KR102138073B1 - Composition comprising sauchinone for preventing or treating of chronic pulmonary disease - Google Patents

Abstract

The present invention is a composition for the prevention or treatment of chronic lung disease containing sochinone, because it can effectively suppress the damage of lung tissue and the expression of IGF2 by smoking, it can be used for the prevention or treatment of various chronic lung diseases Is expected.

Description

Composition comprising sauchinone for preventing or treating of chronic pulmonary disease

The present invention relates to a composition for the prevention and/or treatment of chronic lung diseases, including sochinone as an active ingredient.

Emphysema is one of chronic obstructive pulmonary diseases, which is caused by damage to the alveoli of the peripheral airway area and the space wall between the alveoli of the distal three bronchi is lost and destroyed, resulting in abnormal permanent expansion, thereby reducing the ability of the lung to exchange oxygen-carbon dioxide. It is a condition that causes difficulty breathing. The main cause of emphysema is known as smoking, but it can also be caused by air pollution or exposure to a hazardous working environment, and is known to accelerate the progression rate when accompanied by respiratory infections. The symptoms of alveolar dilation due to tissue destruction, the main symptom of emphysema, are expression of protease such as matrix metalloproteinase (MMP) and neutrophil elastase, while protease is activated. It is known that the expression of the inhibiting α1-antitrypsin (α1-antitrypsin) is reduced. In addition, secretion of various cytokines and chemokines from macrophages and epithelium in the alveoli by smoking leads to the induction of cytokines/chemokines (TNF-α, IL-1β, IL-8, MCP) by cells. -1, IFN-γ, etc.), reactive oxygen species (ROS), etc. are increased, and thus, tissue damage is accelerated due to increased expression and activation of proteolytic enzymes. Such emphysema is a disease in which the lung structure is irreversibly damaged and cannot be fundamentally cured, and currently used treatments include symptomatic treatments such as steroid preparations to relieve the inflammatory response, bronchodilators to relieve dyspnea, and expectorants. Only the method is being used.

Therefore, in order to effectively treat such emphysema, there is an urgent need to develop a therapeutic agent capable of reducing the rate of damage to the lung structure or inhibiting damage to lung tissue.

Domestic registered patent 10-1647029

The present invention has been devised to solve the problems in the prior art as described above, and an object thereof is to provide a composition for the prevention or treatment of chronic lung disease, including sochinon as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.

The present invention provides a pharmaceutical composition for the prevention or treatment of chronic obstructive pulmonary disease, comprising sochinone (sauchinone) as an active ingredient.

In addition, the present invention provides a food composition for preventing or improving chronic obstructive pulmonary disease, comprising sochinone (sauchinone) as an active ingredient.

In another embodiment of the present invention, the chronic obstructive pulmonary disease (COPD) is preferably a chronic obstructive pulmonary disease caused by emphysema, damage to lung tissue, increased IGF2 expression, etc. It is not limited to this.

In another embodiment of the present invention, the composition is characterized in that it maintains or restores lung function by inhibiting damage to lung tissue and expression of IGF2.

In addition, the present invention provides a method of treating chronic obstructive pulmonary disease, comprising the step of administering a composition containing sochinone as an active ingredient to an individual.

In addition, the present invention provides a use for the prevention or treatment of chronic obstructive pulmonary disease of a composition containing sochinone as an active ingredient.

The composition comprising sochinone of the present invention as an active ingredient can effectively be used to effectively prevent and treat chronic obstructive pulmonary disease by effectively inhibiting damage to lung tissue, and the sochinon has a low cytotoxicity, thus causing damage to lung tissue. It is expected to be widely used in the treatment of various lung diseases due to.

1 is a view showing an experimental method according to an embodiment of the present invention.
2 is a view showing the results of confirming the recovery of lung function by Sochinon according to an embodiment of the present invention.
3 is a view showing the results of confirming the effect of inhibiting lung tissue damage by socinone according to an embodiment of the present invention.
4 is a view showing the results of confirming the effect of inhibiting IGF2 expression by Sochinon according to an embodiment of the present invention by RT-PCR.
5 is a view showing the results of confirming the effect of inhibiting IGF2 expression by Sochinon according to an embodiment of the present invention by immunochemical staining.
6 is a view showing the results of confirming the cytotoxicity of Sochinon according to an embodiment of the present invention.

In a situation where there is almost no effective treatment and treatment method for chronic lung diseases including emphysema, the composition for preventing or treating chronic lung diseases including Sochinon of the present invention is expected to be effectively used for the treatment of various lung diseases. .

In the present specification, “chronic obstructive pulmonary disease (COPD)” is also referred to as chronic lung disease, and cough or sputum may be caused by various causes, and in severe cases, lung disease in which dyspnea occurs In other words, preferably, emphysema or lung disease caused by damage to the tissues of the lung is not limited thereto.

In the present specification, "prevention (prevention)" refers to all actions that inhibit or delay the onset of diseases such as chronic obstructive pulmonary disease by administration of the composition according to the present invention.

In the present specification, "treatment (treatment)" refers to all actions in which symptoms such as chronic obstructive pulmonary disease are improved or beneficially changed by administration of the composition according to the present invention.

In the present specification, “individual” refers to a subject to which the composition of the present invention can be administered, and the subject is not limited.

In this specification, "pharmaceutical composition (pharmaceutical composition)" may be characterized in the form of capsules, tablets, granules, injections, ointments, powders or beverages, wherein the pharmaceutical composition is characterized in that it targets humans Can. The pharmaceutical composition is not limited to these, but may be used in the form of oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterile injection solutions, respectively, according to a conventional method. . The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, fragrances, etc., when administered orally, and in the case of injections, buffers, preservatives, and analgesic Agents, solubilizers, isotonic agents, stabilizers, etc. can be used in combination, and for topical administration, bases, excipients, lubricants, preservatives, etc. can be used. The formulation of the pharmaceutical composition of the present invention can be prepared in various ways by mixing with a pharmaceutically acceptable carrier as described above. For example, when administered orally, tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc. may be prepared, and in the case of injections, it may be prepared in unit dosage ampoules or multiple dosage forms. have. Others can be formulated as solutions, suspensions, tablets, capsules, and sustained release preparations.

On the other hand, examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil and the like can be used. In addition, fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like may be further included.

The route of administration of the pharmaceutical composition according to the present invention is not limited to these, but oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical , Sublingual or rectal. Oral or parenteral administration is preferred. As used herein, the term “parenteral” includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, synovial, intrasternal, epidural, intralesional and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of suppositories for rectal administration.

The pharmaceutical composition of the present invention depends on a number of factors, including the activity, age, weight, general health, sex, diet, administration time, route of administration, release rate, drug combination and severity of the particular disease to be prevented or treated, of the specific compound used. It may vary, and the dosage of the pharmaceutical composition varies depending on the patient's condition, weight, disease severity, drug type, administration route, and duration, but can be appropriately selected by those skilled in the art, and 0.0001 to 50 mg per day /kg or 0.001 to 50 mg/kg. Administration may be administered once a day, or may be divided into several times. The above dosage does not limit the scope of the invention in any aspect. The pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions.

In the present invention, the food composition may be used in various foods, for example, beverages, gums, teas, vitamin complexes, health supplements, etc., and may be used in the form of pills, powders, granules, needles, tablets, capsules, or beverages. Can. At this time, the amount of the sochinone in the food or beverage, in general, in the case of the food composition of the present invention can be added to 0.01 to 15% by weight of the total food weight, in the case of a healthy beverage composition, 0.02 to 10 g, preferably based on 100 mL It can be added at a rate of 0.3 to 1 g.

The food composition of the present invention may include food additives conventional in the art, for example, flavoring agents, flavoring agents, colorants, fillers, stabilizers, and the like. The food composition according to the present invention is an essential component, and there is no particular limitation on the components added in addition to the sochinone, and may contain various flavoring agents or natural carbohydrates, etc., as additional components, like ordinary food. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; Disaccharides such as maltose, sucrose, etc.; Polysaccharides such as dextrin, cyclodextrin; And sugars such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatine, stevia extract (e.g., rebaudioside A, glycyrrhizine, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. have. The proportion of the natural carbohydrate is generally about 1 to 20 g per 100 mL of the composition of the present invention, preferably about 5 to 12 g.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and It may contain salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonic acid used in carbonated beverages, and the like. These ingredients can be used independently or in combination. The proportion of these additives is not so critical, but is generally selected from 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the following examples are presented to help the understanding of the present invention. However, the following examples are only provided to more easily understand the present invention, and the contents of the present invention are not limited by the following examples.

Example

Example One: Sochi Discussion Confirmation of the effect on chronic obstructive pulmonary disease

1.1. Lung function Confirmation experiment

To confirm the effect of sochinone (sauchinone) on pulmonary function decline due to chronic obstructive pulmonary disease (COPD) in raw vegetables, Sochinon was administered at a concentration of 30 mg/kg in FVB mice aged 8 weeks. After oral administration for 2 weeks 5 times a week, the tobacco components of nicotine-derived nitrosamine ketone (NNK) and benzoapyrene (benzo[a]pyrene) were each administered with sochinon at a concentration of 3 μmol for 18 weeks. It was administered orally. The overall experimental method is shown in FIG. 1. Afterwards, in order to confirm lung function, lung compliance and elasticity (ability to breathe through the diaphragm muscle contraction) of the mouse were measured using flexiVent RM, and the results are shown in FIG. 2.

As shown in FIG. 2, in the case of the control group (N/B) administered NKK and benzoapyrene, the purity of the lungs was increased, the elasticity was decreased, and the function of the lungs was significantly reduced, but the experimental group administered with socinone In (N/B+Sau), the lung fluid purity decreased again, and the elasticity increased again, confirming that the lung function was restored as in the normal control. Through the above results, it was confirmed that Sochinon has an effect of preventing and treating emphysema.

1.2. H&E dyeing

In order to confirm the effect of sochinone administration on the damage to lung tissue, the lungs of mice treated in the same manner as in Example 1.1 were separated, and the isolated lungs were fixed for 24 hours using 10% formalin. Later, paraffin blocks were prepared using paraffin. Then, the produced paraffin block was cut to a thickness of 4 μm, and then attached to a silane coated slide and dried. The dried slides were put in a dry oven at 65° C. and deparaffins were processed for 16 hours, and then immersed in xylene solution for 5 minutes and washed twice. Then, the fixed lung tissue sections were deparaffinized and hydrated with 100%, 95%, and 70% alcohol, washed with distilled water, and then stained with Harris hematoxylin solution for 1 minute 30 seconds, 1% HCl alcohol After washing by immersing the solution with ammonia, it was dyed for 30 seconds using an eosin solution. Then, dehydration was sequentially performed using 95% ethanol and 100% ethanol, and finally washed with a xylene solution, sealed, and observed under a microscope to obtain a photograph. The results are shown in FIG. 3.

As shown in Figure 3, in the case of the control group (N/B) administered NKK and benzoapyrene, the alveoli expanded and the lung tissue collapsed, but in the experimental group (N/B+Sau) administered with socinone together It was confirmed that alveolar expansion was suppressed and lung tissue damage was not induced.

1.3. IGF2 Expression level measurement

In order to confirm the expression level of IGF2 (insulin-like growth factor 2), which is known as a major factor inducing emphysema, lungs of mice treated with 0.1, 1, and 10 μM Sochinon, respectively, in the same manner as in Example 1.1 The tissues were obtained and subjected to Reverse-Transcriptase polymerase chain reaction (RT-PCR), western blotting, and immunochemical staining (IHC). For RT-PCR, RAN was extracted from the obtained lung tissue using TRIzol reagent (Thermo Fisher Scientific), and then RT-PCR was performed according to the manual using RT-PCR reagent (Bio-Rad). Primer sequences are listed in Table 1 below. And the results are shown in Figure 4.

As shown in Figure 4, it was confirmed that the expression of IGF2 is reduced in the experimental group treated with sochinone.

Genetic name RT- PCR primer order Sequence number IGF2 F: 5'-GCT CAC TTC CGA TTG CTG GCC AT-3' One R: 5'-TCG TGC TGC ATT GCT GCT GCT TAC CG-3' 2 Actin F: 5'-ACT ACC TCA TGA AGA TC-3' 3 R: 5'-GAT CCA CAT CTG CTG GAA-3' 4

In addition, for immunochemical staining, the isolated lung was fixed using 10% formalin for 24 hours, and then paraffin blocks were prepared using paraffin. Then, the produced paraffin block was cut to a thickness of 4 μm, and then attached to a silane coated slide and dried. The dried slides were put in a dry oven at 65° C. to deparaffin for 16 hours, deparaffinized and hydrated with 100%, 95%, and 70% alcohol, washed with distilled water, and then washed with IGF2 antibody After staining using (Cell signaling), it was observed under a microscope. The results are shown in FIG. 5.

As shown in Figure 5, in the control group (N/B) administered NKK and benzoapyrene, the expression of IGF2 was increased, but it was confirmed that the expression of IGF2 was decreased in the experimental group treated with socinone.

1.4. Cytotoxicity check

To confirm the cytotoxicity of socinone, 5X10 ⁴ in each well of a 96-well plate After adding HBE cells or RPE cells, sochinone was treated at a concentration of 0.1, 1, and 10 μM, and cultured for ₂ days in a cell incubator at 37° C. and 5% CO ₂ conditions. Then, a solution of MTT(3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide; Sigma) was added to a concentration of 0.5 mg/ml per well, and cultured at 37°C for 4 hours. After washing with PBS, the supernatant was completely removed, 200 ul of DMSO was added, reacted for 30 minutes, and absorbance was measured at 570 nm. Cell viability was calculated as a percentage of the absorbance of the experimental group relative to the absorbance of the control group (the experimental group treated with only the medium). The results are shown in FIG. 6.

As shown in Figure 6, sochinone does not affect the survival rate of normal cells, through which it was confirmed that socinone is low in cytotoxicity.

Through the above results, the composition containing socinone of the present invention as an active ingredient effectively reduces the damage of lung tissue and IGF2 expression by NKK and benzoapyrene that mimic tobacco components, effectively reducing lung function. It has been confirmed that maintenance and/or recovery, and through this, it was confirmed that sochinone can be used as a composition for the prevention or treatment of chronic lung diseases including emphysema.

The above-described description of the present invention is for illustration only, and a person having ordinary knowledge in the technical field to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. There will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

<110> Seoul National University R&DB Foundation <120> Composition comprising sauchinone for preventing or treating of chronic pulmonary disease <130> MP18-083 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> IGF2 forward primer <400> 1 gctcacttcc gattgctggc cat 23 <210> 2 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> IGF2 reverse primer <400> 2 tcgtgctgca ttgctgctgc ttaccg 26 <210> 3 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> Actin forward primer <400> 3 actacctcat gaagatc 17 <210> 4 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Actin reverse primer <400> 4 gatccacatc tgctggaa 18

Claims (6)

Pharmaceutical composition for the prevention or treatment of chronic obstructive pulmonary disease, comprising sochinone (sauchinone) as an active ingredient. According to claim 1,
The chronic obstructive pulmonary disease is characterized in that the emphysema (emphysema), pharmaceutical composition. According to claim 1,
The composition is characterized in that it inhibits the damage of lung tissue and IGF2 expression, pharmaceutical composition. Food composition for the prevention or improvement of chronic obstructive pulmonary disease, comprising sochinone (sauchinone) as an active ingredient. The method of claim 4,
The chronic obstructive pulmonary disease is characterized in that the emphysema (emphysema), food composition. The method of claim 4,
The composition is characterized in that it inhibits the damage of lung tissue and IGF2 expression, food composition.

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