KR20200052628A - Composition for preventing or treating inflammatory diseases or allergic diseases comprising supercritical ethanol extract of skimmed perilla frutescens powder - Google Patents

Abstract

The present invention relates to a composition for preventing or treating inflammatory diseases or allergic diseases containing a supercritical skimmed Perilla frutescens powder ethanol extract as an active component. According to the present invention, the supercritical skimmed Perilla frutescens powder (Perilla frutescens peel) ethanol extract inhibits the production of nitrogen oxide and the release of hexosaminidase, TNF-α, and IL-4 to exhibit anti-inflammatory and anti-allergic effects, thereby being able to be usefully utilized as a pharmaceutical composition for preventing or treating inflammatory diseases or allergic diseases, a health food composition for preventing or alleviating inflammatory diseases or allergic diseases, a cosmetic composition for preventing or alleviating inflammatory diseases or allergic diseases, etc.

Description

Composition for preventing or treating inflammatory diseases or allergic diseases comprising supercritical ethanol extract of skimmed perilla frutescens powder}

The present invention relates to a composition for the prevention or treatment of an inflammatory disease or an allergic disease, and more specifically, an inflammatory disease containing an extract obtained by extracting supercritical defatted perilla powder, ie supercritical perilla leaf with ethanol, as an active ingredient. Or it relates to a composition for the prevention or treatment of allergic diseases.

Inflammation is the defense response of a living tissue to an injury that occurs in the body. That is, in response to a variety of harmful stimuli (stressor), to remove the injury caused by the stimulus and the biological defense reaction to restore the original state is an inflammatory reaction. Stimulation of inflammation includes infection, chemical or physical stimulation, and bioconstituent factors related to inflammatory reactions include low-molecular or high-molecular chemicals such as free radicals, proteins, sugars, and lipids, plasma, blood cells, blood vessels, and Connective tissue. The process of inflammation is usually divided into two types of acute and chronic inflammation. Acute inflammation is a short-term reaction within a few days. Plasma components or blood cells show microcirculation to remove foreign substances, while chronic inflammation is a long-term reaction. Causes proliferation and the like.

Allergic disease refers to a pathological phenomenon caused by an antigen-antibody reaction, and serum disease, hyperthermia, and bronchial asthma are typical. Nowadays, however, there is a tendency to see it as a disease that can be caused by allergies, so the antigen-antibody response is very similar to the proven disease, but the antigen-antibody response is not proved, or the involvement of the antigen-antibody response is not proved, but the possibility This includes high things. Allergic diseases by organ, for example, bronchial asthma, hyperthermia, vasomotor rhinitis in the respiratory system, hypertrophic rhinitis, allergic bronchitis, Röppler syndrome (transient lung infiltration), etc. Diarrhea, allergic stomatitis, enteric purpura, etc., circulatory system, nodular arterial peritonitis, obstructive arteritis, angina, endocarditis, etc. Ten, sympathetic ophthalmitis, allergic conjunctivitis, and allergic keratitis. Others include diffuse glomerulonephritis and migraine headaches. In addition, allergic diseases are also classified into immediate type (anaphylaxis type) and delayed type (tuberculin type). Immediate forms include hyperthermia, bronchial asthma, hives, quinque edema, digestive tract allergies, and delayed forms include contact dermatitis or drug allergies.

Meanwhile, macrophages play an important role in specific and non-specific immune responses to microbial and viral infections. Macrophages activated by external stimulation include various inflammatory mediators such as nitric oxide (NO), prostaglandin E2 (PGE2), pro-inflammatory cytokines including TNF-α, IL-1 and IL-6. It produces and releases substances.

Mast cells (mast cells) are derived from bone marrow cells, mature and differentiated in tissues, and then activated by antigen-antibodies and various cytokines to participate in inflammation and allergic reactions. Activated mast cells are known to increase the allergic reaction by biosynthesizing eicosanoids such as histamine, lipid mediators prostaglandines and leukotriens, which have already been made and stored in granules.

Prostaglandins, leukotrienes, and platelet activating factors (PAFs), which are key mediators for inducing inflammatory and allergic diseases, include phospholipase A2, cyclooxygenase, and lipoxygenase. It is thereby produced from the precursor arachidonic acid.

Drug therapies currently used to treat allergic diseases include antihistamines, corticosteroids such as cortisone, or epinephrine or ephedrine, and parasympathetic neuroparalysis agents such as atropine sulfate or Lotex. The development of more fundamental therapeutic drugs is urgently needed.

Republic of Korea Patent Publication No. 10-2015-0040423 (published on April 15, 2015)

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory or allergic diseases.

Another object of the present invention is to provide a health food composition for preventing or improving inflammatory or allergic diseases.

Another object of the present invention is to provide a cosmetic composition for preventing or improving inflammatory or allergic diseases.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases or allergic diseases containing supercritical defatted perilla powder ethanol extract as an active ingredient.

In addition, the present invention provides a health food composition for the prevention or improvement of inflammatory diseases or allergic diseases containing the supercritical defatted perilla powder ethanol extract as an active ingredient.

In addition, the present invention provides a cosmetic composition for the prevention or improvement of inflammatory diseases or allergic diseases containing supercritical defatted perilla powder ethanol extract as an active ingredient.

In addition, the first step of crushing perilla; A second step of supercritical extraction of the crushed perilla at 25 to 35 MPa and a temperature of 30 to 50 ° C. to obtain a defatted perilla powder; A third step of adding ethanol aqueous solution to the obtained defatted perilla powder and immersing in the first step; And after the third step, after recovering the ethanol aqueous solution, a fourth step of secondary immersion by adding a new ethanol aqueous solution; provides a method for producing supercritical defatted perilla powder ethanol extract.

According to the present invention, the supercritical defatted perilla powder (perilla) ethanol extract exhibits anti-inflammatory and anti-allergic effects by inhibiting the production of nitric oxide and the release of β-hexosaminidase, TNF-α, IL-4, It may be useful as a pharmaceutical composition for preventing or treating inflammatory diseases or allergic diseases, a health food composition for preventing or improving inflammatory diseases or allergic diseases, a cosmetic composition for preventing or improving inflammatory diseases or allergic diseases, and the like.

Figure 1 confirms the cytotoxicity of perilla extract (pressed perilla extract hot water extract, compressed perilla extract ethanol extract, supercritical perilla extract hot water extract, supercritical perilla extract ethanol extract) in LPS-treated RAW264.7 macrophages.
Figure 2 confirms the inhibitory effect of nitric oxide production of the perilla extract in RAW264.7 macrophages sensitized with LPS.
Figure 3 confirms the inhibitory effect of TNF-α release of the perilla extract in RAW264.7 macrophages sensitized with LPS.
Figure 4 confirms the inhibitory effect of IL-4 release of the perilla extract in RAW264.7 macrophages sensitized with LPS.
Figure 5 confirms the cytotoxicity of the perilla extract from RBL-2H3 mast cells treated with anti-DNP-IgE.
Figure 6 confirms the inhibitory effect of β-hexosaminidase release of the perilla extract on RBL-2H3 mast cells treated with anti-DNP-IgE.

The inventors of the present invention obtained a defatted perilla powder (perilla perilla) from perilla using a supercritical extraction method, and added ethanol to the perilla perilla extract to prepare perilla perilla ethanol extract, wherein the perilla perilla ethanol extract produces nitrogen oxide And by inhibiting the release of β-hexosaminidase, TNF-α, IL-4 confirming that it exhibits anti-inflammatory and anti-allergic effects, the present invention was completed.

Accordingly, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammatory diseases or allergic diseases containing supercritical defatted perilla powder ethanol extract as an active ingredient.

The defatted perilla powder means defatted perilla powder from which the oil component is removed from the perilla, and is also referred to as perilla perilla, perilla jelly, and imjabak.

The extract may be extracted by adding ethanol aqueous solution to the defatted perilla powder obtained by supercritical extraction of crushed perilla and immersing.

The extract inhibits the production of nitric oxide and the release of β-hexosaminidase, TNF-α, IL-4, and may exhibit anti-inflammatory and anti-allergic effects.

The extract may be contained in an amount of 0.01 to 90 parts by weight based on 100 parts by weight of the composition, but is not limited thereto.

The inflammatory disease may be selected from the group consisting of dermatitis, periodontitis, rhinitis, acute and chronic inflammatory disease, but is not limited thereto.

The allergic disease may be selected from the group consisting of hypersensitivity, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis and urticaria, but is not limited thereto.

When the composition of the present invention is a pharmaceutical composition, for administration, it may include a pharmaceutically acceptable carrier, excipient, or diluent in addition to the active ingredients described above. The carrier, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition of the present invention can be formulated and used in the form of an oral dosage form such as a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, external preparation, suppository, or sterile injectable solution, respectively, according to a conventional method. . In detail, when formulated, it may be prepared using diluents or excipients such as fillers, weights, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used. Solid preparations for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like. Such a solid preparation may be prepared by mixing at least one excipient other than the active ingredient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, and the like. In addition, lubricants such as magnesium stearate and talc may be used in addition to simple excipients. In addition to liquids for oral administration and liquid paraffin, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, can be added. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and challenges. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrosol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

The suitable dosage of the pharmaceutical composition of the present invention depends on the patient's condition and body weight, the degree of disease, the drug form, and the time, but may be appropriately selected by a person skilled in the art, and the daily dosage of the composition is preferably It is 0.001 mg / kg to 50 mg / kg, and can be divided and administered once to several times per day as needed.

In addition, the present invention provides a health food composition for the prevention or improvement of inflammatory diseases or allergic diseases containing supercritical defatted perilla powder ethanol extract as an active ingredient.

When the composition of the present invention is a health food composition, various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof , Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonic acid used in carbonated beverages, and the like. In addition, it may contain flesh for the manufacture of natural fruit juices, synthetic fruit juices and vegetable drinks. These ingredients can be used independently or in combination. In addition, the health food composition may be in the form of any one of meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, drink, alcohol and vitamin complex Can be.

In addition, the health food composition may further include a food additive, and whether or not it is suitable as a food additive is related to the item according to the general rules and general test methods of the Food Additives Code approved by the Ministry of Food and Drug Safety unless otherwise specified. Judging by standards and standards.

Chemical additives such as ketones, glycine, potassium citrate, nicotinic acid, cinnamon acid, natural additives such as chromochromate, licorice extract, crystalline cellulose, high cooling pigment, guar gum, and L-glutamic acid. And mixed preparations such as sodium preparations, alkali addition agents for noodles, preservative preparations, and tar color preparations.

At this time, the composition according to the present invention is added to the food in the process of manufacturing a health food composition can appropriately adjust the content as necessary.

In addition, the present invention provides a cosmetic composition for the prevention or improvement of inflammatory diseases or allergic diseases containing supercritical defatted perilla powder ethanol extract as an active ingredient.

When the composition of the present invention is a cosmetic composition, the cosmetic composition may include, in addition to the active ingredient, a conventional adjuvant such as a stabilizer, a solubilizing agent, vitamins, pigments and fragrances, and a carrier. In addition, the cosmetic composition may further include a skin absorption accelerator to enhance its effect.

The formulation of the cosmetic composition may be prepared in any formulation conventionally prepared in the art, for example, the cosmetic composition is a lotion, emulsion, skin, toner, lotion, essence, sunscreen, cream, makeup base, foundation , Powder, pack, gel, shampoo, rinse, spray, makeup remover and detergent, etc., may be formulated, but is not limited thereto.

When the formulation is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. .

When the formulation is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in the case of a spray, additionally chlorofluorohydrocarbon, propane / butane Or propellants such as dimethyl ether.

When the formulation is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3 -Fatty acid esters of butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan.

When the formulation is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline cellulose , Aluminum metahydroxide, bentonite, agar or trakant, and the like.

In addition, the present invention is a first step of crushing perilla; A second step of supercritical extraction of the crushed perilla at 25 to 35 MPa and a temperature of 30 to 50 ° C. to obtain a defatted perilla powder; A third step of adding ethanol aqueous solution to the obtained defatted perilla powder and immersing in the first step; And after the third step, after recovering the ethanol aqueous solution, a fourth step of secondary immersion by adding a new ethanol aqueous solution; provides a method for producing supercritical defatted perilla powder ethanol extract.

The third and fourth steps may be performed for 20 to 30 hours at a temperature condition of 20 to 30 ° C, but it is not limited thereto.

The third and fourth ethanol aqueous solutions may be 50 to 90% ethanol aqueous solutions, but are not limited thereto.

Hereinafter, the present invention will be described in more detail through examples. These examples are only intended to illustrate the present invention more specifically, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example 1: Preparation of supercritical degreasing perilla powder (perilla)

75 kg of crushed perilla was supercritical extracted for 3 hours at a temperature condition of 30 to 31 MPa, 40 ° C. to obtain perilla perilla.

Example 2: Preparation of perilla extract ethanol extract and perilla extract hot water

To 10 g of perilla extract obtained in Example 1, 70% ethanol was added 10 times to perform primary immersion (at room temperature, 24 hours), and after recovery of the extraction solvent, 70% ethanol was added 10 times to perform secondary immersion (at room temperature, 24). Time), followed by filtration and freeze-drying to prepare supercritical perilla ethanol extract.

In addition, the perilla extract obtained after extracting perilla oil by a screw pressure method was prepared using the same method as the above, and the perilla perilla ethanol extract was prepared, and water was added to 10 g of perilla extract obtained by a supercritical extraction and pressing method. Was added 10 times and extracted at 50 ° C. for 3 hours, followed by filtration and freeze-drying to prepare a supercritical perilla hot water extract and compressed perilla hot water extract.

Example 3: Analysis of anti-inflammatory effect of perilla extract

3-1. Cell culture

RAW264.7 macrophages were distributed from the Korean Collection for Type Culture (KCTC, Korea), DMEM medium (Welgene) with 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin added , Korea) was used after subcultured 2-3 times in a 37 ° C, 5% CO ₂ incubator.

3-2. Cytotoxicity analysis of perilla extract

To confirm the cytotoxicity of perilla extract from RAW264.7 macrophages sensitized with LPS (lipopolysaccharide), MTT analysis was performed. The method is based on the conversion of MTT to purple formazan crystals by reductase in mitochondria in living cells.

RAW264.7 macrophages were dispensed into a 96 well plate at a concentration of 3 X 10 ⁴ cells / well and incubated overnight, then extracts of various concentrations were treated in the presence of LPS (final concentration, 500 ng / mL) and incubated for 48 hours. . Then, 10 μL of MTT (5 mg / mL) was added to each well, and further incubated at 37 ° C. for 4 hours, and then 100 μL of DMSO was added to each well to lyse cells. Thereafter, the plate was kept at room temperature for 5 minutes, and absorbance was measured at 550 nm using a microplate reader (multiwell spectrophotometer, Molecular Devices, Sunnyvale, CA).

As a result, referring to FIG. 1, it was confirmed that neither compressed perilla hot water / ethanol extract nor supercritical perilla hot water / ethanol extract exhibited toxicity to macrophages.

3-3. Analysis of nitric oxide inhibitory effect by perilla extract

In order to analyze the anti-inflammatory effect of perilla extract, the production of nitric oxide (NO) in RAW264.7 macrophages sensitized with LPS was measured.

RAW 264.7 macrophages were dispensed in a 96 well plate at a concentration of 1 X 10 ⁵ cells / well and incubated overnight, then extracts of various concentrations were treated in the presence of LPS (final concentration, 500 ng / mL) and incubated for 24 hours. Thereafter, the NO product in the culture medium was measured with a Griess Reagent System. Griess reagent [2.5% phosphoric acid in which 1% sulfanilamide and 0.1% naphthylethylendiamine are dissolved] was added to 100 μL of the medium supernatant in the same amount, mixed for 10 minutes, and then 540. Absorbance was measured at nm and NO concentration was calculated from the NaNO ₂ standard curve.

As a result, referring to FIG. 2, in the case of supercritical perilla extract ethanol extract, concentration-dependent NO production was suppressed from a concentration of 100 μg / mL, whereas in supercritical perilla extract hot water extract and compressed perilla extract hot water extract, NO Production was not inhibited. In the case of compressed perilla ethanol extract, NO production was not inhibited at low concentrations, but NO production was suppressed at concentrations above 500 μg / mL. In the positive control (L-NAME), NO production was suppressed by about 20%.

3-4. Analysis of the inhibitory effect of TNF-α release by perilla extract

In order to analyze the anti-inflammatory effect of perilla extract, the release level of TNF-α was measured in RAW264.7 macrophages sensitized with LPS.

RAW 264.7 macrophages were dispensed in a 96 well plate at a concentration of 1 X 10 ⁵ cells / well and incubated overnight, then extracts of various concentrations were treated in the presence of LPS (final concentration, 500 ng / mL) and incubated for 24 hours. Thereafter, the culture medium was centrifuged to remove impurities, and TNF-α release level was measured using a TNF-α ELISA kit. Briefly, 50 μL of the supernatant of the medium was added to an ELISA plate attached with anti-TNF-α and reacted for 2 hours at room temperature, followed by washing 5 times using a washing buffer. Then, 100 μL of a TNF-α conjugate solution was added to each well, reacted at room temperature for 2 hours, and then washed 5 times using a washing buffer. Thereafter, 100 µL of a substrate solution was added to each well and reacted at room temperature for 30 minutes, and then 100 µL of a stop solution was added to each well to terminate the reaction. Thereafter, the absorbance at 450 nm was measured using an ELISA reader, and a quantitative curve was prepared with a standard solution of TNF-α to calculate the amount of TNF-α released.

As a result, referring to FIG. 3, it was confirmed that the supercritical perilla ethanol extract and the compressed perilla ethanol extract both inhibit the release of TNF-α in a concentration-dependent manner from a concentration of 50 μg / mL. In the positive control (L-NAME), TNF-α release was suppressed by about 35%.

3-5. Analysis of IL-4 release inhibitory effect by perilla extract

In order to analyze the anti-inflammatory effect of perilla extract, the degree of release of IL-4 in RAW264.7 macrophages sensitized with LPS was measured.

RAW 264.7 macrophages were dispensed in a 96 well plate at a concentration of 1 X 10 ⁵ cells / well and incubated overnight, then extracts of various concentrations were treated in the presence of LPS (final concentration, 500 ng / mL) and incubated for 24 hours. Thereafter, the culture medium was centrifuged to remove impurities, and the release degree of IL-4 was measured using an IL-4 ELISA kit. Briefly, 50 μL of the supernatant of the medium was added to the ELISA plate to which anti-IL-4 was attached and reacted at room temperature for 2 hours, followed by washing 5 times using a washing buffer. Thereafter, 100 μL of the IL-4 conjugate solution was added to each well, reacted at room temperature for 2 hours, and then washed 5 times using a washing buffer. Thereafter, 100 µL of a substrate solution was added to each well and reacted at room temperature for 30 minutes, and then 100 µL of a stop solution was added to each well to terminate the reaction. Thereafter, the absorbance at 450 nm was measured using an ELISA reader, and a quantitative curve was prepared with an IL-4 standard solution to calculate the amount of IL-4 released.

As a result, referring to FIG. 4, in the case of supercritical perilla extract ethanol, the concentration-dependent release of IL-4 was inhibited from a concentration of 50 μg / mL, whereas in the case of compressed perilla extract ethanol, IL at low concentration Although -4 release was not inhibited, IL-4 release was inhibited at a concentration of 100 μg / mL or more. In addition, it was confirmed that the supercritical perilla ethanol extract at the same concentration more effectively inhibits the release of IL-4 than the compressed perilla ethanol extract. In the positive control (L-NAME), IL-4 release was suppressed by about 30%.

Example 4: Analysis of anti-allergic effect of perilla extract

4-1. Cell culture

RBL-2H3 mast cells were pre-sold at the Korean Resource Center for Collection (KCTC, Korea), using 10% fetal bovine serum and 1% penicillin / streptomycin-added MEM medium (Welgene, Korea). It was used after culturing 2-3 times in a 5% CO ₂ incubator.

4-2. Cytotoxicity analysis of perilla extract

To confirm the cytotoxicity of perilla extract from RBL-2H3 mast cells sensitized with anti-DNP-IgE, MTT analysis was performed.

RBL-2H3 mast cells were dispensed into a 96 well plate at a concentration of 3 X 10 ⁴ cells / well, treated with anti-DNP IgE (100 ng / mL) overnight, and incubated at 37 ° C overnight, followed by treatment with extracts of various concentrations. Cultured for 48 hours. Then, 10 μL of MTT (5 mg / mL) was added to each well, and further incubated at 37 ° C. for 4 hours, and then 100 μL of DMSO was added to each well to lyse cells. Thereafter, the plate was kept at room temperature for 5 minutes, and absorbance was measured at 550 nm using a microplate reader (multiwell spectrophotometer, Molecular Devices, Sunnyvale, CA).

As a result, referring to FIG. 5, it was confirmed that the compressed perilla perilla hot water / ethanol extract and the supercritical perilla perilla hot water / ethanol extract did not show any toxicity to mast cells.

4-3. Analysis of the inhibitory effect of perilla extract on β-hexosaminidase release

To analyze the anti-allergic effect of perilla extract, the degree of β-hexosaminidase release from RBL-2H3 mast cells sensitized with anti-DNP-IgE was measured.

RBL-2H3 mast cells were distributed in a 24 well plate at a concentration of 2 X 10 ⁵ cells / well, and treated with anti-DNP IgE (100 ng / mL) to incubate overnight at 37 ° C. Thereafter, the cells were washed with Siraganian buffer, and 160 μL of incubation buffer was added, followed by incubation at 37 ° C. for 20 minutes, followed by treatment of 20 μL of various concentrations of perilla extract and reacted for 10 minutes. Then, 20 μL of the antigen (DNP-HSA, 50 ng / mL) was added and reacted at 37 ° C. for 10 minutes to stimulate the cells to form granules, followed by stopping the soaking reaction in an ice bath for 10 minutes. Recover 25 μL of the supernatant, transfer it to a 96 well plate, add 25 μL of 0.1 M citrate buffer (pH 4.5) in which the substrate (1 mM p-nitrophenyl-Nacetyl-β-D-glucosaminide) is dissolved, and add 37 The reaction was carried out for 1 hour at ℃. Thereafter, 200 μL of a stop solution (0.1 M Na ₂ CO ₃ / NaHCO ₃ , pH 10.0) was added to stop the reaction, and absorbance was measured at 405 nm using a microplate reader.

As a result, referring to FIG. 6, the release of β-hexosaminidase was not observed in the negative control group not sensitized to anti-DNP-IgE, and β-hexosami in the control group sensitized to anti-DNP-IgE. The release of Nidaase was observed. In the case of supercritical perilla ethanol extract and extruded perilla ethanol extract, the concentration-dependent release of β-hexosaminidase was suppressed from a concentration of 100 μg / mL, whereas in compressed / supercritical perilla hot water extract β-hex It did not inhibit the release of sosaminidase. In addition, it was confirmed that the supercritical perilla extract ethanol extract more effectively suppressed the release of β-hexosaminidase than the compressed perilla extract ethanol extract, thereby exhibiting better anti-allergic efficacy. The positive control (wortmannin, wort) inhibited β-hexosaminidase release by about 65%.

The specific parts of the present invention have been described in detail above, and it is obvious that for those skilled in the art, these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted to be included in the scope of the present invention.

Claims (11)

A pharmaceutical composition for the prevention or treatment of inflammatory diseases or allergic diseases containing supercritical defatted perilla powder ethanol extract as an active ingredient. The pharmaceutical composition for preventing or treating inflammatory diseases or allergic diseases according to claim 1, wherein the extract is extracted by adding and dipping an ethanol aqueous solution to the defatted perilla powder obtained by supercritical extraction of crushed perilla. The pharmaceutical composition for preventing or treating inflammatory diseases or allergic diseases according to claim 1, wherein the extract inhibits the production of nitric oxide and the release of β-hexosaminidase, TNF-α, IL-4. The pharmaceutical composition for preventing or treating inflammatory diseases or allergic diseases according to claim 1, wherein the extract is contained in an amount of 0.01 to 90 parts by weight based on 100 parts by weight of the total composition. The pharmaceutical composition for preventing or treating inflammatory diseases or allergic diseases according to claim 1, wherein the inflammatory diseases are selected from the group consisting of dermatitis, periodontitis, rhinitis, acute and chronic inflammatory diseases. The inflammatory disease or allergic disease according to claim 1, wherein the allergic disease is selected from the group consisting of hypersensitivity, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis and urticaria. Pharmaceutical composition for the prevention or treatment of. A health food composition for preventing or improving inflammatory diseases or allergic diseases containing supercritical defatted perilla powder ethanol extract as an active ingredient. A cosmetic composition for preventing or improving an inflammatory disease or allergic disease, which contains supercritical defatted perilla powder ethanol extract as an active ingredient. A first step of crushing perilla;
A second step of supercritical extraction of the crushed perilla at 25 to 35 MPa and a temperature of 30 to 50 ° C. to obtain defatted perilla powder;
A third step of adding ethanol aqueous solution to the obtained defatted perilla powder and primary dipping; And
After the third step, after recovering the ethanol aqueous solution, a fourth step of secondary immersion by adding a new ethanol aqueous solution;
Method for producing supercritical defatted perilla powder ethanol extract comprising a. 10. The method of claim 9, wherein the third step and the fourth step are performed for 20 to 30 hours at a temperature condition of 20 to 30 ° C. 10. The method of claim 9, wherein the third and fourth ethanol aqueous solutions are 50 to 90% ethanol aqueous solutions.

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