KR101868507B1

KR101868507B1 - Cosmetic Composition for Anti-oxidation, Anti-inflammatory and Anti-wrinkling Comprising Enzyme-treated Extracts of Annona Muricata and Method for Manufacturing the Same

Info

Publication number: KR101868507B1
Application number: KR1020170135984A
Authority: KR
Inventors: 송창섭; 유마영; 조재응
Original assignee: (주)해피바이오
Priority date: 2017-10-19
Filing date: 2017-10-19
Publication date: 2018-06-19

Abstract

The present invention relates to a cosmetic composition for antioxidant, anti-inflammation and anti-wrinkle containing an enzyme-treated graviola ( Annona Muricata ) extract as an active ingredient and a process for producing the same.
The extract of Annona Muricata , which is provided from the present invention, has excellent antioxidative, anti-inflammatory and anti-wrinkle effects, and thus has excellent effects as a cosmetic composition when it is applied to a cosmetic base as an effective ingredient.

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement containing an enzyme-treated graviola extract as an active ingredient and a method for producing the same. Manufacturing the Same}

The present invention relates to a cosmetic composition for antioxidant, anti-inflammation and anti-wrinkle containing an enzyme-treated graviola ( Annona Muricata ) extract as an active ingredient and a process for producing the same.

Graviola ( Annona Muricata ) belongs to the family Annonacea and contains about 2,300 species, of which about 130 are widely distributed in the United States, Africa, Asia and Brazil. The height of the tree is 6 ~ 7m, the leaves are ovate or long elliptical, 7.5 ~ 15㎝ long, the tip of the leaf is pointed and the petiole part is wedge shaped. Leaves are smooth and shiny, petiole is short. Flowers are strong in fragrance, 0.5 ~ 2.5㎝ long, sepal spaced from petal, 3 ~ 5㎝ in size. The petals are composed of 6 compound leaves. The petals are thick, leathery, broad egg-shaped, with sharp ends and soft hairs. The weight of the fruit is about 2 ㎏, and some go up to 6 ㎏. The fruit is oval, greenish, fragrant, and has a thorn on its surface.

Graviola has attracted attention in recent years because it has been known to have beneficial effects on the human body enough to be praised as a miracle plant of God. A typical effect is that it is used for the treatment of alternative cancer because it has the effect that the cancer cells are self-destructed by blocking the feeding channel so that the cancer cells do not grow due to the anti-cancer effect. Adriamycin has a more potent cytotoxic effect than adriamycin, a chemical currently used in hospitals, and has the effect of inhibiting the growth of all cancer cells such as colon cancer, liver cancer, stomach cancer, pancreatic cancer, breast cancer, prostate cancer and lung cancer I am receiving great attention.

Graviola is also known to have excellent antibacterial activity. Therefore, it can effectively treat bacterial infections and can exert a parasitic killing effect on a wide range of antimicrobial functions throughout the human body.

Techniques that have been applied to various applications using an Annona Muricata extract include Korean Patent Laid-Open Publication No. 10-2017-0047515 entitled " Cosmetic Composition for Promoting Skin Vitality " Containing Antibacterial Extract of Graviola and Method for Producing the Same Korean Patent Registration No. 10-1648148 entitled " Cosmetic Composition for Improving Skin Wrinkle " and Preparation Method Thereof ", Korean Patent Registration No. 10-1763163 entitled " Cosmetic Composition for Improving Wrinkle of Skin Using Graviola Fermentation Liquid as an Active Ingredient &Quot; graviola composition "

Accordingly, the inventors of the present invention found that when an extract of graviola ( Annona Muricata ) is treated with an enzyme, excellent antioxidation, anti-inflammation and anti-wrinkle effects can be imparted.

Korean Patent Publication No. 10-2017-0047515 Korean Patent Publication No. 10-1648148 Korean Patent Registration No. 10-1763163

It is an object of the present invention to provide a cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement containing an enzyme-treated graviola ( Annona Muricata ) extract as an active ingredient.

Yet another object of the present invention is to provide a method for producing a cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement, which comprises an extract of Annona Muricata as an active ingredient.

In order to achieve the above-mentioned object, the present invention provides a cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement comprising an extract of Annona Muricata as an active ingredient.

In the present invention, the enzyme is characterized by being at least one selected from the group consisting of cellulase and pectinase.

In the present invention, the cellulase and the pectinase are mixed in a weight ratio of 1: 1 to 5: 1.

In the present invention, the enzyme-treated graviola ( Annona Muricata ) extract is contained in an amount of 0.001 to 50% by weight based on the total weight of the composition.

In the present invention, the enzyme-treated graviola ( Annona Muricata ) extract is characterized by having DPPH free radical scavenging ability, nitric oxide formation inhibiting ability and elastase inhibiting ability.

In addition, the present invention provides a method for producing an enzyme- linked immunosorbent assay , comprising the steps of: adding an enzyme to graviola ( Annona Muricata ) Adding ethanol to the enzyme treated graviola ( Annona Muricata ) to precipitate and centrifuge the enzyme; And removing the precipitated enzyme and stirring to obtain an enzyme-treated grape extract ( Annona Muricata ). The present invention also provides a method for producing a cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement.

In the present invention, the enzyme reaction is performed at 30 to 55 ° C for 30 to 120 minutes.

The extract of Annona Muricata , which is provided from the present invention, has excellent antioxidative, anti-inflammatory and anti-wrinkle effects, and thus has excellent effects as a cosmetic composition when it is applied to a cosmetic base as an effective ingredient.

1 is a graph showing cytotoxicity evaluation of macrophages (RAW 264.7) of an enzyme-treated graviola extract, graviola extract and graviola fermented extract according to an embodiment of the present invention.
FIG. 2 is a graph showing DPPH free radical scavenging activity of an enzyme-treated graviola extract, graviola extract, and graviola fermented extract according to an embodiment of the present invention.
FIG. 3 is a graph showing the nitric oxide production inhibiting activity of an enzyme-treated graviola extract, graviola extract, and graviola fermented extract according to an embodiment of the present invention.
FIG. 4 is a graph showing the inhibitory activity of the enzyme-treated graviola extract, graviola extract, and graviola fermented extract according to an embodiment of the present invention.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.

Throughout this specification, when an element is referred to as "including " an element, it is understood that the element may include other elements as well, without departing from the other elements unless specifically stated otherwise.

In the present invention, DPPH free radical scavenging activity, nitric oxide production inhibitory activity and elastase inhibitory activity were examined from the extract of Annona Muricata , and it was found that there was an excellent antioxidative, anti-inflammatory and anti-wrinkle effect Respectively.

Accordingly, in one aspect, the present invention relates to a cosmetic composition for antioxidant, anti-inflammation and anti-wrinkle containing an enzyme-treated graviola ( Annona Muricata ) extract as an active ingredient.

In the present invention, the term " Annona Muricata extract" refers to an extract obtained by adding an enzyme to grape extract ( Annona Muricata ) and then extracting the mixture using a solvent.

The enzyme is preferably at least one selected from the group consisting of a cellulase and a pectinase from the viewpoint that the yield of the effective ingredient is optimally increased and the antioxidative, anti-inflammatory and wrinkle-reducing effects are remarkably increased.

The cellulase and the pectinase are mixed at a weight ratio of 1: 1 to 5: 1 to provide DPPH free radical scavenging ability, nitric oxide formation inhibiting ability and elastase inhibiting ability .

When the enzyme is treated with the graviola ( Annona Muricata ), it is preferable that the enzyme is added in an amount of 5 to 15 parts by weight based on 100 parts by weight of graviola ( Annona Muricata ) from the viewpoint that the yield is remarkably increased.

The enzyme-treated graviola ( Annona Muricata ) extract is characterized by DPPH free radical scavenging activity, nitric oxide formation inhibiting activity and elastase inhibiting activity.

According to another aspect of the present invention, there is provided a method for producing an enzyme, comprising the steps of: adding an enzyme to grape violet ( Annona Muricata ) Adding ethanol to the enzyme treated graviola ( Annona Muricata ) to precipitate and centrifuge the enzyme; And removing the precipitated enzyme and stirring to obtain an enzyme-treated grape extract ( Annona Muricata ). The present invention also relates to a method for producing a cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement.

When an enzyme is added to the gravola ( Annona Muricata ) to perform an enzymatic reaction, water may be added to facilitate the enzyme reaction, followed by stirring and reacting.

The enzymatic reaction is preferably carried out at 30 to 55 DEG C for 30 to 120 minutes in terms of conversion and extraction efficiency. If the temperature and the time are out of the above ranges, the enzyme is not converted and thus it is not preferable because it can not exhibit sufficient antioxidative, anti-inflammatory and anti-wrinkle effects.

After removing the precipitated enzyme using typical filtration method of the addition of ethanol to the gras viola (Annona Muricata) processing the enzyme to precipitate the enzyme and centrifugation, then filter, and the like, stirring extraction and enzyme treatment Gras viola (Annona Muricata ) extract.

The stirring extraction is preferably carried out at 80 to 105 ° C for 120 to 240 minutes in view of enhancing the extraction yield and the effective ingredient. If the temperature and the time are out of the above ranges, the active ingredient is not sufficiently dissolved and the antioxidant, anti-inflammation and anti-wrinkle effects can not be exhibited.

The enzyme-treated graviola ( Annona Muricata ) extract may be processed into various forms such as an extract, a powder obtained by drying the extract, and a concentrate obtained by concentrating and filtering the extract.

As described above, the Annona Muricata extract has excellent DPPH free radical scavenging activity, nitric oxide production inhibitory activity and elastase inhibitory activity, and thus can be used as a composition for antioxidant, anti-inflammation and wrinkle reduction Can be used.

The cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement of the present invention can be used as an external ointment for skin, a cream, a softening longevity, a nutritional lotion, pack, essence, hair tonic, shampoo, rinse, hair conditioner, hair treatment, gel, , Skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, eye cream, moisturizing cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream , Body lotions, and body cleansers, but are not limited thereto. The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

In the cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement of the present invention, the enzyme-treated graviola extract ( Annona Muricata ) may be contained in an amount of 0.01 to 50% by weight, preferably 10 to 30% Weight percent of the composition, but is not limited thereto, including effective amounts that can provide desirable antioxidant, anti-inflammatory and anti-wrinkle effects.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not to be construed as limiting the scope of the present invention. It will be self-evident.

Example 1 Preparation of Enzyme-Treated Graviola Extract

10 g of a mixture of 1 g of Cellulase (Novozyme) and 1 g of pectinase (Novozyme) in a ratio of 1: 1 and 300 ml of water were added to 100 g of graviola powder (200 mesh) purchased from Yeongcheon, And extracted for 12 hours. 3 L of ethanol (EtOH) was added to the obtained extract to precipitate the enzyme, and then the centrifugal separator was removed at 8,000 rpm for 20 minutes. The remaining residue and solution were stirred at room temperature for 3 days, and then concentrated under reduced pressure and lyophilized to obtain a final enzyme-treated graviola extract.

Examples 2 to 5 Preparation of enzyme-treated graviola extract

Cellulase and Pectinase were prepared in the same manner as the enzyme-treated graviola extract of Example 1, except that the cellulase and the pectinase were mixed as shown in Table 1 below.

enzyme Amount of enzyme added Example 2 Example 3 Example 4 Example 5 Cellulase (S) 1g 1g 1g 1g Pectinase (P) 2g 3g 4g 5g Enzyme treatment cost (S: P) 1: 2 1: 3 1: 4 1: 5

&Lt; Comparative Example 1 > Preparation of graviola extract

The graviola extract was prepared in the same manner as in Example 1, except that the enzyme was not treated. Namely, 300 ml of water was added to 100 g of graviola powder (200 mesh) purchased from Yeongcheon, Gyeongbuk Province, and the mixture was extracted at 80 캜 for 12 hours, followed by concentration under reduced pressure and lyophilization to obtain final gravola extract.

<Experimental Example 1> Cytotoxicity test: WST assay

Cytotoxicity tests were carried out on macrophages (RAW 264.7) in order to confirm whether the extracts of Examples 1 to 5 and Comparative Example 1 had irritation to cells. RAW 264.7 cells, a type of macrophage, were counted by the same number of 1.0 × 10 ⁴ cells / well on a 24-well plate using DMEM medium containing 1% penicillin / scrap tromycin and 10% FBS (Fetal bovineserum) Followed by incubation at 37 DEG C and 5% carbon dioxide for 24 hours. Each of the cells of Examples 1 to 5 and Comparative Example 1 was mixed with the culture medium for 24 hours, and then 1 ml of the mixture was added to each well. Then, the cells were incubated in an incubator at 37 ° C and 5% CO ₂ for 24 hours. The supernatant of each well was separately taken, and WST-1 assay solution (ez-cytox) was added to each well. After incubation for 2 hours in an incubator, absorbance was measured at 450 nm using an ELISA reader.

Cell viability was calculated using the following equation (1) as compared with the untreated group (untreated group).

[Equation 1]

Cell survival rate (%) = (absorbance of sample treated group / absorbance of untreated sample group) X 100

As a result, as shown in Fig. 1, in the case of cell viability for macrophages (RAW 264.7), all of the extracts of Examples 1 to 5 and Comparative Example 1 did not show toxicity to cell survival and thus could be used as a safe material for cosmetics Respectively.

&Lt; Experimental Example 2 > Antioxidant Efficacy Test: Free radical formation inhibitory activity

The free radical scavenging activity of DPPH was evaluated in order to evaluate the antioxidative effect using the extracts of Examples 1 to 5 and Comparative Example 1. [ Examples 1 to 5 and Comparative Example 1 were diluted to a concentration of 50 (占 퐂 / ml), followed by mixing 250 占 퐇 of 0.1M DPPH (1,1-diphenyl-2-picrylhydrazyl) Lt; / RTI > At the end of 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, ascorbic acid was used as a positive control, and free radical scavenging activity (%) was calculated using the following equation (2) as a control.

&Quot; (2) "

Free radical scavenging ability (%) = [100- (absorbance of sample-treated group / absorbance of sample-treated group X 100)]

As a result, as shown in FIG. 2, the DPPH free radical scavenging ability of Examples 1 to 5 was superior to that of Comparative Example 1, and the free radical scavenging ability was remarkably high in Example 3 among Examples 1 to 5 Respectively.

<Experimental Example 3> Anti-inflammatory activity test: Nitric oxide formation inhibitory activity

In order to measure anti-inflammatory activity using the extracts of Examples 1 to 5 and Comparative Example 1, experiments were conducted to measure the concentration of nitric oxide produced by the inflammation-inducing reaction. RAW 264.7 cells, a type of macrophage, were counted by the same number of 1.0 × 10 ⁴ cells / well on a 24-well plate using DMEM medium containing 1% penicillin / scrap tromycin and 10% FBS (Fetal bovineserum) , And cultured at 37 DEG C and 5% carbon dioxide for 24 hours. Examples 1-5 and Comparative Example 1 extract 24 hours incubation cell 50 (㎍ / ㎖) after concentration of the medium, and mixed with, added in 1ml to each well and the incubator of 37 ℃, 5% CO ₂ conditions, 24 Lt; / RTI > At this time, LPS (Lipo poly saccharide), which is an inflammation inducer for expressing NO, was treated at a concentration of 1 μg / ml and reacted at 37 ° C and 5% carbon dioxide for 24 hours. After removing the supernatant from each well, 100 ml of the culture solution was placed in a 96-well plate using a Nitric Oxide Detection Kit and 50 μl of Griess reagent A (N-1-Naphthylethylenediamine (NEDHC)) and Griess reagent B (Sulfanilamide) And incubated for 10 minutes. Absorbance was measured at 540 nm using an ELISA plate reader.

As a result, as shown in FIG. 3, Examples 1 to 5 were superior to Comparative Example 1 in inhibiting the formation of nitric oxide. In Examples 1 to 5, nitric oxide (Nitric oxide) It was confirmed that the anti-inflammatory effect was excellent because the inhibitory effect was remarkably high.

<Experimental Example 4> Wrinkle-improving test: Elastase inhibitory activity

In order to measure the wrinkle-reducing effect using the extracts of Examples 1 to 5 and Comparative Example 1, the degree of inhibition of elastase was evaluated. Examples 1 to 5 and Comparative Example 1 were diluted to a concentration of 50 μg / ml, and then N-succinyl- (Ala) 3-p-nitroanilide (S4760, Sigma) was incubated at 25 ° C for 5 min. The absorbance at 410 nm was measured in a 96-well plate. The absorbance of the substrate was measured at 410 nm. And the case where no sample was added was used as a control group and calculated using the following equation (3). At this time, Oleanolic acid was used as a positive control.

&Quot; (3) "

(%) = 100- (absorbance of sample-treated group / absorbance of sample-treated group X 100)

As a result, as shown in FIG. 4, Examples 1 to 5 were superior in inhibiting elastase to Comparative Example 1, and Elastase inhibition was remarkably improved in Example 3 among Examples 1 to 5 And it was confirmed that the effect of improving wrinkles was excellent.

&Lt; Preparation Example 1 > Enzyme-treated graviola ( Annona Muricata ) Preparation of cosmetics containing extract

(1) Production of lotion

Lotion was prepared from the composition shown in Table 2 by a conventional method.

ingredient Content (% by weight) Example 3 (Enzyme-Treated Graviola Extract) 3.00 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 68.00 Sum 100.00

(2) Lotion manufacturing

Lotions were prepared from the compositions shown in Table 3 by a conventional method.

ingredient Content (% by weight) Example 3 (Enzyme-Treated Graviola Extract) 3.00 Ascorbic acid-2-phosphate magnesium 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 91.94 Sum 100.0

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereto will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims

Containing an extract of Grape Viola ( Annona Muricata ) as an active ingredient,
The cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement is characterized in that the enzyme is a complex enzyme in which cellulase and pectinase are mixed in a weight ratio of 1: 1 to 1: 5.

delete

The cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement according to claim 1, wherein the enzyme-treated graviola ( Annona Muricata ) extract is contained in an amount of 0.001 to 50 wt% based on the total weight of the composition.

The method according to claim 1, wherein the enzyme-treated graviola ( Annona Muricata ) extract has DPPH free radical scavenging activity, nitric oxide production inhibitory activity and elastase inhibitory activity. A cosmetic composition for improvement.

(EN) An enzyme reaction is carried out by adding a complex enzyme in which graviola ( Annona Muricata ) is mixed with a cellulase and a pectinase in a weight ratio of 1: 1 to 1: 5;
Adding ethanol to the enzyme treated graviola ( Annona Muricata ) to precipitate and centrifuge the enzyme; And
And removing the precipitated enzyme and stirring to obtain an enzyme-treated grape extract ( Annona Muricata ). The method for producing a cosmetic composition for antioxidant, anti-inflammation and wrinkle improvement according to claim 1,

delete

7. The method of claim 6, wherein the enzymatic reaction is carried out at 30 to 55 DEG C for 30 to 120 minutes.