KR101770766B1 - the composition comprising the specific extract isolated from Leonurus sibiricus as an active ingredient for preventing or treating respiratory inflammatory disease - Google Patents

Abstract

The present invention relates to a composition for preventing or treating a respiratory inflammatory disease containing an extract of a motherwort as an effective ingredient. Inhibitory effect of 5-lipoxygenase (5-LO) on basophilic leukocyte cells of the extract of motherwort according to the present invention; Experiments to inhibit the production of NO (nitrite) using RAW 264.7 cells; Inhibitory activity against respiratory acute inflammation using LPS induced lung injury BALB / c mouse model; Experiment of inhibition of allergic asthma using BALB / c mouse model; By confirming that it is safe and effective for prevention or treatment of respiratory inflammatory disease by confirming the excellent therapeutic effect on respiratory inflammatory disease than the water extract of motherwort described in the prior literature through the return mutation toxicity test, And can be useful for treatment.

Description

The present invention relates to a composition for preventing or treating a respiratory inflammatory disease, which comprises, as an active ingredient, a specific extract of motherwort, a composition for preventing or treating respiratory inflammatory disease, comprising an extract of Leonidus sibiricus as an active ingredient,

The present invention relates to a composition for preventing or treating a respiratory inflammatory disease containing an extract of a motherwort as an effective ingredient.

[Literature 1] Minoguchi K and Adachi M. Pathophysiology of asthma. In: Cherniack NS, Altose MD, Homma I, editors. Rehabilitation of the patient with respiratory disease. New York: McGraw-Hill, 1999, pp 97-104.

[Document 2] Maggi E., Immunotechnology, 3, pp. 233-244, 1998; Pawankar R., Curr. Opin. Allergy Clin. Immunol., 1, pp3-6, 2001;

[Literature 3] Barnes PJ, et al., Pharmacol Rev., 50, pp515-596, 1998

[4] Vestbo, J., Hurd, SS, Agusti, AG, Jones, PW, Vogelmeier, C., Anzueto, A., Barnes, PJ, Fabbri, LM, Martinez, FJ, Nishimura, RA, Sin.DD, Rodriguez-Roisin, R., 2013. Global strategy for management, and prevention of chronic obstructive pulmonary disease: GOLD executive summary. Am. J. Respir. Crit. Care Med. 187, 347-365.

[5] Le, A., Zielinski, R., He, C., Crow, M.T., Biswal, S., Tuder, R.M., 2009. Pulmonary epithelial neutrophilin-1 deletion enhancing development of cigarette smoke-induced emphysema. Am. J. Respir. Crit. Care Med. 180, 396-406

Li, Y., Li, SY, Li, JS, Deng, L., Tian, YG, Jiang, SL, Wang, Y., Wang, YY, 2012. A rat model for chronic obstructive pulmonary disease induced by cigarette smoke inhalation and repetitive bacterial infection. Biol. Pharm. Bull. 35, 1752-1760

[7] (Stampfli, M. R., Anderson, G.P., 2009. How cigarette smoke skews immune responses to promote lung disease and cancer Nat. Rev. Immunol. 9, 377-384

[8] Terashima, T., Wiggs, B., English, D., Hogg, JC, van Eeden, SF, 1997. Phagocytosis of small carbon particles (PM10) by alveolar macrophages stimulated the relaease of polymorphonuclear leukocytes from bone marrow . Am. J. Respir. Crit. Care Med. 155, 1441-1447

[Literature 9] O'Donnell, R., Breen, D., Wilson, S., Djukanovi, R., 2006. Inflammatory cells in the airway in COPD. Thrax 61, 448-454

[10] Profita, M., Sala, A., Bonanno, A., Riccobono, L., Ferraro, M., La Grutta, S., Albano, GD, Montalbano, AM, Gjomarkaj, M., 2010. Chronic obstructive pulmonary disease and neutrophil infiltration: role of cigarette smoke and cyclooxygenase products. Am. J. Physiol. Lung Cell. Mol. Physiol. 298, L262-L269.

[11] Hiemstra, P. S., van Wetering, S., Stolk, J., 1998. Neutrophil serine proteinases and defensins in chronic obstructive pulmonary disease: effects on pulmonary epithelium. Eur. Respir. J. 12, 1200-1208

[12] Hoenderdos, K., Condliffe, A., 2013. The neutrophils in chronic obstructive pulmonary disease. Am. J. Respir. Cell Mol. Biol. 48, 531-539.

[Document 13] Hele DJ, Belvisi MG. 2003. Novel therapies for the treatment of inflammatory airway diseases. Expert Opin Investig Drugs 12: 5-18; Fox JC, Fitzgerald MF. 2009. The role of animal models in the pharmacological evaluation of emerging anti-inflammatory agents for the treatment of COPD. Curr Opin Pharmacol. 9: 231-242

[Literature 14] Barnes PJ. 2000. Mechanisms in COPD: differences from asthma. Chest 117: 10S-14S;

[Document 15] Seatta M, Turato G, Maestrelli P, Mapp CE, Fabbri LM. 2001. Cellular and structural basis of chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 163: 1304-1309

[16] Hye-Young Shin, Sang-Hyun Kim, Sin-Myoung Kang, Ik-Jun Chang, Sang-Yong Kim, Hoon Jeon, Kang-Hyun Leem, Yong Shin, 2009Anti-inflammatory activity of Motherwort ( Leonurus sibiricus L.). Immunopharmacology and Immunotoxicology 31: 209-213.

[Literature 17] Information Sourcing, Illustrated Dictionary of Contemporary Art, Yeongrim Publishers, pp.848-849, 1998

Genetic factors determine the contribution of leukotrienes to acute inflammatory response. J. Immunology 164: 169-164. Junger L. Gould, Robert S. Byrum, Mikelle L. Key, MyTrang Nguyen, Victoria A. Wagoner, 4899-4907

[19] Green LC, Wanger DA, Glogowski J, Skipper PL, Wishnok JS, and Tannenbaum SR 1982 Analysis of nitrate, nitrite and [15N] nitrate in biologic fluids. Anual. Biochem 126: 131

[Literature 20] Vernooy JH, Dentener MA, van Suylen RJ, Buurman WA, Wouters EF. Long-term intratracheal lipopolysaccharide exposure in mice results in chronic lung inflammation and persistent pathology. Am J Respir Cell Mol Biol. 2002; 26 (1): 152-9.

[21] Zosky GR, von Garnier C, Stumbles PA, Holt PG, Sly PD, Turner DJ, 2004, The pattern of methacholine responsiveness in mice is dependent on antigen challenge dose. Respir. Res. 2004: 15

[22] Maron, DM and Ames BN (1983) Revised methods for the Salmonella mutagenicity test, Mutat. Res . 113: 173-215.

The present invention relates to a composition for preventing or treating respiratory-tract inflammatory diseases comprising extracts of motherwort-specific extract or compounds isolated therefrom and an effective ingredient thereof.

In general, an inflammatory reaction is a defensive reaction process of a living body that attempts to repair and regenerate a damaged region when an invasion of biological changes occurs in the cell or tissue of the living body. Thus, these series of reactions include localized blood vessels, various tissue cells of body fluids, and immune-mediated cells. With the recent development of molecular biology, inflammatory diseases have been attempted to be understood at the molecular level of cytokine, and factors affecting these diseases are also being clarified one by one.

Allergic reactions can be classified into four types according to the type of reaction: type I, type II, type III, and type IV, or type I, type II according to the time until onset after re- Type III and Type III allergies are called immediate allergies, and Type IV allergies can be classified as delayed allergies.

Among them, type I allergy is a reaction involving IgE antibody, which is called anaphylactic type allergy, and includes allergic rhinitis such as bronchial asthma, atopic diseases (dermatitis, enteritis), hay fever, allergic conjunctivitis, .

Asthma is a disorder characterized by airway hyperresponsiveness to various stimuli. Clinical symptoms such as wheezing, dyspnea, and cough caused by extensive stenosis of the airways can be reversed naturally or by treatment. . Most asthma is allergic and is characterized by chronic airway inflammation and bronchial hyperresponsiveness (Minoguchi K and Adachi M. Pathophysiology of asthma. In: Cherniack NS, Altose MD, Homma I, editors New York: McGraw-Hill, 1999, pp 97-104).

Asthma can be divided into extrinsic asthma and endogenous asthma depending on its cause. In the case of exogenous asthma, it refers to asthma that causes symptoms when exposed to a causative antigen. Skin test or bronchial induction test for the causative antigen is positive and it is common that the onset age is young. House dust, and ticks are the major causative antigens, and pollen, animal epithelium, and fungi also act as causative antigens. In the case of endogenous asthma, upper respiratory infections, exercise, emotional disturbances, changes in cold weather and humidity cause or exacerbate asthma, which is common in adult asthma. Other medications include asthma, exercise-induced asthma, and occupational asthma.

Asthma is also recognized as a chronic inflammatory disease because inflammatory cells are proliferated, differentiated and activated by IL-4, 5, and 13 produced by TH2 (T helper 2) type immune cells, (Elias JA, et al., J. Clin. Invest., 111, pp. 291-297, 2003). Inflammatory cells such as eosinophils, mast cells, and alveolar macrophages that are activated in the bronchi of patients suffering from asthma are involved in strong bronchoconstriction by secretion of various inflammatory mediators (cysteine leukotrienes, prostaglandins, etc.) (Maggi E., Barnes PJ, et al., Pharmacol. Rev., 50, pp. 515-596, 1998. Immunotechnology, 3, pp 233-244, 1998; Pawankar R., Curr. Opin. Allergy Clin. Immunol., 1, pp 3-6, ).

Therefore, the production of cytokines and immunoglobulin E such as IL-4, IL-5, and IL-13, which are involved in inflammatory cell activation, and the cysteine leukotriene biosynthesis secreted from inflammatory cells such as eosinophils, As a major cause of asthma, many studies are under way to develop drugs to suppress their production.

Chronic Obstructive Pulmonary Disease (COPD) is one of the causes of human health risk, increasing every year in the world. Currently, chronic obstructive pulmonary disease has been noted as a major cause of death in many countries, and in 2020, chronic obstructive pulmonary disease is predicted to be the third cause of mortality in humans (Vestbo, J., Hurd, SS, Agusti, AG, Jones, PW, Vogelmeier, C., Anzueto, A., Barnes, PJ, Fabbri, LM, Martinez, FJ, Nishimura, M., Stockly, RA, Sin.DD, Rodriguez-Roisin, R ., 2013. Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease: GOLD executive summary, Am J Respir Crit Care Med 187, 347-365. In general, chronic obstructive pulmonary disease (COPD) inhibits and blocks airflow through the lungs, leading to death. (Le, A., Zielinski, R., He, C., Crow, MT, Biswal, S., Tuder, RM, 2009. Pulmonary epithelial neutrophilin-1 deletion enhances development of cigarette smoke-induced emphysema., Am J Respir Crit Care Med 180, 396-406.). These chronic obstructive pulmonary diseases are caused by various causes such as tobacco smoke, dust, chemicals, air pollution, and bacterial infection (Li, Y., Li, SY, Li, JS, Deng, L., Jiang, SL, Wang, Y., Wang, YY, 2012. A rat model for stable chronic obstructive pulmonary disease induced by cigarette smoke inhalation and repetitive bacterial infection Biol. Pharm Bull., 1752-1760. In particular, smoking is considered to be the leading cause of chronic obstructive pulmonary disease, and more than 80% of patients with COPD have been identified as smokers (Rabe et al., 2007). Tobacco smoke contains a number of toxic chemicals, causing harmful changes in lung tissue during smoking (Stampfli, MR, Anderson, GP, 2009. Nat. Rev. Immunol., 9, 377-384.). These toxic chemicals cause infiltration of various inflammatory cells including neutrophils in lung tissue, resulting in pulmonary inflammation (Terashima, T., Wiggs, B., English, D., Hogg, JC, van Eeden, SF, 1997. Phargocytosis of small carbon particles (PM10) by alveolar macrophages stimulated the relaease of polymorphonuclear leukocytes from bone marrow., Am J Respir Crit Care Med., 155, 1441-1447). In clinical studies, the number of neutrophils and macrophages in Bronchoalveolar lavage fluid (BALF) or Sputum in patients with COPD was significantly increased (O'Donnell, R., Breen, D., et al. , Wilson, S., Djukanovi, R., 2006. Inflammatory cells in the airway in COPD. Thrax 61, 448-454.). These inflammatory cells produce a variety of enzymes that cause reactive oxygen species, inflammatory cytokines, chemokines, and tissue damage (Profita, M., Sala, A., Bonanno, A., Riccobono, L., Ferraro, M., La Grutta, S., Albano, GD, Montalbano, AM, Gjomarkaj, M., 2010. Chronic obstructive pulmonary disease and neutrophil infiltration: role of cigarette smoke and cyclooxygenase products. Physiol., Lung Cell, Mol. Physiol., 298, L262-L269. In particular, neutrophils play an important role in the development of chronic obstructive pulmonary disease (Hiemstra, PS, van Wetering, S., Stolk, J., 1998. Neutrophil serine proteinases and defensins in chronic obstructive pulmonary disease: effects on Pulmonary epithelium, Eur., Respir. J. 12, 1200-1208.). Neutrophils not only produce a number of inflammatory cytokines, chemokines and chemotactic factors, but also secrete elastinase (Elastase), resulting in the destruction of normal alveolar form and eventually Emphysema (Hoenderdos, K., Condliffe, A., 201. The neutrophils in chronic obstructive pulmonary disease, Am. J. Respir. Cell Mol. Biol., 48, 531-539.). Therefore, suppression of infiltration of inflammatory cells, particularly neutrophils, caused by tobacco smoke is recognized as an important therapeutic tool in the treatment of chronic obstructive pulmonary disease.

Therefore, chronic obstructive pulmonary disease should be appropriately treated differently from asthma, which is characterized by reversible airflow obstruction and allergic bronchial inflammatory response, but the present treatment is merely a symptom relief, (Hele DJ, Belvisi MG, 2003. Novel therapies for the treatment of inflammatory airway diseases. Expert Opin Investig Drugs 12: 5-18; Fox JC , Fitzgerald MF. 2009. Curr Opin Pharmacol. 9: 231-242)

Asthma and COPD, in principle, represent different pathological mechanisms. For example, (1) asthma in mast cells, mast cells, eosinophils, CD4 + cells (Th2), macrophages Whereas COPD differs in that neutrophils, CD8 + cells (Tc), etc. mainly act; (2) In terms of inflammatory mediators, asthma is associated with leukotriene B, histamine, IL-4, IL-5, IL-13, Eotaxin, RANTES, oxidative stress , Whereas COPD is different in that TNF-alpha, IL-8, and GRO-alpha are mainly involved; (3) In terms of inflammation, asthma affects the entire airway and is characterized by AHR (hypersensitive hypersensitivity), epithelial shedding, fibrosis, no parenchymal involvement, COPD acts on the peripheral airways, leading to the formation of epithelial metaplasia, histological metaplasia, dyspnea and dyspnea, while muscularis secretion, relatively reversible airflow obstruction, cough, sneezing, It is known that parenchymal destruction, relatively irreversible airflow obstruction, chronic bronchitis and emphysema are different in that they occur mainly in adulthood (Barnes PJ 2000. Mechanisms in COPD: differences Chest 117: 10S-14S; Seatta M, Turato G, Maestrelli P, Mapp CE, Fabbri LM 2001. Cellular and structural base of chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 163: 1304-1309)

In addition, there has been reported an anti-inflammatory activity against motherwort-measured water extract (MW) prepared by a specific extraction method, that is, a transferring-extraction method for motherwort (Hye-Young Shin, Sin- Myoung Kang, 2009Anti-inflammatory activity of Motherwort ( Leonurus , Jung-Yong Shin, Jong-Pil Kim, Hoon Jeon, Kang-Hyun Lee, Won-Hwan Park, sibiricus L.). Immunopharmacology and Immunotoxicology 31: 209-213).

However, none of the above references mention or mention the therapeutic effect of the crude extracts separated from mother crude extract or ethanol, preferably 20-50% alcohol or ethanol, and the purified fraction isolated therefrom in respiratory inflammatory diseases none.

Motherwort ( Leonurus sibiricus is a 1-year or 1-year-old herb belonging to the genus Labiatae, distributed throughout Korea, and its outposts are referred to as mother-of-pearl. The known components are Leonurine, stachydrine, Leucine, linolenic acid, oleic acid, sterol, rutin, and the like, such as leucin, leonuridine, leonurinine and the like, alkaloids such as lauric acid, linolenic acid, oleic acid, Have been used for the treatment of menstrual irregularities, birth bleeding, and the like (from the Information Sourcing, Illustrated Dictionary of Dietary Contamination, Young Lim, pp. 848-849, 1998).

Accordingly, the inventors of the present invention have developed a therapeutic agent using natural material resources effective for respiratory inflammatory diseases. As a result, 5-lipoxygenase (5-lipoxygenase, 5-lipoxygenase, -LO) inhibition experiment; Experiments to inhibit the production of NO (nitrite) using RAW 264.7 cells; Inhibitory activity against respiratory acute inflammation using LPS induced lung injury BALB / c mouse model; Experiment of inhibition of allergic asthma using BALB / c mouse model; The present inventors completed the present invention by confirming that the present invention can be effectively used for prevention or treatment of respiratory inflammatory diseases by confirming the excellent therapeutic efficacy against respiratory inflammatory diseases more safely and safely through the return mutation toxicity test than the motherwort water extract described in the prior art .

In order to accomplish the above object, the present invention provides a method for preparing a crude extract of motherwort 10 to 60% alcohol or ethanol, preferably 20 to 50% alcohol or ethanol, or a purified fraction isolated therefrom, And a pharmaceutical composition for preventing or treating respiratory inflammatory disease.

The purified fraction separated from the mother liquor soluble in 10 ~ 60% of mother liquor or ethanol as defined in the present invention is prepared by suspending the extract in 10 ~ 60% alcohol or mother liquor of motherwort, and then extracting it with hexane, methylene chloride, chloroform or ethyl acetate , Preferably a purified fraction obtained by adding a non-polar solvent such as hexane or an ethyl acetate solvent to a non-polar solvent obtained by fractionation.

The term " respiratory inflammatory disease "as defined herein includes, but is not limited to, any one selected from the group consisting of rhinitis, otitis media, sore throat, tonsillitis, pneumonia, asthma and chronic obstructive pulmonary disease.

As used herein, the term "prophylactic " means any action that inhibits or delays inflammation, allergy or asthma by administration of a composition comprising the extract.

The term "treatment" as used in the present invention means all the actions of improving or alleviating the symptom of the disease upon administration of the composition containing the extract.

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

The extracts of the present invention can be obtained by the following production methods.

For example, the present invention will be described in detail below.

The motherwort extract of the present invention can be prepared as follows. The dried motherworms are washed and sieved, and after 10 to 60% alcohol or ethanol, preferably 20 to 50% alcohol or ethanol is mixed several times, the mixture is incubated at 30 to 150 ° C, preferably 40 to 100 ° C for 30 minutes For about 1 hour to about 12 hours, preferably about 1 to 20 times, preferably about 2 to 10 times, by ultrasonic extraction, hot water extraction, room temperature extraction or reflux extraction, preferably hot water extraction, The crude extract of the present invention can be obtained by filtration, concentration under reduced pressure, and drying.

The polar solvent or the non-polar solvent soluble extract of the present invention may further contain about 0.0005 to 5 times, preferably 0.05 to 0.5 times the volume (v / w%) of the crude extract, preferably 30 to 90% ), Followed by fractionation using n-hexane, methylene chloride, ethyl acetate and butanol to obtain non-polar solvent-soluble extract fractions, which are dissolved in a nonpolar solvent such as n-hexane, methylene chloride or ethyl acetate; And polar solvent-soluble extract fractions soluble in polar solvents such as butanol and water can be obtained.

In addition, the purified fraction used in the non-polar solvent of the present invention is about 0.05 to 50 times, preferably 0.5 to 5 times the weight of the crude extract obtained in the above-described crude extract, preferably 10 to 60% alcohol or ethanol. (v / w%) of water and then adding the suspension to a suspension containing 0.1 to 20 times, preferably 1 to 10 times the volume of the suspension in hexane, methylene chloride, chloroform or ethyl acetate, , Or a non-polar solvent such as an ethyl acetate solvent is added to the reaction mixture to obtain a purified fraction soluble in the non-polar solvent of the present invention.

The inventors of the present invention conducted experiments on inhibition of 5-lipoxygenase (5-LO) using basophilic leukocyte cells and inhibition activity of NO (nitrite) production using RAW 264.7 cells in extracts obtained by the above- ; Inhibitory activity against respiratory acute inflammation using LPS induced lung injury BALB / c mouse model; Test for inhibiting allergic asthma using BALB / c mouse model: By confirming that it has a strong therapeutic effect on respiratory inflammatory disease through a mutation toxicity test and a cytotoxicity test, it is possible to provide a pharmaceutical composition for preventing or treating respiratory inflammatory disease It is confirmed that it is useful as a health functional food.

Therefore, the present invention relates to a method for producing a crude extract of the present invention comprising the crude extract obtained from the above-mentioned production method, which is soluble in 10 to 60% alcohol or ethanol, preferably 20 to 50% alcohol or ethanol, A pharmaceutical composition for preventing or treating inflammatory diseases, and a health functional food.

In addition, motherwort is a medicinal substance that has been used for a long time as an edible or herbal medicine, and the motherwort extract of the present invention also has no toxicity and side effects.

The pharmaceutical composition of the present invention contains 0.1 to 50% by weight of the above extract relative to the total weight of the composition.

The pharmaceutical compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the production of pharmaceutical compositions.

Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The composition containing the extract of the present invention may be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterilized injection solutions, Can be used.

More specifically, when formulating the composition, it can be prepared using a diluent or an excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like. Solid formulations for oral administration include tablets, pills, powders, granules and capsules, which may contain at least one excipient such as starch, calcium carbonate, sucrose, ), Lactose, gelatin and the like.

In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral use include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol and vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, and glycerol gelatin can be used.

The preferred dosage of the extract of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the administration route and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention can be administered in an amount of (0.0001-100) mg / kg, preferably (0.001-100) mg / kg, once or several times a day. In the composition, the extract of the present invention may be formulated in an amount of (0.0001 to 50) wt% based on the total weight of the total composition.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine and intracerebroventricular injections.

In addition, the present invention relates to the prevention and improvement of respiratory inflammatory diseases including crude extracts obtained by extracting mother mushroom 10 ~ 60% alcohol or ethanol, preferably 20 ~ 50% ethanol or alcohol, or purified fraction isolated therefrom And a health functional food.

&Quot; Health functional food "as defined herein means food prepared and processed using raw materials or ingredients having functionality useful to the human body in accordance with Law No. 6727 on Health Functional Foods." Functional " Structure and function of the nutrient to control or physiological effects, such as to obtain a beneficial effect for health is intended to eat.

The health functional food for preventing or ameliorating respiratory inflammatory disease of the present invention contains 0.01 to 95% by weight, preferably 1 to 80% by weight of the extract, based on the total weight of the composition.

For the purpose of preventing or ameliorating respiratory inflammatory diseases, it is also possible to use pharmaceutical dosage forms such as powders, granules, tablets, capsules, pills, suspensions, emulsions and syrups or health functional foods in the form of tea bags, Manufacturing and processing are possible.

In addition, the present invention relates to the prevention and improvement of respiratory inflammatory diseases including crude extracts obtained by extracting mother mushroom 10 ~ 60% alcohol or ethanol, preferably 20 ~ 50% ethanol or alcohol, or purified fraction isolated therefrom Or a food additive.

In addition, the health functional food may further include food additives, and the suitability of the food functional food as a "food additive" Standards and standards.

Examples of the products listed in the above-mentioned "food additives" include natural products such as ketones, chemical products such as glycine, potassium citrate, nicotinic acid and cinnamic acid, coloring matter, licorice extract, crystalline cellulose, guar gum, Sodium laurate, sodium glutamate preparation, noodles-added alkaline agent, preservative agent, tar pigment preparation and the like.

 Examples of the functional food containing the extract of the present invention include confectionery ice creams such as bread, rice cake, dried fruit, candy, chocolate, chewing gum and confectionery, ice cream products such as ice cream, ice cream powder, low fat milk, Processed products such as processed oil, goat milk, fermented oil, butter oil, concentrated oil, yogurt cream, butter oil, natural cheese, processed cheese, milk powder, milk products, meat products such as hamburger meat products, ham , Fish oil products such as sausages, bacon, etc. Fish products such as noodles, noodles, noodles, noodles, noodles, luxury noodles, improved noodles, noodles such as frozen noodles, pasta, vegetable beverages, Seasonings such as beverages such as soy sauce, miso, kochujang, chunchu, chonggukjang, mixed berries, vinegar, sauce, tomato ketchup, curry, dressing, Lean, shortening, and pizza.

The health functional beverage composition of the present invention is not particularly limited to the ingredients other than the above-mentioned purified product or compound as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient . Examples of the above-mentioned natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.); Disaccharide, (e.g., maltose, sucrose, etc.); And polysaccharides (for example, dextrin, cyclodextrin and the like), and sugar alcohols such as xylitol, sorbitol and erythritol. As natural flavors other than those described above, natural flavors (such as tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin)) and synthetic flavors (saccharin, aspartame, etc.) have. The ratio of the natural carbohydrate is generally about (1 to 20) g, preferably about (5 to 12) g per 100 mL of the composition of the present invention.

In addition to the above-mentioned composition, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the extract of the present invention can be added to food or beverage for the purpose of preventing the objective disease. At this time, the amount of the extract in the food or drink may be 0.01 to 15% by weight of the total food, and the health drink composition may be added in a proportion of 0.02 to 5 g, preferably 0.3 to 1 g, .

The extract according to the present invention, which is added to foods containing beverages in the course of manufacturing the health functional food, can be appropriately added or decreased as needed.

5-lipoxygenase (5-LO) inhibitory activity using basophilic leukocyte cells and inhibitory activity of NO (nitrite) production using RAW 264.7 cells in the motherwort extract according to the present invention; Inhibitory activity against respiratory acute inflammation using LPS induced lung injury BALB / c mouse model; Inhibitory activity against allergic asthma using BALB / c mouse model: It can be used for prevention or treatment of respiratory inflammatory disease by confirming the strong therapeutic effect on respiratory inflammatory disease safely through return mutation toxicity test.

Figure 1 is an HPLC chromatogram (210 nm) of the motherwort EA fraction;
Figure 2 is a diagram showing the UV spectrum of Rutin, isoquercitrin;
Figure 3 is a UV spectrum of the major components (Rutin and Isoquercitrin) for the motherwort EA fraction;
FIG. 4 is a chart showing the chromatogram profile at 270 nm of the extract of mother koji 50%.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided to further understand the present invention, and the present invention is not limited by the examples.

Comparative Example 1: Production of motherwort leach extract

After 500 g of Leonurus sibiricus outpost (human herb) was evenly blended, the motherwort extract was prepared by the decoction method described in the prior art (Immunopharmacology and Immunotoxicology, 2009; 31 (2): 209.213) (Hereinafter referred to as " CMW ").

Example 1: Motherwort extract

1-1. 30%  Manufacture of alcohol extracts

Dried motherwort ( Leonurus sibiricus ) 500 g of human herb (100 g) was mixed thoroughly and 10 L of 30% alcohol mixed with distilled water was added and extracted with hot water at 80 ° C for 2.5 hours. The filtrate was collected, and 10 L of 30% alcohol was added to the residue to extract for 2.5 hours. The filtrates were combined and concentrated under reduced pressure with a vacuum concentrator (EYELA, N-2100, JAPAN) until the crude extract was dried at 45 ° C . Finally, about 2 L of distilled water was added to suspend the suspension, and then lyophilized to obtain 67 g of a motherwort extract (hereinafter referred to as LS30E). The yield of the dried mothocyte hay extract was 13.4%.

1-2. 50%  Manufacture of alcohol extracts

Dried motherwort ( Leonurus sibiricus ) 500 g of human herb (100 g) was mixed with 10 L of 50% alcohol mixed with distilled water and extracted with hot water at 80 ° C for 2.5 hours. The filtrate was collected and the residue was extracted with 10 L of 50% alcohol for 2.5 hours. The filtrates were combined and concentrated under reduced pressure with a vacuum concentrator (EYELA, N-2100, JAPAN) until the herbal extract was dried at 45 ° C . Finally, about 2 L of distilled water was added and suspended, followed by freeze-drying to obtain 64 g of a motherwort extract in powder form (hereinafter referred to as LS50E). The yield of the dried meatsuchocarpa extract was found to be 12.8%.

1-3. 70%  Manufacture of alcohol extracts

Dried motherwort ( Leonurus sibiricus ) 500 g of human herb (100 g) was mixed with 10 L of 70% alcohol mixed with distilled water and extracted with hot water at 80 ° C for 2.5 hours. The filtrate was collected, and 10 L of 70% alcohol was added to the residue to extract for 2.5 hours. The filtrates were combined and concentrated under reduced pressure with a vacuum concentrator (EYELA, N-2100, JAPAN) until the crude extract was dried at 45 ° C . Finally, about 2 L of distilled water was added to suspend and then lyophilized to obtain 65 g of a motherwort extract in powder form (hereinafter referred to as LS70E). The yield of the dried meatsuchocarpa extract was found to be 10.3%.

Yield of motherwort extract by alcohol content Extraction yield However, Alcohol 50% 70% alcohol 13.4% 12.8% 10.3%

Example  2: Ethyl acetate Fraction  Produce

20 g of the 50% crude extract of Example 1 was thoroughly mixed with 2 L of primary distilled water, completely dissolved by ultrasonic wave using an ultrasonic machine (Branson 5510), placed in a separatory funnel, mixed with ethyl acetate (EA) solution 2 L, and then left to stand for 3-4 hours until EA and water layer were separated. After obtaining the separated upper layer EA fractions, 1 L of EA was separated from the water layer and diluted. After the 2nd and 3rd repetitions, 3 L of total EA fractions were concentrated under reduced pressure to obtain 3.4 g of EA fraction of pale blue cabbage (extraction yield: 17%, hereinafter referred to as " LSEA "). The dried fractions were stored at 4 ° C and dissolved at the required concentration.

Experimental Example 1: 5-lipoxygenase (5-LO) inhibitory activity

In order to confirm the 5-lipoxygenase (5-LO) inhibitory action of the sample of the above-mentioned example, the following methods were applied in the literature (Jennifer L. Gould, Robert S. Byrum, Mikelle L Key, MyTrang Nguyen, Victoria A. Wagoner, and Beverly H. Koller, 2000 Genetic factors determine the contribution of leukotrienes to acute inflammatory response J. Immunology 164: 4899-4907)

In order to evaluate the inhibitory effect on the leukotriene secretion, which is the major pathogenesis of asthma in the motherwort extract prepared in the Examples, cells were used in the following manner.

Rat Basophilic Leukemia Cells (RBL-2H3 Cell, CRL-2256, ATCC) were stabilized in culture medium (Dulbecco's Modified Eagle Medium, DMEM) for 24 hours at 37 ° C, The motherwort extract (10, 50, 100) μg / mL was added and reacted for 15 minutes.

Here, A23187 (C7522, Sigma) was added as Inophore to induce leukotriene release for 15 minutes. The amount of cystenyl leukotriene produced was measured using an enzyme immunoassay (EIS, ADI-901-070, Enzo life science) kit.

As shown in Table 2 and Table 3, the extracts of mother moths extracts of the present Example showed a concentration-dependent production of leukotriene, which is an inflammation inducing substance, from basophilic leukocyte cells in comparison with the motherwort water extract (CMW) (4-fold to 7-fold), and thus, 5-lipoxygenase (5-LO) inhibitory effect can be confirmed.

Inhibitory effect of leucotriene on the extracts of motherwort Treated group Leukotriene concentration
(% of Control) Inhibition rate
(%) Normal group 6.9 ± 4.3 Control group 100 ± 19.6 - Positive control (Montelukast 100 [mu] M) 39.2 ± 6.8 60.8 Comparative Example CMW - 100 μg / mL 89.3 ± 7.2 10.7 Alcohol 30% Ext - 100 μg / mL 40.4 ± 3.8 59.6 Alcohol 50% Ext - 100 μg / mL 21.0 + 7.7 79.0 Alcohol 70% Ext - 100 μg / mL 42.1 ± 4.5 59.9

Inhibitory effect of leucotriene on the extract concentration of motherwort (50% alcohol) Treated group Leucotriene concentration
(% of Control) Inhibition rate
(%) Normal group 5.2 ± 0.6 Control group 100 ± 3.1 - Positive control (Montelukast 100 [mu] M) 35.6 ± 1.3 64.4 Comparative Example CMW Ext - 100 μg / mL 90.6 ± 1.3 9.4 Motherwort extract 10 μg / mL 82.5 ± 6.2 17.5 Motherwort extract 50 μg / mL 68.2 ± 6.3 31.8 Motherwort extract 100 μg / mL 34.7 ± 2.6 65.3

Experimental Example 2: Inhibition of NO (Nitrite) formation

(Green LC, Wanger DA, Glogowski J, Skipper PL, Wishnok JS, and Tannenbaum SR, 1982 Analysis) to examine the inhibitory effect of NO (nitrite) formation on the samples of the above- of nitrate, nitrite and [15N] nitrate in biologic fluids. Anual. Biochem 126: 131)

RAW 264.7 cells (ATCC) were cultured in RPMI medium (11875-093, GIBCO, USA) supplemented with 10% fetal bovine serum and penicillin (100 units / ml) / streptomycin (100 ㎍ / ℃, and cultured at 5% CO ₂ condition. Cells were grown in dish (70075, 75 cm ² culture dish, SPL, Korea) and subcultured at intervals of 7 days.

Dividing the RAW 264.7 cells, 48-well cell to the culture dish pieces 2 × 10 ^5/250 μL per well and then cultured in a CO ₂ incubator at 37 ℃ for 24 h, LPS 1 μg / mL (Sigma , L2880) and motherwort extract Were cultured for 24 hours. NO in the culture medium was quantified by mixing 50 μL of the culture supernatant with the same amount of Griess reagent (G2930, Promega) and measuring the absorbance at 550 nm. At this time, sodium nitrite (G296A, Promega) was used as a standard substance.

As shown in Table 4, as shown in Table 4, the extract of the motherwort 50% alcohol inhibited the NO production, which is an inflammation inducing substance, from macrophages in a concentration-dependent manner, and it was confirmed that the extract inhibited the inflammation.

Inhibition of Macrophage NO Expression of Mycorrhizae Extract (LS50E) Treated group NO release
(μM) Inhibition ratio
(% of control) Normal group 8 ± 4 - Control (LPS treated with 1 ug / ml) 40 ± 7 - Test group
(LPS + motherwort) 10 μg / ml 35 ± 9 12.5 50 μg / ml 31 ± 6 22.5 100 μg / ml 25 ± 5 37.5

Experimental Example  3: Motherwort EA In the fraction  About Leukotriene  Comparison of inhibitory effect

According to the method of Experimental Example 1, the ability to inhibit leukotriene secretion was evaluated in the mother liquor extract and EA fraction.

As a result of the above tests, inhibitory effects on the secretion of rucotrienes by concentration (10, 25, and 50 μg / mL) on the alcohol extract and EA fraction from motherwort were evaluated. As a result, Leukotriene production was suppressed from basic leukocyte cells, but EA fraction showed about 80% inhibitory activity, which is 3.8 times higher than EtOH extract.

Experimental Example  4: Identification of major components from the lactose-inactivated fraction

The major components were identified by spectral patterns and spectral patterns of the wavelength of the motherwort EA fractions of the above examples.

 1) LC analysis conditions

  - Column: Phenomenex Luna C18 5 m, 250 mm x 4.6 mm

  - Column Temperature: 40 ℃

  - Injection Volume: 10 μL

  - Flow rate: 1.0 mL / min

  - Eluent: gradient

A - 0.1% Formic acid in H 2 O

    B - Acetonitrile

  - Detector: PDA 210 nm

 2) Main ingredient name (analysis result)

As a result, the major components of the EA fraction were investigated by spectral pattern and HPLC. The major constituents present in the EA fraction were rutin and isoquercitrin (see Figures 1, 2, 3 and 4).

Experimental Example 5: Inhibition of leukotriene inhibition on the major components present in the active fractions

To determine the active ingredient for motherwort, the inhibitory effect on the release of rutcorrhizin to the main components, Rutin, and Isoquercitrin, present in the active fractions, was evaluated in the same manner as in Experimental Example 1 .

As shown in Table 6, Rutin and Isoquercitrin, which are the main components of the lactose-containing fraction, inhibit the production of leukotriene, an inflammation-inducing substance, from basophilic leukocyte cells in a concentration-dependent manner . Especially, 50 μg / mL of isoquercitrin was suppressed to a level similar to that of the positive control substance. As a result, it can be confirmed that the main component of motherwort, rutin, and especially isocercitalin, has the effect of inhibiting 5-lipoxygenase.

Inhibitory effect of leucotriene on the major components of active fractions Treated group Leukotriene concentration
(% of Control) Inhibition rate
(%) Normal group 4.7 ± 3.6 Control group 100 ± 1.0 - Positive control (Montelukast 100 [mu] M) 37.2 ± 6.8  62.8 Routine 10 μg / ml 99.9 + - 6.8  0.1 25 μg / ml 94.5 ± 3.1  5.5 50 μg / ml 89.1 ± 7.7  10.9 Isoquercitrin 10 μg / ml 57.9 ± 9.4  42.1 25 μg / ml 48.7 ± 7.2  51.3 50 μg / ml 36.2 ± 3.8  63.8

EXPERIMENTAL EXAMPLE 6 Analysis of Active Ingredient Content by Various Types of Extracts of Motherwort

To confirm the contents of Rutin and Isoquercitrin as active ingredients of the extracts of the present invention, HPLC was carried out under the following conditions.

 1) LC analysis conditions

  - Column: Waters Sunfire C18 Sum, 250 mm * 4.6 mm

  - Column Temperature: 25 ℃

  - Injection Volume: 10 μL

  - Eluent: gradient

A - 0.1% Formic acid in H 2 O

    B - 0.1% Formic acid in ACN

  - Detector: PDA 270 nm (quantitative)

 2) Analysis results

As a result of the above experiment, as shown in FIG. 4 and Table 7, the contents of Rutin and Isoquercitrin, which are main components of the motherwort extract, were 4.3 to 8.0 mg / g, 1.3 to 1.8 mg / g level.

As a result of analyzing the above components, the content of Rutin and Isoquercitrin in the motherwort EA fraction prepared from 20 g of the 50% crude extract was analyzed using ethylacetate (EA) The content of isocercaline was 13.6 mg / g and 9.8 mg / g, respectively. From these results, it was found that the composition containing the lactose, which is the effective component of the motherwort, and the isocuritic acid was increased by 1.7 times and 5.4 times, respectively, through the ethyl acetate fraction.

Experimental Example  7: Respiratory Acute Inflammation Inhibition Experiment

In order to confirm the inhibitory action on the acute inflammation of the respiratory organ of the above example, Vernooy et al. (Vernooy JH, Dentener MA, van Suylen RJ, Buurman WA, Wouters EF Long-term intratracheal lipopolysaccharide exposure in mice results in chronic lung inflammation and persistent pathology. Am J Respir Cell Mol Biol. 2002; 26 (1): 152-9.).

Aerosols were generated by ultrasonication (Nebulizer control 10, BUXCO) in LPS solution (0.5 mg / 3 mL, L2880, Sigma) diluted with physiological saline and then injected into 7-week-old BALB / c male mice (Orient Bio, After 2 days of inhalation, the inflammatory cells were measured in the bronchial washing solution.

Forty-eight hours after induction of inflammation, the lung bronchus was washed with 3.0 mL of phosphate buffer solution (10010-023, GIBCO) to collect the washing solution. The numbers of mononuclear cells (lymphocytes, mononuclear cells) and polymorphonuclear leukocytes (mostly neutrophils, neutophils) in the washings were counted and the number of neutrophils was counted as an indicator of inflammation induction. The motherwort extract (LS50E or CMW, 200 mg / kg BW) was orally administered 3 times for 3 days from the day before LPS inhalation. The normal control group and the control group were orally administered 300 μL of distilled water for 3 days, and the positive control drug was orally administered once at a dose of 10 mg / kg BW 2 hours after LPS inhalation.

As a result of the above experiment, the increase in the number of neutrophils in the bronchial space was measured as an index of LPS-induced acute respiratory inflammation. The results showed that the extract of mother - of - pearl was inhibited acute inflammation of respiratory system. (Table 9)

Reduction of bronchial inflammatory cells (Neutrophil) Treated group Mononuclear cells (%) Neutrophil
(%) % Inhibition of bronchial inflammation Normal group 99 One Control group (LPS treatment) 51 ± 10 41 ± 10 - Comparative Example (LPS + CMW Ext) 63 ± 6 37 ± 6 9.7 Test group (LPS + motherwort LS50E) 71 ± 6 29 ± 6 29.2 positivity
Control group Montelukast 52 ± 8 39 ± 7 4.8 Dexamethasone 71 ± 10 27 ± 10 34.1 Roflumilast 57 ± 8 40 ± 8 2.4

Experimental Example 8: Allergic asthma inhibition test

(Zosky GR, von Garnier C, Stumbles PA, Holt PG, Sly PD, Turner DJ, 2004). In order to confirm the allergic asthma-suppressing activity of the sample of the above example, The pattern of methacholine responsiveness in mice is dependent on antigen challenge dose. Respir . Res. 2004: 15)

To evaluate the airway inflammation inhibitory activity of motherwort (LS50E or CMW, 200 mg / kg BW) extract, we used eosinophil and neutrophil increase response to pulmonary bronchus induced by exposure of sensitized mice to antigen And was performed in the following manner.

(Sigma, A5503) and Alum (Pierce, Pro no. 77161) diluted with PBS to a BALB / c female mouse (6 weeks old, Orient Bio, mL was intraperitoneally administered at 0, 7, and 14 days. At 21, 25, and 28 days after the initial sensitization, 0.7% egg albumin was aerosolized using high pressure compressed air and sprayed for 50 minutes to induce airway inflammation.

Twenty-four hours after inflammation, the lung bronchus was washed with 3 mL of phosphate buffer solution to collect the washing solution. The numbers of mononuclear cells (lymphocytes, monocytes) and polymorphonuclear leukocytes (neutrophil, eosinophil) in the wash liquor in the washings were counted and the amount of neutrophils and eosinophils was used as an indicator of asthma induction .

The motherwort extract was orally administered 8 times on the first sensitization (20-28) days. The normal control group and the control group were orally administered 300 μL of distilled water for 8 days. The positive control drug was administered orally three times at a dose of 10 mg / kg BW 2 hours after inhalation of 0.7% egg albumin at 21, 25 and 28 days after the first sensitization Respectively.

The increase in the number of neutrophils and eosinophils in bronchial asthma as an indicator of allergic asthma induced by albumin (OVA) resulted in a decrease in the ratio of two inflammatory cells by treatment with lactobacillus extract (LS50E) Significant reduction effect was observed. This indicates that the motherwort extract has an inhibitory effect on allergic asthma. (Table 10)

Inhibitory effect of motherwort extract on allergic bronchial asthma Treated group Mononuclear cells (%) Polymorphonuclear cell (%) Bronchial asthma inhibition rate (%) Monocyte Macrophage Neutrophil Eosinophil Normal group 23 ± 8 76 ± 10 0 1 ± 1 Control group 36 ± 9 25 ± 6 9 ± 6 31 ± 8 - Comparative Example CMW Ext 39 ± 9 26 ± 6 10 ± 6 25 ± 6 12.5 Test group
(Mother-of-pearl LS50E) 51 ± 5 24 ± 8 8 ± 3 17 ± 8 35

Experimental Example 9: Return mutation  Toxicity test

In order to confirm the toxicity of the sample according to the return mutation test, the method proposed by Maron and Ames (1983) described in the literature was partially modified and experimented. (Maron, DM and Ames BN (1983) Revised methods for the Salmonella mutagenicity test, Mutat Res . 113: 173-215)

For the mutagenic search for specific components, strains ( Salmonella typhimurium ) TA98, TA100, TA102, TA1535, and TA1537 were used. The test substance was treated by direct plate incorporation with or without the metabolic activation enzyme system (S-9 mix). Salmonella typhimurium used for the return mutation test TA98 strain was purchased from Molecular Toxicology Inc. (USA). The strains were inoculated into 25 mL of liquid culture medium (2.5% Oxoid Nutrient broth No. 2) containing 50 μL of the frozen test strain solution and incubated in a shaking incubator (VS-8480SFN, Vision Science Co.) ℃ for about 10 hours. Minimal glucose agar plate was prepared containing 1.5% Bacto agar (214010, BD Difco), Vogel-Bonner medium E and 2% glucose, top agar was prepared with 0.6% agar and 0.5% NaCl And 0.05 mM histidine (43011, Fluka) -biotin (47868, Supelco) was added to the top agar.

2 mL each of top agar sterilized by high pressure steam sterilization is pre-heated in a dry bath (dry bath, 11-718-4, Fisher Scientific) at 45 ° C. Subsequently, 0.1 mL of the test substance solution and the culture medium 0.1 mL was added to a top agar and immediately shaken for 2 to 3 seconds with a vortex mixer (37600, Thermolyne). The mixture was poured into a minimal glucose agar plate and tilted in various directions to solidify it. The excipient group (negative control) was prepared by adding 0.1 mL of the excipient instead of the test substance solution and a positive control material solution (2-Aminoanthracene (2AA), Spec: Sigma A1381) by the same method. After the top agar was firm, the plate was closed with the lid closed, and the plate was incubated at 37 ° C for about 48 hours, and the colonies were counted.

As a result of the above experiment, it is considered that the motherwort extract does not induce a return mutation in the strains used under the test conditions (Table 11).

Of motherwort extract Return mutation  Toxicity test Strain Metabolic activity
Enzymatic system Return mutation bacterium / plate 0 μg / plate 500 μg / plate 1000 μg / plate TA98 -S9 19 ± 2 15 ± 2 23 ± 4 + S9 19 ± 1 17 ± 3 16 ± 5 TA100 -S9 87 ± 12 109 ± 24 94 ± 3 + S9 79 ± 14 81 ± 3 92 ± 4 TA102 -S9 67 ± 10 59 ± 14 73 ± 18 + S9 83 ± 10 82 ± 13 79 ± 8 TA1535 -S9 7 ± 2 9 ± 3 8 ± 2 + S9 8 ± 1 7 ± 1 8 ± 3 TA1537 -S9 3 ± 1 4 ± 3 5 ± 3 + S9 9 ± 1 7 ± 4 8 ± 1

The preparation examples of the pharmaceutical composition containing the extract of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically explained.

Preparation Example 1. Preparation of powder

LS50E Extract -------------------------------- 20 mg

Lactose --------------------------------------- 100 mg

Talc ---------------------------------------- 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation Example 2. Preparation of tablets

LS30E Extract -------------------------------- 10 mg

Corn starch --------------------------------- 100 mg

Lactose --------------------------------------- 100 mg

Magnesium stearate -------------------------- 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation Example 3. Preparation of capsules

LS50E --------------------------------------- 10 mg

Crystalline cellulose - 3 mg

Lactose ---------------------------------- 14.8 mg

Magnesium Stearate ---------------------- 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation Example 4. Preparation of injection

LS50E Extract -------------------------------- 10 mg

Mannitol ------------------------------------- 180 mg

Sterile sterilized distilled water for injection ------------------------ 2974 mg

Na ₂ HPO ₄ 12H ₂ O --------------------------------- 26 mg

(2 ml) per ampoule in accordance with the usual injection method.

Formulation Example 5. Preparation of a liquid preparation

LSEA Extract -------------------------------- 20 mg

Ising Party ------------------------------------ 10 g

Mannitol --------------------------------------- 5 g

Purified water --------------------------------------

Each component was added and dissolved in purified water according to a conventional liquid preparation method, the lemon flavor was added in an appropriate amount, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 mL, And sterilized to prepare a liquid preparation.

Formulation Example 6. Preparation of Healthy Foods

LS50E Extract ------------------------------ 1000 mg

Vitamin mixture --------------------------------

Vitamin A Acetate ------------------------- 70 g

Vitamin E ----------------------------------- 1.0 mg

Vitamin B ₁ ---------------------------------- 0.13 mg

Vitamin B ₂ ---------------------------------- 0.15 mg

Vitamin B ₆ ----------------------------------- 0.5 mg

Vitamin B ₁₂ ---------------------------------- 0.2 g

Vitamin C ------------------------------------ 10 mg

Biotin -------------------------------------- 10 g

Nicotinic acid amide 1.7 mg

Folic acid ---------------------------------------- 50 μg

Calcium pantothenate ------------------------------ 0.5 mg

Inorganic mixture --------------------------------

Ferrous sulfate - 1.75 mg

Zinc oxide ---------------------------------- 0.82 mg

Magnesium carbonate ------------------------------ 25.3 mg

Potassium phosphate monohydrate - 15 mg

Secondary calcium phosphate --------------------------------- 55 mg

Potassium citrate ---------------------------------- 90 mg

Calcium carbonate - 100 mg

Magnesium chloride ------------------------------ 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Formulation Example 7. Preparation of health drink

LS50E --------------------------------------- 100 mg

Vitamin C -------------------------------------- 15 g

Vitamin E (powder) ------------------------------- 100 g

Lactic Acid Iron ------------------------------------- 19.75 g

Zinc oxide ------------------------------------- 3.5 g

Nicotinic acid amide 3.5 g

Vitamin A ------------------------------------- 0.2 g

Vitamin B ₁ ----------------------------------- 0.25 g

Vitamin B ₂ ------------------------------------ 0.3 g

Water -------------------------------------------- Quantitative

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 L container, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (7)

A crude extract or crude extract obtained from 20 to 50% (w / w) of a Korean mother-of-pearl motherch having a content of Rutin ranging from 4.3 to 8.0 mg / g and Isoquercitrin ranging from 1.3 to 1.8 mg / As an active ingredient, a purified fraction soluble in ethyl acetate, which is obtained by suspending the extract in water and then fractionating it by adding ethyl acetate. The pharmaceutical composition for preventing or treating respiratory inflammatory disease selected from the group consisting of pneumonia, asthma or chronic obstructive pulmonary disease. delete delete A crude extract or crude extract obtained from 20 to 50% (w / w) of a Korean mother-of-pearl motherch having a content of Rutin ranging from 4.3 to 8.0 mg / g and Isoquercitrin ranging from 1.3 to 1.8 mg / Which is obtained by suspending in water and then fractionating it by adding ethyl acetate as an active ingredient, as an active ingredient. The present invention relates to a health functional food for preventing or ameliorating a respiratory inflammatory disease selected from pneumonia, asthma or chronic obstructive pulmonary disease. delete A crude extract or crude extract obtained from 20 to 50% (w / w) of a Korean mother-of-pearl motherch having a content of Rutin ranging from 4.3 to 8.0 mg / g and Isoquercitrin ranging from 1.3 to 1.8 mg / Which is obtained by suspending in water and then fractionating it by adding ethyl acetate. The present invention relates to a health supplement for preventing or ameliorating a respiratory inflammatory disease selected from the group consisting of pneumonia, asthma or chronic obstructive pulmonary disease. delete

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