KR20160098570A - COMPOSITION COMPRISING EXTRACT OF Ainsliaea apiculate HAVING ANTI-OXIDANT AND ANTI-INFLAMMATORY ACTIVITY - Google Patents

Abstract

The present invention relates to a food composition and a composition having anti-inflammatory and antioxidant activities, comprising an Ainsliaea apiculata extract having anti-inflammatory and antioxidant activities. The food composition and the composition having anti-inflammatory and antioxidant activities, comprising an Ainsliaea apiculata extract as naturally derived substances have excellent activities of inhibiting the expression of an inflammatory factor, have excellent antioxidant activities, and have excellent usability as a pharmaceutical composition and a food composition.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a pharmaceutical composition for antioxidative antiinflammatory activity,

The present invention relates to an antioxidant, anti-inflammatory activity pharmaceutical composition and a food composition containing an extract of Ainsliaea apiculate .

The antioxidant function inhibits or slows down the oxidation reaction occurring in the body. Oxidation reaction in the body induces aging and cell transformation to generate cancer cells. Therefore, the antioxidant activity is a preventive measure to suppress the negative reaction in the body, and the public is also interested.

On the other hand, in the body, mitochondria, which are main functions of ATP production, generate free radicals, which are byproducts of respiration, in addition to cell respiration, and they are unstable, And react indiscriminately. Therefore, if we can prevent the generation of these free radicals, we can prevent the oxidation of the body, and indirectly, we can prevent aging and cancer.

These antioxidant effects are difficult to experiment with free radicals occurring in the human body and use DPPH and ABTS radicals as biomarkers. The DPPH free radical scavenging activity assay is one of the methods for measuring the antioxidant activity of a sample. The DPPH free radical scavenging activity assay is a method for measuring the antioxidative activity of DPPH (1,1-diphenyl-2-picyl-hydrazyl) This method of decolorizing antioxidant activity is a widely used method because it can measure antioxidative activity in a relatively short time and has a high correlation with the antioxidant activity of plants.

Inflammation is one of the defenses of the body against physical stimuli, chemical stimuli, bacterial infections, etc. It is one of the mechanisms to remove external substances and regenerate damaged areas. Once an inflammation-related stimulus is applied, vasoactive substances such as histamine, serotonine, bradykinin, prostaglandin, hydroxyeicosatetraenoic acid (HETE), leukotriene And the vascular permeability is increased to cause inflammation.

Monocytes are transformed into macrophages that are activated by bacterial infection and act as phagocytic cells. Macrophages induce inflammation by secretion of NO, prostaglandin E2, and proinflammatory cytokines, and regulatory cells important in both inflammation and immune response In order for macrophages to function in this way, they must undergo an activation process.

LPS (lipopolysaccharide), one of the cell wall components of Gram-negative bacteria and well known as endotoxin, is the most well-known external factor involved in macrophage activation. Particularly, in monocytes and macrophages such as RAW 264.7 cells, proinflammatory cytokines such as TNF-α, IL-6 (interleukin-6) and IL-1β (interleukin- -inflammatory cytokine secreted by LPS (lipopolysaccharide) in the inflamed area.

When LPS stimulates macrophages, nitric oxide (NO) is produced in the process of converting L-arginine into L-citrulline by an enzyme called iNOS (inducible nitric oxide synthase) Phagocytes produce NO.

In mammals, NO is synthesized by three kinds of NO synthase (NOS): nNOS (neuronal NOS), eNOS (endothelial NOS) and iNOS (inducible NOS). Among these, NO produced by nNOS and eNOS is produced for normal biological function, and the concentration in tissues is kept low to a certain level. However, the amount of NO produced by iNOS is excessively high, and it is harmful to living organisms such as pathological vasodilation, cytotoxicity and tissue damage. In addition, prostaglandin E2 (PGE2), an inflammatory factor, stimulates phospholipid, which is a constituent of cell membrane, stimulated by LPS, and arachidonic acid liberated by an enzyme called phospholipase A2 causes catalytic action of an enzyme called COXs (cyclooxygenase) And is induced to induce an inflammatory reaction.

COX is classified as COX-1 and COX-2. COX-1 acts on normal biological functions such as platelet formation, gastric wall protection and maintenance of renal function in the body. COX-2 synthesizes PHE2 as an inflammatory mediator. PHE2 is known to be deeply involved in the development of cancer, such as promoting inflammation (pain, fever, etc.), immune response, and angiogenesis. Inhibition of iNOS is mostly due to its mechanism of action being linked to inhibitors of COXs. Therefore, iNOS inhibition materials that are relatively easy to search are discovered and developed as anti-inflammatory materials.

On the other hand, non-steroidal anti-inflammatory drugs (NSAIDs), including selective COX-2 inhibitors, have anti-inflammatory, antipyretic and analgesic effects. However, the long-term use of these drugs can sometimes lead to serious side effects. For example, secondary anemia due to gastrointestinal peptic ulcer bleeding, platelet function suppression, inhibition of induction of labor, side effects to the kidney, liver damage, hypersensitivity.

In order to overcome the side effects of these drugs, natural products such as plants are more preferable as a safer material. In this regard, researches are being actively carried out to find and use the anti-inflammatory materials in natural products.

Ainsliaea apiculate is a perennial plant of perennial herbaceous perennial herbaceous perennial plant, which is a plant in Korea and Japan (native plant of South Korea, 2005, Jeollanam-do). Studies on the anti-inflammatory and antioxidant effects of this plant Not reported.

It is an object of the present invention to provide a composition containing a plant extract having excellent antioxidative activity and anti-inflammatory activity at the same time, and the component extracted from Ainsliaea apiculate has excellent free radical scavenging activity and inhibitory effect on inflammatory factor expression And to provide a pharmaceutical composition and a food composition containing the same.

The antioxidative composition according to one embodiment of the present invention contains an extract of Ainsliaea apiculate .

The Corynebacterium extract may be one selected from the group consisting of alcohol having 1 to 5 carbon atoms, hexane, chloroform, alcohol, water, and a mixture thereof, and may be extracted by cold precipitation.

The extract of Corynebacterium sativus is obtained by extracting 1 to 5 times by adding a solvent having a dry weight of 3 to 20 times the weight of the dried powder to a dried bean curd and extracting it at an extraction temperature of 20 to 120 ° C for 30 minutes to 10 days An extract or a powder in which the solvent is removed from the extract.

The antioxidant composition is useful as a pharmaceutical or food composition having an antioxidant activity by containing a scent extract having antioxidative activity, which is confirmed to scavenge DPPH free radicals.

In order to achieve the above object, the anti-inflammatory composition according to another embodiment of the present invention contains an extract of Ainsliaea apiculate .

The Corynebacterium extract may be one selected from the group consisting of alcohol having 1 to 5 carbon atoms, hexane, chloroform, alcohol, water, and a mixture thereof, and may be extracted by cold precipitation.

The extract is obtained by extracting 1 to 5 times by adding a solvent having a dry weight of 3 to 20 times by weight to a dry powder of bean curd and extracting it at an extraction temperature of 20 to 120 ° C for 30 minutes to 10 days, And may be a powder from which the solvent has been removed from the extract.

The antiinflammatory composition is useful for the prevention or treatment of inflammatory diseases by containing a mash extract having anti-inflammatory activity which is confirmed to inhibit the production of NO induced by LPS in RAW 264.7 cells.

A food composition according to another embodiment of the present invention contains an extract of Ainsliaea apiculate . The food composition has anti-inflammatory activity and antioxidative activity and can be used as a food composition in the form of a health functional food, a nutritional supplement, a health food, or a food additive.

Hereinafter, the present invention will be described in more detail.

The antioxidant pharmaceutical composition according to one embodiment of the present invention contains an extract of Ainsliaea apiculate .

The scarlet extract may be obtained by extracting scarlet, drying and crushing and pulverizing it with a solvent. The dried and crushed scarlet may be a scarlet leaf, stem, root, flower, fruit, mixture or outpost thereof. Drying may be carried out in a known manner to such an extent that useful components of the scarlet picked up are not destroyed And can proceed, for example, by a method of natural drying in the shade. In addition, it is preferable that the crushing is performed after crushing to such an extent that the useful components of the crushed stalks can be sufficiently extracted in the subsequent extraction process, and pulverized. The drying and crushing steps may be carried out in reverse order or repeatedly as required.

The extraction may be carried out by, for example, hot water extraction, cold extraction, warming extraction, reflux cooling extraction, ultrasonic extraction, or the like, preferably by a cold extraction method.

The extraction is carried out using a solvent, which may be an alcohol of 1 to 5 carbon atoms, hexane, chloroform, alcohol, water and mixtures thereof, preferably water, methanol, ethanol, .

The extraction is carried out at a temperature of from 20 to 100 ° C, preferably from 20 to 70 ° C, for 30 minutes to 10 days, preferably from 3 to 20 times, preferably from 3 to 10 times, , Or a method of extracting for 1 to 2 days of extraction time. The method of the present invention can be applied to the production of the pharmaceutical composition, 5 times, preferably 2 times to 3 times, to obtain a liquid-phase extract.

The liquid morsel extract may be separated from a slightly scratched material by a method such as vacuum filtration, and then subjected to a concentration or drying process. For example, the liquid morsel extract may be concentrated in a vacuum rotary condenser at 20 to 100 ° C, preferably at 40 to 70 ° C, and the liquid morsel extract may be dried to obtain a powdered marshmallow Extracts may also be obtained. The concentrated or powdered morsel extract can be used as needed in water, alcohol or a mixed solvent thereof. In addition, the liquid morsel extract, the concentrate or the powder thereof may be an extract obtained by fractionation by nucleic acid or ethyl acetate or fractional stroke of the fraction.

In the body, mitochondria within the cell, which is the main function of ATP production, generate free radicals, a byproduct of respiration, which is unstable, and is accompanied by other substances such as lipids, nucleic acids and proteins, It reacts to the enemy. In this way, the oxidation reaction that occurs in the body induces aging and cell mutation, and it is necessary to prevent or suppress it. In other words, if we can prevent the generation of these free radicals, we can prevent the oxidation of the body and indirectly prevent aging and cancer. The antioxidant composition of the present invention may further include additional components. For example, the antioxidant composition may further include additional components such as antioxidants, antioxidants, The pharmaceutical composition may further comprise at least one of a carrier, an excipient and a diluent such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium May contain at least one or more selected from the group consisting of phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, have.

The antioxidant composition may have various formulations and may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, external preparations such as syrups and aerosols, and sterilized suppository solutions according to a conventional method . In the case of formulation, it can be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used, and the preparations for non-oral administration include sterilized aqueous solutions, Emulsions, freeze-dried preparations, suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include withexol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The desired dosage of the antioxidant composition may be appropriately selected depending on the condition and body weight of the patient, the degree of disease, the form of the drug, the route of administration and the period of time. However, for the desired antioxidative effect, the extract or the compound of the present invention may be administered in an amount of 0.01 to 500 mg / kg, preferably 0.1 to 200 mg / kg per day, once or several times per day. The dose of the compound may also be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like.

The anti-inflammatory pharmaceutical composition according to another embodiment of the present invention contains an extract of Ainsliaea apiculate . Among the descriptions of the antiinflammatory pharmaceutical composition, the description overlapping with the antioxidant pharmaceutical composition is omitted.

The extract of Corynebacterium sp. Can inhibit the expression of iNOS as an inflammatory mediator and inhibit the expression of NO produced by the enzyme. Thus, the extract can be used as a pharmaceutical composition for antioxidant activity containing a compound useful for the prevention and treatment of inflammation-related diseases Which can be preferably used as a pharmaceutical composition for the prevention or treatment of inflammatory diseases and provides a pharmaceutical composition for preventing or treating inflammation caused by infection from external bacteria.

The food composition according to another embodiment of the present invention contains an extract of Ainsliaea apiculate and has an anti-inflammatory and excellent antioxidative function and can be used as a health food, a nutritional supplement, a health food, Can be utilized as a composition.

Food compositions of this type can be prepared in a variety of forms according to conventional methods. For example, the health food may be prepared by liquefying, granulating, encapsulating, and pulverizing the above-mentioned Corynebacterium extract in the form of tea, juice, and drink so that it can be consumed. In addition, the extract may be mixed with a known scent extract and a known active ingredient known to have anti-inflammatory effects to prepare a composition. Functional foods also include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods such as canned fruits, jars, jam, maare marlade, fish, meat and processed foods (Eg butter, cheeses, etc.), edible vegetable oils (eg, sourdoughs, etc.), breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, , Margarine, vegetable protein, retort food, frozen food, various kinds of seasonings (for example, soybean paste, soy sauce, sauce, etc.). The preferable content of the above-mentioned Corynebacterium sp. Extract in the above food composition is not particularly limited, but is preferably 0.01 to 50% by weight in the finally prepared food.

The antioxidant composition, antiinflammatory composition and food composition of the present invention contain an extract of Ainsliaea apiculate and can be usefully used as a pharmaceutical composition or a functional food composition having an antioxidative activity and an inhibitory effect on the expression of an inflammatory factor.

FIG. 1 shows the results of evaluation of the amount of NO produced from the samples treated with the scarlet extract prepared in Example 1 of the present invention.
FIG. 2 shows the results of evaluating the antioxidant activity of the extract of Corynebacterium sativus, which is prepared in Example 1 of the present invention, using this DPPH free radical scavenging activity assay.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Example 1: Preparation of Chopped Extracts

The outposts of Ainsliaea apiculate , which is native to Korea, were sampled and dried for 14 days in shade and then crushed to a size of 20 mesh or less.

1 liter of ethanol was added to 100 g of the dried beaten egg powder, and the mixture was cooled at 25 ° C for 7 days. The mixture was extracted three times and filtered to obtain a filtered extract by filtration under reduced pressure (Wattmann, USA). The filtrate was subjected to removal of ethanol at 40 ° C using a vacuum rotary condenser to obtain 12-20 g of a slightly scooped extract residue. 10 mg of the sample was used as a sample for the detection of anti-inflammatory and antioxidant activity.

Example 2: Anti-inflammatory activity

The antiinflammatory effect of extracts of Horseradish was confirmed by measuring the amount of NO and using iNOS (inducible nitric oxide synthase) assay.

RAW 264.7 cells were inoculated into a 24-well plate at a density of 10 × 10 ⁴ cells / well using DMEM medium and cultured for 12 hours. The RAW 264.7 cells were inoculated with 20 μl of scarlet extract (AD) 20, 4, or 0.8 μg / ml LPS (1 [mu] g / ml) at the same time, and cultured for 24 hours. From the cultured samples, the amount of NO ^{2 -} present in the cell culture was measured using a Griess reagent, and the amount of NO produced and iNOS activity were measured. Specifically, 500 μl of the cell culture supernatant of the above cultured samples and 50 μl of a grease reagent (2.5% sulfanilamide, 0.25% naphthylethylenediamine dissolved in 10% phosphoric acid) were mixed and reacted on a 24-well plate for 10 minutes, The absorbance at 540 nm was measured using a reader. The standard concentration curve was obtained by serial dilution of DMSO, and the amount of produced NO and the activity of iNOS were evaluated. The results are shown in FIG. 1 and Table 1. In addition, the antiinflammatory activity was compared between the negative control and the positive control using aspirin in Comparative Examples 1 and 2. The aspirin used inhibited iNOS activity by 50% at a concentration of about 3.2 μg / mL , And 100 μg / mL, respectively.

division Concentration (μg / ml) iNOS activity (%) Positive control group
(Aspirin) 3.2 50 Negative control group
(Negative control) - 100 A bitch extract 20 45.25 4 87.10 0.8 94.94

Referring to FIG. 1, it was confirmed that the sacculifera extract effectively reduced NO production in RAW 264.7 c cells induced by LPS, and the degree of reduction was dose-dependent. NO is synthesized from L-arginine by nitric oxide synthase (NOS). Under pathologic conditions, increased NO promotes the development of inflammation, which is synthesized by iNOS, a synergistic agent with other inflammatory mediators. In addition, iNOS is strongly stimulated by exposure to inflammatory cytokines and bacterial endotoxins. Therefore, a substance capable of reducing NO production by iNOS can be utilized as an anti-inflammatory agent.

In addition, referring to Table 1 above, it was also confirmed that the safflower extract showed an inhibitory effect on iNOS in RAW 264.7 cells induced by LPS, which was concentration-dependent in relation to the safflower extract. NO produced by activated iNOS in inflammatory conditions has deleterious effects on the organism such as pathological vasodilation, cytotoxicity, and tissue damage. RAW 264.7 showed a strong inhibitory effect on the effect of LPS on NO and proinflammatory cytokine. The extracts showed a concentration-dependent reduction of LPS-induced NOS production and that the scarlet extract had a strong inhibitory effect on iNOS protein expression in RAW 264.7 cells induced by LPS .

Example 3 Evaluation of antioxidative activity of extracts

The antioxidant activity of the scarlet extract prepared in Example 1 was measured using a free Radical-scavenging activity test (DPPH). Twenty microliters of TBHQ (positive control), methanol (negative control), and methanol extract of chopped extract (2000, 1000, 200, and 125 μg / ml) were injected into each well of a 96-well microplate and incubated with 0.2 mM DPPH Sigma chemical Co. USA) methanol solution was added to the reaction mixture. The plate was wrapped with a foil to block the light, reacted at room temperature for 20 minutes, and the absorbance at 517 nm was measured. The results are shown in FIG.

Referring to FIG. 2, it was confirmed that the scarlet extract extracts scavenged DPPH free radicals in a concentration-dependent manner, and exhibited a remarkably superior antioxidative activity, indicating that the extract had an excellent antioxidative activity equivalent to that of quinone equivalent to that of the conventional antioxidative function.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, Of the right.

Claims (5)

An antioxidant pharmaceutical composition containing an extract of Ainsliaea apiculate . The method of claim 1, wherein
Wherein the extract is at least one selected from the group consisting of alcohols having 1 to 5 carbon atoms, hexane, chloroform, alcohol, water, and mixtures thereof. The method of claim 1, wherein
The extract is obtained by extracting 1 to 5 times by adding a solvent having a dry weight of 3 to 20 times by weight to a dry powder of bean curd and extracting it at an extraction temperature of 20 to 120 ° C for 30 minutes to 10 days, Wherein the solvent is removed from the extract. An antiinflammatory pharmaceutical composition containing an extract of Ainsliaea apiculate . A food composition comprising an extract of Ainsliaea apiculate and having antioxidant activity and anti-inflammatory function.

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