KR20190118019A - the composition comprising the extract of Veronicastrum sibiricum L. Pennell as an active ingredient for preventing or treating inflammation, allergy and asthma and the use there of - Google Patents

Abstract

The present invention relates to a composition for preventing or treating inflammation, allergy, and asthma, including an extract of veronicastrum sibiricum L. pennell as an active ingredient. The extract of veronicastrum sibiricum L. Pennell is a crude extract, a polar solvent-soluble extract, or a non-polar solvent-soluble extract.

Description

The composition comprising the extract of Veronicastrum sibiricum L. Pennell as an active ingredient for preventing or treating inflammation, allergy and asthma and the use there of}

The present invention relates to a composition for the prophylaxis or treatment of inflammation, allergy and asthma, including cold grass extract as an active ingredient and its use.

[Reference 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.

2 Elias JA, et al., J. Clin. Invest., 111, pp 291-297, 2003

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

Barnes PJ, et al., Pharmacol Rev., 50, pp 515-596, 1998

[Document 5] Korean Patent Registration No. 10-860080

[Patent 6] Korean Patent Publication No. 10-2006-125499

Document 7 Flora of China 18; 1-212. 1998

8 Elias, J.A, et al., J Clin Invest, 111, 291-297, 2003

9, Chen M et al, Immunology, p. 376-84, 2011

Document 10 Kay, A.B. The New England Journal of Medicine, 344, 30-37, 2001

Renz H, et al., J Exp Med, 1777, 1175-1180, 1993

12 Kwak YG, et al., J Clin Invest, 111, 1083-92, 2003

Lee KS et al., FASEB J, 20, 455-465, 2006.

In general, an inflammatory response is a defense process of a living body that attempts to repair and repair the damaged area when an invasion that causes some organic change in a cell or tissue of the living body is applied. Therefore, this series of reactions include local blood vessels, various tissue cells of body fluids, immune-involved cells, and the like. Recently, with the development of molecular biology, inflammatory diseases have been tried at the molecular level of cytokines, and factors affecting these diseases have been identified one by one.

Allergic reactions can be categorized into four types, type I, II, III and IV by their response form, or type I, II by time until onset after resensitization by antigen. Type and III allergies are called immediate allergies, and type IV allergies can be classified as delayed allergies.

Among these, type I allergy is a reaction involving IgE antibodies, and it is called anaphylactic allergy and includes allergic rhinitis such as bronchial asthma, atopic disease (dermatitis, enteritis), hay fever, allergic conjunctivitis, food allergy, etc. This includes.

Asthma is a disease characterized by airway hypersensitivity to various stimuli. Clinical symptoms such as wheezing, dyspnea, and cough caused by extensive airway narrowing are reversibly improved naturally or by treatment. Can be. Most asthma are allergic and characterized by chronic airway inflammation and bronchial hyperresponsiveness (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, pp97-104).

Asthma can be divided into exogenous and endogenous asthma depending on the cause. In exogenous asthma, it is asthma that causes symptoms when exposed to the causative antigen. A skin test or bronchial challenge test for the causative antigen is generally positive and is young. House dust and mites are the most common causative agents, and pollen, animal epithelium, and fungus act as causative antigens. In the case of endogenous asthma, upper respiratory tract infections, exercise, emotional instability, cold climate and humidity changes cause or worsen asthma, which is common in adult-type asthma. Other medications include asthma, exercise-induced asthma and occupational asthma.

Asthma is also recognized as a chronic inflammatory disease because interleukin-4, 5, and 13 produced by TH2 (T helper 2) type immune cells proliferate, differentiate, and activate inflammatory cells to migrate and invade the airways and surrounding tissues. (Elias JA, et al., J. Clin. Invest., 111, pp 291-297, 2003). Inflammatory cells, such as eosinophils, mast cells, and alveolar macrophages, activated in the bronchus of patients with asthma, are involved in potent bronchial contraction by secreting various inflammatory mediators (cysteine leukotriene, prostaglandin, etc.) (Maggi E., Immunotechnology, 3, pp233-244, 1998; Pawankar R., Curr. Opin.Allergy Clin. Immunol., 1, pp3-6, 2001; Barnes PJ, et al., Pharmacol Rev., 50, pp515-596, 1998 ).

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 their action, the cysteine leukotriene biosynthesis secreted from inflammatory cells such as eosinophils is associated with inflammatory and allergic reactions. As a major cause of asthma, many studies are being conducted to develop drugs to suppress their production.

To date, the present inventors have focused on developing therapeutic agents using various resources, in particular, natural products known for their safety and effectiveness, as therapeutic agents using antibodies against various types of cytokines and chemokines that are characteristic of inflammation, allergies and asthma diseases. It has been developed.

In Korean Patent Publication No. 10-2006-0125489, Pseudolysimachion ovtum, P. kiusianum, P. kiusianum var. Diamanticum, P. kiusianum var. Villosum , P. dahuricum, P. pyrethrinum, P. linarifolium, P. linarifolium var. Villosulum, P. rotundum var. Subintegrum Inflammation and allergy of plant extracts of Veronica genus such as P. rotundum var.coreanum, P. insulare, P. undulata, Veronica longifolia And asthma disease treatment effects have been published.

On the other hand, Veronicastrum sibiricum L. Pennell is a Veronicastrum plant, which, unlike the herbaceous plants that are native to Korea, throughout the Russian Far East, Japan, and Northeast China, leaves are rotated and flowery twigs It is distinguished from the Veronica genus plants of Veronica longifolia in that it is divided into four branches with a shallow tip shape, and is also referred to as a hideweed or Siberian cold herb. (National Biodiversity Information Sharing System, http://www.kbr.go.kr/home/rsc/rsc01002v.do?data_gbn_cd=BIO&ktsn_no=120000063371&menuKey=448).

However, none of the above literature discloses or teaches a therapeutic agent for inflammation, allergy and asthma disease, which comprises Veronicastrum sibiricum L. Pennell (Veronicastrum) extract as an active ingredient.

Targeting:; (cold zinnia Veronicastrum Veronicastrum sibiricum L. Pennell) extract The present inventors kkoripul genus (genus Pseudolysimachion) plants in ginsan kkoripul (Veronica longifolia) not naengcho is disclosed in Korea Patent Publication No. 10-2006-0125489 existing call Han, leukotriene development inhibitory evaluation test (Experimental Example 1); Effect test on total cell count in total bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) using Balb / c male mouse (Experimental Example 2); Experimental effect on the number of neutrophil cells compared to the total cell count in total bronchoalveolar lavage (BAL) (Experimental Example 3); Effect on the number of Neutrophil + / Gr-1 + absolute cells in BAL fluid (Test Experimental Example 4); Experiments on the effect of CD11b + / Gr-1 + absolute cell number on lung cells (Experimental Example 5); Experiment on the effect on the number of CD4 + / CD3 + absolute cells in lung cells (Experimental Example 6); Experiment on the effect on the number of Macrophage + / CD11b + absolute cells in lung cells (Experiment 7); Effect experiment on the expression level of inflammatory factors in BAL fluid (Experimental Example 8); Influence on lung tissue changes Experimental Example (Experimental Example 9) Through the animal experiments confirmed that the extract showed superior anti-inflammatory, anti-allergic and asthma inhibitory activity than the extract of Veronica longifolia extracted by the conventional method, the present invention was completed .

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammation, allergies and asthma containing cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold grass) extract as an active ingredient.

In addition, the present invention provides a health functional food for the prevention and improvement of inflammation, allergy and asthma containing cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold grass) extract as an active ingredient.

As one aspect for achieving the above object, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammation, allergies and asthma containing cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum) extract as an active ingredient

Cold grass as defined herein is an outpost, root of cold grass native to or grown in Korea, or imported from China, Russia, Japan, etc., preferably, Korean, more preferably, Gyeonggi-do, most preferably, Ansan, Gyeonggi-do. , Stems, or flowers, preferably, outpost material.

Cold grass extract as defined herein is characterized in that the crude extract, polar solvent soluble extract or non-polar solvent soluble extract.

The crude extract as defined herein is a solvent selected from water containing purified water, lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, butanol, or a mixed solvent thereof, preferably methanol, or a water and ethanol mixed solvent, more preferably Is characterized in that the extract available in 60-100% ethanol.

Non-polar solvent soluble extracts as defined herein include extracts soluble in hexane, methylene chloride, chloroform, or ethyl acetate, preferably hexane, methylene chloride or ethyl acetate, more preferably ethyl acetate, or methylene chloride solvent. .

The polar solvent soluble extract as defined herein includes extracts soluble in a solvent selected from water, methanol, butanol or a mixed solvent thereof, preferably water or butanol, more preferably butanol, except for the above non-polar solvent soluble extract. do.

"Inflammation" as defined herein refers to dermatitis, atopic, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia (fibromyalgia) ), Psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendonitis, hay salt, peritonitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases It may be any one, but is not limited thereto.

“Allergy” as defined herein includes hypersensitivity, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis, urticaria, insect allergy, food allergy or drug allergy, preferably allergic rhinitis, Asthma, allergic dermatitis, atopic dermatitis, contact dermatitis, urticaria, food allergy or drug allergy, more preferably atopic dermatitis or contact dermatitis.

Asthma, as defined herein, refers to physical activities such as house dust mites, pollen, animal hair or dandruff, cockroaches, foods, drugs, colds, tobacco smoke and indoor pollution, air pollution, food additives, and exercise, climate change, yellow dust, stress, etc. Contains bronchial asthma caused.

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

Extracts of the present invention can be obtained by the preparation method as follows.

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

The crude extract of the present invention can be prepared as follows. After washing and rinsing the dried cold grass, a solvent selected from water, including purified water, lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, butanol, or a mixed solvent thereof, preferably methanol or a mixed solvent of water and ethanol, more preferably Is mixed several times with 60 to 100% ethanol and then 30 minutes to 150 hours, preferably from room temperature to 100 ° C, more preferably from room temperature to 60 ° C for 30 minutes to 48 hours, preferably 1 hour to 12 hours. Cold extraction of the present invention by filtration, concentration under reduced pressure, and drying the extract obtained by repeatedly performing ultrasonic extraction, hot water extraction, room temperature extraction or reflux extraction, preferably ultrasonic extraction, about 1 to 20 times, preferably 2 to 10 times. Crude extract can be obtained.

In addition, the polar solvent or non-polar solvent soluble extract of the present invention is about 0.0005 to 500 times the volume of the crude extract, preferably 60 to 90% ethanol crude extract, preferably 0.05 to 50 times the volume (v / w% Non-polar solvent soluble extract fractions which are available in non-polar solvents such as n-hexane, methylene chloride, ethyl acetate by carrying out a conventional fractionation process using n-hexane, methylene chloride, ethyl acetate and butanol after addition of water; And polar solvent soluble extract fractions soluble in polar solvents such as butanol, water and the like.

It is also possible to further carry out conventional fractionation processes customary in the art (Harborne J. B. Phytochemical methods: A guideto modern techniques of plant analysis. 3rd Ed. Pp 6-7, 1998).

The inventors of the present invention target the extracts of Veronicastrum sibiricum L. Pennell (Veronicastrum) instead of Veronicas longifolia, a Pseudolysimachion genus plant disclosed in Korean Patent Publication No. 10-2006-0125489. , Leukotriene development inhibitory evaluation test (Experimental Example 1); Effect test on total cell count in total bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) using Balb / c male mouse (Experimental Example 2); Experimental effect on the number of neutrophil cells compared to the total cell count in total bronchoalveolar lavage (BAL) (Experimental Example 3); Effect on the number of Neutrophil + / Gr-1 + absolute cells in BAL fluid (Test Experimental Example 4); Experiments on the effect of CD11b + / Gr-1 + absolute cell number on lung cells (Experimental Example 5); Experiment on the effect on the number of CD4 + / CD3 + absolute cells in lung cells (Experimental Example 6); Experiment on the effect on the number of Macrophage + / CD11b + absolute cells in lung cells (Experiment 7); Effect experiment on the expression level of inflammatory factors in BAL fluid (Experimental Example 8); Influence on lung tissue changes Experimental (Example 9) through the animal experiments, such as the extract of the conventional method extract Veronica longifolia extract showed excellent anti-inflammatory, anti-allergic and asthma inhibitory activity, the composition is inflammation, It has been found to be useful as a pharmaceutical composition or health functional food for the prevention and treatment of allergy and asthma.

Accordingly, the present invention provides a pharmaceutical composition or health functional food for the prevention and treatment of inflammation, allergies and asthma, containing the cold grass extract obtained by the manufacturing method as an active ingredient.

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

As used herein, the term "prevention" means any action that inhibits or delays inflammation, allergy or asthma by administration of a composition comprising said extract. In addition, the term "treatment" used in the present invention means any action that improves or advantageously changes the symptoms of the disease by administration of the composition comprising the extract.

In another embodiment, a therapeutic method for treating inflammation, allergy and asthma comprising administering the cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum) extract according to the present invention to patients with inflammation, allergies and asthma to provide.

In another aspect, there is provided the use of cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum) extract for the manufacture of a therapeutic for treating inflammation, allergy and asthma patients.

Pharmaceutical compositions comprising tablets according to the present invention, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, weights, binders, wetting agents, sealing agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations include at least one excipient such as starch, calcium carbonate, sucrose, and the like in the extracts and fractions. (sucrose), lactose (lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.

In the above formulations, non-aqueous solvents and suspending agents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol gelatin and the like can be used.

The preferred dosage of the pharmaceutical composition comprising the extract of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the pharmaceutical composition comprising the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The extract of the present invention can be administered to mammals such as mice, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or Intracerebroventricular injection.

The pharmaceutical composition of the present invention comprises 0.1 to 50% by weight of the extract, based on the total weight of the composition.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. As the non-aqueous solvent and suspending agent, vegetable oils such as propylene glycol, polyethylene glycol and olive oil, and injectable esters such as ethyloleate may be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter and glycerogelatin may be used.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention may be administered in 0.0001 ~ 100 mg / kg, preferably in an amount of 0.001 ~ 100 mg / kg divided once or several times a day. Extract of the present invention in the composition may be formulated in an amount of 0.0001 to 50% by weight relative to the total weight of the composition.

The pharmaceutical composition of the present invention can be administered to mammals such as mice, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural and intracere broventricular injections.

In addition, the present invention provides a dietary supplement for the prevention and improvement of inflammation, allergies and asthma, containing cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold grass) extract as an active ingredient.

As defined herein, "health functional food" means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the Health Functional Food Act No. 6767, and "functional" means It means ingestion for the purpose of obtaining useful effects on health use such as nutrient control or physiological action on structure and function.

The dietary supplement for the prevention or improvement of inflammation, allergy and asthma of the present invention comprises the extract in an amount of 0.01 to 95%, preferably 1 to 80% by weight, based on the total weight of the composition.

In addition, a health function in the form of a pharmaceutical dosage form such as powders, granules, tablets, capsules, pills, suspensions, emulsions, syrups, or tea bags, leaches, health drinks, etc. for the purpose of preventing or improving inflammation, allergies and asthma. It can be manufactured and processed as food.

In addition, the present invention provides a dietary supplement for the prevention and improvement of inflammation, allergy and asthma containing cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold grass) extract as an active ingredient.

In addition, the present invention provides a food or food additive for the prevention and improvement of inflammation, allergy and asthma containing cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold grass) extract as an active ingredient.

In addition, the health functional food may further include food additives, and the suitability as a "food additive" is applied to the item in accordance with the General Regulations and General Test Act of the Food Additives Authority approved by the Ministry of Food and Drug Safety unless otherwise specified. Determined by the relevant standards and standards.

Items listed in the "Food Additives Code" include, for example, chemical synthetic products such as ketones, glycine, potassium citrate, nicotinic acid, cinnamon acid, natural additives such as navy, licorice extract, crystalline cellulose, guar gum, L- Mixed preparations, such as a sodium glutamate preparation, a noodles addition alkali preparation, a preservative preparation, and a tar pigment preparation, are mentioned.

Functional foods containing the extract of the present invention include bread, rice cakes, dried fruits, candy, chocolates, chewing gum, confectionery such as ice cream, ice cream products such as ice cream, ice cream powder milk, low fat milk, lactose-degraded milk Processed milk, Goat milk, Fermented milk, Buttered milk, Concentrated milk, Milk cream, Butter oil, Natural cheese, Processed cheese, Milk powder, Dairy products such as whey, Meat products, Egg products, Meat products such as hamburger Meat products such as processed meat products such as sausages, sausages, bacon, etc. Noodles, dried noodles, raw noodles, noodle soups, dehydrated noodles, refined noodles, frozen noodles, pasta noodles, fruit drinks, carbonated drinks, soy milk products , Lactic acid bacteria beverages such as yogurt, beverages such as mixed drinks, seasoning foods such as soy sauce, miso, red pepper paste, chunjang, cheongukjang, mixed sauce, vinegar, sauces, tomato ketchup, curry Lean, shortening and pizza, but is not limited thereto.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the extract as an essential ingredient in the indicated proportions, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the aforementioned natural carbohydrates include monosaccharides (eg, glucose, fructose, and the like); Disaccharides (eg maltose, sucrose and the like); And conventional sugars such as polysaccharides (eg dextrin, cyclodextrin, etc.), and sugar alcohols such as xylitol, sorbitol, erythritol. As flavoring agents other than those described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. have. The proportion of the natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from 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 may be added to food or beverages for the purpose of preventing the desired disease. At this time, the amount of the extract purified in the food or beverage may be added in 0.01 to 15% by weight of the total food weight, the health beverage composition may be added in a ratio of 0.02 to 5g, preferably 0.3 to 1g based on 100ml have.

Extracts according to the present invention added to foods, including beverages in the process of producing the health functional food can be appropriately added or reduced its content as needed.

Cold grass extract (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold vinegar) extract according to the present invention targets the extract of the genus Pseudolysimachion genus (Veronica longifolia), which is disclosed in Korean Patent Publication No. 10-2006-0125489 Han, leukotriene development inhibitory evaluation test (Experimental Example 1); Effect test on total cell count in total bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) using Balb / c male mouse (Experimental Example 2); Experimental effect on the number of neutrophil cells compared to the total cell count in total bronchoalveolar lavage (BAL) (Experimental Example 3); Effect on the number of Neutrophil + / Gr-1 + absolute cells in BAL fluid (Test Experimental Example 4); Experiments on the effect of CD11b + / Gr-1 + absolute cell number on lung cells (Experimental Example 5); Experiment on the effect on the number of CD4 + / CD3 + absolute cells in lung cells (Experimental Example 6); Experiment on the effect on the number of Macrophage + / CD11b + absolute cells in lung cells (Experiment 7); Effect experiment on the expression level of inflammatory factors in BAL fluid (Experimental Example 8); Influence on lung tissue changes Experimental example (Experimental Example 9) showed that it showed superior anti-inflammatory, anti-allergic and asthma-inhibiting activity than the extract of Veronica longifolia extracted by the conventional method, inflammation, allergy and asthma It can be widely used as a preventive or therapeutic agent for diseases.

1 is a result of measuring the effect on the lung tissue changes following the example sample.

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples.

Comparative Example 1: Preparation of Ginseng Oat Grass Extract

As disclosed in the prior invention (Korean Patent Laid-Open No. 10-2006-0125489), a crude extract of Ginsan tail grass was prepared as follows.

1.1 kg of bergamone (Veronica longifolia, Pseudolysimachion) was dried and pulverized, and 5 liters of methanol was added to the ginko footwort outpost, followed by stirring at room temperature for 24 hours. The supernatant was recovered by vacuum filtration. After repeating this process twice, the supernatant was collected, and concentrated under reduced pressure with a reduced pressure concentrator (EYELA, N-2100, JAPAN) to obtain 100.5 g of a crude extract of long acid tail glucose methanol, which was used as a comparative sample of the following experimental example. (Hereinafter referred to as “VLM”)

EXAMPLE  One: Cold grass Crude extract  Produce

1-1: Preparation of Cold Herb Methanol Extract

Dried and crushed cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold grass, cultivated (Ansan, Gyeonggi-do)) Dispersed 1.16 kg of starch in methanol, repeated 4 L three times at a time, and ultrasonic extractor (5510R-DTH, Bransonic) at room temperature Cold grass methanol extract was prepared by exposing to ultrasonic at 60 ° C. for 2 hours. Subsequently, 120.68 g of a cold cold methanol extract was dried by completely drying through a reduced pressure concentrator (EYELA, N-2100, JAPAN) and freeze drying using a freeze dryer (FDU-2100, Lab corporation) at 40 ° C. Hereinafter referred to as " VSM "

1-2. Preparation of cold grass water extract

Dried and crushed cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold grass, cultivated (Ansan, Gyeonggi-do)) Dispersed 1.08 kg of distilled water in distilled water three times 4 L at a time and ultrasonic extractor (5510R-DTH, Bransonic) at room temperature Cold grass water extract was prepared by exposure to ultrasonic waves at 80 ° C. for 2 hours. Subsequently, 52.3 g of dried cold grass water extract was obtained by completely drying through a reduced pressure concentrator (EYELA, N-2100, JAPAN) and freeze drying using a freeze dryer (FDU-2100, Lab corporation) at 40 ° C. Hereinafter referred to as "VSW") was stored at -20 ℃.

1-3. Preparation of Cold Herb 25% Ethanol Extract

10.0 g of dried and pulverized cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold vinegar, cultivated (Ansan, Gyeonggi)) was dispersed in 25% aqueous ethanol solution and repeated 3 times at 4 L at an ultrasonic extractor (5510R-DTH, Bransonic) was used for 2 hours at 80 ° C. to prepare a cold vinegar 25% ethanol extract, followed by concentration under reduced pressure (EYELA, N-2100, JAPAN) and freeze dryer (FDU-2100) at 40 ° C. , Lab corporation) was completely dried through lyophilization to obtain 1.6 g of dry cold 25% ethanol extract (hereinafter referred to as "VS25E") and stored at -20 ° C.

1-4. Preparation of Cold Herb 50% Ethanol Extract

10.0 g of dried and pulverized cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold vinegar, cultivated (Ansan, Gyeonggi)) was dispersed in a 50% aqueous ethanol solution and repeated 3 times 4 L at a time in an ultrasonic extractor (5510R-DTH, Bransonic) was used for 2 hours at 80 ° C. to produce a cold vinegar 50% ethanol extract, followed by concentration under reduced pressure (EYELA, N-2100, JAPAN) and freeze dryer (FDU-2100) at 40 ° C. After drying completely through lyophilization using Lab Corporation, 1.7 g of dry cold 50% ethanol extract (hereinafter referred to as “VS50E”) was stored at -20 ° C.

1-5. Preparation of Cold Herb 75% Ethanol Extract

10.0 g of dried and pulverized cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold vinegar, cultivated (Ansan, Gyeonggi)) was dispersed in a 75% aqueous ethanol solution and repeated 3 times at 4 L at an ultrasonic extractor (5510R-DTH, Bransonic) was used to produce 75% ethanol extract of cold grass by exposure to ultrasound at 80 ° C. for 2 hours, then concentrated under reduced pressure (EYELA, N-2100, JAPAN) at 40 ° C. and freeze-dried (FDU-2100). After drying completely through lyophilization using Lab Corporation, 12.9 g of dried cold vinegar 75 % ethanol extract (hereinafter referred to as “VS75E”) was stored at -20 ° C.

1-6. Cold grass  Preparation of Ethanol Extract

10.0 g of dried and ground cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum: cold vinegar, cultivated (Ansan, Gyeonggi)) was dispersed in 100% ethanol and repeated 3 times 4 L at a time in an ultrasonic extractor (5510R-DTH, Bransonic Cold ethanol extract was prepared by exposing to ultrasonic for 2 hours at 80 ° C. After that, the mixture was concentrated under reduced pressure (EYELA, N-2100, JAPAN) and freeze-dried at 40 ° C. (FDU-2100, Lab corporation). Completely dried through lyophilization using) to obtain 0.5 g of dried 100 % ethanol extract (hereinafter referred to as "VSE") and stored at -20 ° C.

EXAMPLE  2: Cold grass Nonpolar Solvent  Preparation of Soluble Extracts

120.68 g of the cold grass methanol extract of Example 1 was dissolved in water. The resultant was fractionated with 4 L of normal hexane to form an aqueous layer, which was then fractionated with 4 L of ethyl acetate, and the ethyl acetate solution layer was collected from cold grass to prepare a cold acetate ethyl acetate extract. After drying at 40 ° C. with a reduced pressure concentrator (EYELA, N-2100, JAPAN) under reduced pressure and freeze drying with a freeze dryer (FDU-2100, Lab corporation), 12.6 g of cold ethyl ethyl acetate soluble fraction was dried. Obtained (hereinafter referred to as “VSEA”) and stored at −20 ° C.

Experimental Example  One: Leukotrien ( leukotriene ) Occur Inhibitory ability  evaluation

In order to confirm the inhibitory activity of leukotriene development of the preparation example, experiments were carried out by applying the method described in the literature (Goulet, JL, et al., J Immunol , 164 (9), 4899-4907 (2000). ).

1-1. Experiment method

In order to confirm the inhibitory action of 5-lipoxygenase (Lipoxygenase) of the sample of the above examples (Goulet, JL, et al., J Immunol , 164 (9), 4899-4907 (2000), etc. The leukotriene was quantified and the results are shown in Table 2. In Table 2, the positive control group montelukast (montelukast, PHR1603, Sigma-Aldrich) had a final concentration of 100 μM in the medium, and the remaining test drug was prepared. Examples 1 to 2 were processed to a final concentration of 10 to 50 μg / ml, and Preparation Examples 3 to 7 to a final concentration of 100 to 200 μg / ml.

The final concentration of the extract was adjusted to 10 to 50 μg / ml or 100 to 200 μg / ml, respectively, and treated in 5 × 10 ⁵ divided RBL-2H3 cells (22256, KCLB). After 10 minutes of sample treatment, 20 μg of calcium ionophore (A23187, Sigma-Aldrich) was treated to induce leukotriene. After 10 minutes, the supernatant was collected and leukotriene was quantified at 405 nm using an ELISA reader (Powerwave XS, Biotek) using an ELISA method (enzyme-linked immunosorbent assay, ADI-900-070, Ezno lifescience).

1-2. Experiment result

The results of measuring the amount of leukotriene generation for the samples are shown in Tables 1 and 2 below. In Examples 1 and 2, both of them showed no inhibitory effect at 10 μg / ml, but at 50 μg / ml, the sample of Example 2 was more effective in inhibiting the development of leukotriene than the samples of Example 1 Was found to be more effective in suppressing bronchial inflammation (Table 1). Example 1 In the sample, 100 μg / ml was effective in inhibiting the development of leukotriene except for Examples 1-2 to 1-4, and particularly, the effect was excellent in the Example 1-6 sample. At 200 μg / ml, Examples 1-3 to 1-6 samples except Example 1-2 were effective for suppressing bronchial inflammation, and in particular, Example 1-6 samples were more effective for inhibiting bronchial inflammation ( Table 2).

Leukorrien ( leukotriene ) Occur Inhibitory ability  Experiment result division Leucoterien concentration (pg / ml) Inhibition rate (caused criteria) Normal 136.5 ± 7.9 Triggering group 2603.5 ± 400.9 Positive control group 100 μM 684.4 ± 26.8 74% Example 1-1 10 μg / ml 2794.6 ± 627 - 30 μg / ml 2279.1 ± 441.8 12% 50 μg / ml 2557.3 ± 280.7 2% Example 2 10 μg / ml 2689.9 ± 115.4 - 30 μg / ml 2503.9 ± 293.8 4% 50 μg / ml 1288.7 ± 216.8 51%

Leukorrien ( leukotriene ) Occur Inhibitory ability  Experiment result division Leucoterien concentration (pg / ml) Inhibition rate (caused criteria) Normal 161.7 ± 3.0 Triggering group 2656.8 ± 30.1 Positive control group 100 μM 167.9 ± 0.5 94% Example 1-2 100 μg / ml 2557.7 ± 285.3 4% Example 1-3 2723.8 ± 466.3 - Example 1-4 2645.1 ± 369.4 - Example 1-5 2125.9 ± 112.0 20% Example 1-6 14930 ± 393.0 44% Example 1-2 200 μg / ml 2910.9 ± 467.8 - Example 1-3 1797.5 ± 336.6 32% Example 1-4 1550.8 ± 192.2 42% Example 1-5 764.5 ± 55.5 70% Example 1-6 708.1 ± 83.4 73%

Experimental Example 2: Determination of total cell number in total bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage)

In order to determine the effect on the total cell count in the total bronchial alveolar lavage (BAL) of the sample, the experiment described in the literature was applied as follows (Schins et al., Toxicol Appl Pharmacol . 195 (1), 1-11 (2004) and Smith et al., Toxicol Sci , 93 (2), 390-399 (2006)).

2-1. Experiment method

Six groups of Balb / c male mice were used in each group, except for the normal group, in 0.25 mg / ml coal, 10 mg / ml fly ash, and 0.25 mg / ml diesel exhaust particle (DEP) mixture. The final concentration of Alum is mixed to 8%, and the fine dust mixture is injected into the airway and nose of the experimental animal using the Intra-Nazal-Trachea (INT) injection method described in the literature (Lim et al., Free Radic). Biol Med . 25 (6), 635-644. (1998)) were injected directly at 50 μl on the 3rd and 6th day of the experiment. The positive control, Example samples were orally administered daily (10 days) diluted with 0.5% sodium carboxymethyl cellulose (CMC, 419273, Sigma-Aldrich) solution at a concentration of 200 mg / kg in each group. BAL fluid was recovered by autopsy on day 11 after the experiment.

2-2. Experiment result

Table 3 shows the results of measuring the effect on the total cell number in the total bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) for the samples. The total BAL cell number was decreased compared to the induction group, and it was confirmed that all the samples contributed to the reduction of inflammation level, and Example 1-1 showed a lower total BAL cell number than Comparative Example 1. The total BAL cell number of the Example 2 treatment group was lower than that of Example 1-1, indicating that the Example 2 sample had higher bronchial inflammation inhibitory activity.

Effect of total bronchoalveolar lavage (BAL; bronchoalveolar lavage) on total cell count division Total BAL cell (× 10 ⁵ cells / ml) Inhibition rate (caused) Normal 11.3 ± 4.48 Triggering group 231.0 ± 30.48 Positive control group 117.3 ± 21.35 49% Example 1-1 147.5 ± 30.38 36% Example 2 121.3 ± 19.47 47% Comparative Example 1 206.5 ± 18.71 11%

Experimental Example 3 Measurement of Neutrophil Cell Ratio to Total Cell Number in Total Bronchoalveolar Lavage (BAL)

In order to determine the effect of the total bronchial alveolar lavage (BAL; bronchoalveolar lavage) on the neutrophil cell number to the total cell number of the sample, the experiment described in the literature was applied as follows (Schins et al., Toxicol Appl Pharmacol . 195 (1), 1-11 (2004) and Smith et al., Toxicol Sci , 93 (2), 390-399 (2006)).

3-1. Experiment method

It proceeded in the same manner as in Experimental Example 2. Diff-Qick staining in recovered bronchial alveolar lavage (BAL) (Takano et al., Am J Respir Crit Care Med , 156 (1), 36-42. (1997), Hemacolor Rapid staining of blood smear, 1.11661) .0001, Merck) was observed after staining with neutrophil.

3-2. Experiment result

Table 4 shows the results of measuring the effect of the total bronchoalveolar lavage (BAL; bronchoalveolar lavage) on the ratio of neutrophil cells to total cell numbers. Total neutrophil cell ratio is shown to decrease as compared to the induced group was confirmed that all the samples that contribute to the inflammation levels decreased, and further compared to a lower total BAL in Examples 1-1 than the Example 1 cleaning fluid (BAL; bronchoalveolar The ratio of neutrophil cell to total cell number in lavage was shown, and the ratio of neutrophil cell to total cell count in bronchoalveolar lavage (BAL) was lower in Example 2 than in Example 1-1. The samples showed higher bronchial inflammation inhibitory activity.

Effect of total bronchoalveolar lavage (BAL) on neutrophil cell ratio to total cell count division Cell rate (%) Inhibition rate (caused) Normal 5.3 ± 1.0 Triggering group 99.0 ± 9.0 Positive control group 27.5 ± 7.9 72% Example 1-1 64.5 ± 6.3 36% Example 2 39.0 ± 6.4 61% Comparative Example 1 92.2 ± 5.7 9%

Experimental Example 4 Measurement of Neutrophil + / Gr-1 + Absolute Cell Count in BAL Fluid

In order to determine the effect of Neutrophil + / Gr-1 + absolute cell number in BAL fluid of the sample of the above example, the experiment described in the following literature was applied (Beutner EH., Bacteriological Reviews ., 25 (1): 49-76, ((1961)).

4-1. Experiment method

Bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) was carried out in the same manner as in Experimental Example 2 except for measuring the number of cells. Specific fluorescence fluorescent antibody staining method was performed on the recovered BAL fluid using Gr-1 antibody (553128, BD Biosciences, San Jose, CA, USA) with fluorescent label binding. Neutrophil + / Gr-1 + absolute cell counts in total leukocytes were measured by FACS (Fluorescence-activated cell sorting, BD Biosciences, San Jose, CA, USA).

4-2. Experiment result

Table 5 shows the results of measuring the number of Neutrophil + / Gr-1 + absolute cells in the BAL fluid of the samples. All of them showed a decrease in the number of Neutrophil + / Gr-1 + absolute cells as compared to the induction group. Furthermore, the results showed lower Neutrophil + / Gr-1 + absolute cell numbers in the Example 2 sample treatment group compared to the Example 1-1 sample. Example 2 It was confirmed that the sample showed high bronchial inflammation inhibitory activity.

Effect of Neutrophil + / Gr-1 + Absolute Cell Count on BAL Fluid division Cell number (× 10 ⁴ cells / ml) Inhibition rate (caused) Normal 1.6 ± 0.7 Triggering group 122.0 ± 25.5 Positive control group 86.9 ± 16.9 29% Example 1-1 62.8 ± 8.5 49% Example 2 67.8 ± 14.2 44%

Experimental Example 5 Measurement of CD11b + / Gr-1 + Absolute Cell Number in Lung Cells

In order to confirm the effect on the CD11b + / Gr-1 + absolute cell number in the lung cells of the sample, the experiment described in the following literature was applied (Beutner EH., Bacteriological Reviews ., 25 ( 1): 49-76, ((1961)).

5-1. Experiment method

Bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) was carried out in the same manner as in Experimental Example 2 except for measuring the number of cells. Specific recovery of lungs was performed using fluorescently bound CD11b antibody (553310, BD Biosciences, San Jose, CA, USA) and Gr-1 antibody (553128, BD Biosciences, San Jose, CA, USA). A specific fluorescence fluorescent antibody staining method was performed and CD11b + / Gr-1 + absolute cells in the total lung cell counts were performed using FACS (Fluorescence-activated cell sorting, BD Biosciences, San Jose, CA, USA). The number was measured.

5-2. Experiment result

The results of measuring CD11b + / Gr-1 + absolute cell numbers in lung cells of the samples are shown in Table 6 below. In all of them, the CD11b + / Gr-1 + absolute cell number was reduced compared to the induced group. Furthermore, the sample of Example 2 showed a lower CD11b + / Gr-1 + absolute cell number in Example 2 than in Example 1-1. It was confirmed to exhibit high bronchial inflammation inhibitory activity.

Total lung Cell count  medium CD11b + / Gr -1+ absolute On the cell count  Impact Experiment Results division Cell number (× 10 ⁵ cells / ml) Inhibition rate (caused) Normal 34.5 ± 3.35 Triggering group 81.5 ± 0.96 Positive control group 27.6 ± 8.95 66% Example 1-1 71.8 ± 0.39 12% Example 2 37.7 ± 8.82 54%

Experimental Example 6 Measurement of CD4 + / CD3 + Absolute Cell Count in Lung Cells

In order to confirm the effect on the CD4 + / CD3 + absolute cell number in the lung cells of the sample of the Example was tested by applying the method described in the literature (Beutner EH., Bacteriological Reviews ., 25 (1): 49-76, ((1961)).

6-1. Experiment method

Bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) was carried out in the same manner as in Experimental Example 2 except for measuring the number of cells. Specific recovery of lungs was performed using fluorescently bound CD4 antibody (550280, BD Biosciences, San Jose, CA, USA) and Gr-1 antibody (554829, BD Biosciences, San Jose, CA, USA). A specific fluorescence fluorescent antibody staining method was performed and total lung cell counts were performed using FACS (Fluorescence-activated cell sorting, BD Biosciences, San Jose, CA, USA). Medium CD4 + / CD3 + absolute cell numbers were measured.

6-2. Experiment result

The results of measuring CD4 + / CD3 + absolute cell numbers in lung cells of the samples are shown in Table 7 below. In all of them, the number of CD4 + / CD3 + absolute cells was decreased in comparison with the induction group. Moreover, the CD4 + / CD3 + absolute cell numbers were lower in Example 2 than in Example 1-1. It confirmed that it was shown.

Total lung Cell count  CD4 + / CD3 + absolute On the cell count  Impact Experiment Results division Cell number (× 10 ⁵ cells / ml) Inhibition rate (caused criteria) Normal 75.6 ± 3.14 Triggering group 132.6 ± 7.21 Positive control group 81.4 ± 0.13 39% Example 1-1 103.6 ± 5.26 22% Example 2 80.2 ± 0.34 40%

Experimental Example 7: Determination of Macrophage + / CD11b + absolute cell number in lung cells

In order to determine the effect on the macrophage + / CD11b + absolute cell number in the lung cells of the sample of the Example was tested by applying the method described in the literature (Beutner EH., Bacteriological Reviews ., 25 (1): 49-76, ((1961)).

7-1. Experiment method

Bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) was carried out in the same manner as in Experimental Example 2 except for measuring the number of cells. The recovered lungs were subjected to a specific fluorescence fluorescent antibody staining method using a CD11b antibody (553310, BD Biosciences, San Jose, CA, USA) with fluorescent labels. Fluorescence-activated cell sorting (BD Biosciences, San Jose, Calif., USA) was used to determine the macrophage + / CD11b + absolute cell count among the total lung cell counts.

7-2. Experiment result

Macrophage + / CD11b + absolute cell number in the lung cells of the samples was measured in Table 8 below. All of them showed a decrease in the number of Macrophage + / CD11b + absolute cells compared to the induction group. Furthermore, the Example 2 sample showed a lower number of Macrophage + / CD11b + absolute cells compared to the Example 1-1 sample. It was confirmed that it exhibits inhibitory activity on inflammation.

Total lung Cell count  Macrophage + of CD11b + absolute On the cell count  Impact Experiment Results division Cell number (× 10 ⁵ cells / ml) Inhibition rate (caused criteria) Normal 35.2 ± 3.04 Triggering group 88.8 ± 0.22 Positive control group 35.4 ± 8.10 60% Example 1-1 79.2 ± 0.74 11% Example 2 45.6 ± 9.96 49%

Experimental Example 8: Measurement of inflammatory factor expression in BAL fluid

In order to confirm the effect on the expression level of inflammatory factors in the BAL fluid of the sample of the Example was tested by applying the method described in the literature (Brandt EB et al., J. Allergy Clin. Immunol. , 132 (5): 1194-1204, (2013).

Evaluation tests were performed using ELISA to measure expression of inflammatory factors such as IL-17A, TNF-α MIP2, and CXCL-1 in BAL fluid.

8-1. Experiment process

Bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) was carried out in the same manner as in Experimental Example 2 except for measuring the number of cells. Levels of IL-17A, TNF-α MIP2, and CXCL-1 in BAL fluid were measured by ELISA. IL-17A Antibody (M1700, R & D Systems, Minneapolis, USA), TNF-α Antibody (MTA00B, R & D Systems, Minneapolis, USA) MIP2 Antibody (MM200, R & D Systems, Minneapolis, USA), and CXCL-1 Antibody (MKC00B, R & D Systems, Minneapolis, USA) was diluted with buffer and coated on micro wells and then incubated at 4 ° C. for 16 hours. Each well was washed three times with washing buffer and then 100 μl of serum diluted 10-fold was dispensed.

After being left at room temperature for 1 hour, the cells were washed twice and treated with 100 μl of Avidin-HRP-bound antibody (DY007, R & D Systems, Minneapolis, USA), and washed again after standing at room temperature for 1 hour. Dispense 100 μl of TMB substrate (DY007, R & D Systems, Minneapolis, USA), leave for 30 minutes in the dark, treat 50 μl of stop solution (DY007, R & D Systems, Minneapolis, USA) and measure absorbance at 450 nm. It was.

8-2. Experiment result

As can be seen in Table 9, the inflammatory factors (IL-17A, TNF-α, MIP2, CXCL-1) of the sample administration group was reduced compared to the induction group. Compared with Example 1-1, Example 2 sample treatment group showed lower IL-17A, TNF-α, MIP2, CXCL-1 expression, and it was confirmed that Example 2 sample showed high bronchial inflammation inhibitory activity.

Bronchoalveolar Lavage  ( BAL  in fluid) Inflammatory factor  Effect on Expression Experimental Results division Concentration (pg / ml) /% inhibition based on trigger group IL-17A TNF-α MIP2 CXCL-1 Normal 10.45 ± 3.16 87.53 ± 12.73 79.01 ± 10.38 98.73 ± 11.53 Triggering group 25.11 ± 5.62 166.00 ± 34.77 136.13 ± 9.19 605.36 ± 87.17 Positive control group 19.48 ± 1.89 / 22% 124.57 ± 5.74 / 25% 118.75 ± 9.26 / 13% 229.27 ± 23.21 / 62% Example 1-1 12.53 ± 2.83 / 50% 78.16 ± 9.48 / 53% 121.46 ± 19.77 / 11% 463.24 ± 105.91 / 23% Example 2 9.94 ± 2.33 / 60% 72.43 ± 15.35 / 56% 102.92 ± 9.10 / 24% 373.88 ± 62.31 / 38%

Experimental Example  9: Lung tissue  Inspection Analysis

In order to confirm the effect on the lung tissue change of the sample of the Example was tested by applying the method described in the existing literature as follows.

To isolate lung tissue and identify histopathological abnormalities, the lung cell wall was observed by H & E staining using the method described in the literature (Nandedkar SD et al., Blood ., 112 (6): 2529-2538, ( 2008)).

9-1. Experiment process

Bronchoalveolar lavage fluid (BAL; bronchoalveolar lavage) was carried out in the same manner as in Experimental Example 2 except for measuring the number of cells. The extracted lungs were immediately fixed in a fixed solution of 10% formaldehyde solution (F8775, Sigma-Aldrich, USA), rinsed in washed water for 8 hours, and embedded in epoxy (A3183, Sigma-Aldrich, USA). Sections were prepared by microtome (microtome, Leica RM2265, Wetzlar, Germany) and stained with Hematoxylin & Eosin by standard methods, and observed with a 400 × optical microscope (Bright contrast Microscope, Tokyo, Japan).

9-2. Experiment result

As shown in Figure 1, when seen in the respiratory injury causing group it can be seen that the lung cell wall thicker than the normal group. However, in the group administered with Examples 1-1 to 2, it was confirmed that the lung cell wall was thinned to the normal group level.

Claims (15)

A pharmaceutical composition for the prevention or treatment of inflammation, allergies and asthma, comprising cold extract (Veronicastrum sibiricum L. Pennell; Veronicastrum) as an active ingredient. The method of claim 1,
The cold grass is a pharmaceutical composition, characterized in that the root, stem, or flower of Korea, or imported from China, Russia, Japan. The method of claim 1,
The cold grass extract is a pharmaceutical composition, characterized in that the crude extract, polar solvent soluble extract or non-polar solvent soluble extract. The method of claim 3,
The crude extract is a pharmaceutical composition, characterized in that the extract is soluble in a solvent selected from a lower alcohol having 1 to 4 carbon atoms, such as purified water, methanol, ethanol, butanol, or a mixed solvent thereof. The method of claim 3,
The non-polar solvent soluble extract is a pharmaceutical composition, characterized in that the extract soluble in hexane, methylene chloride, chloroform, or ethyl acetate. The method of claim 1,
The inflammation includes dermatitis, atopic dermatitis, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia, psoriatic arthritis, Pharmaceuticals characterized by a disease selected from the group consisting of osteoarthritis, rheumatoid arthritis, periarthritis, tendinitis, hay salt, peritonitis, myositis, hepatitis, cystitis, nephritis, Sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases Composition. The method of claim 1,
The allergy is a pharmaceutical composition characterized in that it is hypersensitivity, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis, urticaria, insect allergy, food allergy or drug allergy disease. The method of claim 1,
The asthma is bronchial asthma due to dust mites, pollen, animal hair, dandruff, cockroaches, food, drugs, cold, tobacco smoke, indoor pollution, air pollution, food additives, physical activity, climate change, yellow dust, or stress Pharmaceutical composition. A dietary supplement for the prevention or improvement of inflammation, allergies and asthma, containing cold extract (Veronicastrum sibiricum L. Pennell; Veronicastrum) as an active ingredient. A dietary supplement for the prevention or improvement of inflammation, allergies and asthma, containing cold extract (Veronicastrum sibiricum L. Pennell; Veronicastrum) as an active ingredient. The method of claim 10,
The health functional food is a health functional food in the form of powders, granules, tablets, capsules, pills, suspensions, emulsions, syrups, tea bags, leach teas, or health drinks. A dietary supplement for the prevention or amelioration of inflammation, allergies and asthma, containing cold extract (Veronicastrum sibiricum L. Pennell; Veronicastrum) as an active ingredient. A dietary supplement for the prevention or amelioration of inflammation, allergies and asthma, containing cold extract (Veronicastrum sibiricum L. Pennell; Veronicastrum) as an active ingredient. Foods for the prevention or improvement of inflammation, allergies and asthma, containing cold grass (Veronicastrum sibiricum L. Pennell; Veronicastrum) extract as an active ingredient. Food additive for the prevention or improvement of inflammation, allergies and asthma, containing cold extract (Veronicastrum sibiricum L. Pennell; Veronicastrum) as an active ingredient.

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