KR20180075092A - Composition for preventing, improving or treating chronic obstructive pulmonary disease comprising Bupleurum falcatum extract as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating or treating chronic obstructive pulmonary disease comprising a Bupleurum falcatum extract as an effective component. The Bupleurum falcatum extract can reduce the expression levels of CXCL8, CXCL5, ALDH3A1 and ALDH1A1 which are increased due to chronic obstructive pulmonary disease or lung cancer in the EMT signaling pathway. Therefore, the composition of the present invention can be useful for medicines or health functional food for the prevention, alleviation or treatment of chronic obstructive pulmonary disease or lung cancer.

Description

[0001] The present invention relates to a composition for preventing, ameliorating or treating chronic obstructive pulmonary disease,

The present invention relates to the use of Bupleurum falcatum ) extract as an active ingredient for the prevention, amelioration or treatment of chronic obstructive pulmonary disease.

Since COPD is a major source of smoking, smoking cessation is very important, and it can occur not only in smoking cessation but also in occupational groups that deal directly with environmental pollutants such as cadmium (Cd) and silica (Si), and rarely in chronic occupational diseases like chronic obstructive pulmonary disease RTI ID = 0.0 > a1-antitrypsin < / RTI > gene.

Chronic obstructive pulmonary disease (COPD) is caused by excessive production of mucus to protect the tissue from inflammation in the lungs, resulting in clearance problems resulting in closure of the airways and destruction of alveolar cells Is a fatal disease that can lead to death due to induced hypoxia.

Chronic obstructive pulmonary disease is classified into chronic bronchitis and emphysema depending on the tissue and mechanism of development. The most important feature of chronic bronchitis is overexpression of overexpressed mucus in airway epithelial cells, resulting in accumulation The emphysema is characterized by the destruction of the alveolar cells, resulting in hypoxia or heart disease, and irreversible bronchiole obstruction.

Such chronic obstructive pulmonary disease may increase the incidence of lung cancer, particularly small cell lung cancer, by 4 to 5 times. Thus, chronic obstructive pulmonary disease and lung cancer are closely related to the epithelial mesenchymal transition (EMT) signaling system (EBioMedicine 2 (2015) 1578-1579, Clinical and Translational Medicine 2014, 3:33).

Meanwhile, bronchodilators and corticosteroids are known to help relieve the symptoms of chronic obstructive pulmonary disease. In particular, direct administration of corticosteroids and bronchodilators to the airway, the target tissue of the inflammatory response, is known to be a preferred method of avoiding systemic side effects rather than systemic administration of the drug. Other drugs such as Theophylline are also known to be beneficial, but most of the therapies that have been developed so far can not cure chronic obstructive pulmonary disease or improve it fundamentally.

On the other hand, Shijo is also called Shijo-no-Sho, Shun-minari. It grows in the grass and is 40 ~ 70cm high. There are no hairs in the whole aeration, and the branches are divided on the thin long stem. Roots are thick and short. The leaves are staggered, line-shaped or bar-shaped, the edges are flat, and the veins are parallel. The leaf on the root is like petiole with its lower part narrowed, and its length is 10 ~ 30cm. The leaves on the stem are 4 ~ 10cm long, 5 ~ 15mm long and the tip is pointed. Flowers bloom from August to September in yellow, with a double-inflorescence at the end of stem and branch. There are 2 ~ 7 peduncle, 5 ~ 10 flowers each. The gun flakes are narrow, narrow, split into 5 segments, 15 mm long, and the small guns are long elliptical or wider, 2.5-4 mm long. There are 5 petals and stamen and the ovary is lower (lower). Fruit is elliptical with division, about 3mm in length, ripened in September-October. Reproduction is either giving up or giving up seeds. The root contains saponin and fatty oil, and it is used as a medicinal herbal medicine for fever, analgesic, tonic, respiratory, digestive and circulatory diseases. They are distributed in Korea, Japan, China, Mongolia, Siberia, Caucasus and Europe.

Prior art related to Seoh is a pharmaceutical composition for the prevention and treatment of inflammatory diseases including a triterpenoid saponin compound of Korean Patent Laid-Open Publication No. 2011-0061883, and Korean Patent Publication No. 2012-0038613 discloses a pharmaceutical composition for treating lung cancer A pharmaceutical composition for preventing or treating chronic obstructive pulmonary disease is disclosed, but a composition for preventing, ameliorating or treating chronic obstructive pulmonary disease containing the extract of the present invention as an active ingredient has not been disclosed.

Accordingly, the present invention provides a composition for preventing, ameliorating or treating chronic obstructive pulmonary disease, wherein the effective ingredient of the extract of the present invention is effective for preventing or treating chronic obstructive pulmonary disease The present inventors have completed the present invention by confirming that the expression level of the protein involved in EMT signaling, which is known to be associated with chronic obstructive pulmonary disease, is controlled in the in vitro cell model.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention or treatment of chronic obstructive pulmonary disease containing Bupleurum falcatum extract as an active ingredient.

In addition, the present invention relates to the use of Bupleurum falcatum ) extract as an active ingredient for preventing or ameliorating chronic obstructive pulmonary disease.

The present invention relates to a composition for preventing, ameliorating or treating chronic obstructive pulmonary disease containing an extract of Aspergillus oryzae as an active ingredient, wherein the extract has an increased or decreased amount of expression by the chronic obstructive pulmonary disease or lung cancer in the EMT signaling pathway The amount of protein expression can be controlled, and thus it can be usefully used for medicines or health functional foods for the prevention, improvement or treatment of chronic obstructive pulmonary disease or lung cancer.

1 is a microarray result showing changes in the expression levels of CXCL8, CXCL5, ALDH3A1 and ALDH1A1 genes in normal cells, chronic obstructive pulmonary disease and lung cancer cell lines. Normal is the expression level of the gene obtained from normal cells, COPD is the expression level of the gene obtained in the patient with chronic obstructive pulmonary disease, and Cancer is the expression level of the gene obtained from the lung cancer cell.
FIG. 2 is a model system for in vitro COPD. FIG. 2 is a graph showing the changes in the amounts of CXCL8 and CXCL5 expressed in cells treated with tobacco smoke extract and the amounts of CXCL8 and CXCL5 expressed in cells treated with tobacco smoke extract (A, B), and the expression level of CXCL8 in the A549 cells, a lung cancer cell line, was examined by treatment with the extract of Seikosugi.
FIG. 3 shows the expression levels of E-cadherin, N-cadherin, vimentin, and ALDH3A1 protein, which are EMT signaling-related proteins, according to the treatment of the extract of the present invention, in a cell model in which chronic obstructive pulmonary disease is induced by treating tobacco smoke extract It is the result of Western blot which confirmed change.

The present invention relates to the use of Bupleurum The present invention relates to a pharmaceutical composition for the prevention or treatment of chronic obstructive pulmonary disease containing an extract of falcatum as an active ingredient.

The seed extract may be prepared by a method including, but not limited to, the following steps:

1) extracting Bupleurum falcatum with an extraction solvent;

2) filtering the extract of step 1); And

3) Concentrating the filtrate obtained in step 2) under reduced pressure and drying to prepare an extract.

In step 1), the extraction solvent is preferably water, a C ₁ -C ₄ lower alcohol or a mixture thereof, more preferably an ethanol extract, more preferably a 70% (v / v) ethanol extract Not limited.

In the above production method, the extraction of Seo can be carried out by any conventional method known in the art such as filtration, hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction. It is preferable that the extraction solvent is added by 1 to 20 times the volume of the reference volume, more preferably 3 to 10 times. The extraction temperature is preferably 20 to 50 DEG C, but is not limited thereto. The extraction time is preferably 10 to 100 hours, more preferably 24 to 96 hours, most preferably 72 hours. In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

The composition of the present invention is characterized by inhibiting the development of lung cancer caused by chronic obstructive pulmonary disease. The above-mentioned chronic obstructive pulmonary disease is preferably emphysema or chronic bronchitis, but is not limited thereto.

The composition of the present invention may contain a pharmaceutically acceptable carrier, excipient or diluent in addition to the above-mentioned active ingredient. The composition may be oral or parenteral, and may be in the form of a capsule, a powder, a granule, Emulsions, syrups, and aerosols. When formulating the composition, it is prepared by using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are generally used. Solid formulations for oral administration include capsules, powders, granules, tablets, pills, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used. Liquid formulations for oral administration include suspensions, emulsion syrup aerosols and the like. Various excipients such as wetting agents, sweetening agents, fragrances, preservatives and the like may be included in addition to water and liquid paraffin which are commonly used simple diluents. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of suppositories include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like. Intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine, intracerebral, or intracerebral injection methods are preferably selected for parenteral administration.

The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The dosage of the composition of the present invention varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of disease, , Preferably 30 to 500 mg / kg, more preferably 50 to 300 mg / kg, and can be administered 1 to 6 times a day. The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

In addition, the present invention relates to the use of Bupleurum The present invention relates to a health functional food composition for preventing or ameliorating chronic obstructive pulmonary disease.

The health functional food of the present invention can be used as it is or in combination with other food or food ingredients, and can be suitably used according to conventional methods. There is no particular limitation on the kind of the health food. Examples of the food to which the extract of the present invention can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums and ice cream, various soups, drinks, tea, , An alcoholic beverage and a vitamin complex, and includes all the health foods in a conventional sense. The health drink containing the composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as a conventional beverage. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention.

The health functional food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, Glycerin, alcohols, carbonating agents used in carbonated beverages, and the like. And may further contain flesh for the production of fruit juice or vegetable beverages. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 2 parts by weight based on 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Example  1. Preparation of Seiho Extract

In order to prepare the extract of the present invention, Sehho was purchased from Kwang Myung Dang Pharmaceutical. And extracted with 70% (v / v) ethanol corresponding to a volume of 10 times the total weight of the cells. Thereafter, the filtrate obtained by using the filter paper was subjected to removal of the solvent by using a reduced pressure concentrator to prepare a Seiho extract.

Example  2. Chronic obstructive pulmonary disease ( COPD ) And lung cancer (common cancer) Marker  Expression level analysis

MRNA was obtained from a normal human lung cell line (n = 3), a lung cell line derived from COPD patients (n = 3) and a lung cancer cell line (n = 3) in order to identify a biomarker associated with chronic obstructive pulmonary disease and lung cancer, Affymetrix Human Gene ST array 2.0 was analyzed using this. The information on the cell line used in Example 2 is shown in Table 1.

Affymetrix Human Gene ST array Cell line used for analysis Cell name Cell type Classification Where to buy NHLF Normal human lung fibroblast normal Lonza BSMC Bronchial smooth muscle cell normal Lonza NHBE Normal human bronchial epitheilal cell normal Lonza DHLF-COPD Diseased human lung fibroblast-COPD Chronic obstructive pulmonary disease Lonza DBSM-COPD Diseased bronchial smooth muscle cell-COPD Chronic obstructive pulmonary disease Lonza DHBE-COPD Diseased human bronchial epithelial cell-COPD Chronic obstructive pulmonary disease Lonza HLA-CAF Cancer associated fibroblast human lung adenocarcinoma Lung cancer Neuromics A549 Adenocarcinomic human alveolar basal epithelial cell Lung cancer ATCC NCI-H460 Non-small lung cancer epithelial cell Lung cancer ATCC

As a result, various gene expression changes were observed. Among them, COPD and gene products (CXCL8, CXCL5, ALDH3A1 and ALDH1A1) showing progressive expression patterns in lung cancer were observed compared to normal cells (FIG. 1) (epithelial Mesenchymal transition) signaling.

Example  3. Treatment by Seiho extract EMT  Involved in signal transduction CXCL8  And CXCL5  Inhibitory effect of protein on expression

Smoking is known to be a major cause of COPD and lung cancer, and a modified lung cell line model through long-term cigarette-smoke condensate (CSC) treatment has been widely used as an in vitro model for the development and progression of COPD and lung cancer . In this example, BEAS-2B cells, normal human bronchial epithelial cells, were treated with 50 μg / ml of tobacco smoke extract for 2 to 4 weeks to establish a chronic obstructive pulmonary disease model in vitro . The BEAS-2B cells and the CSC-treated BEAS-2B cells were treated with 10% fetal bovine serum (FBS), 100 units / ml penicillin, 100 μg / ml streptomycin The cells were cultured in an incubator maintained at 37 ° C and 5% (v / v) CO ₂ using the DMEM medium for subsequent in vitro experiments.

BEAS-2B cells and BEAS-2B cells treated with tobacco smoke extract were divided into 1 × 10 ⁴ cells per well on a 96-well culture plate, cultured for 24 hours, and cultured at 0, 25, 50, 100 μg / Seoho extract was treated. Forty-eight hours after the treatment, the supernatant of the culture was collected and the amount of CXCL8 and CXCL5 released into the cell was measured using an enzyme-linked immunosorbent assay (ELISA).

When the BEAS-2B cells were treated with a cigarette smoke condensate, the amount of CXCL8 and CXCL5 protein released into the cell culture medium was significantly increased. Thus, in vitro COPD model system showed overactivation of the EMT signal transduction system Respectively. When BEAS-2B cells treated with the tobacco smoke extract-treated seed extract of the present invention were treated, the amount of CXCL8 and CXCL5 protein released into the cell culture medium was recovered to a normal level in a concentration-dependent manner (Fig. 2 (A) and 2 (B)).

On the other hand, when A549 lung cancer cell line was treated with Seiho extract for 48 hours, it was confirmed that the amount of CXCL8 protein released into the cell culture was decreased (Fig. 2 (C)).

Example  4. By Seiho Extract EMT  Identification of changes in signal transduction-related protein expression

In Example 4, 50 μg / ml of tobacco smoke extract was added to the normal bronchial epithelial cells, BEAS-2B cells, for 2 to 4 weeks in order to examine changes in the expression level of EMT signaling- In vitro chronic obstructive pulmonary disease cell model was used.

2x10 < ⁵ > BEAS-2B and CSC-treated BEAS-2B cells were seeded in a 60 mm cell culture dish and cultured for 24 hours. Each dish was treated with 0, 25, 50, 100 μg / ㎖ of Seiho extract for 48 hours, and then cell lysate was obtained by using RIPA buffer. Using these results, various protein expression changes related to EMT signaling Respectively.

As a result, as shown in FIG. 3, the expression of N-cadherin, vimentin and ALDH3A1 proteins associated with EMT signal transduction increased when the tobacco smoke extract was treated, but the tobacco smoke extract . The expression of E-cadherin protein was decreased when the tobacco smoke extract was treated. It was confirmed that the concentration of E-cadherin protein was increased when the tobacco smoke extract and quercetin extract were treated together.

Claims (8)

Shiho ((Bupleurum A pharmaceutical composition for preventing or treating chronic obstructive pulmonary disease containing an extract of falcatum as an active ingredient. The pharmaceutical composition for preventing or treating chronic obstructive pulmonary disease according to claim 1, wherein the solvent of said extract is water, a C ₁ -C ₄ lower alcohol or a mixture thereof. The pharmaceutical composition for preventing or treating chronic obstructive pulmonary disease according to claim 1, wherein the occurrence of lung cancer caused by chronic obstructive pulmonary disease is suppressed. The pharmaceutical composition according to claim 1, wherein the chronic obstructive pulmonary disease is emphysema or chronic bronchitis. The pharmaceutical composition for preventing or treating chronic obstructive pulmonary disease according to claim 1, which further comprises a pharmaceutically acceptable carrier, excipient or diluent. The pharmaceutical composition for preventing or treating chronic obstructive pulmonary disease according to claim 1, wherein the composition is prepared from any one of capsules, powders, granules, tablets, suspensions, emulsions, syrups and aerosols. Shiho ((Bupleurum falcatum ) extract as an active ingredient for the prevention or amelioration of chronic obstructive pulmonary disease. [Claim 7] The composition according to claim 7, wherein the active ingredient is at least one selected from the group consisting of a nutrient, a vitamin, an electrolyte, a flavoring agent, a colorant, a pectic acid and a salt thereof, an alginic acid and a salt thereof, an organic acid, a protective colloid thickening agent, , And a carbonating agent used in a carbonated drink. ≪ RTI ID = 0.0 > 18. < / RTI >

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