently, research on antioxidant substances for the removal of active oxygen associated with skin aging has been conducted. However, some of the side effects of the substances developed by these studies are questionable.

It is required to develop a substance which can be used in cosmetics without any adverse effect as well as its content without any side effects or cytotoxic risks and being excellent in antioxidative and antiinflammatory effects with little or no risk for such side effects or cytotoxicity. In recent years, in order to meet the needs of consumers, research and development of cosmetics using natural raw materials have been actively conducted.

Licorice is a perennial herb that belongs to legume plants and is known as a medicinal plant widely used as an oriental medicinal herb in Asia. Glycyrrhizin is known as the main component of licorice, flavonoid glycosides such as liquiritin and isoliquiritin, and licoricidin, an isoflavonoid. Licorice has been reported to have antioxidant, immunity and antibacterial effects.

Patent Document 10-0530669 B1 (cosmetic composition for skin whitening comprising lactate and licorice extract) relates to a cosmetic composition for whitening skin, and more particularly to a cosmetic composition for skin whitening comprising 10-20% by weight of lactate and 0.0001- To 15% by weight of a cosmetic composition for skin whitening.

Patent No. 10-1466551 B1 (cosmetic composition for skin whitening containing Lactuca sativa extract prepared by using licorice) relates to a cosmetic composition for skin whitening containing an extract of Lactobacillus sp. It inhibits the production of melanin and has excellent skin whitening effect and can be used as natural cosmetics.

Patent No. 10-1017922 B1 (a whitening cosmetic composition containing a licorice extract and a natural thickening agent) contains a licorice extract as an active ingredient and does not use a polymer thickening agent as a synthetic polymer, and a natural thickening agent such as bentonite And xanthan gum which is an organic point-of-care tablet, thereby providing an excellent whitening effect.

Patent No. 10-1072133 B1 (cosmetic composition for enhancing skin whitening efficacy) is directed to a cosmetic composition which promotes the inhibition of melanin overproduction and promotes the release of melanin produced on the skin, thereby improving the whitening effect. , A birch extract, and a papain enzyme to improve skin whitening effect.

Patent Application No. 10-1081914 B1 (composition for external application for skin containing an extract extracted from a crude drug mixture) includes Gakyung, licorice, Kidney, allium, gardenia, yellowish white, golden, yellowtail, The present invention relates to a composition for external application for skin, which comprises an extract obtained from a crude drug mixture composed of an active ingredient and an antioxidant, To an extract having an effect of inflammation.

However, none of the above documents disclose a method for producing licorice extract and a cosmetic composition containing the same, which demonstrate the ARE enhancing effect associated with maintenance of skin homeostasis. Accordingly, the inventors of the present invention have studied the possibility of application to cosmetics in various materials not yet known, and as a result, it has been made possible to produce a licorice extract excellent in maintaining skin homeostasis.

KR 10-0530669 B1 (Nov. 17, 2005) KR 10-1466551 B1 (2014.11.23.) KR 10-1017922 B1 (Feb. 21, 2011) KR 10-1072133 B1 (April 4, 2011.) KR 10-1081914 B1 (2011.11.03.)

The present invention also provides a method for extracting licorice extract which minimizes the damage of main components of licorice root extract, and provides a licorice root extract having Nrf2-related ARE activity and a cosmetic composition having excellent skin homeostasis.

SUMMARY OF THE INVENTION The present invention has been made in order to solve the above-mentioned problems and problems of the prior art,

The present invention provides a composition for promoting Nrf2 activity using an extract of licorice as an active ingredient.

In addition, the present invention provides the Nrf2 activity enhancing composition according to the present invention, wherein the licorice extract is fractionated with water and ethyl acetate to exclude cytotoxic or skin irritant components and water fractions.

Extracting the licorice into an alcohol solvent having a carbon number of C ₁ to C ₄ ;

Lyophilizing the extract to obtain a powder;

Obtaining the powder with water and ethyl acetate to obtain a fraction; And

To obtain a water fraction of said fraction.

Also, the present invention provides a skin homeostasis-maintaining cosmetic composition, which comprises the licorice extract of the present invention in an amount within a range of 0.001 to 10% by weight based on the total weight of the composition.

The present invention has the effect of enhancing the ARE activity associated with Nrf-2, and by applying it to the skin, it is always exposed to the outside to maintain the homeostasis of the stressed skin and to prevent skin aging by active oxygen. Can develop advanced concept of hypo-skin-friendly anti-aging cosmetics.

1 is a graph showing the results of Nrf2 induction activity test for licorice extract prepared in Examples and Comparative Examples.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention is a Nrf2 activity enhancing composition comprising an licorice extract as an active ingredient.

Here, 'Nrf2' is an abbreviation of 'NF-E2-related factor-2' and is an essential transcription factor that binds to an antioxidant-response element (ARE) and regulates the expression of various antioxidant genes. Since the term " Nrf2 " is widely used by a person skilled in the art, the definition of the term " Nrf2 " is generally used, and a detailed description thereof will be omitted.

Preferably, the licorice extract is fractionated with water and ethyl acetate to exclude cytotoxic components and water fractions. As described below, the present inventors have surprisingly found that there is a great difference in the efficacy between the water fraction and the ethyl acetate fraction, and also the cytotoxicity or the skin irritation component, and a more detailed explanation will be given later May be accomplished through embodiments.

According to another aspect of the present invention,

Extracting licorice into an alcohol solvent having a carbon number of C ₁ to C ₄ ;

Lyophilizing the extract to obtain a powder;

Obtaining the powder with water and ethyl acetate to obtain a fraction; And

And obtaining a water fraction of the fraction.

Here, the alcohol solvent is preferably a 75-90% ethanol solvent, more preferably an 85% ethanol solvent. As will be understood from the following examples, there may be differences in effect between the main action and the side effect, for example, depending on the ethanol concentration, the degree of Nrf2 activity may be different and the extent of skin irritant component extraction may be different.

The water and ethyl acetate are preferably used in an amount of 1 to 10% by weight based on the weight of the powder. If the content is less than 1% by weight, the fraction may not be well-sorted and cytotoxic components may not be removed. When the content is more than 10% by weight, the solvent is unnecessarily excessive, Which is undesirable because excessive time and cost are involved.

Yet another aspect of the present invention is a skin homeostasis-maintaining cosmetic composition comprising the licorice extract of the present invention described above.

It is preferable that the licorice extract is contained within the range of 0.001 to 10% by weight based on the weight of the whole composition. If the content is less than 0.001% by weight, the desired effect may be insufficient. If the content is more than 10% by weight, And the like may occur.

The cosmetic composition of the present invention is not particularly limited in the form of the formulation, and can be manufactured in all the formulations well known and obvious to those of ordinary skill in the art. As a non-limiting example, there may be mentioned one or more formulations selected from cosmetics, essences, creams, packs, patches, skin adhesive gels, foundations, and makeup bases.

Hereinafter, the present invention will be described in more detail with reference to examples. It is to be noted, however, that the following examples are for the purpose of illustration only and are not intended to limit the scope of the present invention.

Example 1

Preparation of licorice extract

Licorice of Jecheon country of origin was extracted with 40% (w / v) 85% ethanol solvent for 10 ~ 15 hours. The extracted licorice extract was filtered, concentrated under reduced pressure and lyophilized to obtain a licorice powder. The fractions were fractionated with purified water and ethyl acetate using 1 ~ 10% of the weight of licorice extract powder. After the fractionation, the mixture was divided into an ethyl acetate fraction and a water fraction layer to obtain a water layer, followed by freeze-drying to obtain a water fraction powder. The thus obtained product was called "Example 1" and used in the following test examples.

Example 2

Preparation of licorice extract

Licorice of Jecheon country of origin was extracted with 40% (w / v) 85% ethanol solvent for 10 ~ 15 hours. The extracted licorice extract was filtered, concentrated under reduced pressure and lyophilized to obtain a licorice powder. The fractions were fractionated with purified water and ethyl acetate using 1 ~ 10% of the weight of licorice extract powder. After the fractionation, the ethyl acetate layer was obtained by dividing the ethyl acetate fraction layer and the water fraction layer by the licorice extract and lyophilized to obtain an ethyl acetate fraction powder. The thus obtained product was called "Example 2" and used in the following test examples.

Comparative Example 1

Preparation of licorice extract

Licorice of Jecheon country was extracted with 20% (w / v) 20% ethanol solvent at 40 ℃ for 10 ~ 15 hours. The extracted licorice extract was filtered, concentrated under reduced pressure and lyophilized to obtain a licorice powder. The licorice extract thus obtained was called "Comparative Example 1" and used in the following test examples.

Comparative Example 2

Preparation of licorice extract

The liquorice of Jecheon origin was extracted with 40% (w / v) 50% ethanol solvent at 40 ℃ for 10 ~ 15 hours. The extracted licorice extract was filtered, concentrated under reduced pressure and lyophilized to obtain a licorice powder. The licorice extract thus obtained was referred to as "Comparative Example 2" and used in the following test examples.

Comparative Example 3

Preparation of licorice extract

The liquorice of Jecheon origin was extracted with 40% (w / v) 70% ethanol solvent at 40 ℃ for 10 ~ 15 hours. The extracted licorice extract was filtered, concentrated under reduced pressure and lyophilized to obtain a licorice powder. The licorice extract thus obtained was referred to as "Comparative Example 3" and used in the following test examples.

Comparative Example 4

Preparation of licorice extract

The liquorice of Jecheon, the country of origin, was extracted at 40 ° C for 10 to 15 hours using a 95% ethanol solvent at 20 times (w / v) dry weight. The extracted licorice extract was filtered, concentrated under reduced pressure and lyophilized to obtain a licorice powder. The licorice extract thus obtained was called "Comparative Example 4" and used in the following test examples.

Test Example One

ARE-inducible activity in HaCaT-ARE-GFP-luciferase cells

In order to examine the inducible activity of Nrf2 activity of the licorice extract of the present invention, HaCaT-ARE-GFP-luciferase cells were used. HaCaT-ARE-GFP-luciferase cells are cells in which the anti-oxidant response element (ARE) that binds Nrf2 protein is inserted into HaCaT cell, a human keratinocyte cell line. When ARE is activated by drug, GFP and Luciferse are simultaneously expressed .

The luciferase extracts prepared in Example 1 and Comparative Example 1 were treated with 200 ppm (ug / mL) for 24 hours, and the respective luciferase values were measured and analyzed by Student t-test on HaCaT-ARE-GFP-luciferase cells Activity was statistically treated to compare ARE induction.

As shown in FIG. 1, the Nrf2 transcription factor expression was increased in a concentration-dependent manner, and it was found that the treatment with 200 ppm increased the Nrf2 transcription factor by 12-fold.

Example 3 to 4, and Comparative Example 5 to 8

Cream formulation containing licorice extract (formulation)

The composition of the cream containing the licorice extract prepared in the above Examples and Comparative Examples was prepared as shown in Table 1 below.

number ingredient Example 3 Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Comparative Example 8 One Pro-type glycerin monostearate 2.0 2.0 2.0 2.0 2.0 2.0 2 Stearic acid 1.5 1.5 1.5 1.5 1.5 1.5 3 Cetearyl alcohol 2.2 2.2 2.2 2.2 2.2 2.2 4 Wax 1.0 1.0 1.0 1.0 1.0 1.0 5 Squalane 3.0 3.0 3.0 3.0 3.0 3.0 6 Hardened vegetable oil 1.0 1.0 1.0 1.0 1.0 1.0 7 Sorbitan stearate 0.6 0.6 0.6 0.6 0.6 0.6 8 Mineral oil 5.0 5.0 5.0 5.0 5.0 5.0 9 Polysorbate 60 1.5 1.5 1.5 1.5 1.5 1.5 10 Dimethicone 1.0 1.0 1.0 1.0 1.0 1.0 11 Trioctanoin 5.0 5.0 5.0 5.0 5.0 5.0 12 Betaine 3.0 3.0 3.0 3.0 3.0 3.0 13 Triethanolamine 1.0 1.0 1.0 1.0 1.0 1.0 14 glycerin 5.0 5.0 5.0 5.0 5.0 5.0 15 Sodium hyaruronate 3.0 3.0 3.0 3.0 3.0 3.0 16 Example 1 5 - - - - - Example 2 - 5 - - - - Comparative Example 1 - - 5 - - - Comparative Example 2 - - - 5 - - Comparative Example 3 - - - - 5 - Comparative Example 4 - - - - - 5 17 Distilled water Balance Balance Balance Balance Balance Balance 18 Preservative, incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount

The aqueous components of the raw materials 12 to 15 and 17 were completely dissolved by heating to 80 ° C among the components constituted by the composition of Table 1 and then the raw materials 1 to 11 and 16 were heated to 80 ° C to obtain the raw materials 12 to 15 and 17 (Homo Mixer Mark II, Primix, JPN) for 15 minutes at 3,000 rpm. Thereafter, the raw material 18 was added, stirred for 5 minutes, and then cooled to room temperature.

Test Example 2

Assessment of primary skin irritation

To evaluate the degree of primary skin irritation using the cosmetic compositions of the above Examples and Comparative Examples, the following evaluation was made. The subjects were selected from more than 30 adults who did not meet the selection criteria and who were not eligible under the exclusion condition. The subjects were flattened except for the vertebrae and applied to areas where there was no pigmentation or skin damage. Skin irritation was assessed by the Frosch & Kligman, CTFA (Cosmetic, Toiletry and Fragrance Association) guideline, and the skin reaction was evaluated using the following formula (1) according to the criteria of Table 2 below. The skin irritation information of the test substance was also determined by referring to the skin irritation index table of Table 3 using the following equation (2).

sign Rating Criteria + One Some erythema, spots or diffuse ++ 2 Uniform erythema of the degree of increase +++ 3 Severe erythema with edema ++++ 4 Severe erythema with edema and small bubbles

Skin irritation index division Remarks 0.00 to 0.25 Non-magnetic 6.25% 0.26 to 1.00 Weak irritation 25.0% 1.01 to 2.50 Moderate irritancy 62.5% 2.51 to 4.00 Strong irritant 100.0%

As shown in the following Table 4, the creams of Examples and Comparative Examples were applied to the skin for 24 hours, and the skin reactions at 1 hour and 24 hours after the removal of the skin patches were visually evaluated by the researchers and skin irritation index and skin irritation degree were observed. Example 3 was judged to be non-magnetic and comparative to weakly irritant.

Test Products Skin irritation index Degree of skin irritation Example 3 0.03 Non-irritant Example 4 0.37 Weak irritation Comparative Example 5 0.85 Weak irritation Comparative Example 6 0.77 Weak irritation Comparative Example 7 0.68 Weak irritation Comparative Example 8 0.61 Weak irritation

[Formulation Example 1] Flexible longevity

number ingredient content(%) One glycerin 5.00 2 1,3-butylene glycol 3.00 3 PEG 1500 1.00 4 Allantoin 0.10 5 DL-Panthenol 0.30 6 EDTA-2Na 0.02 7 Benzophenone-9 0.04 8 ethanol 10.00 9 Octidodeces-16 0.20 10 Polysorbate 20 0.20 11 Licorice extract 1.0 12 Preservative, fragrance, pigment a very small amount 13 Distilled water Balance

[Formulation Example 2] Convergent lotion

number ingredient content(%) One glycerin 2.00 2 1,3-butylene glycol 2.00 3 Allantoin 0.10 4 DL-Panthenol 0.30 5 EDTA-2Na 0.02 6 Benzophenone-9 0.04 7 ethanol 15.00 8 Polysorbate 20 0.20 9 Licorice extract 1.0 10 Citric acid a very small amount 11 Preservative, fragrance, pigment a very small amount 12 Distilled water Balance

[Formulation Example 3] Nutritional lotion

number ingredient content(%) One Cetearyl alcohol 1.00 2 Glyceryl stearate 0.50 3 Polysorbate 60 1.00 4 Sorbitan sesquioleate 0.30 5 Cetyl octanoate 6.00 6 Squalane 4.00 7 Shapower Oil 4.00 8 Butylene glycol 4.00 9 glycerin 4.00 10 Carbomer 0.10 11 Triethanolamine 0.10 12 Licorice extract 3.00 13 Preservative, fragrance, pigment a very small amount 14 Distilled water Balance

[Formulation Example 4] Essence

number ingredient content(%) One glycerin 10.00 2 Betaine 5.00 3 PEG 1500 2.00 4 Allantoin 0.10 5 DL-Panthenol 0.30 6 EDTA-2Na 0.02 7 Benzophenone-9 0.04 8 Hydroxyethylcellulose 0.10 9 Carboxyvinyl polymer 0.20 10 Triethanolamine 0.18 11 Octyldodecanol 0.30 12 Octyldodec-16 0.40 13 ethanol 6.00 14 Licorice extract 3.00 15 Preservative, fragrance, pigment a very small amount 16 Distilled water Balance

[Formulation Example 5] Pack

number ingredient content(%) One Polyvinyl alcohol 15.00 2 Cellulose sword 0.15 3 glycerin 3.00 4 PEG 1500 2.00 5 Sheik Destrin 0.15 6 DL-Panthenol 0.30 7 Allantoin 0.10 8 Monoammonium glycyrrhizin acid 0.20 9 Nicotinamide 0.40 10 ethanol 5.00 11 PEG 40 hardened castor oil 0.30 12 Licorice extract 3.00 13 Preservative, fragrance, pigment a very small amount 14 Distilled water Balance

Claims

A composition for promoting Nrf2 activity comprising an extract of licorice as an active ingredient.

[Claim 2] The composition for promoting Nrf2 activity according to claim 1, wherein the licorice extract is fractionated with water and ethyl acetate to exclude cytotoxic or skin irritant components and water fractions.

Extracting licorice into an alcohol solvent having a carbon number of C ₁ to C ₄ ;
Lyophilizing the extract to obtain a powder;
Obtaining the powder with water and ethyl acetate to obtain a fraction; And
The method of claim 1, comprising obtaining a water fraction of the fraction.

4. The method of claim 3, wherein the alcohol solvent is a 75-90% ethanol solvent.

[4] The method of claim 3, wherein the water and ethyl acetate are used in an amount of 1 to 10% by weight based on the weight of the powder.

A cosmetic composition for skin homeostasis maintenance, comprising the licorice extract of claim 1 in an amount of 0.001 to 10% by weight based on the total weight of the composition.

The skin homeostasis-maintaining cosmetic composition according to claim 6, wherein the cosmetic composition is at least one selected from the group consisting of lotion, essence, cream, pack, patch, skin adhesive, foundation, and makeup base.