KR20200021740A - Composition comprising Viola tokubuchiana extract for preventing, improving or treating respiratory disease - Google Patents

Abstract

The present invention relates to a pharmaceutical composition or a health functional food composition for preventing, alleviating or treating respiratory diseases, containing an extract of Viola tokubuchiana. Therefore, the composition containing the extract of Viola tokubuchiana of the present invention can significantly inhibit the proliferation of bronchial smooth muscle cells in patients with respiratory diseases such as asthma, thereby being effective as the composition for preventing, alleviating or treating respiratory diseases.

Description

Composition comprising Viola tokubuchiana extract for preventing, improving or treating respiratory disease

The present invention relates to a pharmaceutical composition or a nutraceutical composition for preventing, ameliorating or treating a respiratory disease, including an extract of Viola tokubuchiana.

Respiratory disease mainly refers to diseases related to the lungs and airways, and the airways are largely composed of mucous membranes and bronchial smooth muscles. There are many secretory glands in the mucosa that continue to secrete the necessary secretions, and when the bronchial smooth muscle contracts, the airways become narrow. Bronchial smooth muscle is known to play an important role in the invention of airway obstruction such as asthma, chronic bronchitis and emphysema among respiratory diseases.

Respiratory diseases associated with bronchial smooth muscle are characterized by airway remodeling, airway obstruction, airway inflammation and airway hyperresponsiveness that may be irreversible. Airway remodeling is a structural change in the airways that results in subepithelial fibrosis, globult cell hyperplasia, myofibroblast to smooth muscle cells, and neovascularization. And cause irreversible airway obstruction and airway hypersensitivity. The increase of bronchial smooth muscle cells is expected to be the mechanism by which thickening, proliferation and migration of bronchial smooth muscle cells, increased division of bronchial smooth muscle cells, reduction of apoptosis, migration of mesenchymal cells and bronchial smooth muscle This may be due to increased differentiation into cells.

Minolamo violet (Viola tokubuchiana) is a violet and plant distributed in Gwangneung and Jirisan areas of Gyeonggi-do. It is mainly used for ornamental purposes, but its pharmaceutical efficacy is not known.

Republic of Korea Patent Publication No. 10-2012-0042083 relates to a composition for the prevention and treatment of diseases induced by respiratory apoptosis, including red garlic extract, the red garlic extract inhibits the decrease in the concentration of glutathione in bronchial smooth muscle cells It is disclosed that there is an effect of suppressing the death.

However, there is a need for research on natural extracts that can be used stably for respiratory diseases caused by various causes in addition to the death of bronchial smooth muscle cells. There is no bar.

Accordingly, the present inventors have confirmed that the extract of the Mindong Moe violet flower is effective in the prevention, improvement or treatment of respiratory diseases such as asthma by inhibiting the proliferation of bronchial smooth muscle cells, and completed the present invention.

Accordingly, the present inventors have completed the present invention by developing a composition for the prevention, improvement or treatment of respiratory diseases, including the extract of Mindong Moe violet (Viola tokubuchiana).

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for preventing or treating respiratory diseases, including extracts from Viola tokubuchiana.

Still another object of the present invention is to provide a dietary supplement for preventing or improving respiratory diseases, including extract of Mindong Mobi (Viola tokubuchiana).

Still another object of the present invention is to provide a health functional food composition for improving respiratory function, including extract of Mindong Mo violet (Viola tokubuchiana).

In order to achieve the above object, the present invention can provide a pharmaceutical composition for preventing or treating respiratory diseases, including Mindong Moo violet (Viola tokubuchiana) extract.

According to a preferred embodiment of the present invention, the extract may be obtained from the outpost of Mindong Moe violets.

According to a preferred embodiment of the present invention, the extract may be extracted by using water, an organic solvent or a mixture thereof as a solvent.

According to a preferred embodiment of the present invention, the extract may be included in a concentration of 0.3 to 1000 ㎍ / ㎖.

According to a preferred embodiment of the present invention, the respiratory disease is any one selected from the group consisting of cold, chronic bronchitis, emphysema, cystic fibrosis, inflammatory airway disease, bronchiectasis, bronchial stenosis, asthma, pneumonia and obstructive pulmonary disease It may be abnormal.

According to a preferred embodiment of the present invention, the composition may be to inhibit the proliferation of bronchial smooth muscle cells.

The present invention may also provide a dietary supplement for preventing or improving respiratory diseases, including extracts from Mindong Moe violet (Viola tokubuchiana).

According to a preferred embodiment of the present invention, the respiratory disease is any one selected from the group consisting of cold, chronic bronchitis, emphysema, cystic fibrosis, inflammatory airway disease, bronchiectasis, bronchial stenosis, asthma, pneumonia and obstructive pulmonary disease It may be abnormal.

The present invention may also provide a health functional food composition for respiratory function improvement comprising a extract of Mindong Mo violet (Viola tokubuchiana).

Therefore, the composition comprising the extract of Viola tokubuchiana of the present invention can significantly inhibit the proliferation of bronchial smooth muscle cells in patients with respiratory diseases such as asthma, and thus is effective as a composition for preventing, improving or treating respiratory diseases. .

FIG. 1 shows that the extract of Mindong Moe Violet Outpost does not show cytotoxicity against bronchial smooth muscle cells of asthma patients. The control group was treated with DMSO.
Figure 2 shows that the treatment of bronchial smooth muscle cells of asthma patients significantly reduced the proliferation of bronchial smooth muscle cells when treated with Mindong Moe violet outpost. The control group was treated with DMSO.

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

As described above, most of the drugs used in the conventional treatment of respiratory diseases are focused on regulating immune to inflammatory responses, and therefore, there are limitations when used in respiratory diseases caused by causes other than inflammation. In order to overcome this, it was necessary to develop a composition derived from a natural extract that can be used for the treatment of respiratory diseases caused by various causes including inflammation.

The composition comprising the extract of Mindong Moe violet (Viola tokubuchiana) according to the present invention significantly inhibits the proliferation of bronchial smooth muscle cells and thus is effective as a composition for preventing, improving or treating respiratory diseases caused by an increase in the volume of bronchial smooth muscle.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating respiratory diseases, including extracts from Viola tokubuchiana.

Preferably, the extract of the Mindong Moe violet flower of the present invention is obtained from the outpost of Mindong Moe violet, but is not limited thereto.

The extract of the present invention is preferably extracted with water, an organic solvent or a mixture thereof as a solvent, more preferably water, a lower alcohol of C ₁ to C ₄ or a mixture thereof as a solvent. Most preferably, the solvent is extracted using methanol as a solvent, but is not limited thereto.

The extract of the present invention is preferably contained in a concentration of 0.3 to 1000 ㎍ / ㎖, more preferably contained in a concentration of 1 to 300 ㎍ / ㎖, most preferably in a concentration of 3 to 30 ㎍ / ㎖ Most preferably included. The concentration range corresponds to the concentration range that can be significantly inhibited proliferation of the extract of Mindong Moe violet flowers without showing toxicity to bronchial smooth muscle cells (Fig. 1).

The respiratory disease can be applied without limitation as long as it is a disease associated with bronchial smooth muscle, preferably, cold, chronic bronchitis, emphysema, cystic fibrosis, inflammatory airway disease, bronchiectasis, bronchial stenosis, asthma, pneumonia and obstructive pulmonary disease It is preferably at least one selected from the group.

Asthma of the present invention is atopic asthma, non-atopic asthma, allergic asthma, atopic bronchial IgE-mediated asthma, bronchial asthma, essential asthma, intrinsic asthma, endogenous asthma caused by pathophysiological disturbance, environment Exogenous asthma caused by enemy factors, essential asthma with unknown or unclear causes, non-atopic asthma, bronchitis asthma, emphysema asthma, asthma caused by exercise, asthma caused by allergens, cold In the group consisting of air-induced asthma, occupational asthma, infectious asthma, non-allergic asthma, early asthma, whistle infant syndrome and bronchiolitis caused by bacterial, fungal, protozoa or viral infections At least one selected asthma, but is not limited thereto.

The bronchial smooth muscle cells of the respiratory disease patients are thicker than those of normal bronchial smooth muscle cells. This is caused by an increase in cell number or a decrease in apoptosis, and the increase in cell number is caused by the proliferation or migration of bronchial smooth muscle cells. The composition comprising the extract of the present invention may be able to relieve airway remodeling symptoms of patients with respiratory diseases by inhibiting the proliferation of bronchial smooth muscle cells. In particular, the growth inhibitory effect of bronchial smooth muscle cells appears more significantly when the concentration of the extract of Mindong Moe violet flower is treated at 30 μg / ml or more (FIG. 2).

The compositions of the present invention may be in various oral or parenteral formulations. In formulating the composition, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants are usually used.

Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or lactose (at least one compound). lactose) and gelatin. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions or syrups, and include various excipients such as wetting agents, sweeteners, fragrances, or preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Can be.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations or suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used.

The composition of the present invention may be administered orally or parenterally, and when parenteral administration, it is preferable to select an external skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injection mode.

The composition of 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 an effective dose level refers to the type, severity, and activity of the patient's disease. , Drug sensitivity, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of drugs, and other factors well known in the medical arts. The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with a minimum amount without side effects, which can be readily determined by one skilled in the art.

The dosage of the composition of the present invention varies depending on the weight, age, sex, health status, diet, time of administration, administration method, excretion rate and severity of the disease of the patient, the daily dosage of the extract 0.01 to 1000 mg / kg, preferably 30 to 500 mg / kg, more preferably 50 to 300 mg / kg, based on the amount, and may be administered 1 to 6 times per day. However, the dosage may be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, age, etc., and the above dosage does not limit the scope of the present invention in any way.

The compositions of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.

Therefore, the extract of Viola tokubuchiana significantly inhibits the proliferation of bronchial smooth muscle cells and thus is effective as a pharmaceutical composition for preventing or treating respiratory diseases.

The present invention also provides a dietary supplement for preventing or improving respiratory diseases, including extracts from Viola tokubuchiana. The Mindong Moe violet extract is the same as the Mindong Moe violet extract contained in the pharmaceutical composition, and as a description for this.

The respiratory disease of the present invention is preferably at least one selected from the group consisting of cold, chronic bronchitis, emphysema, cystic fibrosis, inflammatory airway disease, bronchiectasis, bronchial stenosis, asthma, pneumonia and obstructive pulmonary disease, bronchial smooth muscle Any disease caused by the proliferation of cells can be applied without limitation.

The present invention also provides a health functional food composition for respiratory function improvement comprising a extract of Mindong Mo violet (Viola tokubuchiana). The Mindong Moe violet extract is the same as the Mindong Moe violet extract contained in the pharmaceutical composition, and as a description for this.

The "improved breathing function" means any effect that the respiratory function is improved qualitatively or quantitatively.

The extract of the present invention may be added as it is or may be used together with other food or food ingredients, and may be appropriately used according to a conventional method. The health food of the present invention includes ingredients that are commonly added in food production, and include, for example, proteins, carbohydrates, fats, nutrients and seasonings.

There is no particular limitation on the kind of the food. Examples of foods to which the extract may be added include extracts, meat, sausages, breads, biscuits, rice cakes, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream products, including ice cream, Various soups, beverages, alcoholic beverages and vitamin complexes, dairy products and dairy products, etc., and includes all the health functional foods in the conventional sense.

Mindong Moe violet extract of the present invention can be added to the food as it is, or used in conjunction with other food or food ingredients, and may be suitably used in accordance with conventional methods. The mixing amount of the active ingredient can be suitably determined according to the purpose of use (prevention or improvement). In general, the amount of the extract in the health food can be added to 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for health and hygiene or health control purposes, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

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

In addition to the above, the Mindong Moe violet extract of the present invention is a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, such as colorants and neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like. In addition, the extract may contain a flesh for the production of natural fruit juice and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

Therefore, the extract of Viola tokubuchiana (Viola tokubuchiana) significantly inhibits the proliferation of bronchial smooth muscle cells and is effective as a dietary supplement composition for preventing or improving respiratory diseases or improving respiratory function.

Hereinafter, the present invention will be described in more detail with reference to Examples and Preparation Examples. These examples and preparations are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not interpreted to be limited by these examples and preparations. .

Preparation of Mindong Mo violet Extract

In order to confirm whether the extract of the Mindong Mobi flower has an effect of improving respiratory function, an extract of the Mindong Mo violet outpost was prepared.

Specifically, in order to prepare extracts of the Mindong Mobi flower outpost, add the Mindong Mobis outpost raw material to 99.9% of methanol (Methanol, Methyl alcohol) for 3 days at 45 ℃ (2 hours after 15 minutes sonication, 10 times / 1 day Extraction was carried out, and the resultant was filtered using a non-fluorescent cotton as a filter and concentrated at 45 ° C.

Culture of Bronchial Smooth Muscle Cells (DBSMC-AS)

In order to determine whether the bronchial smooth muscle cell proliferation of asthma patients is effectively inhibited when the extract of the Mindong Mo violet flower is treated, the bronchial smooth muscle cells of asthma patients were cultivated.

Bronchial smooth muscle cells of asthma patients (DBSMC-AS (Diseased Bronchial Smooth Muscle cell-Asthma); Lonza, 194850) were treated with 5% fetal bovine serum (FBS), 0.1% rhEGF, 0.1% insulin SMBM medium containing% GA-1000, 0.2% rhFGF-B was incubated in a 37 ° C. incubator with 5% CO ₂ . Cells between 3 and 20 generations were used for the experiment.

Confirmation of Cytotoxicity of Mindong Moe violet Extract

MTT cytotoxicity experiments were performed to determine whether the extract of Mindong Mobi flowers outpost is toxic to bronchial smooth muscle cells of asthma patients.

Specifically, in order to investigate the survival rate of bronchial smooth muscle cells of asthma patients against the extract of the Mindong Mobi flower outpost, 5 × 10 ⁵ cells / well in 24 wells, and after 24 hours , 10 or 30 μg / ml) and treated with 0.1% DMSO as a control (CTL) and incubated for 24 hours. After that, remove the culture medium, dissolve MTT (3- (4,5-Dimethylthiazol-2-yl) -2) in 1XPBS to make 0.005 g / mL, and add 500 μl of each well at 37 ° C for 5% CO ₂ incubator. Reaction was carried out for 1 hour. Subsequently, the supernatant was removed, dissolved by adding DMSO, and then, 100 µl was dispensed into 96 wells, followed by measurement of absorbance at 570 nm using a Microplate Autoreader ELISA (Bio-Tek Instruments Inc., Winooski, VT). Confirmed.

As a result, as shown in FIG. 1, it could be confirmed that the extract of Mindong Mobi Outpost did not show toxicity to bronchial smooth muscle cells of asthma patients.

Measurement of Bronchial Smooth Muscle Cell Proliferation in Asthmatic Patients

In order to determine whether the proliferation of bronchial smooth muscle cells of asthma patients is effectively inhibited when the outpost extract of the small-angled mote violet is treated, the outgrowth of the cells is measured by treating the extract of the small-headed mote violet to the bronchial smooth muscle cells of asthma patients. Was intended.

Specifically, bronchial smooth muscle cells of asthma patients cultivated in 96 wells (starvation) after 12 hours of cultivation (starvation) in a medium with 0.1% FBS added to the medium prepared in Example 1 Treatment was incubated with μg / ml. After 24 hours, the cell number of each group was measured by measuring ELISA using BrdU kit. The control group (CTL) was treated with 0.1% DMSO.

As a result, the proliferation of bronchial smooth muscle cells of asthma patients treated with Mindong Moe violet Outpost extract as shown in FIG. 2 was significantly reduced compared to the control group.

Preparation Example 1-1 A Tablet Comprising a Branch Flower Extract

Starch----------50 mg

Lactose----------50 mg

Magnesium Stearate---5 mg

Birch Extract----100 mg

The tablets were prepared by mixing the components and using a wet granulation method or a dry granulation method according to a conventional method for preparing a tablet.

Preparation Example 1-2 Health Beverage Containing Branches Extract

Liquid fructose---------------0.5%

Oligosaccharide----------------2%

Sugar------------------2%

Salt------------------0.5%

Water-------------------75%

Birch Extract------------20%

The ingredients were mixed to a total volume of 50 ml, and the final mixture was filtered through a 3 μm filter and sterilized for 20 seconds at 90-93 ° C. for 20 seconds, and then filled into a 50 ml bottle for 20 minutes at 80-85 ° C. Secondary sterilization produced a health beverage product.

Claims (9)

Pharmaceutical composition for the prevention or treatment of respiratory diseases, including the extract of Viola tokubuchiana.
According to claim 1, wherein the extract is a pharmaceutical composition for the prevention or treatment of respiratory diseases, characterized in that obtained from the outpost of Mindong Moe violet.
The pharmaceutical composition for preventing or treating respiratory diseases according to claim 1, wherein the extract is extracted using water, an organic solvent or a mixture thereof as a solvent.
According to claim 1, wherein the extract is a pharmaceutical composition for preventing or treating respiratory diseases, characterized in that it comprises a concentration of 0.3 to 1000 ㎍ / ㎖.
The method of claim 1, wherein the respiratory disease is any one or more selected from the group consisting of cold, inflammatory airway disease, bronchiectasis, bronchial stenosis, asthma, pneumonia and obstructive pulmonary disease Composition.
According to claim 1, wherein the composition is a pharmaceutical composition for preventing or treating respiratory diseases, characterized in that to inhibit the proliferation of bronchial smooth muscle cells.
Health functional food composition for the prevention or improvement of respiratory diseases, including Mindong Moe violet (Viola tokubuchiana) extract.
The method of claim 7, wherein the respiratory disease is any one or more selected from the group consisting of cold, inflammatory airway disease, bronchiectasis, bronchial stenosis, asthma, pneumonia and obstructive pulmonary disease Food composition.
Health functional food composition for respiratory function improvement including Mindong Mo violet (Viola tokubuchiana) extract.

Images