KR101569876B1 - Pharmaceutical Composition for Preventing or Treating Respiratory Disease Containing Mixed Herbal Extract - Google Patents

Abstract

There is provided a pharmaceutical composition for the prevention and / or treatment of respiratory diseases, comprising a mixture of two or more herbal extracts selected from the group consisting of Saururus chinensis, Saururus chinensis, Bacillus subtilis,

Description

TECHNICAL FIELD The present invention relates to a pharmaceutical composition for preventing or treating respiratory diseases containing mixed herbal medicine extracts,

There is provided a pharmaceutical composition for the prevention and / or treatment of respiratory diseases, comprising a mixture of two or more herbal extracts selected from the group consisting of Saururus chinensis, Saururus chinensis, Bacillus subtilis,

Asthma, allergic rhinitis, bronchitis, and chronic obstructive pulmonary disease are the most common respiratory diseases. They are all inflammatory diseases and are characterized by airway damage.

Asthma, a representative respiratory disease, is a disease characterized by nonspecific airway hyperresponsiveness due to chronic airway inflammation and reversible airway obstruction. In about 30% of asthmatic patients, about 80% of asthmatic patients show asthma symptoms at the age of 4 to 5 years old. In Korea, about 80% of asthmatic patients have asthma symptoms. And the incidence rate is around 10%. Asthma is a disease that causes airway inflammation, airway hyperresponsiveness, and mucous membrane protein secretion. It is immunologically characterized by chronic infiltration of eosinophils, increase of Th2 cells, and increase of activated mast cells. Inflammatory disease.

Allergic rhinitis is a disease that causes symptoms such as runny nose, sneezing, nose and itchy eyes due to allergic inflammation reaction and it is a disease that worsens quality of life. In most industrialized countries, 8-24% of the population is one of the most common chronic diseases in adults and children, with such a disease. Rhinitis can also be caused by other mechanisms other than allergies, but allergies constitute about two-thirds of all rhinitis. Allergic rhinitis is often accompanied by asthma and shares much of the genetic tendency and pathophysiology.

Chronic Obstructive Pulmonary Disease (COPD) is a respiratory disease characterized by an abnormal inflammatory response of the airways and lungs caused by the ingestion of harmful particles or gases, and in addition, a non-reversible airflow limitation. The prevalence of chronic obstructive pulmonary disease is 10% in the whole population, and it can be up to 50% in smokers and the prevalence and mortality rate in developed countries is continuously increasing. Symptoms of chronic obstructive pulmonary disease include cough, sputum and dyspnea during exercise, frequently accompanied by acute exacerbation of symptoms and clinically including chronic bronchitis. In the airway, inflammatory cell infiltration and thickening of the airway wall narrow the airway diameter, resulting in increased airway resistance. In alveoli, inflammatory infiltration of the alveolar wall and alveolar rupture caused by alveolar destruction lead to a decrease in elastic pressure, resulting in a decrease in aerobic flow.

Acute bronchitis is acute inflammation of the trachea and bronchi, usually associated with respiratory tract infections. Acute bronchitis can be classified into acute infectious bronchitis and acute irritant bronchitis according to the cause. Most of the causes of infectious bronchitis are caused by the virus. Acute irritant bronchitis may be due to irritation by various metallic substances, volatile solvents, substances such as nitrogen dioxide or tobacco, or allergic reactions caused by inhalation of allergens. Chronic bronchitis is defined as inflammation of the mucous membranes of the chronic bronchial epithelium, usually with coughing and excessive phlegm almost every day for two consecutive years over a period of 3 months or more per year. Patients with chronic bronchitis have an increased cough, , Difficulty in breathing, and so on. In many countries chronic bronchitis and its worsening mortality rate are increasing.

Currently, there is no fundamental treatment for respiratory diseases, and many methods and drugs are used to prevent seizures and prevent complications, but they do not have satisfactory effects. It is mainly used for inhalation bronchodilator, oral or injectable bronchodilator (sympathetic stimulant and theophylline medicines), steroid preparations (for inhalation, oral use, injecting use), leukotriene antagonist (montelukast, trirukast, ), Anti-allergic drugs (disodium chloroclicate, cromolyn sodium, ketotifen), etc. have been used as therapeutic agents. Antihypertensive agents such as anticholinergic agents and beta2 receptor agonists are not effective for inflammation that exacerbates the disease, and only mere symptoms are alleviated. Therefore, there is a fear of drug resistance and aggravation of symptoms during long-term use. Steroids, which are known to be effective in inflammation, are a problem for long-term use due to serious side effects. Therefore, the combination of the two is often prescribed in the form of an inhaler than the oral antibiotics due to the side effects of steroids are difficult to take the dose is poor compliance.

Therefore, it is necessary to overcome the limitations of such current drug treatment, to develop a new therapeutic agent capable of fundamentally treating the cause and effectively improving the symptoms.

Accordingly, the present inventors have studied to develop a preventive and therapeutic agent for respiratory diseases such as asthma, allergic rhinitis, bronchitis, and chronic obstructive pulmonary disease. As a result, they have found that a combination of two or more of Saururus chinensis extract, 5-lipoxygenase (5-LO) inhibitory activity, phosphodiesterase (PDE) -4 inhibitory activity, adrenergic beta 2 receptor activation, cysteinyl leukotriene (CysLT1 receptor) inhibitory activity and Muscarinic receptor (eg, Muscarinic M3 receptor) inhibitory activity, and these activities were ascertained to be enhanced by the addition of each extract alone Thus completing the present invention.

Accordingly, one example of the present invention provides a pharmaceutical composition for preventing and / or treating respiratory diseases, which comprises, as an active ingredient, at least two kinds of herbal medicine extracts selected from the group consisting of Saururus chinensis, In the above pharmaceutical composition, an optimal combination ratio between each herbal medicine ingredient having the best pharmacologically active synergistic effect is also provided.

Another example of the present invention provides a health functional food composition for improving respiratory diseases, comprising at least two herbal extracts selected from the group consisting of Saururus chinensis, Bacillus thuringiensis, Bacillus subtilis, and Bacillus.

Another example provides a method for preparing a composition for preventing and / or treating a respiratory disease, comprising the step of mixing two or more selected from the group consisting of Saururus chinensis, Bacillus subtilis, Bacillus subtilis, and Baculus.

There is provided a pharmaceutical composition for the prevention and / or treatment of respiratory diseases, comprising a mixture of two or more herbal extracts selected from the group consisting of Saururus chinensis, Saururus chinensis, Bacillus subtilis, The pharmaceutical composition containing the mixed herbal medicine extract has an anti-inflammatory activity and an inhibitory activity against the airway contraction factor (for example, 5-lipoxygenase (5-LO) inhibitory activity, A phosphodiesterase (PDE) -4 inhibitory activity, an adrenergic beta 2 receptor activation, a cysteinyl leukotriene receptor (CysLT1 receptor) inhibitory activity, a muscarinic receptor (eg, Muscarinic M3 receptor) inhibitory activity, etc.), and is characterized by showing an advantageous effect for prevention and / or treatment of respiratory diseases such as asthma, allergic rhinitis, bronchitis, chronic obstructive pulmonary disease, cold and cough.

In addition, in the case of mixed herbal medicine extracts obtained by mixing two or more kinds of Saururus chinensis extract, Bacillus thuringiensis extract, Bacillus subtilis extract and Bacillus subtilis extract at a certain ratio, the remarkably elevated respiratory tract It is confirmed that it has the effect of prevention and treatment of disease, and provides the optimum mixing ratio of herbal medicine extract to obtain the best synergistic effect.

Saurururi Herba is a perennial plant of the Saururianaceae ( Saururus) Chinensis (Laureiro) Baillon is known to have pharmacological actions such as antiinflammation, cell protection, anticancer, and hypoglycemia, and it is used for edema, jaundice, gonorrhea, gonorrhea and dysuria.

Chelidonid Herba is a plant of the genus Papaveraceae, Chelidonium I have Majus Linne. asiaticum Ohwi), and when the stem is cut, the juice of the crustacean comes out, and it is named as the baby grasshopper, also called the seed poop, the grasshopper. It has been used in oriental medicine and civilian medicine to treat skin diseases, various cancers, eye diseases, arthritis, athlete's foot, gastritis, stomach cramps, gastric ulcers, hepatitis, abdominal pain and digestive diseases.

Solani Nigri Herba is a prey of Solanum nigrum Linne belonging to Solanaceae. It is one year early in the field or roadside of each place in Korea. It is used for anti-inflammation, cell protection, central nervous system suppression, It is known to have an action such as anticancer, and it is used as fever, diuretic, gangjeong, and tonic medicine.

Trichosanthis Semen is a perennial plant belonging to the genus Cucurbitaceae, Trichosanthes kirilowii Maxim.) or ssangbyeon gwalru (as the seeds of Trichosanthes rosthronii Harms), parenthesis ruja, also called the gwaru is used as a fruit, root, foliage, herbs etc. rind.

Korean Patent Laid-Open Publication No. 2013-0142638 discloses that the purified product and fractions of Saururus chinensis having toxic substances are effective for the prevention and treatment of asthma and allergic diseases. Also disclosed in Korea Patent Publication 2006-0111790 is Saururus chinensis extract, It is disclosed that there is a preventive and therapeutic effect on sex diseases. However, there is no known effect of preventing and / or treating respiratory diseases such as asthma, allergic rhinitis, bronchitis, chronic obstructive pulmonary disease, etc. of two or more mixed herbal medicine extracts selected from Saururus chinensis,

One example is a pharmaceutical composition comprising, as an active ingredient, at least two herbal extracts selected from the group consisting of Saururus chinensis, Bacillus thuringiensis, Bacillus subtilis, and Bacillus.

The mixed herbal extract may be used to inhibit 5-lipoxygenase (5-LO) inhibitory activity, phosphodiesterase (PDE) -4 inhibitory activity, adrenergic beta 2 receptor activation, leukotriene receptor (CysLT1 receptor) inhibitory activity, muscarinic receptor (eg, muscarinic M3 receptor) inhibitory activity, etc., and can exert excellent effects on prevention and / or treatment of various respiratory diseases.

Accordingly, the pharmaceutical composition may be a pharmaceutical composition for preventing and / or treating respiratory diseases.

As used herein, " treatment " refers to a reduction or amelioration of symptoms, a reduction in the extent of disease, a delay or alleviation of disease progression, an improvement, alleviation or stabilization of a disease state or symptom, partial or complete recovery, Treatment results, and the like.

The respiratory disease may be any disease accompanied by inflammation. Specifically, the respiratory diseases include 5-lipoxygenase (5-LO), phosphodiesterase (PDE) -4, adrenergic beta 2 receptor (in the case of inactivation) May be all respiratory diseases having symptoms caused by at least one selected from the group consisting of a cysteinyl leukotriene receptor (CysLT1 receptor), a muscarinic receptor (eg, Muscarinic M3 receptor), and the like. For example, the respiratory diseases include asthma such as bronchial asthma, atopic asthma, atopic bronchial IgE mediated asthma, non-atopic asthma, allergic asthma, and non-allergic asthma; Allergic rhinitis; Bronchitis such as chronic bronchitis or acute bronchitis; Chronic obstructive pulmonary disease; cold; Cough, and the like.

In the present invention, the term " mixed herbal medicine extract or mixed extract " refers to a mixture of herbal medicine extracts obtained by mixing two or more selected from among four herbal medicine extracts obtained by extracting the above four herbal medicines, Or a mixture of two or more selected herbal medicines selected from the group consisting of herbal medicines. The 'extract' means a crude extract or a specific solvent-soluble extract (fraction), which may be in the form of solution, concentrate or powder.

5-LO inhibitory activity, phosphodiesterase (PDE) -4 inhibitory activity, beta- 2- (2-ethylhexyl) -1,2,3,4-tetrahydronaphthalene A synergistic effect on adrenergic beta 2 receptor activation, cysteinyl leukotriene receptor inhibitory activity, and muscarinic receptor (eg, muscarinic M 3 receptor) inhibitory activity.

In one example, a mixing ratio between each herbal ingredient is provided to maximize such synergistic effect. When the mixed herbal medicine extract is a mixture of herbal medicine extracts, the herbal medicine ingredient means each herbal medicine extract. When the mixed herbal medicine extract is an extract of the mixed herbal medicine, the herbal medicine ingredient means each herbal medicine in the mixed herbal medicine. In addition, the mixing ratios between the herbal medicine ingredients described in the present specification are expressed on the basis of weight, and the herbal medicine component (for example, "Saururus chinensis" Quot; saury "). When the mixed herbal medicine extract is a mixture of herbal medicine extracts, the extraction solvent or fraction solvent of each herbal medicine extract is independently selected and may be the same or different from each other.

The mixing ratio of each herbal medicine ingredient in the mixed herbal medicine extract may be 1:10 to 10: 1, 1: 5 to 5: 1, or 1: 3 to 3: 1 for the two mixed herbal extracts, 1 to 3: 1 to 3: 1 to 3: 1 to 10: 1 to 10: 1 to 5: 1 to 5: 1 to 10: 1 to 10: 1 to 5: 1 to 5: 1 to 5: 1 for four herbal extracts, 1 to 3: 1 to 3: 1 to 3: 1 to 3, 1: 3 to 1: 3 to 1: 3, 1: : 1: < / RTI > The mixing ratio between the herbal medicine ingredients described in the present specification is calculated on the basis of the weight of solids removed (in the case of a mixture of herbal extracts) or the weight of the herbal medicine itself (in the case of mixed herbal extracts). As used herein, the term " to " between two numerical values means the interval between numerical values including the numerical values mentioned before and after.

In one embodiment, the mixed herbal extract may be a mixed herbal extract of Saururus chinensis and White herring. At this time, the mixing ratio of each herbal ingredient (sow wheat: white oak or white oak: three hundred oak) may be 1:10 to 10: 1, 1: 5 to 5: 1, or 1: 3 to 3:

In another example, the mixed herbal extract may be a mixed herbal extract of Saururus chinensis and Lycopersicon esculentum. In this case, the mixing ratio of the herbal medicine components (Sambucoptera: Yugu or Yongsu: Sambucoptera) may be 1:10 to 10: 1, 1: 5 to 5: 1, or 1: 3 to 3: 1.

In another example, the mixed herbal extract may be a mixed herbal extract of Saururus chinensis and gingiva. At this time, the mixing ratio between the herbal medicine components (Sambucchus: gangrene or gangrene) may be 1:10 to 10: 1, 1: 5 to 5: 1, or 1: 3 to 3: 1.

In another example, the mixed herbal extract may be a mixed herbal extract of white oyster and egg white. At this time, the mixing ratio between the herbal medicine components (white lid: lily or lily: white lily) may be 1:10 to 10: 1, 1: 5 to 5: 1, or 1: 3 to 3:

In another example, the mixed herbal extract may be a mixed herbal extract of bagred herb and gull root. At this time, the mixing ratio between the herbal medicine components (white ginseng: ginseng or ginseng: white ginseng) may be 1:10 to 10: 1, 1: 5 to 5: 1, or 1: 3 to 3: .

In another example, the mixed herbal extract may be a herbal extract and a mixed herbal extract of guluth. In this case, the blending ratio between each of the herbal medicine components (reference: cochlein or cochlein) may be 1:10 to 10: 1, 1: 5 to 5: 1, or 1: 3 to 3:

In another example, the mixed herbal medicine extract may further comprise a herbal extract, a gherkin extract, or a herbal extract and a mixed herbal extract of ghuline in a mixed herbal medicine extract of Saururus chinensis and White herring. In another example, the mixed herbal medicine extract may further comprise a white herring extract, a gherkin extract, or a mixed herbal extract of white gherkin and ghuline in a mixed herbal medicine extract of Saururus chinensis and Saccharomyces cerevisiae. In another example, the mixed herbal medicine extract may further comprise a white herring extract, a herb extract, or a mixed herbal extract of white herb and herb in a mixed herbal medicine extract of Saururus chinense and Manduro. In another example, the mixed herbal medicine extract may further comprise a Saururus chinensis extract, a Bacillus subtilis extract, or a mixed herbal extract of Saururus chinensis and Bacillus subtilis in a mixed herbal medicine extract of white irritants and herbs. In another example, the mixed herbal medicine extract may further comprise a Saururus chinensis extract, a mandarin extract, or a mixed herbal extract of Saururus chinensis and Saccharomyces sp. In another example, the mixed herbal medicine extract may further comprise a herbal extract of Saururus chinensis, an extract of Bacillus thuringiensis, or a mixed herbal medicine extract of Saururus chinensis and Bacillus thuringiensis in a mixed herbal medicine extract of the gut and gut.

Specifically, the mixed herbal medicine extract may be a mixed herbal medicine extract of Saururus chinensis, White mackerel, and Yonggi. At this time, the mixing ratio of each herbal medicine ingredient (Sambucchus: white gruel: Yonggu, Sambuccho: Yukyu: white gruel, white gruel: 1: 3 to 1: 3 to 1: 3 to 1: 3 to 1: 3 to 1: 3 to 1: 3 to 1: Or 1: 1: 1 to 3.

In another example, the mixed herbal extract may be a mixed herbal extract of Saururus chinensis, Bacillus thuringiensis, and Baculus. At this time, the mixing ratio of each herbal medicine ingredient (Sambucchus: Sambucchus: Sanguinea, Sambuccho: Safflower: Saffacea, Safflower: Safflower: Safflower, Safflower: Safflower: Safflower, 1 to 10: 1 to 10, 1 to 5: 1 to 5: 1 to 5, 1 to 3: 1 to 3: 1 to 3, 1: 1 to 3: 3: 1 to 3, or 1: 1: 1 to 3.

In another example, the mixed herbal extract may be a mixed herbal extract of Saururus chinensis, Yonggyu, and Manduro. At this time, the mixing ratio of each herbal medicine ingredient (Sambucchus: Yugyu: guluin, samsungcho: guluin: yuchu, yuchyu: guluin, gulgyu: guluin: samsungcho, guluin: 1: 3 to 1: 3 to 1: 3 to 1: 3 to 1: 3 to 1: 3 to 1: 3 to 1: Or 1: 1: 1 to 3.

In another example, the mixed herbal extract may be a herbal extract, a mixture of herbs, and a mixed herbal extract of gingiva. At this time, the mixing ratio of each herbal medicine ingredient (white liquor: Yukyu: guluin, white gulch: guluin: yonggu, yongyu: white gulch: guluin, 1: 1 to 5: 1 to 5: 1 to 5: 1 to 3: 1 to 3: 1 to 3, 1: 1 to 1: 3: 1 to 3, or 1: 1: 1 to 3.

In another example, the mixed herbal extract may be a mixed herbal extract of Saururus chinensis, Bacillus thuringiensis, Bacillus subtilis, and Baculus. At this time, the mixing ratio of each herbal medicine ingredient (Sambuche: Baekgukje: Yukyu: 루 인,, Sambuchecho: Baekukchae: Sangyecho: Gwangju: Yonggyu, Yonggyu: Sangbyeon: Gwangyu: Yonggyu: Yonggyu: Yonggyu: Yonggyu: Yonggyu: Yonggyu: Yonggyu: Sanyakchecho, Yonggyu: Sanyakchecho: Baekryeochae: gwakuin, yongyu: Sanyakchecho: gwaine: white gilt, white gilt: Yoon-gyu: Baekgulchae, Yonggyu: Gwacheon: Baekgukbyeon: Sauryweed, Gwacheon: Sauryweed: Baekwolchae: Yonggyu, 규:: 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 삼 ) Is from 1 to 10: 1 to 10: 1 1 to 3: 1 to 3: 1 to 3: 1 to 3, 1: 3 to 1: 3 to 1: 3 to 1: 1 to 5: 1 to 5: 1: 1 to 3: 1 to 3, or 1: 1: 1: 1 to 3.

The extracts of at least two kinds of herbal medicines selected from the group consisting of Saururus chinensis extract, Bacillus thuringiensis extract, Bacillus subtilis extract, Bacillus subtilis extract or Saururus chinensis, Bacillus thuringiensis, Bacillus subtilis, Each of which is obtained by extracting at least two kinds of mixed medicines selected from the group consisting of Saururus chinensis, Bacillus thuringiensis, Bacillus subtilis, and Baculus with at least one selected from the group consisting of water and a straight chain or branched alcohol having 1 to 4 carbon atoms Or (2) a solvent-soluble extract obtained by adding a linear or branched alcohol having 1 to 6 carbon atoms or a non-polar solvent to the crude extract. Saururus chinensis, Bacillus thuringiensis, Bacillus subtilis, and Bacillus can be used as they are, or they can be used by cutting or grinding them into appropriate sizes.

In a preferred embodiment, the solvent used in the preparation of the crude extract is water, or a mixture of water, or 10-95% (v / v), 20-95% (v / v), 30-95% (V / v), 50 to 95% (v / v), 60 to 95% (v / v), 70 to 95% (v / v), 10 to 90% (V / v), 30 to 90% (v / v), 40 to 90% (v / v), 50 to 90% (v / v), 60 to 90% (V / v), 10 to 85% (v / v), 20 to 85% (v / v), 30 to 85% (v / v), 40 to 85% (V / v), 60 to 85% (v / v), 70 to 85% (v / v), 10 to 80% (v / v), 20 to 80% The number of carbon atoms in the molecule is 80% (v / v), 40-80% (v / v), 50-80% (v / v), 60-80% (v / And the alcohol aqueous solution may be at least one selected from the group consisting of methanol aqueous solution, aqueous ethanol solution, isopropanol aqueous solution, and water saturated butanol in the concentration range described above. The solvent used for preparing the solvent-soluble extract may be selected from i) a linear or branched alcohol having 1 to 6 carbon atoms (e.g., 3 to 5 carbon atoms) such as propanol, isopropanol, and butanol Or ii) at least one selected from the group consisting of nonpolar solvents such as hexane, dichloromethane, chloroform, and ethyl acetate.

In the case of allergic diseases such as asthma, when an allergic antigen enters the living body, an IgE antibody specific to an allergic antigen is generated and adheres to the surface of the allergen-induced cell. When the allergen enters the living body again, IgE Allergen-stimulated cells that bind to and stimulate the antibody release various mediators including histamine, causing an inflammatory reaction by increasing the number of inflammatory cells including eosinophils. When this inflammatory reaction occurs, it causes an acute inflammatory reaction such as bronchoconstriction, vasodilation, sensory neuropathy, and cholinergic bronchoconstriction in the airway.

Among respiratory diseases such as asthma, chronic obstructive pulmonary disease, bronchitis, and allergic rhinitis, cysteinyl leukotrienes are very powerful anti-inflammatory mediators among various mediators involved in airway inflammation. Leukotriene is involved in allergic and allergic reactions such as asthma, chronic bronchitis, obstructive airways disease, obstructive pulmonary disease, allergic rhinitis, contact dermatitis and allergic conjunctivitis, inflammatory diseases such as arthritis or inflammatory bowel disease and pain (Dahlen SE, Eur. J. Pharmacol., 533: 40-56, 2006). Leukotriene is involved in airway smooth muscle contraction, mucus secretion, mucosal edema, and induction of neutrophils and eosinophils in the airways in allergic or inflammatory respiratory diseases. The synthesis of leukotrienes includes three enzymes: 5-lipoxygenase (5-LO), 5-lipoxygenase activating protein, and leukotriene C4 (LTC4) Lt; / RTI > 5-LO is the most important enzyme in the biosynthesis of leukotrienes, which catalyzes the oxidation of arachidonic acid and is involved in the production of leukotriene B4 (LTB4) or cyteinyl leukotrienes (Cysteinyl-LT) through subsequent metabolism (ACS Med Chem. Lett., 1, 170-174, 2010). Leukotrienes mainly act on two receptors, the cysteinyl leukotriene receptor 1 (Cys LTR1) and the leukotriene receptor 2 (Cys LTR2), which are involved in the pathophysiology of allergic and inflammatory diseases The symptoms are mainly related to Cys LTR1 (Korean Journal of Pediatrics Vol. 52, No. 6, 2009). Cys LTR1 antagonists inhibit the binding of these leukotrienes to Cys LTR1 and inhibit airway constriction by inhibiting airway constriction, excessive mucus secretion, and migration of eosinophils (Therapeutics and Clinical Risk Management, Vol. 3, No.5 , 885-892, 2007). The herbal extracts of the present invention were confirmed to have an inhibitory activity of 5-LO enzyme action (see Example 3.1, Tables 1 to 7) and an inhibitory activity of the action of Cys LTR1 (see Example 3.4, Tables 22 to 28) , Which demonstrates the prevention and / or therapeutic effect of respiratory diseases of the mixed herbal medicine extract.

Phosphodiesterases (PDEs) have so far been identified in 11 subtypes, involved in the regulation of signal transduction by participating in the degradation of cyclic nucleotides (cAMP and / or cGMP). Among these PDEs, PDE4, PDE7, PDE8 and the like are specific for cAMP. Among them, PDE4 is known as a major enzyme involved in cAMP metabolism in immune and inflammatory cells. PDE4 inhibitors exhibit antiallergic and antiinflammatory effects by inhibiting the degradation of cAMP in lymphocytes, eosinophils, neutrophils, monocytes, and reducing histamine and leukotrienes and various cytokine releases (J Allergy Clin Immunol Vol 108, No. 4, 530 -6, 2001). Thus, selective PDE4 inhibitors have been noted as an alternative to anti-inflammatory drugs in place of steroids and have been shown to exhibit anti-inflammatory effects in patients with various inflammatory diseases such as asthma, allergic diseases, chronic obstructive pulmonary disease or bronchitis (JPET 308, 555-563, 2004). The mixed herbal medicine extract of the present invention was found to have the inhibitory activity of the action of PDE4 enzyme (see Example 3.2, Tables 8 to 14), demonstrating the effect of preventing and / or treating respiratory diseases of the mixed herbal medicine extract.

Beta receptor is a single polypeptide glycoprotein embedded in the cytoplasm, and three (β1, β2, β3) subtypes have been identified to date. Beta receptors in the bronchial smooth muscle of humans are wholly β2 subtypes.

The beta 2 agonist binds to the beta 2 receptor and activates the beta 2 receptor, thereby activating the adenyl cyclase of the airway smooth muscle cells, thereby lowering the intracellular calcium ion concentration, thereby relaxing the contracted airway smooth muscle. Beta 2 agonists not only act directly on the airway smooth muscle but also indirectly by the release of mediators involved in bronchoconstriction from inflammatory cells and the secretion of neurotransmitters that cause bronchoconstriction from airway nerves . In addition, beta-2 agonists inhibit the release of mast cells and suppress the release of microbes and leaks after exposure to chemical media such as histamine and leukotriene, thereby suppressing the development of bronchial mucosal edema and increasing the ion transport of airway epithelium Increased mucociliary clearance and decreased cough may also be achieved (Tuberculosis and Respiratory Diseases Vol. 60, No. 3, Mar. 2006). The herbal extracts of the present invention were found to bind to the beta-2-adrenergic receptor (see Example 3.3, Tables 15-21) and to have an action similar to that of the beta 2 agonist (i.e. activation of the beta-2-adrenergic receptor) , Which demonstrates the preventive and / or therapeutic effect of respiratory diseases of the mixed herbal medicine extract.

In addition, when the parasympathetic nerve is stimulated, acetylcholine (ACh) is secreted into the nerve conduction material to stimulate muscarinic (M) receptors in the airway smooth muscle, thereby activating the intracellular enzyme GTP, converting it into cGMP and contracting the airway smooth muscle . In addition to nerve stimulation, ACh is secreted from various cells of the airway, and inflammatory stimulation increases choline acetyltransferase expression and increases ACh. Humans have at least five closely related muscarinic receptor genes, three of which (M1, M2, M3) are expressed in the lungs. These subtypes appear to have different physiological functions in different tissues of the lungs. The M3 receptor is distributed in the smooth muscle and submucosa of the airway and promotes smooth muscle contraction and mucus secretion, thus inhibiting the action of the M3 receptor can suppress these symptoms. The mixed herbal medicine extract of the present invention has been found to have an inhibitory activity of muscarinic M3 receptor function (see Example 3.5, Tables 29 to 35), which demonstrates the prevention and / or therapeutic effect of respiratory diseases of the mixed herbal extract will be.

The content of the mixed herbal medicine extract as an active ingredient in the pharmaceutical composition may be appropriately adjusted depending on the mode and purpose of use, patient condition, symptom type, and the like, and is 0.001 to 99.9% by weight, 0.01 to 70% by weight, Or 0.1 to 50% by weight.

The pharmaceutical compositions can be administered to mammals, including humans, by a variety of routes. The mode of administration may be any conventionally used mode and may be, for example, oral administration or parenteral administration such as rectal, intravenous, intramuscular, subcutaneous, intrauterine, or intracerebral injection. The pharmaceutical composition may be formulated into parenteral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, transdermal preparations, suppositories and sterilized injection solutions according to a conventional method Can be used.

In addition to the mixed herbal extract, the pharmaceutical composition may further comprise a pharmaceutically acceptable and physiologically acceptable carrier, an excipient, and / or an adjuvant such as a diluent. Examples of the carrier, excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, At least one selected from the group consisting of microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, one or more diluents or excipients selected from the group consisting of commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants can be used. The solid preparations for oral administration include at least one selected from the group consisting of tablets, pills, powders, granules, capsules, etc. These solid preparations may contain at least one excipient such as starch, calcium carbonate , Sucrose, lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. As a liquid preparation for oral administration, at least one selected from the group consisting of a suspension, a solution, an emulsion, and a syrup may be used. In addition to water and liquid paraffin which are commonly used diluents, various excipients such as a wetting agent, , A perfume, a preservative, and the like. Formulations for parenteral administration include one or more selected from the group consisting of sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and transdermal preparations. Examples of the suspending agent may include at least one selected from the group consisting of propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, at least one selected from the group consisting of witepsol, macrogol, tween 61, kakarose, laurin, glycerogelatin and the like can be used.

In embodiments where the pharmaceutical composition is applied to humans, the mixed herbal extract composition may be administered alone, but may be administered in admixture with a pharmaceutical carrier selected in view of the general mode of administration and standard pharmaceutical practice . For example, the mixed extract-containing compositions of the present invention may be in the form of tablets containing starch or lactose, in the form of capsules containing the active ingredient alone or as an excipient, or as elixirs containing chemicals that taste or color It can be administered orally, buccally or sublingually in the form of a suspension. Such liquid preparations may be formulated with suspending agents (e. G., Mixtures of PEG-6 esters with semisynthetic glycerides such as methylcellulose, withexol or apricot kernel oil or with PEG-8 and caprylic / capric glycerides Such as a glyceride mixture, such as a mixture.

The dose of the pharmaceutical composition may be varied depending on the patient's age, weight, sex, dosage form, health condition, and disease severity, and may be administered once or several times a day, It is possible. For example, the daily dosage may be 0.5 to 500 mg / kg or 1 to 300 mg / kg, based on the active ingredient content, but is not limited thereto. The above-mentioned dosage is an average case, and the dose may be high or low depending on individual differences.

Another example of the present invention is a method for producing a pharmaceutical composition having a preventive and / or therapeutic activity for respiratory diseases, which comprises the step of preparing two or more mixed herbal extracts selected from the group consisting of Saururus chinensis, ≪ / RTI > The respiratory disease may be selected from the group consisting of asthma (such as bronchial asthma, atopic asthma, atopic bronchial IgE mediated asthma, nonatopic asthma, allergic asthma or non-allergic asthma), allergic rhinitis, bronchitis Bronchitis, etc.), chronic obstructive pulmonary disease, cold, cough (such as chronic cough), and the like.

In one embodiment, the method comprises:

At least one member selected from the group consisting of Saururus chinensis, Bacillus thuringiensis, Bacillus subtilis, and Bacillus each is independently selected from the group consisting of water and at least one selected from the group consisting of water and straight chain or branched alcohols having 1 to 4 carbon atoms, (V / v), 30-95% (v / v), 40-95% (v / v), 50-95% (V / v), 70 to 95% (v / v), 10 to 90% (v / v), 20 to 90% (v / v), 30 to 90% (V / v), 60 to 90% (v / v), 70 to 90% (v / v), 10 to 85% (V / v), 30 to 85% (v / v), 40 to 85% (v / v), 50 to 85% (v / v), 60 to 85% (V / v), 10 to 80% (v / v), 20 to 80% (v / v), 30 to 80% Aqueous solution of a linear or branched alcohol having 1 to 4 carbon atoms in an amount of 80 to 80% (v / v), 60 to 80% (v / v), or 70 to 80% (v / v) Aqueous solution, and blister At least one member selected from the group consisting of butanol) and the extract, to afford a Houttuynia cordata extract, back extract gulchae, yonggyu parenthesis extract and the herbal extract of two or more selected from the group consisting of ruin extract; And

Mixing the obtained two or more crude drug extracts

. ≪ / RTI >

In another example,

Preparing a herbal mixture by mixing at least two kinds of herbal medicines selected from the group consisting of Saururus chinensis; And

Water or 10 to 95% (v / v), 20 to 95% (v / v) of water, and 1 to 4 linear or branched alcohols having 1 to 4 carbon atoms, , 30 to 95% (v / v), 40 to 95% (v / v), 50 to 95% (v / v), 60 to 95% V / v), 30 to 90% (v / v), 40 to 90% (v / v), 50 to 90% (v / v) , 60 to 90% (v / v), 70 to 90% (v / v), 10 to 85% (v / v), 20 to 85% (V / v), 60-85% (v / v), 70-85% (v / v), 10-80% , 20 to 80% (v / v), 30 to 80% (v / v), 40 to 80% (v / v), 50 to 80% , Or 70 to 80% (v / v) aqueous solution of a linear or branched alcohol having 1 to 4 carbon atoms (for example, at least one selected from the group consisting of an aqueous methanol solution, an aqueous ethanol solution, an aqueous isopropanol solution and a water saturated butanol) Three hundred Step of obtaining at least two kinds of herbal medicine extracts selected from the group consisting of herbal extracts, herbal extracts,

. ≪ / RTI >

When the herbal or herbal medicine extracts are mixed, the mixing ratio between them is as described above.

At this time, each of the herbal medicine extract preparation step or the extract preparation step of the herbal medicine mixture may be carried out by adding at least one selected from the group consisting of linear or branched alcohols having 1 to 6 carbon atoms, such as propanol, isopropanol and butanol, Or extracting a solvent-soluble extract obtained by adding at least one selected from the group consisting of hexane, dichloromethane, chloroform and ethyl acetate to a nonpolar solvent, and then purifying the extract.

The extraction step may be carried out after each herbal drug has been cut or pulverized (for example, in the case of a coarse) to an appropriate size.

The extraction process (extraction of crude extract and / or solvent-soluble extract) used in the above method may be performed by any extraction method commonly used. For example, the extraction process may be performed by a method comprising the steps of cold-pressing, hot water extraction, ultrasonic extraction, , But the present invention is not limited thereto.

In one embodiment, the process for preparing the extract of Saururus chinensis extract, White irrigation extract, Radix extract, Radix extract, or a mixture of two or more herbal medicines according to the present invention will be described in more detail as follows: In order to obtain an effective extraction, a mixture of saururus chinensis, white cucurbits, millet, crushed gulurin or two or more herbal medicine mixtures is added in an amount of 5 to 30 times (mL or L) of the weight of the herbal medicine (each herbal medicine or herbal medicine) 5 to 25 times, 5 to 15 times, 7 to 30 times, 7 to 25 times, 7 to 10 times, for example, 0.5 to 3 L per 100 g of herbal medicine material) Water containing 10 to 20 vol., 7 to 15 vol., 10 to 30 vol., 10 to 25 vol., 10 to 20 vol., Or 10 to 15 vol., And a linear or branched alcohol having 1 to 4 carbon atoms One selected from the group (V / v), 30-95% (v / v), 40-95% (v / v), 50-95% (v / v), 60 to 95% (v / v), 70 to 95% (v / v), 10 to 90% (v / v), 20 to 90% (v / v), 40 to 90% (v / v), 50 to 90% (v / v), 60 to 90% (v / v), 70 to 90% (v / v), 20 to 85% (v / v), 30 to 85% (v / v), 40 to 85% (v / v), 50 to 85% (v / v), 70 to 85% (v / v), 10 to 80% (v / v), 20 to 80% (v / v), 30 to 80% (v / v), 50-80% (v / v), 60-80% (v / v), or 70-80% (v / v) methanol, aqueous solution, ethanol aqueous solution, isopropanol aqueous solution, Butanol can be used for the reflux extraction using at least one extraction solvent selected from the group consisting of ethanol, ethanol, and butanol. The extraction temperature may be suitably adjusted according to the extraction method, and may be, for example, 25 to 110 ° C, 40 to 110 ° C, 25 to 90 ° C, or 40 to 90 °.

In order to increase the yield, the extraction process (primary extraction) is performed to filter the obtained extract, and the filtrate is collected. The residue is re-extracted (secondary extraction) in the same manner and then filtered to obtain an extract can do. The obtained extract can be produced by a conventional method such as filtration and / or concentration (for example, concentration under reduced pressure) to obtain a final extract of Saururus chinensis, an extract of Bombyx mori, a extract of Bombyx mori, a extract of Bombyx mori, or a mixed herbal medicine. The extraction efficiency can be increased by mixing the extraction solution obtained after the second extraction and the filtrate obtained after each extraction, but it is not limited to the extraction number of the extract of the present invention.

In order to purify impurities such as unnecessary proteins, polysaccharides and fatty acids contained in the filtrate after filtering and / or concentrating the extract (crude extract) obtained by the primary and / or secondary extraction as described above, the obtained crude extract (For example, a filtrate of crude extract or a dried or softened concentrate) or a crude extract (e.g., a dried or softened concentrate) is mixed with water (e.g., 1 to 10 times by volume, 2 To 8 vol. Vol., Or 3 to 6 vol. Vol. (ML or L)) is added 1 to 10 vol, 1 to 7 vol or 1 to 5 vol of lower alcohols or suspension of the crude extract or suspension, A step of purifying impurities by adding a nonpolar solvent and performing a layer separation of 1 to 10 times, 1 to 6 times, or 2 to 4 times to obtain a solvent-soluble fraction. For example, when the crude extract is a filtered solution after filtration, a fraction can be obtained by adding a fraction solvent (lower alcohol or non-polar solvent) of 1 to 5 times the volume of the filtrate to the filtrate. In case of the soft-tune concentrate, the volume of the suspension is 1 to 10 times the volume of the suspension suspended in water (distilled water) of 1 to 10 times, 2 to 8 times, or 3 to 6 times, for example about 5 times, , 1 to 7 vol. Vol. Or 1 to 5 vol. Vol. Fraction may be added to obtain a fraction. The lower alcohol may be at least one selected from the group consisting of linear or branched alcohols having 1 to 6 carbon atoms such as propanol, isopropanol, and butanol. The nonpolar solvent may be at least one selected from the group consisting of hexane, dichloromethane, chloroform and ethyl acetate And may be at least one selected from the group consisting of When the amount of the lower alcohol or the non-polar solvent used as the fraction solvent is less than that of the crude extract (for example, the filtrate or the dried or softened concentrate) or the suspension, fine particles are formed due to unnecessary components such as fatty acids, The amount of the solvent used may be equal to or more than the volume of the crude extract or suspension, for example, 1 to 10 times by volume, 1 to 7 times by volume, or 1 to 5 times by volume, as the extraction efficiency of the active ingredient may be lowered, It can be used in volumetric volume.

The step of concentrating the obtained lower alcohol or the non-polar solvent soluble fraction by vacuum filtration to 40 to 80? To remove the solvent remaining in the sample to obtain a concentrate.

The crude extract, fraction or concentrate thereof (dried or softened concentrate) thus obtained may be added to the crude extract, fraction or concentrate to adjust the content of the remaining solvent so as to be suitable for use as a raw material for the medicament. 5 to 25 times, 5 to 20 times, 10 to 30 times, or 10 to 25 times, or 10 to 20 times by weight of water in an amount of 1 to 5 times or 2 to 3 times Adding water of the same amount again, homogeneously suspending it, and concentrating it.

In the above production method, the mixing ratio of each herbal extract (based on the weight of the solid) or the herbal medicine (based on the weight of the herbal medicine) in the mixing step of two or more herbal medicine extracts or the mixing step of two or more herbal medicines is as described above. That is, when two herbal medicine extracts or two herbal medicines are mixed, the mixing step may be carried out by mixing the two herbal medicine extracts or two herbal medicines at a mixing ratio of 1:10 to 10: 1, 1: 5 to 5: 1 , Or 1: 3 to 3: 1. When the three herbal medicine extracts or the three herbal medicines are mixed, the mixing step is carried out such that the mixing ratio of the three herbal medicine extracts or the three herbal medicines is 1 to 10: 1 to 10: 1 to 10, 1 to 5: 1 To 5: 1 to 5: 1 to 3: 1 to 3: 1 to 3, 1: 1 to 3: 1 to 3, or 1: 1: 1 to 3. When mixing four kinds of herbal medicine extracts or four kinds of herbal medicines, the mixing step is carried out so that the mixing ratio of the four herbal medicine extracts or four kinds of herbal medicines is 1 to 10: 1 to 10: 1 to 10: 1 to 10, 1 1 to 3: 1 to 3: 1 to 3: 1: 1: 1 to 5: 1 to 5: 1 to 5: 1 to 5 or 1 to 3: To 3: 1 to 3, or 1: 1: 1: 1 to 3.

In order to homogeneously mix each of the extracts in the mixed extract, water may be added to the mixed extract in an amount of about 2 to 3 times by weight, followed by concentration under reduced pressure at a temperature of 40 to 60 ° C to prepare a composition.

In another aspect, the present invention provides a health functional food for preventing and / or improving respiratory diseases, which contains two or more mixed herbal extracts selected from the group consisting of Saururus chinensis, Bacillus thuringiensis, . The respiratory diseases include asthma (such as bronchial asthma, atopic asthma, atopic bronchitis IgE mediated asthma, non-atopic asthma, allergic asthma or non-allergic asthma), allergic rhinitis, bronchitis (chronic bronchitis or acute bronchitis, Chronic obstructive pulmonary disease, and the like. The health functional food may be various foods, beverage compositions, food additives, and the like.

The content of the mixed herbal medicine extract contained in the health functional food may be, for example, 0.001 to 20% by weight or 0.01 to 15% by weight of the whole food, without any particular limitation, depending on the form of the food, But may be, but is not limited to, 0.001 to 15 g, 0.02 to 10 g, or 0.3 to 1 g based on 100 ml for a health beverage composition.

There is no particular limitation on the liquid ingredient other than the mixed herbal medicine extract contained in the content range of the health beverage composition, and further ingredients such as various flavors and / or natural carbohydrates such as ordinary beverages may be included. The above-mentioned natural carbohydrates include monosaccharides such as glucose; Disaccharides such as fructose; Polysaccharides such as maltose, sucrose and the like; Conventional sugars such as dextrin, cyclodextrin and the like; Sugar alcohols such as xylitol, sorbitol and erythritol; And the like. The flavoring agent may be at least one selected from the group consisting of natural flavoring agents (tau martin, stevia extract (for example, rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharin, aspartame, etc.) The ratio of the natural carbohydrate may generally be about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the beverage composition, but is not limited thereto.

In addition to the above, the health functional food may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors or natural flavors, coloring agents, thickening agents (cheese, chocolate etc.), pectic acid or its salts, A salt thereof, an organic acid, a protective colloid thickener, a pH adjuster, a stabilizer, a preservative, a glycerin, an alcohol, and a carbonating agent used in a carbonated beverage. In addition, the health functional food, particularly the health functional beverage composition, may contain flesh for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These components may be used independently or in combination. The ratio of such additives is generally selected from the range of 1 to about 20 parts by weight per 100 parts by weight of the total composition, but is not limited thereto.

The herbal extracts of two or more kinds selected from the group consisting of Saururus chinensis, Saururus chinensis, Bacillus subtilis, Bacillus subtilis, and Bacillus subtilis, which are proposed in the present invention, can be used for the treatment of 5-lipoxygenase (5-LO) inhibitory activity, phosphodiesterase PDE) -4 inhibitory activity, adrenergic beta 2 receptor activation, cysteinyl leukotriene receptor (CysLT1 receptor) inhibitory activity, muscarinic receptor (eg, muscarinic M3 receptor) Therefore, it is expected to exert more favorable effects on respiratory diseases such as asthma, allergic rhinitis, bronchitis, chronic obstructive pulmonary disease, cough and cold since it exerts superior effects than each herbal medicine extract. In addition, the mixed herbal medicine extract has an advantage in that, in view of the general characteristics of natural extracts, there is less concern about adverse effects such as side effects and toxicity when administered to human than other synthetic drugs.

Hereinafter, the present invention will be described in detail with reference to preferred embodiments. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

[ Reference example : Preparation of single crude drug extract]

Reference example  1: Water Crude extract  Produce

200 g of dried and crushed ginseng (dried ginseng) were prepared. Each ginseng was treated with 2.6 L of distilled water at 80 ℃ for 3 hours The filtrate was collected by filtration to collect the filtrate, and the residue was re-extracted in the same manner. The filtrate was combined with the previously obtained filtrate and concentrated under reduced pressure to obtain 27.2 g (SC-W) of Saururus chinensis extract and 43.0 g (W-W), 33.6 g of water extract (SN-W) and 19.4 g of water extract (TK-W) were obtained.

Reference example  2: Ethanol Crude extract  Produce

200 g each of dried and cut Saururus chinensis, Bacillus thuringiensis, Yukyu, dried and pulverized guluth were prepared. The mixture was extracted with 2.6 L of 80% (v / v) ethanol at reflux for 3 hours at 80 ° C and then filtered to collect the filtrate , And the residue was re-extracted by the same method, followed by filtration, and the filtrate was combined with the previously obtained filtrate and concentrated under reduced pressure. In the state where most of the solvent was evaporated, about 10 times as much weight of distilled water was added to the concentrate for azeotropic concentration, and the concentration process was repeated twice more to obtain 24.6 g (SC-E) of Saururus chinensis ethanol extract, 41.4 (SN-E) and 34.8g (TK-E) of extracts of gangrene were obtained, respectively.

Reference example  3: Isopropanol Crude extract  Produce

200 g each of dried and cut Saururus chinensis, Bacillus thuringiensis, Yukyu, dried and pulverized guluth were prepared, and the mixture was extracted with 2.6 L of 80% (v / v) isopropanol at reflux for 3 hours at 80 ° C., , And the residue was re-extracted by the same method, followed by filtration, and the filtrate was combined with the previously obtained filtrate and concentrated under reduced pressure. In the state where most of the solvent was evaporated, about 10 times as much weight of distilled water as the weight was added to the mixture for azeotropic concentration, and the concentration was further repeated twice to obtain 22.2 g (SC-I) of Saururus chinensis isopropanol extract, 33.8 , 27.6 g (SN-I) of isopropanol extract and 32.6 g (TK-I) of extract of Bundang isopropanol were obtained, respectively.

Reference example  4: Solvent-soluble extract ( Butanol Fraction )

Five times as much distilled water as weight was added to each of the crude extracts of Saururus chinensis, Saururus thunbergii, Bacillus thuringiensis, Bacillus subtilis and Bacillus subtilis prepared according to the production methods of Referential Examples 1 to 3, and then suspended in the same volume of water saturated with butanol , And the obtained butanol-soluble fraction was concentrated under reduced pressure. After evaporating most of the solvent, about 10 times by weight of distilled water was added to the mixture to azeotropically concentrate the mixture, and then the same was repeated twice to obtain the following butanol fraction:

12.0 g (SC-I-B, butanol fraction of isopropanol extract), 12.5 g (SC-E-B, butanol fraction of ethanol extract), 9.8 g (SC-W-B, butanol fraction of water extract)

4.6 g (CM-W-B, butanol fraction of water extract), 9.6 g (CM-E-B, butanol fraction of ethanol extract), 8.8 g (butanol fraction of CM-I-B, isopropanol extract);

12.6 g (SN-E-B, butanol fraction of ethanol extract), 11.6 g (SN-I-B, butanol fraction of isopropanol extract), 9.8 g (SN-W-B, butanol fraction of water extract) And

27.0 g (TK-EB, butanol fraction of ethanol extract), 29.0 g (butanol fraction of TK-IB, isopropanol extract), 10.4 g (TK-WB, butanol fraction of water extract)

[ Example  2: Preparation of mixed extract]

Manufacturing example  1: Saururus chinensis extract and Bag  Mixture of extracts

(SC-W) and white clover extract (CM-W) of Reference Example 1, Saururus chinensis ethanol extract (SC-E) and White cucumber ethanol extract (CM-E) of Reference Example 2, Reference Example 3 Butanol fraction (SC-WB) and water-extracted butanol fraction (CM-WB) of the water extract of Saururus chinensis in Reference Example 4, Reference Example (SC-I) Butanol fraction (SC-IB) of the Saururus chinensis extract and the butanol fraction (SC-IB) of the Saururus chinensis extract of Reference Example 4 and the butanol fraction (SC-IB) of the Isopropanol extract 1: 3, 1: 3, 1: 5, 1: 3, 1: 1, 10: 1, 15: 1, and 20: 1. In order to obtain homogeneous mixing, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C. to prepare a mixed herbal medicine extract of Saururus chinensis and Bacillus thuringiensis for each solvent.

Manufacturing example  2: Saururus chinensis extract and The  Mixture of extracts

(SC-W) and water extract (SN-W) of Reference Example 1, the Saururus chinensis ethanol extract (SC-E) and the reference ethanol extract (SN-E) of Reference Example 2, the Saururus chinensis Butanol fraction (SC-WB) of the water extract of Saururus chinensis and the butanol fraction (SN-WB) of the water extract of Reference Example 4 of Reference Example 4, Saururus chinensis extract of Reference Example 4 (SN-EB) of the ethanol extract, butanol fraction (SC-IB) of the ethanol extract, the butanol fraction (SN-EB) of the ethanol extract of the reference ethanol, the butanol fraction 1: 3, 1: 3: 1, 5: 1, 10: 1, 1: 15: 1, and 20: 1. To obtain a homogeneous mixture, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C to prepare a herbal medicine extract of Saururus chinensis and Yonggusi for each solvent.

Manufacturing example  3: Saururus chinensis extract and Gulouin  Mixture of extracts

(SC-W) and bamboo plant extract (TK-W) of Reference Example 1, the Saururus chinensis ethanol extract (SC-E) of Reference Example 2 and the coelu ethanol extract (TK-E) of Reference Example 2, The butanol fraction (SC-WB) and the butanol fraction (TK-WB) of the bamboo extract of water extract of Saururus chinensis (SC-WB) and Reference Example 4 of Referential Example 4 in Reference Example 3, , The butanol fraction (SC-EB) of the Saururus chinensis ethanol extract and the butanol fraction (TK-EB) of the coelua ethanol extract of Reference Example 4, the butanol fraction (SC-IB) of the Saururus isopropanol extract of Reference Example 4 and the coeluin isopropanol extract 1: 1, 1: 5, 1: 3, 1: 1, 3: 1 (by weight) of the butanol fraction (TK-IB) , 5: 1, 10: 1, 15: 1, and 20: 1. To obtain a homogeneous mixture, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 DEG C to prepare a mixed herbal medicine extract of Saururus chinensis and Wurlauin for each solvent.

Manufacturing example  4: Bag  Extract and The  Mixture of extracts

(CM-W) and water-soluble extract (SN-W) of Reference Example 1, the white oyster ethanol extract (CM-E) and the reference ethanol extract (SN-E) of Reference Example 2, Reference Example 3 Butanol fraction (CM-WB) and butanol fraction (SN-WB) of the water extract of white irrigation water (CM-WB) and reference water extract (Reference Example 4) The butanol fraction (CM-EB) of the white oyster ethanol extract of Example 4 and the butanol fraction (SN-EB) of the ethanol extract of the oyster extract, the butanol fraction (CM-IB) of the white oyster isopropanol extract of Reference Example 4 and the butanol 1: 5, 1: 3, 1: 3, 1: 5, 1: 5, 1: : 1, 10: 1, 15: 1, and 20: 1. To obtain a homogeneous mixture, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C to prepare a mixed herb extract of white oyster and egg white for each solvent.

Manufacturing example  5: Bag  Extract and Gulouin  Mixture of extracts

(CM-W) and bamboo plant extract (TK-W) of Reference Example 1, the white oyster ethanol extract (CM-E) and the ghoulhu ethanol extract (TK-E) of Reference Example 2, (CM-WB) and the butanol fraction (TK-WB) of the bamboo shoot extract of Reference Example 4, , The butanol fraction (CM-EB) of the white irrigation ethanol extract of Reference Example 4 and the butanol fraction (TK-EB) of the extract of the blanched ethanol, the butanol fraction (CM-IB) of the white irrigation isopropanol extract of Reference Example 4, 1: 1, 1: 5, 1: 3, 1: 1, 1: 2, 1: 3: 1, 5: 1, 10: 1, 15: 1, and 20: 1. To obtain a homogeneous mixture, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C to prepare a mixed herbal extract of white clover and gangrene.

Manufacturing example  6: The  Extract and Gulouin  Mixture of extracts

(TK-E), Reference Example 3 (Reference Example 2), and Reference Example 3, respectively, in the same manner as in Reference Example 1, except that the water extract (SN-W) Butanol fraction (SN-WB) of a water-extracted extract of Reference Example 4 and a butanol fraction (TK-WB) of a versatile human extract, Reference Example 4 (SN- (SN-EB) and ethanol extract of butanol fraction (TK-EB) of ethanol extract of butanol fraction, butanol fraction (SN-IB) of reference isopropanol extract of reference example 4 and butanol fraction of alcohol extract of isobutanol 1: 5, 1: 3, 1: 1, 3: 1, 5: 1 (by weight of extract extract: weight of extract extracts: TK-IB) , 10: 1, 15: 1, and 20: 1. To make homogeneous mixing, water was added to each mixture in an amount of 3 times by weight, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C to prepare a mixed herbal medicine extract for each solvent.

Manufacturing example  7: Saururus chinensis extract, Bag  The extract and The  Mixture of extracts

(SC-E) of the Saururus chinensis extract (SC-W), the white irrigation water extract (CM-W) and the water extract (SN-W) Reference Example 2 of Reference Example 1, (SC-I), white islet isopropanol extract (CM-I) and canine isopropanol extract (SN-I) of Reference Example 3, Reference Example 4 (SC-WB), butanol fraction (CM-WB), and butanol fraction (SN-WB) of water extract of Saururus chinensis and water extract, butanol fraction (SC-IB) of the extract of Saururus chinensis Isopropanol (SC-IB) and Isobacterium Isopropanol Extract of Reference Example 4 in the same manner as in Example 1 except that the butanol fraction (CM-EB) and the butanol fraction The butanol fraction (CM-IB) and the butanol fraction (SN-IB) of the isopropanol extract, 1: 1: 3, 1: 3: 1, 3: 3: 1: 1, 3: 3: 1, 1: 1 as the weight basis (Saururus chinensis extract weight: 5: 1, 5: 5: 1, 1: 5: 5, and 5: 1: 5 . To obtain a homogeneous mixture, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C. to prepare a mixed herbal medicine extract of Saururus chinensis, Bacillus thuringiensis and Bacillus subtilis for each solvent.

Manufacturing example  8: Saururus chinensis extract, Bag  The extract and Gulouin  Mixture of extracts

(SC-W), white cedar water extract (CM-W), and water extract (TK-W) in the same manner as in Reference Example 1, Saururus chinensis extract (SC-E) (SC-I) and white isoleucine isopropanol extract (CM-I) and ginsenoside isoleucanol extract (TK-I) of Reference Example 3, (SC-WB), the butanol fraction (CM-WB) and the butanol fraction (TK-WB) of the water extract of Bacillus subtilis, the extract of Saururus chinensis extract of Reference Example 4 Butanol fraction (SC-EB) of butanol fraction (SC-EB) and white algae ethanol extract, butanol fraction (TK-EB) of gangrene ethanol extract, butanol fraction (SC-IB) of extract of Saururus chinensis isoflavone The butanol fraction (CM-IB) of isopropanol extract of white irrigation and the butanol 1: 1: 3, 1: 3: 1, 3: 1: 1: 1: 1: 5: 1, 1: 5: 5, and 5: 1, 3: 3: 1, 3: 3: 3, 3: 1: 3, 1: 5: 1: 5. To obtain homogeneous mixing, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C to prepare a mixed herbal medicine extract of Saururus chinensis, Bacillus subtilis, and Grapevine for each solvent.

Manufacturing example  9: Saururus chinensis extract, The  extract, Gulouin  Mixture of extracts

(SC-W), water extract (SN-W), water extract (TK-W), water extract of Saururus chinensis (SC-E) and water extract (TK-I), Reference Example 3 (SC-I) and Reference Isopropanol Extract (SN-I), Reference Example 3 (SC-WB) of the water extract of Saururus chinensis, the butanol fraction (SN-WB) of the water extract of canola, the butanol fraction (TK-WB) of the water extract of umbellinae, the butanol fraction (SC-IB) of the extract of Saururus chinensis isopropanol extract (SC-IB) and the extract of the isopropanol extract of Reference Example 4, butanol fraction (SN-EB) The fraction (SN-IB) and the extract of Bundang isopropanol (TK-IB) 1: 1: 3, 1: 3: 1, 3: 3: 1, 3: 3: 1, 1: 3: 5: 1, 5: 5: 1, 1: 5: 5, and 5: 1: 5. To obtain a homogeneous mixture, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C to prepare a mixed herbal medicine extract of Saururus chinensis, Yonggi, and Kwangju.

Manufacturing example  10: Bag  extract, The  extract, Gulouin  Mixture of extracts

(CM-W), a water extract (SN-W), a water extract (TK-W), a water extract of Bacillus (CM-E) in Reference Example 2, (SN-I) and gangrene isopropanol extract (TK-I) of Reference Example 3, the extracts of SN-E and SN-E, Butanol fraction (SN-WB) of the water extract, the butanol fraction (TK-WB) of the water extract of Wuchuan, the white elutriation ethanol extract of Reference Example 4 (CM-WB) Butanol fraction (CM-EB) of the isobutanol fraction (CM-EB) and the ethanol extract of canola extract, the butanol fraction (SN-EB) and the butanol fraction Butanol fraction (SN-IB) and gangrene isopropanol extract (TK-IB) of isopropanol extract of Yonggi 1: 1: 3, 1: 3: 1, 3: 3: 1, 3: 3: 1, 1: 3: 5: 1, 5: 5: 1, 1: 5: 5, and 5: 1: 5. To obtain a homogeneous mixture, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C to prepare a mixture of herbal extracts of Bacillus thuringiensis, Bacillus subtilis, and Baculaine for each solvent.

Manufacturing example  11: Saururus chinensis extract, Bag  extract, The  The extract and Gulouin  Mixture of extracts

(SC-W), white cedar water extract (CM-W), water extract (SN-W) and water extract (TK-W) in the same manner as in Reference Example 1 and Saururus chinensis ethanol extract (SC-I) of the Reference Example 3 and the white algae extract (SC-E), the white algae ethanol extract (CM-E) (SC-WB) of the water extract of Saururus chinensis (Saururus chinensis) and the butanol fraction of the water extract of white mulberry (CMR-I), the isopropanol extract Butanol fraction (SC-EB) of the saururic acid ethanol extract of Reference Example 4 and the ethanol fraction of sucrose (CM-WB), the butanol fraction (SN-WB) of the water extract and the butanol fraction (TK- The butanol fraction (CM-EB) of the extract, the butanol fraction (SN-EB) of the ethanol extract and the butanol (SN-IB) of extracts of butanol fraction (SC-IB) and isobutanol fraction (CM-IB) of isobutanol isopropanol extract and isobutanol fraction (SN-IB) of isobutanol fraction The butanol fraction (TK-IB) of the isopropanol extract was diluted 1: 1: 1: 1, 1: 1: 1: 3, 1: 1: 3: 1, 1: 3: 1, 1: 3: 1, 3: 3: 1, 3: 3, 3: 1, 3: 1: 1: 1, 1: 1: 5: 5, 1: 5: 1, 1: 5: 5: 1, 5: 5: 5: 1, 5: 1: 5: , 5: 1: 5: 5, and 1: 5: 5: 5. To obtain homogeneous mixing, 3 parts by weight of water was added to each mixture, and the mixture was concentrated under reduced pressure at a temperature of 50 ° C to prepare a mixed herbal medicine extract of Saururus chinensis, Bacillus thuringiensis, Bacillus subtilis and Baculaine.

[ Example  3: pharmacological activity test]

3.1: Lipoxygenase  (5- lipoxygenase ; 5- LO ) Inhibitory activity

Each of the mixed herbal medicine extracts prepared in Preparation Examples 1 to 11 and the respective herbal medicine extracts prepared in Reference Examples 1 to 4 (hereinafter referred to as "test drug") were mixed with 5 (Rufahl RA et al., Ana. Biochem. 2007, 15; 364 (2), 204-12). 5-LO metabolizes arachidonic acid, a substrate, to produce various leukotrienes that are inflammatory and asthmatic inducers. Thus, by measuring the amount of leukotrienes (e.g., LTB4) produced from arachidonic acid, the degree of activity of 5-LO can be assessed. In addition, evaluation of the 5-LO activity inhibition of the test drug can confirm the prevention and / or improvement and / or therapeutic effect of respiratory diseases.

Specifically, 3.0 μM arachidonic acid was prepared as a substrate, and each test drug was dissolved in dimethylsulfoxide (DMSO) to prepare a final concentration of 100 μg / ml. Human recombinant 5-LO (Cayman Chemical 60402) was dissolved in a pH 7.4 Tris buffer at a concentration of 10 U / ml, and 100 μl of the prepared test drug and fluorescent probe dihydrorhodamine-1,2,3 (DHR123) (5-LO) at 25 < 0 > C for 5 minutes. After the addition of 25 μM arachidonic acid, the reaction was allowed to proceed for 20 minutes and the amount of LTB4 produced after the reaction was measured as fluorescence intensity at excitation and emission of 485 nm and 530 nm using a spectrophotometer (SpectraMax, Molecular Devices) . The 5-LO inhibition rate of the test drug was evaluated by the following equation:

5-LO inhibition rate (%) = [1- (LTB4 production amount in each test drug and 5-LO treated sample) / (amount of LTB4 production in untreated drug treated with 5-LO only)] * 100

The obtained results are shown in Tables 1 to 7 below.

[Table 1]

[Table 2]

[Table 3]

[Table 4]

[Table 5]

[Table 6]

[Table 7]

3.2: Phosphodiesterase  ( phosphodiesterase ; PDE ) -4 inhibitory activity

Each of the mixed herbal medicine extracts prepared in Preparation Examples 1 to 11 and each of herbal medicine extracts prepared in Reference Examples 1 to 4 (hereinafter referred to as "test drug") was added to the phosphodiesterase- 4 (PDE-4) inhibitory activity (see Nicholson CD et al., Trends Pharmacol. Sci. 12 (1); 19-27, 1991).

PDE-4 hydrolyzes cAMP and depletes intracellular cAMP, resulting in bronchoconstriction. Therefore, it is possible to evaluate the activity of PDE-4 by measuring the amount of the substance obtained in the course after cAMP degradation. By evaluating inhibition of PDE-4 activity of the test drug, prevention and / or improvement of respiratory diseases and / .

Specifically, 1.01 μM [ ³ H] cAMP + cAMP (ARC0163) was prepared as a substrate, and each test drug was dissolved in DMSO to prepare a final concentration of 100 μg / ml. Human monocytic cell line U937 cell (ATCC CRL 1593.2) was cultured in RPMI 1640 medium (Gibco) containing 100 units / ml penicillin-streptomycin and 10% (w / v) fetal bovine serum at 37 ° C in a 5% CO ₂ incubator Lt; / RTI > The cells (1 x 10 ⁹ cells) were washed with PBS and resuspended in buffer (20 mM Tris-HCl pH 7.1, 3 mM 2-mercaptoethanol, 1 mM MgCl 2, 0.1 mM EGTA, 1 μM PMSF, 1 ug / ml leupeptin) After dissolving and homogenizing, the supernatant obtained by centrifugation was separated into PDE4 using a Sephacryl S-200 column (GE Healthcare Life Science). PDE-4 derived from human U937 cells obtained above was dissolved in Tris-HCl buffer (pH 7.5) at a concentration of 2.5 μg / ml, and the prepared test drug was added with 100 μl of the enzyme (PDE-4) After 15 minutes of reaction, the substrate was treated and allowed to react for 20 minutes. The obtained reaction product was boiled at 100 ° C to terminate the reaction, and 5'-nucleotidase (Sigma Aldrich) was added thereto. The reaction was carried out on an AG1-X2 resin column (Bio-Rad) and [ ³ H] adenosine Perkin Elmer MicroBeta 1450). The inhibition rate of PDE-4 of the test drug was evaluated by the following equation:

PDE-4 inhibition rate (%) = [1- (each test drug and PDE-4 in the treated sample ^[3 H] Adenosine generation amount) / (untreated test drug (the processing man PDE-4) ^[3 H in the sample ] Adenosine production amount)] * 100

The obtained results are shown in Tables 8 to 14 below.

[Table 8]

[Table 9]

[Table 10]

[Table 11]

[Table 12]

[Table 13]

[Table 14]

3.3: beta-2-adrenergic receptor (beta 2- adrenergic  receptor Ligand  Liver-binding inhibition activity

Each of the mixed herbal medicine extracts prepared in Preparation Examples 1 to 11 and the respective herbal medicine extracts prepared in Reference Examples 1 to 4 (hereinafter referred to as " test drug " Experiments were performed to assess the activity of the 2-adrenergic receptor and ligand binding (McCrea KE and Hill SJ, Br. J. Pharmacol. 110: 619, 1993). When the beta-2-adrenergic agonist binds to the receptor, it activates the beta-2-adrenergic receptor and increases cAMP, thereby inducing expansion of the airway smooth muscle, thereby preventing / treating respiratory diseases. In this example, similar to the beta-2-adrenergic agonist, in order to determine whether the test drug can bind to the beta-2-adrenergic receptor and induce activation, Adrenoceptor binding inhibition rate of the competitive substance in the binding (rate of inhibition of receptor binding of the competitive substance), the test drug binds to the beta-2-adrenergic receptor to induce activation of the receptor and prevention of respiratory diseases And / or treatment.

Each test drug was dissolved in DMSO and prepared to a final concentration of 100 ug / ml. The [ ³ H] CGP-12177 ligand (Perkin Elmer, NET-1061; competitor), an antagonist specific for the human recombinant beta-2-adrenergic receptor (Perkin Elmer, ES-034-M) expressed in CHO- The degree of binding was measured. in Tris-HCl buffer, pH7.4 human recombinant beta-2-adrenoceptor 50 ug and 0.2 nM of ^[3 H] CGP-12177, and was 60 minutes of reaction the test drug prepared in 100 ul 25 ℃, specific binding them . The obtained reaction mixture was filtered with a glass fiber filter paper (GF / B, Whatman), and the residue containing the ligand-receptor complex bound to the filter paper was washed three times with Tris-HCl buffer , And the binding of the labeled ligand ([ ³ H] CGP-12177) to the receptor was measured. The labeled ligand-receptor binding was quantified by measuring the labeled ligand level in the residue after filtration using a scintillation counter (Counter-Microbeta, Perkin Elmer).

To determine the non-specific binding of the ligand [ ³ H] CGP-12177 used as the competitive material, 10 μM (micromole) of ICI-118551 (3- (isopropylamino) -1 - [(7-methyl-4-indanyl) oxy ] butan-2-ol). (Treatment with [ ³ H] CGP-12177, filtration, and measurement of binding between the labeled ligand and the receptor). ICI-118551 is also a ligand that specifically binds to the beta-2 adrenergic receptor and is used in an unlabeled state, unlike [ ³ H] CGP-12177. Since the unlabeled ICI-118551 is used at a very high concentration (10 uM) compared to the labeled [ ³ H] CGP-12177 (0.2 nM), all of the beta-2 adrenergic receptors are associated with unlabeled ICI-118551 In this case, if [ ³ H] CGP-12177 labeled with the residue obtained as a result of the filtration is detected, the detected [ ³ H] CGP-12177 is not a beta-2 adrenergic receptor And a non-specific combination. Thus, the binding of the labeled ligand ([ ³ H] CGP-12177) when not labeled (ICI-118551) (total binding: labeled ligand and beta-2 adrenergic receptor binding ([ ³ H] CGP-12177) when unlabeled ligand (ICI-118551) was used in the binding between a labeled ligand and a site other than a beta-2 adrenergic receptor (ie, non-specific binding) Specific binding of the labeled ligand to the beta-2 adrenergic receptor can be obtained by subtracting the binding (non-specific binding: binding between the labeled ligand and sites other than the beta-2 adrenergic receptor). The binding between the labeled ligand and the receptor can be obtained by measuring the level of the labeled ligand (for example, the signal level of the labeling substance) in the residue that has not passed through the filter after filtration as described above.

Therefore, the percent inhibition of receptor binding of the competitor by the test drug was evaluated by the following equation:

(%) = [1- (specific binding in the presence of the test drug) / (specific binding when the test drug is not treated)] * 100

In the above formula, the specific binding is defined as the value obtained by subtracting the non-specific binding from the total binding between the labeled ligand and the receptor, as described above.

The obtained results are shown in Tables 15 to 21 below.

[Table 15]

[Table 16]

[Table 17]

[Table 18]

[Table 19]

[Table 20]

[Table 21]

3.4: Leukotriene  ( cysteinyl leukotriene ; CysLT1 ) Receptor binding inhibitory activity

The mixed herbal medicine extracts prepared in Preparation Examples 1 to 11 and the single herbal medicine extracts prepared in Reference Examples 1 to 4 (hereinafter referred to as "test drug") were mixed with leucotriene cysteinyl leukotriene LT1; CysLT1) receptor binding (see Martin V et al., Biochem. Pharmacol. 62 (9): 1193, 2001).

The CysLT1 receptor binds with the ligand leukotriene to induce respiratory diseases such as asthma by inducing bronchoconstriction, increased vascular permeability, airway infiltration of inflammatory cells, edema formation, promotion of mucous secretion, inflammation, hypersensitivity, Inhibition of ligand-to-ligand binding may inhibit such respiratory diseases. Therefore, by evaluating the inhibitory activity of the test drug between the CysLT1 receptor and the ligand, the preventive and / or therapeutic effect of respiratory diseases can be confirmed.

Each test drug was dissolved in DMSO and prepared to a final concentration of 100 ug / ml.

The degree of binding of [ ³ H] LTD4 ligand (Perkin Elmer, NET-1019) specific for the human recombinant CysLT1 receptor (Perkin Elmer, ES-470-M) expressed in CHO-K1 cells was measured. In a Tris-HCl buffer of pH 7.4, 5 ug of human recombinant CysLT1 receptor, 0.3 nM [ ³ H] LTD4 and 100 μl of the prepared test drug were reacted at 25 ° C for 30 minutes and used for the measurement of specific binding . The obtained reaction mixture was filtered with a glass fiber filter paper (GF / B, Whatman), and the residue containing the ligand-receptor complex bound to the filter paper was washed three times with Tris-HCl buffer The binding between the labeled ligand ([ ³ H] LTD4) and the receptor was measured. The labeled ligand-receptor binding was quantified by measuring the labeled ligand level in the residue after filtration using a scintillation counter (Counter-Microbeta, Perkin Elmer).

To measure the nonspecific binding of the labeled ligand [ ³ H] LTD4, the above procedure was performed using the unlabeled ligand LTD4 with the labeled ligand in an amount of 300 nM.

The specific binding of the labeled ligand to the CysLT1 receptor can be determined by non-specific binding (binding between the labeled ligand and the receptor obtained when no unlabeled ligand is used) (labeled ligand obtained when unlabeled ligand is used And receptor binding), and the specific method is as described in Example 3.3 above.

The inhibitory activity of CysLT1 receptor and ligand binding of each test drug was evaluated by the following equation:

(%) = [1- (specific binding in the presence of the test drug) / (specific binding when the test drug is not treated)] * 100

In the above formula, the specific binding is defined as the value obtained by subtracting the non-specific binding from the total binding between the labeled ligand and the receptor, as described above.

The obtained results are shown in Tables 22 to 28 below.

[Table 22]

[Table 23]

[Table 24]

[Table 25]

[Table 26]

[Table 27]

[Table 28]

3.5: Muscarin  Receptor Muscarinic M3 receptor ) Binding inhibitory activity

The mixed herbal medicine extracts prepared in Preparation Examples 1 to 11 and the herbal medicine extracts prepared in Reference Examples 1 to 4 (hereinafter referred to as "test drug") were mixed with muscarinic (Buckley NJ et al., Mol. Pharmacol. 35 (4): 469, 1989) for evaluating the activity for binding to the receptor (Muscarinic M3 receptor). Activation of muscarinic M3 receptors promotes contraction and mucus secretion of the airway smooth muscle, thus inhibiting the activity of muscarinic M3 receptors can prevent and / or treat respiratory diseases. Therefore, by evaluating the inhibitory activity of the test drug against muscarinic M3 receptor and ligand, prevention and / or therapeutic effect of respiratory diseases such as asthma and chronic obstructive pulmonary disease can be confirmed.

Each test drug was dissolved in DMSO and prepared to a final concentration of 100 ug / ml.

The degree of binding of [ ³ H] N-Methylscopolamine ligand (Perkin Elmer, NET-636), a specific antagonist to human recombinant muscarinic M3 receptor (Perkin Elmer, RBHM3M) expressed in CHO-K1 cells was measured. 12 μg of human recombinant muscarinic M3 receptor, 0.8 nM of [ ³ H] N-Methylscopolamine and 100 μl of the prepared test drug were reacted at 25 ° C. for 120 minutes in Tris-HCl buffer at pH 7.4, . The reaction mixture thus obtained was filtered with a glass fiber filter paper (GF / B, Whatman), and the residue containing the ligand-receptor complex bound to the filter paper was repeatedly washed with Tris-HCl buffer, The binding between the labeled ligand ([ ³ H] N-Methylscopolamine) and the receptor was measured. The labeled ligand-receptor binding was quantified by measuring the labeled ligand level in the residue after filtration using a scintillation counter (Counter-Microbeta, Perkin Elmer).

To measure the nonspecific binding of the labeled ligand [ ³ H] N-Methylscopolamine, the unlabeled atropine, another ligand of the muscarinic M3 receptor, along with the labeled ligand, in an amount of 1 uM, Respectively.

Specific binding of a labeled ligand to a muscarinic M3 receptor can result in non-specific binding (binding between the labeled ligand and the receptor, obtained when no unlabeled ligand is used) (label obtained when unlabeled ligand is used Binding between the ligand and the receptor), and a specific method thereof is as described in Example 3.3 above.

The inhibitory activity of muscarinic M3 receptor and ligand binding of each test drug was evaluated by the following equation:

(%) = [1- (specific binding in the presence of the test drug) / (specific binding when the test drug was not treated)] * 100

In the above formula, the specific binding is defined as the value obtained by subtracting the non-specific binding from the total binding between the labeled ligand and the receptor, as described above.

The obtained results are shown in Tables 29 to 35 below.

[Table 29]

[Table 30]

[Table 31]

[Table 32]

[Table 33]

[Table 34]

[Table 35]

[ Example  4: Preparation of tablets]

200 mg of the crude herbal medicine extracts prepared in Preparation Examples 1 to 11 were mixed with 10 mg of light silicic anhydride, 2 mg of magnesium stearate, 50 mg of microcrystalline cellulose, 25 mg of starch glycolic acid sodium, 113 mg of corn starch and an appropriate amount of anhydrous ethanol And then tableted by a conventional method.

[ Example  5: Preparation of health food]

1000 mg of any of the herbal extracts (any of Preparation Examples 1 to 11)

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

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 quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

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

[ Example  6: Preparation of health drink]

1000 mg of any of the herbal extracts (any of Preparation Examples 1 to 11)

Citric acid 1000 mg

Oligosaccharide 10 g

Plum concentrate 2 g

Taurine 1 g

Purified water was added and the entire 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the resulting solution was stirred and heated at 85 DEG C for about 1 hour. The resulting solution was filtered to obtain a sterilized 2L container, sealed sterilized and refrigerated, Of healthy beverage composition.

The composition ratio is merely an example of a component suitable for a favorite drink, and the blending ratio may be arbitrarily changed in accordance with the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (17)

Wherein the herb extract comprises two herbal extracts selected from the group consisting of Saururus chinensis,
The mixed herbal medicine extract may be a mixture of herbal medicine extracts mixed at a mixing ratio of 1:10 to 10: 1 on the weight basis, or a mixture of herbal medicine extracts of Saururus chinensis, Saururus chinensis, Which is an extract of a mixed herbal medicine mixed at a mixing ratio of 1:10 to 10: 1 on the basis of weight, two kinds of selected from the group consisting of white clover,
A pharmaceutical composition for preventing or treating respiratory diseases. [3] The method of claim 1, wherein the extract of Saururus chinensis extract, Bacillus thuringiensis extract, Bacillus subtilis extract, and Bacillus subtilis extract or mixed herbal medicine comprises at least one solvent selected from the group consisting of water and a straight chain or branched alcohol having 1 to 4 carbon atoms Which is obtained by adding at least one nonpolar solvent selected from the group consisting of linear or branched alcohols having 1 to 6 carbon atoms or hexane, dichloromethane, chloroform, and ethyl acetate to the obtained crude extract or the crude extract, Gt; delete The pharmaceutical composition according to claim 1, wherein the mixing ratio of the two species is 1: 5 to 5: 1 by weight. The pharmaceutical composition according to claim 1, wherein the mixing ratio of the two species is 1: 3 to 3: 1 by weight. A herbal extract of three kinds selected from the group consisting of Saururus chinensis,
The mixed herbal medicine extract may be a mixture of herbal medicine extracts mixed with three kinds selected from the group consisting of Saururus chinensis extract, Bacillus thuringiensis extract, Bacillus subtilis extract, and Bacillus subtilis, or a mixture of three herbal extracts selected from the group consisting of Saururus chinensis, Wherein the mixture of the three species is 1 to 3: 1 to 3: 1 to 3 on the basis of weight, the pharmaceutical composition for preventing or treating respiratory diseases. Including herbal extracts of Saururus chinensis, Bacillus thuringiensis,
The mixed herbal medicine extract is an extract of a mixture of herbal medicine extracts mixed with Saururus chinensis extract, Bacillus thuringiensis extract, Bacillus subtilis extract, and Bacillus subtilis or a mixed herbal medicine mixed with Saururus chinensis, Bacillus thuringiensis, Bacillus subtilis, Wherein the mixing ratio is 1 to 3: 1 to 3: 1 to 3: 1 to 3 on a weight basis. The method of claim 1, wherein the mixed herbal medicine extract is a mixture of Saururus chinensis extract and White irrigation extract, or a mixed herbal medicine comprising Saururus chinensis and White irritant, and the mixture ratio of Saururus chinense extract and White irrigation extract, Wherein the ratio is 1: 10 to 10: 1 by weight. 8. The pharmaceutical composition according to claim 8, wherein the mixing ratio of the Saururus chinensis extract to the Bacillus thuringiensis extract or the mixture ratio of Bacillus thuringiensis and Bacillus thuringiensis is 1: 5 to 5: 1 by weight. [Claim 9] The pharmaceutical composition according to claim 9, wherein the mixing ratio of the Saururus chinensis extract to the White Ibis extract or the mixture ratio of Saururus chinensis and White Ibis is 1: 3 to 3: 1 by weight. 11. The pharmaceutical composition according to any one of claims 8 to 10, wherein the mixed herbal extract extract further comprises a herbal extract, a gherkin extract, or a herbal extract and a mixed herbal extract of ghuline. 12. The method according to claim 11, wherein the mixed herbal medicine extract is a mixture of herbal medicine extracts mixed with Saururus chinensis extract, White mulberry extract, and Mulberry extract or a mixed herbal medicine mixed with Saururus chinensis, Wherein the mixing ratio of the extract, the extract, and the extract is in the range of 1 to 3: 1 to 3: 1 to 3. 12. The method according to claim 11, wherein the mixed herbal medicine extract is a mixture of herbal medicine extracts mixed with Saururus chinensis extract, Bacillus thuringiensis extract, and Bacillus subtilis, or a mixed herbal medicine mixed with Saururus chinensis, Wherein the blend ratio of Bacillus thuringiensis extract, Bacillus thuringiensis extract, and Bacillus subtilis extract or the mixture ratio of Bacillus thuringiensis, Bacillus thuringiensis, and Baculus is 1: 3: 1 to 3: 1 to 3. 12. The method according to claim 11, wherein the mixed herbal medicine extract is a mixture of herbal medicine extracts mixed with Saururus chinensis extract, Bacillus thuringiensis extract, Bacillus subtilis extract, and Bacillus subtilis or an extract of mixed herbal medicine mixed with Saururus chinensis, Wherein the mixing ratio of the Saururus chinense extract, Saururus chinensis extract, Bacillus subtilis extract, and Bacillus subtilis or the mixing ratio of Saururus chinensis, Bacillus subtilis, Bacillus subtilis and Baculus is from 1: 3: 1 to 3: 1 to 3: , ≪ / RTI > 11. A pharmaceutical composition according to any one of claims 1, 2, and 4 to 10 wherein the respiratory disease is asthma, allergic rhinitis, bronchitis, chronic obstructive pulmonary disease, cold or cough. 10. A herbal composition comprising the mixed herbal extract according to any one of claims 1, 2, and 4 to 10,
Health functional food for prevention or improvement of respiratory diseases. 12. The pharmaceutical composition according to claim 11, wherein the respiratory disease is asthma, allergic rhinitis, bronchitis, chronic obstructive pulmonary disease, cold or cough.