KR101286463B1 - A composition comprising extract of Cnidii Rhizoma for treating or preventing respiratory disease - Google Patents

Abstract

The present invention relates to a prophylactic or therapeutic agent for respiratory diseases containing Cnidii Rhizoma extract as an active ingredient, and a method for preparing the same, wherein the extract is 5-lipoxygenase inhibitory activity, airway contraction inhibitory activity, airway inflammation inhibitory effect. And it is confirmed that the anti-edema effect is excellent, such as asthma, chronic obstructive pulmonary disease, acute bronchitis, allergic rhinitis, cough, sputum, acute lower respiratory tract infection (bronchitis and bronchiolitis), sore throat, tonsillitis, laryngitis It is found to be useful for the prevention or treatment of respiratory diseases.

Description

A composition comprising extract of Cnidii Rhizoma for treating or preventing respiratory disease}

The present invention relates to a composition for the prophylaxis or treatment of respiratory diseases containing Cnidii Rhizoma extract as an active ingredient, more specifically, the extract of the uterus, 5-lipoxygenase inhibitory activity, airway contraction inhibitory activity, airway inflammation inhibition And by confirming the excellent anti-edema effect of ear edema, acute upper respiratory infections such as asthma, chronic obstructive pulmonary disease, acute bronchitis, allergic rhinitis, cough, phlegm, acute lower respiratory tract infection (bronchitis and bronchiolitis), sore throat, tonsillitis, laryngitis Provided are compositions useful for the prevention or treatment of respiratory diseases.

Asthma, chronic obstructive pulmonary disease, allergic rhinitis, cough, sputum, acute bronchitis, bronchiolitis, pharyngitis, tonsillitis, laryngitis are typical diseases of respiratory diseases.

Asthma refers to chronic inflammation of the airways, especially the bronchus. Inflammation caused by asthma can be exacerbated by a wide variety of predispositions, including soot, allergens, cold winds, exercise, and respiratory infections. Persistent inflammation causes airway deformity and hyper-responsiveness. These causes include general symptoms such as wheezing (a narrowing of the airways, wheezing or crotch breathing), a shortness of breath, coughing, and excessive sputum.

Asthma can be summarized as four pathological symptoms: markedly increased influx of eosinophils in the airways, excessive secretion of mucus, edema observed, and above all, airway It becomes characteristic characteristic that) narrows.

The respiratory tract consists of muscles called mucosa and bronchial smooth muscle, and there are many secretory glands in the mucosa, which continue to secrete the necessary secretions. When the bronchial smooth muscle contracts, the respiratory tract becomes narrow. Inflammation, caused by a variety of predispositions such as fumes, allergens, cold air, exercise, and respiratory infections, increases the secretions from the secretory glands, which block the airways, causing the mucous membrane to swell into the airways, narrowing the airways. . Because of this, seizure cough and dyspnea with severe wheezing appear severely, and during the seizure, a dry cough occurs and you feel chest compressions. Many people with asthma have difficulty breathing without knowing the cause of chronic cough and chest compressions. These symptoms tend to appear suddenly in everyday life.

Currently, the concept of asthma has been newly established from bronchial stenosis to chronic bronchial inflammatory disease, and it is important to alleviate symptoms when there is a symptom, but it is important to manage the inflammation in the long term.

Asthma is a pathological / physiological cause of airway inflammation, airway-hyperresponsiveness (AHR), mucin hypersecretion, and immunologically invasive eosinophils. It is a chronic airway inflammatory disease, characterized by an increase in the number of Th2 cells and an increase in the number of activated mast cells. Asthma symptoms are characterized by the infiltration of eosinophils, granulocyte lineages, into the airways, and eosinophils play a very important role in the pathology / physiology of asthma by producing various triggers that promote airway inflammation and bronchial contraction. To perform. Asthma-causing antigens cause differentiation of T cells into Th2 cells, and Th2 cells, such as IL (interleukin) -5, granulocyte-macrophage stimulating factor (GM-CSF), IL-3, IL-13, and IL-4 Cytokines are secreted, and IL-4 acts on B cells to promote the production of IgE and to activate mast cells. When inflammatory cells, such as mast cells, are activated, various inflammatory mediators are advantageous and cause acute inflammatory reactions such as bronchial contraction, vasodilation, sensory nerve sensitization, and cholinergic bronchoconstriction. Most asthma is reversible, but in some patients, asthma progresses, remodeling occurs due to fibrosis of the upper and lower cells, an increase in the number of blood vessels and mucous secretions, and changes in the airway structure due to thickening of airway smooth muscle, and chronic obstructive pulmonary disease. Airway obstruction may also occur in Chronic Obstructive Pulmonary Disease (COPD).

Chronic obstructive pulmonary disease refers to a group of diseases in which airflow obstruction occurs and the air velocity decreases without causing lung disease or heart disease. Clinically, when chronic bronchitis with cough with chronic sputum and alveolar alveoli below the end-bronchial bronchi are abnormally expanded and the emphysema with which the alveolar septum is destroyed is difficult to distinguish, these are collectively referred to as chronic obstructive pulmonary disease. Chronic obstructive pulmonary disease, similar to asthma, symptoms of respiratory distress, cough, sputum and other airway disease symptoms worsen lung function eventually leading to death. The main cause of the disease is smoking, in addition to pollution and congenital diseases, respiratory infections, and the cause is caused by neutrophils (macrophage) and macrophages (macrophage) is known. Neutrophils are representative inflammatory cells that secrete several types of proteases, causing the destruction of lung parenchyma and chronic mucus secretion. Macrophages are important inflammatory cells of IFN-γ (interferon-γ) and IL-13 overexpression. In addition to secreting mediators that cause tissue damage (reactive oxygen species [ROS], NO [nitric oxide] metabolites, etc.), they are also involved in wound healing (TGF-β [transforming growth factor-β], FGF2 [fibroblast growth factor 2], VEGF [endothelial growth factor], etc.) is a cell that causes the chronic inflammation to secrete. Irreversible airway obstruction is characterized by small airway fibrosis (obstructive bronchiolitis), which is characterized by fibrosis due to pulmonary emphysema caused by destruction of the alveoli and inflammation of the small airways and subsequent repetitive damage. There is this. Most patients with these diseases are accompanied by emphysema and inflammation of the small airways and fibrosis.

Both asthma and chronic obstructive pulmonary disease show symptoms of respiratory distress, cough, and sputum, and there are no common causes of the cure for the disease itself. Currently, there are only two treatments for asthma, which are inflammation inhibitors and relief agents that alleviate respiratory distress. This is because the appearance of asthma is similar to that caused by a wide variety of pathogens and triggers. In addition, it is thought that it is difficult to study the disease itself. In particular, the majority of severe asthma, which accounts for a large portion of asthma-related total mortality and economic burden, is treated with high-dose steroids, but there is still a problem that asthma is not well controlled. Asthma treatment should be combined with drug therapy, environmental therapy (hemotherapy), and immunotherapy, and if left for asthma for a long time, a scar on the mucous membrane of the bronchus will be unrecoverable, and as a result, asthma is often worsened. It is important to treat it thoroughly, and inflammatory treatment of the bronchial mucosa continues, even if symptoms disappear, resulting in bronchial damage. Asthma can be chronically and temporarily improved and then recur, and even worse, even during treatment, especially when pediatric asthma is actively treated early, reducing the number of asthma attacks and reducing the degree of seizures. Asthma is an allergic disease, so it is not easy to treat, and unless it is fundamentally treated, it will easily recur when exposed to the triggering environment, so thorough environmental management such as drug therapy is required.

Another typical respiratory disease, allergic rhinitis, also known as nasal allergy or allergic rhinitis, symptoms suddenly sneezing in succession, clear runny nose, large amounts of nasal congestion, and at the same time the head is heavy and tears. The symptoms are very similar, but the antigen is not clear is called vasomotor rhinitis. In other words, when you wake up in the morning of the day and the body temporarily cools, the symptoms as described above may occur and heal in a few hours. It is often confused with a cold, which is different from the common cold, often accompanied by asthma or hives.

Allergic reactions of allergic rhinitis include a kind of antigen and antibody hypersensitivity, release of histamine from the membranes of mast cells and basophils, release of arachidonic acid, and cyclooxygenase (COX) and 5-lipoxygenase. Prostaglantines and leukotrienes are produced by (5-lipoxygenase, 5-LO) to mediate the initial response between 2 to 90 minutes of antigen exposure and the late response after 4 to 8 hours. The initial response is mainly due to the mediator and the late response is mainly due to the cell infiltration. In addition, allergic and nonallergic rhinitis both act as risk factors for the development of asthma. Treatment of allergic rhinitis involves desensitization when the antigen is clear, and there are other drug therapies, surgical therapies, and physical therapies, but it is difficult to cure them completely.

Jinhae refers to stopping coughing, and expectoration means removing phlegm. Coughing and sputum may occur due to physicochemical factors such as cold air, external foreign substances including pathogenic microorganisms, air pollutants and allergens. Coughing is a reflexive defense mechanism caused by airway mucosal stimulation that, when prolonged coughing caused by excessive stimulation, reduces and worsens the quality of life of the patient. In addition, when dust or irritants are introduced from the outside in the same principle as coughing, the body's bronchus pushes the muscles out with saliva along with saliva, which is the cause of sputum (sputum) formation and inflammation caused by bronchial inflammation such as the lungs. There is a thick purulent sputum.

Acute bronchitis is not a separate disease but is accompanied by other upper and lower extremities. Acute bronchitis often comes with upper respiratory infections such as nasopharyngitis and influenza, whooping cough, measles, typhoid fever, diphtheria, scarlet fever infections, and all but bacterial diseases are due to viral infections. Occasionally, sputum is found in pneumococci, Staphylococcus aureus, Haemophilus influenzae, and various types of research, but this does not mean that bacterial infections are the cause, and antibiotic treatment does not affect the course of the disease. The main symptoms are redness, swelling, and dryness in the bronchial mucosa, mucus or purulent secretions can be seen in the bronchus. It usually recovers without causing complications, but when it is chronically developed, it causes swelling, thickening, and atrophy of the mucous membrane, which can lead to proliferation of fibers, bronchial stenosis, and emphysema. .

Chronic bronchitis is a sputum that lasts for more than three months a year for two consecutive years and coughs persist. Stimulation of smoking, air pollution, occupational exposure, etc. causes bronchial damage, which is thought to cause chronic bronchitis. Chronic coughing, sputum, and difficulty breathing are the main symptoms. As the disease progresses, respiratory distress gradually develops over several months and years, resulting in difficulty in breathing even with a slight activity.

Laryngitis is caused by inflammation of the larynx itself by infection with viruses or bacteria, or by inflammation of surrounding tissues such as pharyngitis and tonsillitis spread to the larynx. This is often a partial symptom of a cold, most often accompanied by a cold (acute rhinitis) or pharyngitis, coughing, and voice changes. Laryngitis is an infectious disease commonly referred to as upper respiratory tract disease, and it is difficult to clearly distinguish it from pharyngitis, laryngitis and bronchitis.

Bronchiolitis (bronchiolitis) is an inflammation of the lungs causing dyspnea, also known as bronchitis. Inflammation of the lining of the bronchioles causes the inner wall cells to clump or swell, resulting in narrowing of the lumen and thus shortness of breath, contagious and often prevalent. The bronchioles carry air from the bronchus to the alveoli and the alveoli supply oxygen to the blood. The cause may be a virus or bacteria infection or both. In some cases, the disease develops with each cold. Risk factors include, among others, diseases that reduce resistance, particularly respiratory infections, family history of allergic diseases, and obesity. Complications include permanent pulmonary disease, chronic bronchitis, partial collapse of the lungs, bronchiectasis, recurrence of pneumonia, and rarely chronic obstructive pulmonary disease.

Sore throat (laryngopharyngitis) is a common cold, throat and upper respiratory tract infection. Beta hemolytic streptococci, staphylococci, pneumonia, bacteria such as Haemophilus and anaerobic strains or viruses such as influenza virus, herpes simplex virus, parainfluenza virus, coxsackie virus, echo virus, etc. Inflammation of the upper respiratory tract mucosa, including acute and chronic. Acute is caused by rapid temperature changes, colds, febrile illnesses, overwork, weak constitution, bacterial infections, and chronic recurrences of acute sore throat, excessive smoking, drinking, overwork, irritating food intake, neck overuse, This may be caused by a sore throat reflux disease. Rarely, it may be caused by inhaling irritating gases, chemicals, or chemical vapors, or by spreading inflammation from adjacent areas such as the sinuses.

As described above, respiratory diseases such as asthma, chronic obstructive pulmonary disease, allergic rhinitis, acute bronchitis, bronchiolitis, pharyngitis, tonsillitis, laryngitis have some differences in their causes and symptoms, but they have commonalities in terms of inflammatory diseases. Among the drugs currently in use, airway dilators have no effect on inflammation that worsens the disease, and thus only relieve symptoms. In addition, steroids, which are known to be effective against inflammation, are problematic for long-term use due to serious side effects. Therefore, a combination of the two is often prescribed, but due to the side effects of steroids developed in the form of inhalants than the oral drug is difficult to take, there is a problem in taking compliance.

Therefore, there is an urgent need to develop a new therapeutic agent that can overcome the limitations of these current drug therapies, thereby fundamentally treating the cause and effectively improving the symptoms. However, respiratory diseases are involved in various leukocyte cells and various cytokines and inflammatory mediators that are released here. Therefore, it is thought that effective treatment with single-component synthetic drugs is difficult. It seems to be an effective treatment.

To this end, the present invention provides a cheongung extract as a composition for the prevention or treatment of respiratory diseases described above.

Cnidii Rhizoma is a perennial herb belonging to the umbelliferae , a medicinal resource plant using the root stem of Ligusticum chuanxiong Hort. Or Cnidium officinale Makino. Main ingredients include essential oils such as nidlide, ligustilide, butylidenephthalide and senkyunolide (Chem. Pharm. Bull., 1984, 32, 3770) Phenolic substances (Yakugaku zasshi, 1989, 109, 402-406), such as Korean J. Phamacogn., 1990, 21, 69-73), ferulic acid, chlorogenic acid, etc. This is known. Cheongung is known for its pharmacological effects such as jingyeong, sedation, blood pressure lowering, vasodilation, antibacterial action, and antifungal action (Korean Society Tabacco Sci., 1994, 16, 20-25). It is used for diseases such as menstrual irregularity, tonic, coldness, and anemia (National Institute of Pharmacology, Modern Herbal Medicine, 1994, Pp. 338-340, Hakchangsa).

In addition, the cheongung extract of the present invention can prevent or treat respiratory diseases due to the activity of inhibiting the activity of 5-lipoxygenase (5-lipoxygenase, 5-LO). 5-lipoxygenase is known to be the major action enzyme required to produce leukotrienes (LTs) from arachidonic acid (Science, 1983, 220, 568-75). The leukotriene thus produced is known to be a major cause of airway contraction and inflammation, which is a serious symptom in respiratory diseases including asthma, bronchitis, and the like. The production of these leukotrienes by inhibiting 5-lipoxygenase It is thought to be able to cure respiratory diseases mainly by blocking airway contraction and relieving inflammation.

The production of leukotrienes by 5-lipoxygenase is as follows. First, IgE-mediated activation of mast cells (immune cells that are the main causes of allergy, surface factors to which IgE antibodies can attach to the surface of mast cells) is derived from IgE-mediated activation. By phospholipase A2 is activated by the phospholipidase A2, the phospholipidase generates arachidonic acid (arachidonic acid) from the phospholipids (phospholipids) in the cell membrane. In addition, as a signal for stimulation of mast cells, 5-lipoxygenase is transferred from the cytoplasm to the nuclear membrane, and 5-lipoxygenase activating protein in the cytoplasmic nuclear membrane is used to inhibit 5-lipoxygenase. Activated (Science, 1983, 220, 568-75). The activated 5-lipoxygenase oxidizes arachidonic acid, thereby converting arachidonic acid to hydroperoxyeicosatetraenoic acid (HPETE), which is produced by LTA synthase (LTA synthase, Rukotriene A synthase). To LTA4 (Lucotriene A4) and then to LTB4 (Lucotriene B4) or LTC4 (Lucotriene C4) (Science, 1983, 220, 568-75).

LTA4 (Lucotriene A4) is converted to LTB4 by LTA4 hydroxylase (LTA4 hydroxylase). Alternatively, LTA4 (Lucotrienne A4) is converted to LTC4 (Lucotrienne C4). ) Is converted to LTC4 (rucotriene C4) by synthetase. The LTC4 (rucotriene C4) thus produced is transported out of cells and metabolized to LTD4 (rucotriene D4) and LTE4 (rucotriene E4) (Science, 1983, 220, 568-75).

LTB4 (Luctorien B4) is a powerful chemoattractant such as neutrophils, eosinophils, and monocytes, attaching phagocytes to the vessel wall and degranulating neutrophils. It is known as a major factor that causes airway inflammation by producing superoxide anions. LTC4 (Lucotriene C4) and its metabolite LTD4 (Lucotriene D4) are potent bronchoconstrictors known to increase vascular permeability and increase mucous secretion of the airways (Science, 1983, 220). , 568-75).

LTC4 (rucotriene C4), LTD4 (rucotriene D4) and LTE4 (rucotriene E4) all have cysteine residues, collectively called cysLTs, most of which are mast cells Produced in neutrophils, eosinophils, and alveolar macrophage, acts on cysLT1 (cysLT receptor type 1) to stimulate airway smooth muscle, causing airway contraction, and inflammatory reactions in respiratory diseases such as mucus hypersecretion and asthma Are known to be the major contributing factors (Science, 1983, 220, 568-75).

Inhibition of 5-lipoxygenase effectively blocks the production of these leukotrienes and can be used to treat respiratory diseases mainly by suppressing airway contraction, suppressing mucous secretion and reducing inflammation (Curr Med Chem). ., 2007, 14, 1966-77; Prostaglandins & Lipid Mediators, 2007, 83, 188-97; Pediatrics, 2008, 122, e1249-55). The present invention was completed by confirming that there is a contraction inhibitory activity, an airway inflammation inhibitory effect, and an edema anti-inflammatory effect, and preparing a extract of the uterus showing the synergistic effect of the highest efficiency.

Korean Patent Laid-Open Publication No. 2006-128153, which discloses ethanol extract of cheongungung in the prior art, has an anti-inflammatory effect by inhibiting 5-lipoxygenase activity, but it is an invention related to a composition containing cheongung extract excellent in treating acne, The use of the composition is limited to cosmetics, and the extract has a therapeutic effect against inflammation of respiratory diseases by inhibiting 5-lipoxygenase activity and the effect of fractions fractionating the ethanol extract with butanol. It is not disclosed in the description.

In addition, it has been disclosed that water extracts of herbal medicines containing the uterus inhibit bronchial spasms induced by histamine and acetylcholine in the guinea pig model, and are effective in allergic infections (Zhongguo Zhong Xi Yi). Jie He Za Zhi, 1994, 14 [8], 465-8), the extract only discloses water extracts.

In addition, the water extracts containing some of the uterus and the ethyl acetate fraction thereof have the effect of preventing or treating skin diseases by inhibiting inflammation by inhibiting ear edema and infiltration of inflammatory cells in mice. Although disclosed, the herbal extracts are water and organic solvent extracts thereof, and also through the inhibitory activity of 5-lipoxygenase to inhibit airway contraction, respiratory tract inflammation and anti-edema anti-inflammatory effect of respiratory diseases. It is not confirmed anywhere in the detailed description of Korean Patent Publication No. 2010-4215.

An object of the present invention is to provide a composition for the prevention or treatment of respiratory diseases containing the extract as an active ingredient.

Still another object of the present invention is to confirm that the extract of the uterus is remarkably excellent in 5-lipoxygenase inhibitory activity, airway contraction inhibitory activity, airway inflammation inhibitory effect and ear edema anti-inflammatory effect.

The present invention relates to a five-lipoxygenase inhibitory activity, airway contraction inhibitory activity, airway inflammation inhibitory effect, uterine edema anti-inflammatory effect and a method for preparing the same.

The cheongung extract according to the present invention means a fraction which is a crude extract or a specific solvent soluble extract of the crude extract, which may be in solution, concentrate or dry powder.

The present invention confirms that the Rhizome extract has a remarkable effect on 5-lipoxygenase inhibitory activity, airway contraction inhibitory activity, airway inflammation inhibitory effect and ear edema anti-inflammatory.

The cheongung extract according to the present invention concentrates the crude extract (primary extract) obtained by extraction with a lower alcohol having 1 to 4 carbon atoms or an aqueous solution thereof, and then re-fractions with a lower alcohol having 1 to 4 carbon atoms or an aqueous solution thereof or a nonpolar solvent. One fraction (secondary extract).

The lower alcohol having 1 to 4 carbon atoms may be preferably selected from ethanol, methanol, propanol, isopropanol, sec-butanol and butanol. The nonpolar solvent may be selected from the group consisting of ethyl acetate, dichloromethane, chloroform and butanol. The mixing ratio of the alcohol in the aqueous alcohol solution of 1 to 4 carbon atoms may be mixed in 10 to 95% (v / v), preferably 30 to 95% (v / v). When using butanol as an extraction solvent, it is preferable to use saturated butanol (aqueous solution of 75 to 85% butanol). At this time, the extraction temperature is preferably 40 to 120 ℃, preferably 60 to 90 ℃, extraction time is 2 to 24 hours, preferably 4 to 12 hours.

In addition, the crude extract of the uterus (primary extract) is concentrated and again added to the lower alcohol having 1 to 4 carbon atoms, their aqueous solution or non-polar solvent individually (or in combination) to further fractionate it to obtain a fraction (secondary extract). Can be. The lower alcohol having 1 to 4 carbon atoms may be preferably selected from ethanol, methanol, propanol, isopropanol and butanol. The nonpolar solvent may be selected from the group consisting of ethyl acetate, dichloromethane, chloroform and butanol. At this time, the extraction temperature is preferably 40 to 120 ℃, preferably 60 to 90 ℃, extraction time is 2 to 24 hours, preferably 4 to 12 hours.

In addition, to the concentrate of the crude extract (primary extract), suspended by adding 5 to 20 times (v / w) of purified water, followed by ethyl acetate, dichloromethane, chloroform and butanol (including saturated butanol). Fractions (secondary extracts) can be obtained by adding them separately (or sequentially) from the organic solvent group and further extracting them. At this time, the extraction temperature is preferably 40 to 120 ℃, preferably 60 to 90 ℃, extraction time is 2 to 24 hours, preferably 4 to 12 hours.

In addition, the crude extract may be suspended by adding purified water and then hexane, ethyl acetate and butanol may be sequentially added to obtain an organic solvent fraction (secondary extract).

Extract of the present invention is not limited to the number of extraction depending on the extraction efficiency. When the amount of the solvent used in the preparation of the cheonunggong extract is too low, the solubility of the extract is not only low extraction efficiency, but also a problem that the filter is clogged during filtration, if too large, the amount of lower alcohol used is not economical because Since handling problems may occur, the amount of the solvent to be used is preferably within the above range.

In the present invention, a method of re-fractionation after the first extraction was selected. This is because when the extract of the herbal medicine is extracted in a large amount, even though the extract is filtered after the first extraction, the extraction efficiency is increased because the content of the extraction solvent is high in the herbal medicine itself. This is to prevent falling. In addition, as a result of verifying the extraction efficiency of each step, it was confirmed that about 90% of the total extraction amount was extracted by the first and second extraction, and the third or more extraction method was inefficient due to economic efficiency compared to the yield. Judging.

The cheongung extract thus obtained is 2 to 10 times (v / w), preferably 4 to 6 times (v / w) water relative to the weight of the concentrate to remove the remaining lower alcohol to be suitable for use as a pharmaceutical raw material After azeotropically concentrating 2 to 8 times and again adding water to homogeneously suspend, the powdery Cheongung extract can be prepared using conventional drying methods such as reduced pressure drying, spray drying, or freeze drying.

As the extraction method used in the present invention, all conventionally used methods may be used, and for example, immersion (cold and warm), hot water extraction, ultrasonic extraction, or reflux cooling extraction may be used, but is not limited thereto.

In another aspect of the present invention, the extract of the uterine extract of the present invention fractionated cheonryung crude extract, 5-lipoxygenase inhibitory activity, airway contraction inhibitory activity, airway inflammation inhibitory activity and ear edema significantly higher than that of crude extract Since it has an anti-inflammatory effect, it can be usefully used for preventing or treating respiratory diseases or symptom relief.

Therefore, the present invention provides a composition for the prevention or treatment of respiratory diseases containing the extract as an active ingredient.

The respiratory disease may be any disease causing inflammation and airway contraction, generally acute bronchitis, catarrhal bronchitis, obstructive bronchitis, inflammatory bronchitis, bronchial asthma, atopic asthma, non-atopic asthma, atopic IgE mediated asthma, Allergic asthma, non-allergic asthma, chronic bronchial contraction, acute bronchial contraction, chronic obstructive pulmonary disease, bronchial adenomas, pulmonary tuberculosis, emphysema, lung abscess and the like.

The content of the cheongung extract as an active ingredient in the composition according to the present invention may be appropriately adjusted according to the use form and purpose, the severity of the patient, etc., and may be 0.01 to 99.9% by weight, preferably 0.1 to 50% by weight, based on the weight of the solids. However, the present invention is not limited thereto.

In addition, the above composition may be administered in various formulations of oral and parenteral administration in actual clinical administration. In the case of formulation, a diluent or excipient such as a filler, a weight agent, a binder, a wetting agent, a disintegrant, Can be used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which solid preparations comprise at least one excipient in crude extracts of celestial and certain solvent soluble extracts of crude extracts, or compounds isolated therefrom. For example, starch, cellulose, calcium carbonate, crospovidone, light silicic anhydride, lactose, magnesium stearate, talc, enzymatically treated stevia, microcrystalline cellulose, magnesium stearate, gelatin and the like can be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, and freeze-drying agents. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like.

The dosage of the extract-containing composition of the present invention may vary depending on the patient's worm, age, sex, health status, diet, time of administration, administration method, excretion rate and severity of the disease, and for a predetermined time depending on the judgment of the doctor or pharmacist It may be administered once or several times a day at intervals. For example, the daily dose may be 0.001 to 500 mg / kg, preferably 0.1 to 200 mg / kg, based on the active ingredient content.

In another aspect, the present invention provides a health functional food for the prevention or amelioration of respiratory diseases such as asthma, acute bronchitis, chronic obstructive pulmonary disease containing cheongung extract. The health functional food may be various foods, beverages, food additives and the like.

The content of the extract as an active ingredient contained in the health functional food may be appropriately contained according to the form of the food and the intended use thereof, for example, may be added to 0.01 to 50% by weight of the total food weight, preferably It may be added at a ratio of 0.1 to 10% by weight. In addition, the health beverage composition may be added at a ratio of 0.02 to 10 g, preferably 0.1 to 5 g based on 100 ml.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient, as long as it contains the extract as an essential ingredient at the indicated ratio, and there is no particular limitation to the liquid ingredient. Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like Sugar, and sugar alcohols such as xylitol, sorbitol, and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above 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 thickeners, 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 can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0.001 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

In particular, the cheongungung of the present invention, when administered to the human body as a herbal medicine that has been used in herbal medicine since ancient times, it is considered that there is no concern of side effects compared to other synthetic medicines, and the results of toxicity tests on the herbal composition actually standardized through animal experiments. It turned out to have no effect on me.

The cheongung extract according to the present invention was confirmed that the 5-lipoxygenase inhibitory activity, airway contraction inhibitory activity, airway inflammation inhibitory effect and ear edema anti-inflammatory effect, these activities are in the fraction state than the crude extract of the uterus It was confirmed that the effect was significantly increased in the extract. In addition, the present invention shows a good effect in animal models of various anti-asthmatic diseases, unlike the conventionally used therapeutics, asthma (bronchial asthma, atopic asthma, atopic bronchial IgE mediated asthma, non-atopic asthma, allergic asthma) And non-allergic asthma), acute bronchitis, as well as chronic obstructive pulmonary adenoma, pulmonary tuberculosis, empyema, lung abscess, cough, sputum, allergic rhinitis, acute lower respiratory tract infection (bronchitis and bronchiolitis), catarrhal bronchitis and obstructive or It is thought to be very useful for respiratory diseases such as inflammatory bronchial disease, sore throat, tonsillitis and laryngitis.

1 shows bronchial contraction inhibitory activity after treatment of the extract bronchus of 0.3 mg / ml of Examples 1 to 33 and Comparative Examples 1 to 3 with a bronchial bronchus of guinea pigs.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the invention to those skilled in the art.

Example 1 Preparation of Butanol Fraction of 50% Aqueous Ethanol Solution

100 g of the uterus was extracted by reflux cooling with 1000 ml of 50% ethanol aqueous solution twice at 88 ° C. for 4 hours, and the extract was filtered and concentrated under reduced pressure. The obtained concentrate was further suspended by adding distilled water five times the weight of the concentrate, followed by lyophilization, to obtain 23.4 g of a crude extract of the celestial powder.

Thereafter, 10 times (v / w) purified water was suspended in the crude extract, and the same amount of saturated saturated butanol (80% butanol aqueous solution) was added thereto to separate the layers two times, and only butanol was collected by filtration and concentrated under reduced pressure. Most of butanol and purified water were evaporated, and about 100 ml of purified water was added thereto to concentrate azeotropically, and then repeated twice to completely remove butanol. The concentrate thus obtained was further suspended by adding distilled water five times the weight of the concentrate, and then lyophilized to obtain 3.4 g of a powdery fraction.

Examples 2 to 33 and Comparative Examples 1 to 3 were performed under the conditions of Table 1 below, but other conditions were performed in the same manner as in Example 1. However, in the case of water extract was extracted with reflux for 8 hours, and then immediately lyophilized.

< Experimental Example  1: 5- Lipoxygenase  Inhibitory effect>

The following method was performed by using the cells to evaluate the activity inhibitory ability against the leukotriene producing enzyme, which is the main pathogenesis of asthma, of the astronomical extracts prepared by Examples 1 to 33 and the extracts prepared by Comparative Examples 1 to 3. The results were as shown in Table 2.

To this end, rat basophilic leukemia cells (Rat basophilic leukemia cells, RBL-1) were stabilized at 37 ° C. using RPMI medium without serum, and then extracts were added and arachidonic acid (arachidonic acid) was added as a substrate. Lucotriene was produced for a minute. The amount of produced rucotrienes was measured using a cysteinyl leukotriene EIA kit. Positive controls were zileuton and montelukast, inhibitors of 5-lipoxygenase.

As shown in Table 2, it was found that the cheongung extract of Examples 1 to 33 of the present invention is excellent in inhibiting 5-lipoxygenase.

< Experimental Example  2: bronchial contraction inhibition test ( in vitro )>

In order to evaluate the bronchial contraction inhibitory activity of the 17 examples and the extracts of Comparative Examples 1 to 3 having the best 5-lipoxygenase inhibitory activity in the extracts of Examples 1 to 33, the following method ( Nature, 1977, 270, 256-257; Jpn. J. Pharmacol., 1996, 72, 1-8; Kor. J. Pharmacogn., 1999, 30 [4], 377-383). 1 is shown. The results are shown in Fig. In Figure 1 positive control rolipram (rolipram) was 20μM, each extract was treated with 0.3mg / ㎖, the inhibition rate of bronchial contraction (%) by concentration for the eight examples of the excellent effect of which is shown in Table 3 It was.

To this end, Hartley male guinea pigs (400-500 g, BGI, Korea) were sensitized by intravenous injection of anti-ovalbumin anti-serum at a volume of 2 ml / kg. 48 hours after sensitization, guinea pigs were bled and the bronchus was removed. Thereafter, the other tissues attached to the bronchus were removed from the thyroid liquid, and the cartilage was cut in a ring shape so that two to three cartilages were included. After the incision, the cartilage of the ring was cut while preserving the bronchial muscles, the threads were connected to both sides, and then suspended in an organ bath. After stabilization of a certain time, 10 μg / ml of carbachol was added. Maximum contraction was induced, and the bronchus was washed and stabilized with tyrode fluid. After this, 5 μM of indomethacin (indomethacin) was added and the extract was added after 1 minute, and after 5 minutes, 20 μg / ml of ovalbumin was added to cause contraction. Shrinkage (%) was calculated by comparing the contraction by carbacol and egg albumin. Bronchoconstriction was measured using a physiological activity meter (powerlab 8/30, ADInstument) connected to a force transducer (WPI).

As shown in Figure 1, the cheongung extract of Examples 1 to 33 has an excellent bronchial contraction inhibitory activity, it can be seen that it shows a good dose-dependent activity, and also that the fraction has a significant inhibitory effect than crude extract Could.

Experimental Example 3 Airway Shrinkage Inhibition Test in vivo )>

In order to evaluate the airway contraction inhibitory activity of the extract of Example 8 and the extracts of Comparative Examples 1 to 3 excellent in bronchial contraction inhibitory activity in Experimental Example 2 using the respiratory volume measurement in the guinea pig (European Journal of Pharmacology, 2000, 403, 169-179; Br. J. Pharmac., 1983, 78, 67-74), and the results are shown in Table 4 below.

Hartley male guinea pigs (400-450 g, SLC, Japan) were sensitized by intravenous injection of anti-albumin antiserum at a volume of 1.5 ml / kg. 48 hours after sensitization, the preparation to be tested for guinea pigs was administered orally. Thirty minutes after oral administration, pyrilamine maleate (0.5 mg / kg) and propranolol (propranolol (0.05 mg / kg) were injected subcutaneously and pretreated. ) Was placed in a double chamber plethysmograph box type 855 (HSE, German) and the basic airway resistance (RxV) was measured. After 30 minutes of pretreatment, 1% egg albumin was aerosolized using high pressure compressed air, sprayed for 2 minutes, and air containing albumin was flowed out for 30 seconds. Airway resistance was then measured in a pletismometer box for 15 minutes and airway resistance was calculated by AUC (area under the curve), and is shown as percent inhibition in Table 4 compared to solvent control. Montelukast was used as a positive control in this experiment.

As shown in Table 4, it was found that the cheongung extract of Examples 1 to 33 of the present invention showed a dose-dependent excellent activity in the airway contraction inhibitory effect of sensitized guinea pigs. In addition, it can be seen that the fraction has a significant inhibitory effect than the crude extract.

< Experimental Example  4: bronchial inflammation inhibition test>

In order to determine whether the extract of Example 8 and the extracts of Comparative Examples 1 to 3 having excellent bronchial contraction inhibitory activity in Experimental Example 2 have an inhibitory effect on pulmonary bronchial inflammation in the mouse asthma model, the antigen was exposed to sensitized mice. Leukocyte increase response of the induced bronchioles was confirmed (Pharmacological Research, 2010, 61, 288-297; Biochemical Pharmacology 2010, 79, 888-896), and the results are shown in Table 5.

To this end, BALB / c female mice (6.5 weeks old, SLC, Japan) were mixed with 0.2 ml of 10 μl of egg albumin (Ovalbumin, OVA, sigma) and Alum (Pierce) 1: 1 mixture for 0, 7 and 14 days. Intraperitoneal administration to sensitization. After 8 and 10 days of final sensitization, 1.0% egg albumin was aerosolized using high-pressure compressed air and sprayed for 50 minutes to induce airway inflammation. 10 mg / kg of rolipram (sigma) was used as a positive control drug, and the administration of the positive control drug and the test substance was administered orally twice a day from 21 to 23 days after sensitization. Morning administration on the day of challenge was performed 1 hour before challenge. 24 hours after the last inflammation, the waste broth was washed with 1.5 ml of phosphate buffer (pH 7.2) to collect the washing solution. Leukocyte cell counts in the washes were fractionated using a Hematology analyzer (Hematology analyzer, Drew Scientific Inc., HEMAVET® HV950FS, M-950HV). The results are shown in Table 5 below to calculate the inhibition rate (%) based on the leukocyte cell count of the negative control group (1% CMC [carboxymethyl cellulose] administration group).

As shown in Table 5, it was confirmed that the leukocyte increase in the bronchial bronchus caused by exposing the antigen to the sensitized mouse of the horoscope extracts of Examples 1 to 33 of the present invention was dose-dependently suppressed. In addition, it can be seen that the fraction has a significant inhibitory effect than the crude extract.

< Experimental Example  5: On ear edema  Anti-inflammatory inhibitory effect test for>

To evaluate the anti-inflammatory activity of the extract of Example 8 and the extracts of Comparative Examples 1 to 3 excellent in bronchial contraction inhibitory activity in Experimental Example 2, mice using edema induced by croton oil were used. The results are shown in Table 6. Croton oil, when applied to the skin, causes inflammatory effects of redness, swelling and blisters on the surface of application (Archives of Pharmacal Research, 1993, 16 [1], 18-24).

As a test animal, male ICR mice (Oriental Bio, Korea) having a body weight of 20-23 g were used in each group. The mice fasted the previous day were orally administered with a sample and a positive control group COX-2 inhibitor celecoxib (100 mg / kg, Celecoxib, Pfizer), and 3% croton oil dissolved in acetone after 1 hour in the right ear of the mouse. It was applied evenly on the inside and outside to cause edema of the ear. Four hours after the induction of edema, the mouse was anesthetized with ether, and both ears were measured for the degree of edema of the mouse by a speed change method using a thickness gauge, and edema of the negative control group (1% CMC-administered group). Based on the degree of inhibition (%) was calculated in Table 6 below.

As shown in Table 6, it was found that the cheongung extract prepared as Examples 1 to 33 of the present invention showed excellent activity in the inhibitory effect in the edema inflammation animal model of mice induced with croton oil, Fractions had a significant inhibitory effect than crude extracts.

Experimental Example 7. Toxicity Test

7-1. Acute toxicity

The extract of Example 1 of the present invention was performed to determine the toxicity of the single oral administration to ICR mice. The extracts were administered at doses of 1,000, 2,000 and 4,000 mg / kg in a single dose (10 males and 5 males) per group, followed by mortality, general symptoms, body weight and necropsy findings compared to the excipient controls.

the results are as follow. No dead animals were observed during the test period, and weight change observations showed no changes in body weight associated with the administration of the test substance. Abnormal findings were not observed in the autopsy findings.

As a result, the extract of Example 1 of the present invention, when administered orally to a single ICR mouse, the minimum lethal dose (MLD: Minimum Lethal Dose) is determined to be more than 4,000 mg / kg in both sexes, indicating that it is a safe substance. there was.

7-2. Organ organs toxicity test in experimental group and control group

Repeated dose toxicity test

The extract of Example 1 of the present invention was carried out to determine the schematic toxicity by repeated oral administration for 2 weeks. A test group in which doses of 1,000 and 2,000 mg / kg / day of test substance were administered to ICR mice was set, and an excipient control group in which only excipients were administered was set. The number of animals was 5 male and female in each group. Test items were mortality, general symptoms, weight change, feed and water intake, urinalysis, hematology and blood biochemistry, autopsy findings, organ weight and histopathological findings.

The test results are as follows. No dead animals related to the toxicity of the test substance were observed during the study period, and no change in general symptoms related to the administration of the test substance was observed. As a result of weight change, no significant change was observed by the administration of test substance, and no significant change by the administration of test substance was observed. The results of urinalysis did not show any toxicological abnormalities associated with the administration of the test substance. Hematological and blood biochemical tests showed no toxicological abnormalities associated with the administration of the test substance. Long-term weight and gross necropsy findings showed no gross abnormalities associated with the administration of the test substance, and histopathological findings showed no toxicological abnormalities associated with the administration of the test substance.

In view of the above results, the extract of Example 1 of the present invention was confirmed to be a safe substance as a non-toxic amount (NOAEL) was determined to be 2,000 mg / kg / day in both male and female as a result of repeated two-week oral administration test for ICR mice.

Preparation Example 1 Preparation of Tablet

Extract of Example 1 ............................................ 200 mg

Corn starch ......................................... 50 mg

CELLACTOOSE ......................................... 100 mg

Crospovidone ......................................... 20 mg

Light silicic anhydride ............................................ 5 mg

Lactose ... ... 70 mg

Magnesium Stearate ......................................... 5 mg

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

Preparation Example 2 Preparation of Powder

Extract of Example 1 ............................................ 200 mg

Corn starch ........................... 50 mg

Lactose ... .. 100 mg

Talc ........................ ... 10 mg

The powders were prepared by mixing the above components and filling the airtight cloth according to a conventional powder preparation method.

Preparation Example 3 Preparation of Granules

Extract of Example 1 ... 300 mg

Corn starch ..................... 1000 mg

Lactose ... 1000 mg

Enzyme Treatment Stevia ... 50 mg

Talc ........................ .. 10 mg

The powders were prepared by mixing the above components and filling them in airtight cloths according to a conventional granulation method.

Preparation Example 4 Preparation of Capsule

Extract of Example 1 ... 300 mg

Microcrystalline cellulose ............... 150 mg

Lactose ......................... 30 Mg

Magnesium Stearate ....................... 0.5 mg

Capsules were prepared by mixing the above ingredients and filling the gelatin capsules according to a conventional capsule preparation method.

Preparation Example 5 Preparation of Injection

Extract of Example 1 ...................................... 50 mg

Mannitol ... 100 mg

Sterile Distilled Water for Injection ......................................... 3500 ml

Na ₂ HPO _{4 ,} 12H ₂ O ......................................... .. 20 mg

pH Adjuster ... Quantity

Injectables were prepared by dissolving the active substance and the rest of the ingredients in sterile distilled water for injection, adjusting the pH to about 7.5 according to a conventional injection method, and filling a 2 ml ampoule with sterilized distilled water for injection.

Preparation Example 6 Preparation of Liquid

Extract of Example 1 ... 500 mg

Isomerization sugar ... 10 g

Mannitol ... .. 5 g

Purified water ................................................. Appropriate amount

According to the conventional method of preparing a liquid solution, each component is added and dissolved in purified water, orange micron is added to the appropriate amount, the above components are mixed, purified water is added to adjust the total amount to 100 ml, and the container is filled and sterilized to prepare a liquid solution. do.

< Formulation example  7: Manufacture of Healthy Foods>

Extract of Example 1 ... 500 mg

Vitamin Blend ...........................

Vitamin A Acetate ......................... 70 μg

Vitamin E ........................................ 1.0 mg

Vitamin B1 ............................................... 0.13 mg

Vitamin B2 ......................................... 0.15 mg

Vitamin B6 ......................................... 0.5 mg

Vitamin B12 ......................... 0.2 μg

Vitamin C ............................................... 10 Mg

Biotin ... 10 μg

Nicotinic Acid Amide ......................................... 1.7 mg

Folic Acid ... 50 μg

Calcium Pantothenate ......................................... 0.5 mg

Mineral mixture ...............

Ferrous Sulfate ....................................................... 1.75 Mg

Zinc Oxide ......................................... 0.82 mg

Magnesium Carbonate ......................................... 25.3 mg

Potassium monophosphate ......................................................... 15 Mg

Dibasic Calcium Phosphate ... Mg

Potassium Citrate ......................................... 90 mg

Calcium Carbonate ... 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.

Preparation Example 8 Preparation of Healthy Drinks

Extract of Example 1 ............................................ 200 mg

Citric Acid ... Mg

Oligosaccharide ............................................... 100 g

Plum Concentrate ... 2 g

Taurine ... ... 1 g

Purified water is added ..................... 900 ml total

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, &Lt; / RTI &gt;

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

Claims (5)

Crude extract obtained by extracting cheongung with 1-4 carbon atoms or 10-95% alcohol aqueous solution thereof contains soluble purified fraction as an active ingredient selected from the group consisting of ethyl acetate, dichloromethane and butanol as an active ingredient. A composition for preventing or treating asthma, chronic obstructive pulmonary disease, allergic rhinitis, cough, phlegm, bronchitis, sore throat, tonsillitis and laryngitis. delete The method of claim 1,
The asthma is asthmatic, chronic obstructive pulmonary disease, allergic rhinitis, characterized in that the bronchial asthma, atopic asthma, atopic bronchial IgE-mediated asthma, non-atopic asthma, allergic asthma and non-allergic asthma Cough, sputum, bronchitis, sore throat, tonsillitis and laryngitis. Composition for the prevention or treatment of respiratory diseases selected from the group consisting of. The method of claim 1,
The bronchitis is asthma, chronic obstructive pulmonary disease, allergic rhinitis, cough, sputum, bronchitis, sore throat, tonsillitis, characterized in that selected from the group consisting of acute bronchitis, chronic bronchitis, bronchiolitis, catarrhal bronchitis and obstructive and inflammatory bronchial diseases A composition for preventing or treating respiratory diseases selected from the group consisting of laryngitis. Crude extract obtained by extracting cheongung with 1-4 carbon atoms or 10-95% alcohol aqueous solution thereof contains soluble purified fraction as an active ingredient selected from the group consisting of ethyl acetate, dichloromethane and butanol as an active ingredient. Health functional food for the prevention or improvement of respiratory diseases selected from the group consisting of asthma, chronic obstructive pulmonary disease, allergic rhinitis, cough, sputum, bronchitis, sore throat, tonsillitis and laryngitis.

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