KR20190094261A - Anti-inflammatory cosmetic composition comprising Acacetin from the enzymatic extract of viola mandshurica as an active ingredient and preparation method of the same - Google Patents

Abstract

The present application relates to a method for preparing an enzymatic extract of whole plant body of Viola mandshurica, an enzymatic extract of whole plant body of Viola mandshurica prepared by the method, and an anti-inflammatory functional cosmetic composition comprising the enzymatic extract of whole plant body of Viola mandshurica containing acacetin as an active ingredient. The preparation method includes the steps of obtaining a concentrate; obtaining an enzyme additive; inactivating an enzyme; and obtaining an enzymatic extract of whole plant body of Viola mandshurica.

Description

Anti-inflammatory cosmetic composition comprising Acacetin from the enzymatic extract of viola mandshurica as an active ingredient and preparation method of the same}

The present application relates to an anti-inflammatory functional cosmetic composition containing acetin (Acacetin) of violet starch enzyme extract as an active ingredient and a method of manufacturing the same.

Viola mandshurica is a perennial plant belonging to the family Violaceae . It grows in the mountains and wild places, and is wild in Jeju, Gyeongnam (Namdo), Chungbuk, and Gyeonggi-do. Young plants are edible, and all plants, including nets, are called 地 丁 (designated) and are medicinal. Dongbobom, according to the herbaceous tree, violets are effective in antibacterial, purulent skin disease, acne, arthritis, etc.

Inflammation is an immune response of biological tissues caused by external stimuli. When inflammation occurs, hyperpigmentation usually occurs. Inflammation occurs due to UV rays or bacterial infections, and the color of the face becomes black, and blemishes and blemishes occur. . At this time, cells involved in inflammatory reactions such as lymphocytes, macrophages, endothelial cells, and fibroblasts, which are cells in the skin, are involved. Exposure to stimulants such as -γ (INF-γ), interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) increases iNOS expression. As a result, the production of NO increases the inflammatory response as well as the ROS, which may exacerbate the disease by increasing pigmentation, skin aging and tissue damage [S. N. Park, et al., 2010, Bu Hee Jung, 2016]. Therefore, in order to prevent this phenomenon and maintain healthy skin, studies have been conducted to maintain skin's intrinsic function and activate skin cells by using cosmetics enhanced with bioactive substances obtained from natural products.

Linarin is mainly contained in chrysanthemum plants such as gamguk, gujeolcho and thistle, and is flavonoid glycoside combined with rutinose in acacetin. Linarin is a flavonoid, a colorless needle-like crystal (including one molecule of crystal water) that has a dark green color in iron (III) chloride solution and is insoluble in water and most other organic solvents. Linarin is hydrolyzed with concentrated hydrochloric acid to produce one molecule of acetin, glucose and rhamnose.

Aceacetin is called Robinia pseudo - acacia As a glycoside akashine on the leaves of L. (legume), Wisteria thistle ( Cirsium) It is contained in the leaves of purpuratum (Maxim. ) or Asteraceae as glycoside linalin. Acecetin is an almost colorless needle-like crystal that dissolves in iron (III) chloride as a brownish-brown color and a vivid yellow in alkali. Insoluble in water, chloroform, benzene and petroleum ether, easily soluble in warm alcohol.

In the prior art related to the present application, the antioxidant activity and the inhibitory activity of α-Amylase and α-Glucosidase (Journal of the Korean Society of Food Science and Nutrition v.37 no.4, pp. 405-409, 2008, 1226-331) and antioxidant activity and whitening effect of violet fermented extract (Korean Society of Agronomic Crops, 2009) are known, but have not been disclosed or implied for the cosmetic composition including an active ingredient exhibiting the anti-inflammatory effect of violet starch enzyme extract. In addition, although the Republic of Korea Patent Publication No. 10-2010-0090371 is known, which is a prior document regarding a composition containing a violet leaf extract having a blood sugar lowering effect as an active ingredient, Republic of Korea Patent No. 10-1107614 is known but violet Only the whitening effect, the antioxidant effect and the antimicrobial effect of the extract are disclosed, and the active ingredient or cosmetic composition exhibiting the anti-inflammatory effect of the violet extract has not been disclosed or implied in any of the above-mentioned prior documents.

Under these backgrounds, the present inventors have conducted a intensive research effort to develop a natural cosmetic composition having an excellent anti-inflammatory effect, and as a result, the present invention has developed a cosmetic composition comprising a violet starch enzyme extract which has increased the content of an active ingredient having an anti-inflammatory effect by enzymatic hydrolysis of the violet starch extract. This application was completed.

An object of the present application is to provide a method for producing violet starch enzyme extract and violet starch enzyme extract prepared by the above method.

Another object of the present application is to provide an anti-inflammatory functional cosmetic composition comprising a violet starch enzyme extract containing acetin as an active ingredient.

Other objects and advantages of the present application will become more apparent from the following detailed description taken in conjunction with the appended claims and drawings. Content not described herein is omitted because it can be sufficiently recognized and inferred by those skilled in the art or similar fields of the present application.

Hereinafter, the content of the present application will be described in detail. In addition, description and embodiment of one embodiment disclosed by this application can be applied also to description and embodiment of other aspect about common matter. In addition, all combinations of the various elements disclosed in the present application are within the scope of the present application. In addition, the scope of the present application is not to be limited by the specific description described below.

In order to achieve the object of the present application, the present application provides a violet starch enzyme extract and a method of manufacturing the same as an embodiment.

Violet starch enzyme extract of the present application is to concentrate the filtrate of violet starch extract to obtain a concentrate, and to add the complex hydrolase to the concentrate to obtain an enzyme additive and hydrolysis by stirring the enzyme additive After the reaction, the enzyme is inactivated, and the inactivated enzyme additive is centrifuged and the lower layer is concentrated to obtain a violet starch enzyme extract.

According to the exemplary embodiment of the present application, the extract of the violet starch extract may be obtained by dipping at 10 to 40 times 30% to 70% ethanol of the dried violet starch dry weight (kg) and extracting at room temperature. More preferably, 20 to 35 times 40% to 60% ethanol of the weight of the violet outpost may be used, and most preferably, the extract may be obtained by dipping in 30 times 50% ethanol and extracting at room temperature.

The concentrate of the filtrate of the violet starch extract may be concentrated using a vacuum suction suction filtration method, a freeze pressure reduction method, a high temperature evaporation concentration method, a low temperature pressure reduction method, but preferably a pressure suction suction filtration method.

The hydrolase of the present application requires water in chemical reactions and is an enzyme that acts on various hydrolysis reactions in vivo, such as ester hydrolase, glycoside hydrolase (amylase, nucleosidase, etc.), peptide Hydrolase, pyrophosphate hydrolase, complex hydrolase may be used and 5% to 50% of the weight of the violet outpost (kg) may be used. In the present application, biscozyme, which is most preferably a complex hydrolase, is used, and the amount of the enzyme is preferably 10% to 30% of the weight of the violet outpost (kg), and most preferably 20 of the weight of the violet outpost (kg). Use%. Stirring of the enzyme additive of the present application may be carried out at 30 ° C. to 60 ° C. for hydrolysis reaction, more preferably at 40 ° C. to 55 ° C., most preferably at 50 ° C. for hydrolysis reaction.

Violet starch enzyme extract of the present application may be contained in the anti-inflammatory functional cosmetic composition and violet starch enzyme extract containing Acacetin in a concentration of 10ppm to 100ppm may be used as an active ingredient of the cosmetic composition.

As described above, the present application not only has an effect of providing a method of preparing a violet starch enzyme extract and a violet starch enzyme extract, but also provides an anti-inflammatory functional cosmetic composition comprising the violet starch enzyme extract containing acetin as an active ingredient. Excellent effect.

1 is a schematic diagram showing the separation process of the active ingredient of violet starch extract.
2 is a structural diagram of Compound 1 (Linarin, Acetin 7-O-rutinoside).
3 is a structural formula of Compound 2 (Acacetin, 4'-Methoxyapigenin).
Figure 4 is a schematic diagram showing the process of obtaining a violet starch enzyme extract from dry violet starch 50% ethanol extract.
5 is a graph showing the HPLC chromatogram of violet outpost 50% ethanol extract.
6 is a graph showing the HPLC chromatogram of violet starch enzyme extract.
7 is a graph showing the effect of inhibiting nitrogen monoxide (NO) production according to the concentration of Acacetin.
8 is a graph showing the effect of inhibiting the production of nitric oxide (NO) according to the concentration of 50% ethanol extract violet flowers.
9 is a graph showing the effect of inhibiting nitrogen monoxide (NO) production according to the concentration of violet starch enzyme extract.

Hereinafter, the present invention will be described in more detail through examples according to the present application. However, the following examples of the present application are merely examples of the present application. These examples are intended to illustrate the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention as set forth in the appended claims is not limited by these examples. something to do.

Experimental Example  Violet outpost  Isolation and Structure Identification of Active Ingredients of Extracts

The dried violet outposts were extracted at room temperature for 24 hours using 20% to 80% ethanol. The obtained extract was concentrated using only a vacuum suction filter, suspended in 100% distilled water and then fractionated in the order of hexane, chloroform, ethyl acetate, butanol, distilled water. Next, a precipitate which is not dissolved in methanol is produced from the Ethyl acetate and butanol fractionation layers, and the precipitates are washed separately with methanol to obtain Compound 1, and the chloroform fractionation layer is chloroform in an open column filled with normal silica gel. Compound 2 was obtained by column chromatography using methanol as a mobile phase (FIG. 1).

Compound 1 and Compound 2 were obtained by using ¹ H & ¹³ C-NMR spectroscopy (nuclear magnetic resonance spectroscopy) to identify the chemical structure of each compound in each 400 MHz DMSO-D6 and 400MHz CDCl ₃ conditions (Table 1 And Figure 2, Table 2 and Figure 3). Compound 1 was identified as Linarin, Acacetin 7-O-rutinoside, and Compound 2 was identified as Acacetin, 4'-Methoxyapigenin.

Example  Violet outpost  Ethanol Extracts and Violets outpost  Obtaining Enzyme Extracts

The dried violet outposts were soaked in 30% 50% of the dry weight (kg) in 50% ethanol and extracted at room temperature for a time to obtain a violet outpost ethanol extract.

The filtrate of the extract obtained by filtering the violet starch ethanol extract was concentrated to the extent that ethanol is completely removed, and then added to the concentrate biscozyme, a complex hydrolase of 20% by weight of the violet starch (kg). The additive was hydrolyzed with stirring at 50 ° C. and then treated at 100 ° C. for 1 hour to inactivate the enzyme. The deactivated additives were then centrifuged and the lower layer was concentrated to give a violet starch enzyme extract (FIG. 4).

Experimental Example  2. violets outpost  Ethanol Extracts and Violets outpost  Of enzyme extract HPLC  analysis

HPLC analysis was carried out to confirm the change in the content of Acacetin of the violet starch ethanol extract obtained by Example 1 and the violet starch enzyme extract obtained by the enzyme treatment.

The following conditions were used for HPLC analysis (Table 3).

Detector: PDA UV 330nm

Flow rate: 1.0ml / min

Column Temperature 30.0 ℃

Sample Temperature 30.0 ℃

Running Time: 60.0min

Column: Kromasil C18 5㎛, 4.6x250mm

Injection volume: 10 μl

Solvent condition: acetonitrile: water (0.1% Acetic acid) = 10: 90-100% acetonitrile

HPLC analysis of the violet starch enzyme extract obtained by treatment with each violet starch ethanol extract, it was confirmed that the acacetin content of violet starch enzyme extract (Fig. 6) was significantly increased than violet starch ethanol extract (Fig. 5).

Experimental Example  3. violets outpost  Of extract Active ingredient Acacetin, violet outpost  Ethanol Extracts and Violets outpost  Anti-inflammatory Effects of Enzyme Extracts

In order to measure the anti-inflammatory effects of the active ingredient Acacetin and the violet starch ethanol extract obtained in Example 1 and the violet starch enzyme extract obtained by the enzyme treatment obtained in Example 1 to inhibit the NO production in the following manner Measured.

RAW 264.7 cells were dispensed into 96-well plates at 2 x 10 ⁵ cells per well, and then incubated in 37 ° C., 5% CO ₂ incubator for 24 hours. After culturing the cells overnight, the cells were replaced with fresh medium, and LPS (1㎍ / mL), an inflammation-inducing substance, and 5, 25, 50ppm of the active ingredient Acacetin, and each extract were administered at a concentration of 10,50,100ppm for 24 hours. Incubated for After incubation, the supernatant of the medium was taken and centrifuged at 13,000 rpm for 3 minutes to collect only the supernatant, and then reacted 1: 1 with Griess reagent and the absorbance was measured at 570 nm.

As a result, Acacetin at 44ppm low concentration showed a good NO inhibitory effect of 44% (Fig. 7). In addition, violet ethanol extract did not show the NO production inhibitory effect (Fig. 8), but the violet starch enzyme extract with increased acacetin content by enzyme treatment

90% NO production inhibitory effect was confirmed at the concentration of 100ppm (Fig. 9).

Hereinafter, various cosmetic formulations are preferred for use in the present invention, but the scope of the present invention is not limited to the disclosed formulations.

Preparation Example 1. toilet water

The present invention, the lotion containing the present invention violet starch enzyme extract obtained in Example 1 was prepared as shown in Table 4.

Raw material Raw material Content (% by weight) One Invention Violet Outpost Enzyme Extract 10.0 2 glycerin 3.0 3 Butylene Glycol 2.0 4 Propylene glycol 2.0 5 Polyoxyethylene Cured Castor Oil 1.0 6 ethanol 10.0 7 Triethanolamine 0.1 8 antiseptic a very small amount 9 Pigment a very small amount 10 Spices a very small amount 11 Purified water Remaining amount

Preparation Example 2. Lotion

The lotion containing the present invention violet starch enzyme extract obtained in Example 1 was prepared as shown in Table 5.

Raw material Raw material Content (% by weight) One Invention Violet Outpost Enzyme Extract 10.0 2 Beeswax 1.0 3 Polysorbate 60 1.5 4 Solvidan Sesquioleate 0.5 5 Floating paraffin 10.0 6 Sorbidate stearate 1.0 7 Lipophilic Monostearine Glycerin 0.5 8 Stearic acid 1.5 9 Glyceryl Stearate / Pig-400 Stearate 1.0 10 Propylene glycol 3.0 11 Carboxy Polymer 0.1 12 Triethanolamine 0.2 13 antiseptic a very small amount 14 Pigment a very small amount 15 Spices a very small amount 16 Purified water Remaining amount

Preparation Example 3. cream

The present invention The cream containing the present invention violet starch enzyme extract obtained in Example 1 was prepared as shown in Table 6.

content Raw material Content (% by weight) One Invention Violet Outpost Enzyme Extract 10.0 2 Stearic acid 2.0 3 Cetyl alcohol 2.0 4 Glyceryl Monostearate 2.0 5 Polyoxyethylene sorbitan monostearate 0.5 6 Solvanesesquioleate 0.5 7 Glyceryl Monostearate / Glyceryl Stearate / Polyoxyethylene Stearate 1.0 8 Wax 1.0 9 Floating paraffin 4.0 10 Squalane 4.0 11 Caprylic / Capric Triglycerides 4.0 12 Carboxy Vinyl Polymer 0.3 13 Butylene glycol 5.0 14 glycerin 3.0 15 Triethanolamine 0.5 16 Purified water Remaining amount

Preparation Example 4 . Essence manufacturer

The essence containing the present invention violet starch enzyme extract obtained in Example 1 was prepared as shown in Table 7.

content Raw material Content (% by weight) One Invention Violet Outpost Enzyme Extract 10.0 2 Cytostolol 1.7 3 Polyglyceryl 2-oleate 1.5 4 Ceramide 0.7 5 Steares-4 1.2 6 cholesterol 1.5 7 Dicetylphosphate 0.4 8 Concentrated glycerin 5.0 9 Macadamia oil 15.0 10 Carboxy Vinyl Polymer 0.2 11 Xanthan Gum 0.2 12 antiseptic a very small amount 13 Spices a very small amount 14 Purified water Remaining amount

From the above description, those skilled in the art will appreciate that the present application can be implemented in other specific forms without changing the technical spirit or essential features. In this regard, it should be understood that the embodiments described above are exemplary in all respects and not limiting. The scope of the present application should be construed that all changes or modifications derived from the meaning and scope of the following claims and equivalent concepts rather than the detailed description are included in the scope of the present application.

The present application provides an anti-inflammatory functional cosmetic composition comprising a method for producing violet starch enzyme extract and violet starch enzyme extract prepared by the above method, and the violet starch enzyme extract containing acecetin as an active ingredient, which is very effective in the natural cosmetic industry. It is a useful invention.

Claims (6)

Concentrating the filtrate of the violet starch extract to obtain a concentrate;
Adding a complex hydrolase to the concentrate to obtain an enzyme additive;
Inactivating the enzyme after the hydrolysis reaction by stirring the enzyme additive;
Centrifuging the inactivated enzyme additives and the lower layer is concentrated to obtain a violet starch enzyme extract method of producing a violet starch enzyme extract.
The method according to claim 1, wherein the complex hydrolase is biscozyme and is added to 20% (W / W) of the weight of the starch and stirred at 30 ° C to 60 ° C for hydrolysis.
Violet starch enzyme extract prepared according to the method of claim 1 or 2 The extract of claim 3 is a violet starch enzyme extract characterized by containing Acacetin Anti-inflammatory functional cosmetic composition comprising a violet starch enzyme extract of claim 3 or 4 as an active ingredient The cosmetic composition of claim 5 is an anti-inflammatory functional cosmetic composition, characterized in that the formulation of any one selected from lotion, lotion, cream and essence.

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