KR20200039357A - Pharmaceutical composition comprising fermented extract of peppermint for preventing or treating of respiratory diseases - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating, or treating respiratory diseases, comprising a mixture obtained by mixing a fermented extract of Mentha piperita with an extract of Forsythia velutina or an extract of Maianthemum dilatatum. In addition, the present invention relates to a pharmaceutical composition for antitussive or expectorant use, comprising a mixture obtained by mixing a fermented extract of Mentha piperita with an extract of Forsythia velutina or an extract of Maianthemum dilatatum. The compositions of the present invention suppress migration of airway smooth muscle in patients with asthma, and suppress mucous secretion in mucosal epithelial cells, and thus are effective as compositions for preventing, alleviating, or treating respiratory diseases. Moreover, the compositions of the present invention suppress bronchoconstriction, and suppress formation of phlegm and the like, and thus are also effective as a pharmaceutical composition for antitussive or expectorant use. In particular, the fermented extract of Mentha piperita is even more effective when fermented using lactic acid bacteria of genus Lactobacillus.

Description

Pharmaceutical composition comprising fermented extract of peppermint for preventing or treating of respiratory diseases

The present invention relates to a composition for preventing, improving or treating respiratory disease, comprising a mixture of a peppermint fermented extract and a longevity-rich flower extract or a large crane flower extract.

In addition, the present invention relates to a pharmaceutical composition for Jinhae or Geodam, comprising a mixture of a peppermint fermentation extract and a longevity-rich flower extract or a large crane flower extract.

Peppermint ( Mentha piperita ) is a hybrid mint mixed with water mint (Mentha aquatica) and spearmint (Mentha spicata). It has a very characteristic scent due to the volatile oil present throughout the plant. Peppermint oil is traditionally used in the East and West to treat cancer, colds, cramps, indigestion, or toothache, and has also been used as a fragrance or preservative.

Forsythia velutina is a species of ash family that is distributed in Jangsu-san, Hwanghae-do, and is a Korean species. It is resistant to air pollution and pests and is used for ornamental or fence purposes, but its pharmaceutical efficacy is unknown.

The large crane flower ( Maianthemum dilatatum ) is a family of lilies and is a Korean species distributed in Ulleung-gun, North Gyeongsang Province. The upper part of the large crane flower is used to treat bladder bleeding and excessive menstruation.

Mucus production in the airways is an important defense mechanism to maintain lung health. However, excessive mucus secretion can cause airway obstruction or airway hypersensitivity, and hypersecretion of mucus in asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis (CF), and pneumonia It is a pathological feature.

Mucin is a glycoprotein that imparts viscoelasticity to mucus and is encoded by the mucin group gene (MUS). Among the 18 human MUSs known to date, MUC2, MUC4, MUC5AC, and MUC5B are found in the airway, and among them, MUC5AC is mainly expressed in airway epithelial cells or airway goblet cells. It is known that MUC5AC increases secretion as symptoms of patients with bronchial disease worsen, and is responsible for abnormal mucus to mucus cell proliferation. However, the overproduction of mucin and the proliferation of mucus cells are related, but they are not synonymous and can follow different signal or gene regulation pathways.

Inflammation of the bronchus, which is common among smokers or the elderly, not only reduces the quality of life, but also increases the risk of secondary infectious diseases. In addition, the elderly have difficulty in draining mucus (sputum) due to poor physical fitness, so there is a possibility of progressing to chronic obstructive pulmonary disease, so it must be improved early.

Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death in the United States and Europe. COPD, such as asthma or cystic fibrosis, is characterized by the accumulation of inflammatory mucus in the narrow airway lumen. Therefore, lung function may be reduced and the risk of infection or mortality of the respiratory system may be increased. The biggest cause of COPD is cigarette smoke, and acrolein contained in the cigarette smoke induces MUC5AC production or mucus cell proliferation.

Asthma is a heterogeneous disease (heterogenouse disease) caused by a variety of triggers, allergens, certain drugs (such as aspirin), exercise, bacterial or viral infections or obesity. Asthma increases the irritability of the respiratory system, especially the bronchial smooth muscle, and is accompanied by away remodeling and excessive contraction. Airway remodeling refers to structural changes in the airways due to repeated inflammatory reactions, which are caused by damage to the airway epithelium, increased goblet cell hyperplasia or mucus secretion, bronchosmooth muscle proliferation, airway vessels and nerve proliferation. , Causing airway obstruction and airway irritability. It is known that airway smooth muscle of chronic asthma patients can promote airway remodeling similar to the migration of vascular endothelial cells caused by arteriosclerosis.

Republic of Korea Patent Publication No. 10-2017-0118613 relates to a pharmaceutical composition for the prevention or treatment of respiratory diseases, including the extract of rat tail, the rat tail forgetting extract is abnormal hyperproliferation of spleen cells, secretion or airway of allergic inflammatory cytokines It has been disclosed that there is an effect of suppressing shrinkage.

However, the effect of the mixture of the peppermint fermented extract with the longevity-rich flower or the large crane flower extract on respiratory diseases or respiratory function has not been studied.

Thus, the present inventors tried to provide a natural-friendly plant extract material that can be used as a preventive or therapeutic agent for respiratory diseases such as asthma, as a result of mixing peppermint fermented extract with peppermint fermented extract and longevity-riched extract or large crane flower extract The mixture inhibits migration of airway smooth muscle of asthma patients and suppresses mucus secretion of mucus epithelial cells, thus confirming that it can be usefully used as a preventive or therapeutic agent for respiratory diseases, and completed the present invention.

Therefore, the object of the present invention is peppermint ( Mentha piperita ) fermentation extract; Forsythia velutina extract or large crane flower ( Maianthemum dilatatum ) further comprising an extract; It is to provide a composition for preventing, improving or treating respiratory diseases.

Another object of the present invention, peppermint ( Mentha piperita ) fermented extract and; Forsythia velutina extract or large crane flower ( Maianthemum dilatatum ) further comprising an extract; It is to provide a pharmaceutical composition for antitussive or expectorant.

In order to achieve the above object, the present invention is peppermint ( Mentha piperita ) fermentation extract; Further it includes a longevity manrihwa (Forsythia velutina) flower extract, or a large crane (Maianthemum dilatatum) extract; It may provide a pharmaceutical composition for preventing or treating respiratory diseases.

According to a preferred embodiment of the present invention, the extract may be extracted from any one site selected from the group consisting of peppermint, longevity, or outpost of large crane flowers, flowers, leaves, fruits, stems, shells, and branches. .

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

According to a preferred embodiment of the present invention, the peppermint fermentation extract is from the group consisting of peppermint extract of the Bacillus sub-blocks bus (Bacillus subtilis), Lactobacillus ramno suspension (Lactobacillus rhamnosus) or MRS three Levy cyano (Saccharomyces cerebisiae) in Saccharomyces It may be performed using any one or more strains selected.

According to one preferred embodiment of the present invention, the respiratory disease is asthma, bronchiectasis, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), cystic fibrosis, emphysema, pulmonary tuberculosis sequelae, chronic bronchitis, allergic rhinitis, expectorant expectation, Acute lower respiratory tract infection, bronchitis, bronchiolitis, acute upper respiratory infection, inflammatory lung disease, sinusitis, pharyngitis, tonsillitis and laryngitis may be any one or more selected from the group consisting of.

In addition, the present invention is peppermint ( Mentha piperita ) fermentation extract; Forsythia velutina extract or large crane flower ( Maianthemum dilatatum ) further comprising an extract; It can provide a health functional food composition for preventing or improving respiratory diseases.

In addition, the present invention is peppermint ( Mentha piperita ) fermentation extract; Forsythia velutina extract or large crane flower ( Maianthemum dilatatum ) further comprising an extract; It can provide a pharmaceutical composition for antitussive or expectorant.

Therefore, in the case of the composition comprising the peppermint fermentation extract of the present invention and the longevity flower extract or large crane flower extract, it suppresses migration of bronchial smooth muscle of asthma patients and suppresses mucus secretion of mucus epithelial cells, thereby preventing and improving respiratory diseases. Or it is effective as a therapeutic composition. In addition, the composition of the present invention is effective as a pharmaceutical composition for antitussive or expectorant because it suppresses the narrowing of the bronchus and suppresses the production of sputum.

In particular, the peppermint fermentation extract is more effective when fermented using lactobacillus lactic acid bacteria.

1 is a secretion amount of MUC5AC when fermenting peppermint extract with Bacillus subtilis , Lactobacillus rhamnosus , or Saccharomyces cerebisiae , respectively , and then treating mucus epithelial cells. Is confirming the change. It was confirmed that the effect of inhibiting secretion of MUC5AC was increased when fermentation was performed with each strain than when the peppermint extract was not fermented (NF), and in particular, the effect of inhibiting secretion was significantly increased when fermented with lactic acid bacteria in Lactobacillus. .

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

As described above, in patients with respiratory diseases such as asthma, airway obstruction or airway improvement are the main symptoms. Accordingly, there is a need to develop a composition that suppresses migration of bronchial smooth muscle using natural plant materials to prevent airway narrowing and significantly reduces mucus secretion.

The mixture of the longevity-rich flower extract or the large crane flower extract, respectively, mixed with the peppermint fermentation extract according to the present invention has the effect of inhibiting the migration of bronchial smooth muscle of asthma patients and suppressing the expression of genes related to mucus secretion in mucus epithelial cells. , It is effective as a composition for preventing, improving or treating respiratory diseases to a composition for improving respiratory function.

Therefore, the present invention is a peppermint ( Mentha piperita ) fermentation extract; Additionally comprising a Forsythia velutina extract or a Maianthemum dilatatum extract; Provided is a pharmaceutical composition for preventing or treating respiratory disease.

The peppermint fermentation extract, longevity flower extract or large crane flower extract of the present invention is from any one selected from the group consisting of peppermint, longevity flower or large crane flower outpost, flowers, leaves, berries, stems, shells, and branches. It is preferably extracted.

As the extraction method of the extracts, a filtration method, hot water extraction, immersion extraction, reflux cooling extraction or ultrasonic extraction may be used, but is not limited thereto.

The extracts of the present invention are preferably extracted with water, an organic solvent or a mixture thereof as a solvent, and more preferably, it is preferably extracted with water, a lower alcohol of C ₁ to C ₄ or a mixture thereof as a solvent. And, most preferably, it is most preferably extracted with water (DW) as a solvent, but is not limited thereto.

The peppermint fermentation extract of the present invention extracts the peppermint extract extracted and prepared using water, an organic solvent or a mixture thereof as a solvent, Bacillus subtilis , Lactobacillus rhamnosus or Saccharomyces three It is preferably prepared by fermentation using any one or more strains selected from the group consisting of Levisia (Saccharomyces cerebisiae ).

Increasing the thickness of airway smooth muscle in asthma may be caused by an increase in cell division, a decrease in cell death, apoptosis, migration of mesenchymal cells, or differentiation into airway smooth muscle cells. The mechanism of proliferation and migration of airway smooth muscle is known to have many similar parts.

Mucus secretion induction of mucus epithelial cells in the present invention stimulated mucus epithelial cancer cells with PMA to induce the expression or secretion of MUC5AC. PMA (phorbol 12-myristate 13-acetate) is a type of phorbol ester, and is known to cause various inflammatory reactions through PKC activation. Recently, it has been known that PMA induces the expression of the MUC2 gene, and it is widely used for mucin protein induction in mucus.

The respiratory disease of the present invention may be any target if it is a respiratory disease in which hypersensitivity reactions occur to bronchial smooth muscle and mucus epithelial cells, but preferably asthma, bronchiectasis, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), Cystic fibrosis, emphysema, pulmonary tuberculosis sequelae, chronic bronchitis, allergic rhinitis, antitussive expectora, acute lower respiratory tract infection, bronchitis, bronchiolitis, acute upper respiratory infection, inflammatory lung disease, sinusitis, pharyngitis, tonsillitis and laryngitis It may be, but is not limited to this.

The pharmaceutical composition of the present invention may be various oral or parenteral formulations. When formulating the composition, one or more buffers (e.g. saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g. EDTA or glutathione), fillers, extenders, binders, adjuvants (e.g., Aluminum hydroxide), suspending agents, thickener wetting agents, disintegrating agents or surfactants, diluents or excipients.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in one or more compounds such as starch (corn starch, wheat starch, rice starch, potato Starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethyl -It is prepared by mixing cellulose or gelatin. For example, tablets or dragees can be obtained by mixing the active ingredient with a solid excipient and then grinding it and adding a suitable adjuvant to the granule mixture.

In addition, lubricants such as magnesium stearate, talc and the like are used in addition to simple excipients. Liquid preparations for oral administration include suspending agents, intravenous solutions, emulsions or syrups, etc. In addition to the commonly used simple diluents, water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents or preservatives are included. You can. In addition, if necessary, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and may further include an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, and a preservative. .

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

The composition of the present invention may be administered orally or parenterally, and when applied parenterally, external to the skin; Injectable injections in the intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine epidural or cerebrovascular; Transdermal administration; Alternatively, nasal inhalants may be formulated according to methods known in the art.

The injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi. For injections, examples of suitable carriers include, but are not limited to, solvents or dispersion media comprising water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and / or vegetable oils. You can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanol amine or sterile water for injection, isotonic solutions such as 10% ethanol, 40% propylene glycol and 5% dextrose. Etc. can be used. In order to protect the injection from microbial contamination, it may further include various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In addition, the injection may additionally include an isotonic agent such as sugar or sodium chloride in most cases.

In the case of inhalation dosages, the compounds used according to the invention may be used in the form of pressurized packs, using suitable propellants, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. It can be conveniently delivered in the form of an aerosol spray from a nebulizer. For pressurized aerosols, the dosage unit can be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges used in inhalers or insufflators can be formulated to contain a powder mixture of a compound and a suitable powder base such as lactose or starch. Formulations for parenteral administration are described in Remington's Pharmaceutical Science, 15th Edition, 1975.Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a prescription generally known to all pharmaceutical chemistries.

The composition of the present invention is administered in a pharmaceutically effective amount. "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 is the patient's disease type, severity, drug activity, and sensitivity to the drug. , Administration time, route of administration and rate of discharge, duration of treatment, factors including co-drugs and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. That is, the total effective amount of the composition of the present invention can be administered to a patient in a single dose, and can be administered by a fractionated treatment protocol that is administered for a long time in multiple doses. . Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The dosage of the pharmaceutical composition of the present invention may vary depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and disease severity. As a daily dosage, when administered parenterally, it is preferably administered in an amount of 0.01 to 50 mg per kg body weight per day, more preferably 0.1 to 30 mg per day, based on peppermint fermentation extract, longevity flower extract or large crane flower extract. When possible, and when administered orally, the peppermint fermentation extract, longevity flower extract, or large crane flower extract of the present invention is preferably administered in an amount of 0.01 to 100 mg per kg of body weight per day, more preferably 0.01 to 10 mg per day. It can be divided into 1 to several times as much as possible. However, since the dosage may be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, and age, the dosage is not limited to the scope of the present invention in any way.

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

In addition, the present invention is a peppermint ( Mentha piperita ) fermentation extract; Additionally comprising a Forsythia velutina extract or a Maianthemum dilatatum extract; Provides a health functional food composition for preventing or improving respiratory disease. The peppermint fermentation extract, longevity flower extract, or large crane flower extract is the same as that used in the pharmaceutical composition, and thus the description is replaced by the above description.

The food composition according to the present invention can be prepared in various forms according to conventional methods known in the art. Drinks (including alcoholic beverages), fruits and processed foods (e.g. canned fruits, canned foods, jams, marmalades, etc.), fish, meat and processed foods (e.g. ham, sausages) Corn beef, etc.), breads and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), juice, various drinks, cookies, syrup, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine , Plant protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauce, etc.) can be prepared by adding the longevity flower extract or large crane flower extract together with the peppermint fermentation extract of the present invention. In addition, it is not limited to the nutritional supplement, but may be prepared by adding the peppermint fermentation extract, longevity flower extract or large crane flower extract of the present invention to capsules, tablets, pills, and the like. In addition, the health functional food is not limited to this, for example, peppermint fermentation extract, longevity flower extract, or large crane flower extract itself of the present invention is prepared in the form of tea, juice and drink so that it can be consumed (health drink). It can be taken by liquefaction, granulation, encapsulation and powdering. In addition, in order to use the peppermint fermentation extract, longevity flower extract, or large crane flower extract of the present invention in the form of food additives, it may be prepared and used in the form of a powder or a concentrate. In addition, the peppermint fermentation extract of the present invention, the longevity flower extract or large crane flower extract can be prepared in the form of a composition by mixing with a known active ingredient known to have an effect of preventing and improving inflammatory diseases.

0.04 g, 바람직하게는 약 0.02

0.03 g 이다.When the peppermint fermentation extract, longevity flower extract, or large crane flower extract of the present invention is used as a health drink, the health drink composition may contain various flavoring agents or natural carbohydrates, etc., as additional ingredients, like a normal drink. The natural carbohydrates described above include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin and cyclodextrin; Sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as taumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrate is generally about 0.01 per 100 mL of the composition of the present invention.

0.04 g, preferably about 0.02

0.03 g.

In addition, the peppermint fermentation extract, longevity flower extract, or large crane flower extract of the present invention may be contained as an active ingredient in the food composition for preventing and improving inflammatory diseases, the amount of which is effective to achieve an inflammatory disease prevention and improvement action It is not particularly limited, but is preferably 0.01 to 100% by weight based on the total weight of the total composition. The food composition of the present invention may be prepared by mixing peppermint fermentation extract, longevity flower extract or large crane flower extract together with other active ingredients known to be effective in the composition for preventing and improving inflammatory diseases.

In addition to the above, the health food of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acids, alginic acids, salts of alginic acids, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, Glycerin, alcohol or carbonic acid. In addition, the health food of the present invention may contain flesh for the manufacture of natural fruit juice, fruit juice beverage, or vegetable beverage. These ingredients may be used independently or in combination. The proportion of these additives is not critical, but is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention is a peppermint ( Mentha piperita ) fermentation extract; Additionally comprising a Forsythia velutina extract or a Maianthemum dilatatum extract; Provided is a pharmaceutical composition for antitussive or expectorant.

The peppermint fermentation extract, longevity flower extract, or large crane flower extract is the same as that used in the pharmaceutical composition, and thus the description is replaced by the above description.

The "jinhae" means to stop coughing, and the "gudam" means to discharge sputum (sputum). The composition has an effect capable of alleviating airway obstruction or airway hypersensitivity by inhibiting migration of bronchial smooth muscle or inhibiting secretion of mucus from airway mucus epithelial cells. Therefore, by narrowing the narrow airways, it is possible to alleviate cough caused by it or to facilitate sputum discharge, and it is effective as a pharmaceutical composition for antitussive or expectorant sputum because it can suppress the occurrence of excessive sputum.

Since the pharmaceutical composition has the same meaning as the pharmaceutical composition in the 'pharmaceutical composition for preventing or treating respiratory disease', the description is substituted for the description.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

Preparation of extract

<1-1> peppermint extract, Longevity  Preparation of extract or large crane flower extract

A peppermint extract, a longevity flower extract or a large crane flower extract was prepared by extracting the raw material of peppermint outpost, longevity stem or large crane flower outpost in distilled water (DW) for 2.5 hours at 100 ° C.

<1-2> Preparation of Peppermint Fermentation Extract

Peppermint extract is fermented, microbe Bacillus sub-blocks in a sterile's Peppermint Extract (Bacillus subtilis), Lactobacillus Caucasus ramno (Lactobacillus rhamnosus) or as Saccharomyces Celebi Asia (Saccharomyces cerebisiae ) , and then fermented for 5 days under shaking at 140 rpm in an environment of 25 ° C. and 80% humidity to inoculate a peppermint fermentation extract.

Bronchial smooth muscle cell (DBSMC-AS) culture of asthma patient

When the peppermint fermentation extract and the longevity flower extract or the large crane flower extract are treated together, the asthmatic patient may check whether the movement of the bronchial smooth muscle cells (Asthma, DBSMC-AS) of asthma patients is effectively inhibited. We wanted to cultivate bronchial smooth muscle cells.

Specifically, 5% fetal bovine serum (FBS), 0.1% rhEGF, 0.1% insulin (Insulin), 0.1% GA-1000, 0.2% rhFGF-B in bronchial smooth muscle cells (Lonza, 194850) of asthma patients It was cultured in a 37 ° C incubator with 5% CO ₂ using SMBM medium containing. Cells between 3 and 20 generations (passage) were used for the experiment.

Culture of human lung mucus epithelial cancer cell line (NCI-H292)

When the peppermint fermentation extract and the longevity flower extract or the large crane flower extract are treated together, the mucus of the human lung is checked to determine whether the mucus secretion of human pulmonary mucoepidermoid cells (NCI-H292) is inhibited. We wanted to cultivate epithelial sheep cancer cell lines.

Specifically, 10% fetal bovine serum (FBS), 1% penicillin and 1% streptomycin were administered to the mucus epithelial cancer cell line of the human lung (American Type Culture Collection, Manassas, VA, USA; CRL-1848). Using RPMI 1640 medium containing it was cultured in a 37 ℃ incubator with 5% CO ₂ present. Cells between 10 and 30 generations (passage) were used for the experiment.

Inhibitory effect of bronchial smooth muscle cells (DBSMC-AS) migration in asthmatic patients

When the peppermint fermentation extract and the longevity flower extract or the large crane flower extract were treated together in bronchial smooth muscle cells of asthma patients, it was attempted to confirm whether the migration was effectively suppressed.

Specifically, the DBSMC-AS of <Example 2> was cultured in a 24-well plate and starvated in a medium containing 0.1% FBS for 12 hours, and then wounded in cells using a 100 µl tip. Came out. Thereafter, the peppermint fermentation extract of <Example 1> and the longevity flower extract or the large crane flower extract were treated together and incubated for 80 hours to observe the degree of migration with an electron microscope. The control group (CTL) was not treated.

As a result, when the peppermint fermentation extract and the longevity flower extract or the large crane flower extract were treated together, it was confirmed that the migration of bronchial smooth muscle cells of asthma patients was significantly inhibited.

Inhibitory effect of mucus epithelial cells

When the peppermint fermentation extract and the longevity flower extract or the large crane flower extract were treated together with mucus epithelial cells, the secretion of MUC5AC was suppressed to effectively inhibit mucus secretion through RT-PCR.

Specifically, NCI-H292 of <Example 3> was dispensed into a 24-well plate and cultured in a 37 ° C incubator with 5% CO ₂ . When the cells grew to 80%, they were replaced with serum-free RPMI 1640 medium and cultured in an incubator for 12 hours. After pre-treatment with the peppermint fermentation extract of <Example 1> and the longevity flower extract or large crane flower extract, the supernatant was obtained, diluted 10 times with PBS, plated in 96 wells and dried at 37 ° C until drying. It was dried. Then, after washing 3 times with PBS, it was blocked with 2% BSA for 1 hour at room temperature, and then washed 3 times with PBS. The MUC5AC antibody was treated at room temperature for 1 hour and then washed with PBS. The HRP goat anti-mouse IgG conjugate was treated at room temperature for 1 hour, and then washed 3 times with PBS. The absorbance was taken at 450 nm by adding a TMB solution and treating a stop solution. The control group (CTL) was not treated, and the solvent control group (Veh) was treated with 10 μg / ml of DMSO.

As a result, it was confirmed that the fermentation effect of MUC5AC was increased when fermented with each strain compared to the case where the peppermint extract was not fermented. In particular, the peppermint fermentation extract fermented with Lactobacillus rhamnosus significantly increased the inhibitory effect of MUC5AC secretion.

Claims (7)

Peppermint ( Mentha piperita ) fermentation extract;
Further it includes a longevity manrihwa (Forsythia velutina) flower extract, or a large crane (Maianthemum dilatatum) extract;
Pharmaceutical composition for the prevention or treatment of respiratory diseases.
The prevention of respiratory diseases according to claim 1, wherein the extract is extracted from any one site selected from the group consisting of peppermint, longevity flower or outpost of large crane flower, flower, leaf, fruit, stem, shell, and branch. Or a therapeutic pharmaceutical composition.
According to claim 1, The extract is water, an organic solvent or a pharmaceutical composition for preventing or treating respiratory diseases, characterized in that extracted by using a mixture of them as a solvent.
According to claim 1, wherein the peppermint fermentation extract is peppermint extract Bacillus subtilis ( Bacillus subtilis ), Lactobacillus rhamnosus ( Lactobacillus rhamnosus ) or Saccharomyces cerevisiae ( Saccharomyces) cerebisiae ) A pharmaceutical composition for the prevention or treatment of respiratory diseases, characterized in that it is carried out using any one or more strains selected from the group consisting of.
According to claim 1, The respiratory disease is asthma, bronchiectasis, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), cystic fibrosis, emphysema, pulmonary tuberculosis sequelae, chronic bronchitis, allergic rhinitis, antitussive expectorant disease, acute respiratory infection, A pharmaceutical composition for the prevention or treatment of respiratory disease, characterized by at least one selected from the group consisting of bronchitis, bronchiolitis, acute upper respiratory infection, inflammatory lung disease, sinusitis, pharyngitis, tonsillitis and laryngitis.
Peppermint ( Mentha piperita ) fermented extract;
Further it includes a longevity manrihwa (Forsythia velutina) flower extract, or a large crane (Maianthemum dilatatum) extract;
Health functional food composition for preventing or improving respiratory disease.
Peppermint ( Mentha piperita ) fermented extract;
Further it includes a longevity manrihwa (Forsythia velutina) flower extract, or a large crane (Maianthemum dilatatum) extract;
Pharmaceutical compositions for antitussive or expectorant.

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