KR102151860B1 - Composition for Improving Pulmonary Fibrosis Using an Extract of Astilbe rubra - Google Patents

Abstract

The present invention suppresses EMT in a concentration-dependent manner, inhibits the expression of E-cadherin and Vimentin, which are involved in EMT activation, without cytotoxicity to the lung cancer cell line A549, and inhibits the expression of the EMT-related transcription factor Snail. Disclosed is a composition for improving pulmonary sclerosis using an extract of Noru urine, which inhibits the production of collagen, which is a fibrous substance, and suppresses oxidative stress applied to A549 cells.

Description

Composition for Improving Pulmonary Fibrosis Using an Extract of Astilbe rubra}

The present invention relates to a composition for improving pulmonary sclerosis using a noru urine ( Astilbe rubra ) extract.

Pulmonary fibrosis or idiopathic pulmonary fibrosis (IPF) is an unexplained disease characterized by fibrosis of the pulmonary interstitial in which excessive adherence of connective tissue, especially collagen, to the alveolar wall. Pulmonary fibrosis is a fatal disease that progresses to respiratory failure due to a slow progression of lung function, and the average survival period after diagnosis is within 2-3 years. The prevalence of pulmonary fibrosis varies from report to report, but it is known to be 2-29 out of 100,000 population, and it is designated as a rare and intractable disease in Korea.

In addition, radiation therapy has been used for various chest cancers such as breast cancer and lung cancer, and about 70-80% of all patients receiving radiation therapy for lung organs have mild or severe lung organ side effects. One of its representative side effects is pulmonary fibrosis, which causes loss of lung function, reduces lung capacity of the patient, negatively affects the quality of life, and in severe cases leads to death.

The causes of pulmonary fibrosis include lung damage, exposure to toxic substances or toxic environments such as fine or ultrafine dust, anticancer drugs, autoimmune diseases, or idiopathic interstitial pneumonia (Proc Am Thorac Soc). 2006, 3:285-92; Am J Respir Crit Care Med 2002, 165:277-304). The basic histological changes of pulmonary fibrosis are inflammatory changes such as cell infiltration, swelling, and effusion in the alveolar septum, and extracellular matrix such as collagen, proteoglycan, fibronectin, and glycoprotein. It is the destruction of the lung parenchyma due to infiltration and fibrosis (Am J Respir Crit Care Med 2002, 165:277-304; N Engl J Med 2001, 345:517-25).

In the past, the etiology of pulmonary fibrosis was regarded as inflammation, and anti-inflammatory treatment was attempted, but the effect was insignificant (Annals of Internal Medicine 2001, 134:136-51; Chest 1996, 110:1058-67), so recently inflammation Epithelial-mesenchymal transition (EMT) is being treated as a new etiology and treatment target (Annals of Internal Medicine 2001, 134:136-51; Chest 2007, 132:1311-21; The Journal of Clinical Investigation 2009, 119:213-24).

In particular, E-cadherin is reported to be an important molecule of EMT along with Vimentin, as it is known to promote EMT and cause pulmonary fibrosis by being converted to N-cadherin at the beginning of EMT, which is associated with the TGF-β pathway. Snail1, Snail2 (Slug) and Snail3 are known as EMT-related transcription factors in the nucleus (The Journal of Clinical Investigation 2009, 119:213-24; Nat Rev Mol Cell Biol 2006, 7:131-42; Cell Biol Int 2002, 26 :463-76).

In recent years, studies have been reported that oxidative stress causes pulmonary fibrosis (Am J Respir Crit Care Med. 2005, 172(4):417-22; Biochim Biophys Acta. 1832(7):1028-40) ), there is active research to apply antioxidants to the treatment of pulmonary fibrosis. For example, Korean Patent Publication No. 2006-0030786, which discloses curcumin derivatives having antioxidant activity, can be used in the treatment of pulmonary fibrosis, or WO 2008/098196, which discloses treprostinil.

The present invention discloses an activity of improving pulmonary fibrosis of a noru urine extract having EMT inhibitory activity and antioxidant activity.

It is an object of the present invention to provide a composition for improving pulmonary fibrosis using an extract of Noru urine having EMT inhibitory activity, antioxidant activity, and the like.

Other or specific objects of the present invention will be presented below.

As confirmed in the Examples and Experimental Examples below, the present inventors found that the extract of Nolu urine was not cytotoxic to A549, a lung cancer cell line, but inhibited EMT in a concentration-dependent manner and expressed E-cadherin and Vimentin involved in EMT activation. It was confirmed that it suppressed the expression of EMT-related transcription factor Snail, etc., as well as inhibiting the production of collagen, a lung fibrosis material, and suppressing oxidative stress applied to A549 cells.

The present invention is provided on the basis of these experimental results, and the present invention can be understood as a composition for improving pulmonary fibrosis comprising a noru urine extract as an active ingredient in one aspect, and for inhibiting metastasis of lung cancer cells in the other aspect. It can be recognized by the composition.

In the present specification, "Noru urine extract" refers to a stem, branch, leaf, root, outpost, or a mixture thereof, which is an extraction target, water, lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), methylene chloride, Ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), methyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof It refers to an extract obtained by leaching by using (ie, an extract soluble in the extraction solvent), an extract obtained using a supercritical extraction solvent such as carbon dioxide, pentane, or a fraction obtained by fractionating the extract, and the extraction method includes the polarity of the active substance, In consideration of the degree of extraction and preservation, arbitrary methods such as cold sedimentation, reflux, warming, ultrasonic radiation, and supercritical extraction can be applied. In the case of a fractionated extract, a fraction obtained by suspending the extract in a specific solvent and then mixing and policing it with a solvent having a different polarity, the crude extract adsorbed on a column filled with a polar or non-polar solid phase such as silica gel, and then a hydrophobic solvent, a hydrophilic solvent, or It is meant to include a fraction obtained by using these mixed solvents as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or a solid extract from which the extraction solvent has been removed by a method such as freeze drying, vacuum drying, hot air drying, or spray drying. Preferably, it means an extract obtained by using water, ethanol or a mixed solvent thereof as an extraction solvent, more preferably an extract obtained by using a mixed solvent of water and ethanol as an extraction solvent.

In addition, in the present specification, "inhibition of metastasis of lung cancer cells" means inhibiting metastasis and invasion of lung cancer cells to other organs such as the other stomach and liver.

In addition, in the present specification, the term "active ingredient" refers to a component capable of exhibiting a desired activity alone, or capable of exhibiting activity together with a carrier that is not itself active.

In the composition of the present invention, the active ingredient may be included in an arbitrary amount (effective amount) depending on the use, formulation, etc., as long as it can exhibit the effect of improving pulmonary fibrosis and inhibiting metastasis of lung cancer cells, and the usual effective amount is the entire composition. It will be determined in the range of 0.001% to 15% by weight on a weight basis. Herein, "effective amount" refers to the intended medical and pharmacological effects, such as the effect of improving pulmonary fibrosis, when the composition of the present invention is administered to a mammal, preferably a person, to which the composition of the present invention is administered during the administration period according to the recommendations of a medical expert, etc. It refers to the amount of the active ingredient contained in the composition of the present invention. Such effective amounts can be determined empirically within the range of ordinary skill in the art.

The composition of the present invention can be grasped as a food composition in a specific aspect.

The food composition of the present invention may be prepared in any form, such as beverages such as tea, juice, carbonated beverages, ionized beverages, processed oils such as milk and yogurt, gums, rice cakes, Korean confectionery, bread, Foods such as confectionery and noodles, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jelly, and health functional food preparations such as bars can be prepared. In addition, the food composition of the present invention can be classified as a product as long as it conforms to the enforcement regulations at the time of manufacture and distribution in terms of legal and functional classification. For example, it is a health functional food in accordance with the Korean "Health Functional Food Act", or confectionery, bean, tea, etc. according to each food type according to the food code of the Korean "Food Sanitation Act" (“Food Standards and Standards” notified by the Ministry of Food and Drug Safety). It may be beverages, special purpose foods, etc.

The food composition of the present invention may contain food additives in addition to the active ingredients. Food additives can generally be understood as substances that are added to food and mixed or infiltrated in manufacturing, processing, or preserving food. Since they are consumed daily and for a long time with food, their safety must be ensured. In the Food Additive Code (“Food Sanitation Act” in Korea) governing the manufacture and distribution of food, food additives with guaranteed safety are limited in terms of ingredients or functions. In the Korean Food Additives Code (KFDA notice "Food Additive Standards and Specifications), food additives are classified into chemical synthetic products, natural additives, and mixed preparations in terms of ingredients. These food additives are sweeteners and flavoring agents in terms of function. , Preservatives, emulsifiers, acidulants, thickeners, etc.

Sweeteners are used to impart a suitable sweetness to foods, and both natural and synthetic ones can be used in the food composition of the present invention. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents are used to improve taste or flavor, and both natural and synthetic flavors can be used. Preferably, it is the case of using a natural one. In the case of using natural ingredients, the purpose of nutrient enhancement can be combined in addition to flavor. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or may be obtained from green tea leaves, roundtails, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, you can use those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo. The natural flavoring agent may be a liquid concentrate or a solid extract. In some cases, synthetic flavoring agents may be used. As the synthetic flavoring agents, esters, alcohols, aldehydes, terpenes, and the like may be used.

As a preservative, calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, pectin Etc. may be used, and as the acidulant, arithmetic, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like can be used. The acidulant may be added so that the food composition has an appropriate acidity for the purpose of suppressing the growth of microorganisms in addition to the purpose of enhancing taste. As the thickening agent, a suspending agent, a settling agent, a gel-forming agent, a swelling agent, and the like may be used.

In addition to the food additives described above, the food composition of the present invention may contain a physiologically active substance or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutritional properties and ensuring stability as a food additive.

Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoyl thiamine, etc., and minerals include calcium preparations such as calcium citrate, magnesium stearate. Magnesium preparations, such as magnesium preparations, iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, and zinc.

In the food composition of the present invention, the food additive as described above may be included in an appropriate amount to achieve the purpose of addition according to the product type.

With regard to other food additives that may be included in the food composition of the present invention, reference may be made to the food code or food additive code according to the laws of each country.

The composition of the present invention may be understood as a pharmaceutical composition in other specific embodiments.

The pharmaceutical composition of the present invention may be prepared in an oral dosage form or a parenteral dosage form according to an administration route by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the meaning of "pharmaceutically acceptable" means that the application (prescription) does not have toxicity beyond adaptable without inhibiting the activity of the active ingredient.

When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, suspensions, wafers according to a method known in the art together with a suitable carrier It can be prepared in such a formulation. Examples of suitable pharmaceutically acceptable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, corn starch, potato starch, wheat starch, and other starches, cellulose, methylcellulose, ethylcellulose, Celluloses such as sodium carboxymethylcellulose and hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable Yu, etc. are mentioned. In the case of formulation activity, if necessary, it may be formulated including diluents and/or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

When the pharmaceutical composition of the present invention is prepared in a parenteral dosage form, it may be formulated in the form of eye drops, injections, transdermal administration, nasal inhalation, and suppositories according to a method known in the art together with a suitable carrier. When formulated as an eye drop, suitable carriers may include sterile water, saline, and isotonic solutions such as 5% dextrose, and benzalkonium chloride, mefilparaben, ethylparaben, etc. may be added for preservative purposes if necessary. When formulated as an injection, sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof may be used as suitable carriers, preferably Ringer's solution, PBS (phosphate buffered saline) containing triethanol amine, or sterilization for injection Water, isotonic solutions such as 5% dextrose can be used. When formulated into a transdermal dosage form, it can be formulated in the form of an ointment, cream, lotion, gel, external solution, pasta, liniment, air roll, and the like. In the case of nasal inhalants, suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide can be used to form an aerosol spray.When formulated as a suppository, the base is Withepsol ( witepsol), tween 61, polyethylene glycols, cacao butter, laurin paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, and the like.

Regarding the specific formulation of the pharmaceutical composition is known in the art, for example, Remington's Pharmaceutical Sciences (19th ed., 1995) and the like can be referred to. These documents are considered as part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, patient severity, and route of administration. May be in the /kg range. Administration can be made once a day or divided into several times. These dosages should not be construed as limiting the scope of the invention in any aspect.

As described above, according to the present invention, it is possible to provide a composition for improving pulmonary fibrosis using roe deer urine extract. The composition of the present invention can be commercialized as foods such as health functional foods or drugs such as natural medicines.

Figure 1 is a graph showing the cell proliferation rate of A549 cells according to the concentration (0, 2, 8, 31, 125, 500 ㎍ / ㎖) of the extract of Noruojum extract.
Figure 2 is a graph showing the dose-response relationship of the EMT inhibitory effect after treatment of the extract of Noruojum outpost at various concentrations (25, 50, 75, 100, 125 ㎍ / ml).
3 is a graph showing the inhibitory effect on the expression of TGF-β1 mediated Snail/Smad2/3 after treatment of the extract of Noruojum extract at various concentrations (25, 50, 75, 100, 125 μg/ml).
Figure 4 is a graph showing the effect on the migration of cells in A549 cells when treated with EMT inducers TGF-β1 and Noruojum outpost extract by concentration.
5 is a graph showing the effect on the invasion of cells when treated with EMT inducers TGF-β1 and Noruozum outpost extract by concentration in A549 cells.
Figure 6 is a graph showing the antioxidant effect of DPPH assay at various concentrations (25, 50, 75, 100, 125, 150 ㎍ / ml) of the extract of Noruojum outpost.
7 is a graph showing the inhibitory effect of active oxygen by DCF-DA assay at various concentrations (50, 75, 100, 125 μg/ml) of the extract of noruo zoom.
8 is a graph showing the effect of inhibiting collagen production at various concentrations (25, 50, 75, 100, 125 μg/ml) of the extract of Noruo Zum outpost.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these Examples and Experimental Examples.

Example 1. Preparation of ethanol extract of Noruo Zum outpost

The noru-o-zum outpost was shredded in the shade, then warmed three times in a 60°C water bath with 70% ethanol, and concentrated to obtain a 70% ethanol extract of Noru-o-zum outpost.

Experimental Example 1. Stability evaluation of Noru urine extract

The presence or absence of toxicity of the extract of Noru urine was analyzed.

As an experiment to quantify cell proliferation or viability, the production of a color developing substance called formazan in tetrazolium salts (WST-1) by mitochondrial dehydrogenase in cells was measured using a microplate reader. Human lung epithelial cells (A549) were seeded with 5×10 ³ densitiy in 100 µl/well and stored at 5% CO ₂ and 37°C for 24 hours. After removing the medium with the cells, 100 µl per well of Noru urine extract prepared at various concentrations of 2-500 µg/mL was treated, and after 24 hours, 1/10 of WST-1 was added to the medium for 1 hour incubation. And then measured in a microplate reader.

All data were expressed as mean ± standard error, and the statistical difference between the control and treatment groups was determined by using Student’s t-test analysis. When the P value was less than 5%, it was judged to be statistically significant.

As shown in Figure 1, the extract of Noru urine showed more than 80% cell viability at a concentration of 125 μg/ml. That is, it could be confirmed that the extract of Noruo Zum outpost showed almost no cytotoxicity at a relatively high concentration.

Experimental Example 2. Evaluation of EMT inhibitory activity of Noru urine extract and study of its mechanism

Western blot analysis was performed to investigate the EMT inhibitory activity and its mechanism according to the concentration of the Noru Ozum extract (25, 50, 75, 100, 125 μg/ml).

Western blot analysis is an experimental method to confirm the presence or expression of a specific protein, and an antigen-antibody reaction was used. First, as a protein extraction method, cells were seeded in 1 ml/well at a density of 6.25×10 ⁴ for 24 hours and then cultured at 5% CO ₂ and 37°C for 24 hours. After removing the medium that had been with the cells, 1 mL per well of the extract of Noru urine at various concentrations was treated. After 48 hours, lipa buffer appropriate for the cell concentration was added and stored for 5 minutes on ice, and then cell lysate was transferred to a 1.5 mL tube using a scrapper. After 30 minutes by vortexing every 10 minutes, it was centrifuged at 14,000g for 15 minutes at 4℃, and only the supernatant was collected and transferred to a new 1.5mL tube, and then protein was quantified using a BCA kit. 10 ㎍ protein was calculated, a mixture was made with distilled water and sample buffer, and denaturation was performed, and 80V 20min, 120V 60min were loaded on the gel. After the electrophoresis was completed, the gel was transferred to the membrane and blocked in skim milk for 1 h. After that, the first and second antibodies were sequentially attached, and then ECL was sprayed to detect light on the X-ray film.

Fig. 2 shows the results of the EMT inhibitory activity of the extract of Noru Ozum, and Fig. 3 shows the results of the inhibition mechanism.

E-cadherin, N-cadherin, and Vimentin induced by TGF-β1 as an EMT inducing standard were used as markers to evaluate EMT inhibitory action. As shown in FIG. 2, it was confirmed that EMT was induced by increasing N-cadherin and Vimentin and decreasing E-cadherin by TGF-β1. GAPDH was used as a housekeeping gene. After treating the extract of Noruosum at various concentrations (25, 50, 75, 100, 125 ㎍/ml), the dose-response relationship of the EMT inhibitory effect was confirmed. As a result, N-cadherin and Vimentin were reduced in a concentration-dependent manner. , It was confirmed to increase E-cadherin, and therefore, it was confirmed that the extract of Noru urine has an inhibitory effect on TGF-β1-mediated EMT.

In order to explore the mechanism of EMT inhibition of the extract of Noruosum extract, the effect on the expression of TGF-β1 mediated Snail/Smad2/3 was confirmed. As shown in FIG. 3, Snail increased due to TGF-β1 (2 ng/ml) and The expression of p-smad2/3 protein was inhibited in a concentration-dependent manner by the extract of Noru urine.

Experimental Example 2. Evaluation of effect on migration and invasion of Noru urine extract

In human epithelial cells, A549 cells, TGF-β1 5 ng/ml, an EMT inducer, and various concentrations of hepatitis urinary extract were treated to confirm the effect on cell migration and invasion.

For migration, A549 cells were seeded at 3×10 ⁴ density in a 96 well plate, and cultured at 37°C for 24 hours at 5% CO ₂ . Using a wound maker, the cells were scratched, washed twice with media, and treated with 100 µl per well of Noru urine extract. After stabilizing for 15 minutes or more in an Incucyte zoom machine, images were taken every 2 hours until 48 hours.

For invasion, lay matrigel on a 96 well plate and overnight, seed A549 at a density of 3×10 ⁴ , incubate for 24 hours at 5% CO ₂ , 37°C, and scratch the cells using a wound maker. Washed twice with. After matrigel was diluted in an appropriate amount in the medium to make various concentrations of Noru urine extract, it was treated with 50µl per well. After 15 minutes of stabilization in the Incucyte zoom machine, images were taken every 2 hours until 72 hours.

The results are shown in Figs. 4 and 5, and Figs. 4 and 5 show that when A549 cells were treated with EMT-inducing substances, TGF-β1 and Noru urine extract by concentration, the Noru urine extract was concentration dependent on migration and invasion of A549 cells. Show suppression.

Experimental Example 3. Evaluation of Antioxidant Effect of Noru Uzum Extract

In order to confirm the antioxidant effect, DPPH assay was performed at various concentrations (25, 50, 75, 100, 125 μg/ml) of the extract of Noru urine.

After diluting the sample in methanol by concentration, 1 ml of the sample and 0.25 ml of a 0.2 mM DPPH solution were mixed and stored at room temperature for 30 minutes in a light-shielding state, and then the absorbance was measured at 517 nm using a microplate reader. The DPPH inhibition efficiency was calculated by the following formula, where A0 is the control and A1 is the absorbance of the sample.

(Inhibition rate %) = [(A0-A1)/A0] X 100

In addition, in order to check whether active oxygen is suppressed in living cells, A549 cells were pretreated with Noru urine extract at different concentrations (50, 75, 100, 125 μg/ml) for 48 hours, and then H ₂ O ₂ as a positive control was added. After treatment with 5 mM for 30 minutes, DCF-DA assay was performed. A549 cells were seeded at a density of 5×10 ^{4 in} a 48 well plate, and cultured at 37°C for 24 hours, 5% CO ₂ , and then treated with 500 µl per well of Noru urine extract. After 48 hours, H ₂ O ₂ 5 mM was treated for 30 minutes, DCF-DA reagent was treated for 1 hour, and washed once with PBS, and then the cells were lysed with 0.1N NaOH and fluorescence values were measured.

The DPPH assay results are shown in Figure 6, and the DCF-DA assay results are shown in Figure 7.

As can be seen in Figure 6, Noru urine extract (GL0456) showed DPPH scavenging activity in a concentration-dependent manner, and as confirmed in FIG. 7, in order to confirm whether it inhibits active oxygen in living cells, noru urine extract in A549 cells After pre-treatment of the extract at different concentrations (50, 75, 100, 125 μg/ml) for 48 hours, the positive control H ₂ O ₂ was treated with 5 mM for 30 minutes, and DCF-DA assay was performed. Was shown to decrease the intracellular active oxygen caused by H ₂ O ₂ in a concentration-dependent manner.

Experimental Example 4. Noru urine Evaluation of collagen inhibitory effect of extract

Western blot analysis was performed at various concentrations (25, 50, 75, 100, 125 μg/ml) in order to confirm the collagen inhibitory effect of the Noru Ozum extract. The Western blot analysis method was the same as above, but the protein amount was 30 μg, and after denaturation was performed at 100° C. for 13 minutes, the TGX 10 precast gel was loaded at 120 V for 60 minutes.

The results are shown in Fig. 8, and it was confirmed that the extract of Noru urine inhibited the production of collagen in a concentration-dependent manner.

Claims (9)

A composition for improving pulmonary fibrosis, comprising a noru urine extract as an active ingredient.
The method of claim 1,
The extract is a composition, characterized in that the extract of water, methanol, ethanol or a mixed solvent thereof of Noruojum outpost.
The method of claim 1,
The composition, characterized in that the pulmonary fibrosis is caused by fine dust or ultrafine dust.
The method according to claim 1 or 2,
The composition is a composition, characterized in that the food composition.
The method according to claim 1 or 2,
The composition is a composition, characterized in that the pharmaceutical composition.
The method of claim 1,
The extract is a composition, characterized in that the mixed solvent extract of water and ethanol of Noru ojum outpost.
The method of claim 1,
The extract is a composition, characterized in that the 70% ethanol extract of Noruo Zum outpost.

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