KR20200127730A - Composition comprising extracts of Houttuynia cordata for acute gastritis and gastric mucosal protection - Google Patents

Abstract

The present invention relates to a composition for acute gastritis and gastric mucosa protection containing a Houttuynia cordata extract. The Houttuynia cordata extract reduces severe hemorrhagic mucosa in the gastric mucosa, increases a physiologically active substance (PGE_2), and recovers the swelling of epithelial cells and the boundary between broken cells in an HCl/Ethanol-induced gastritis animal model, thereby being able to be usefully used as a pharmaceutical composition for preventing and treating acute gastritis, and protecting gastric mucosa, and a food composition for preventing or alleviating acute gastritis, and protecting gastric mucosa.

Description

Composition comprising extracts of Houttuynia cordata for acute gastritis and gastric mucosal protection}

The present invention relates to a composition for acute gastritis and gastric mucosa protection comprising Eoseongcho extract, wherein the Eoseongcho extract reduces severe hemorrhagic mucosa in the gastric mucosa in an HCl/Ethanol-induced gastritis animal model and increases physiologically active substance (PGE2). It can be usefully used as a pharmaceutical composition for preventing and treating acute gastritis, protecting gastric mucosa, preventing and improving acute gastritis, and protecting gastric mucosa because it restores the boundary between the swelling of the epithelial cells and the broken cells.

The stomach is an organ that stores food that has entered through the esophagus, breaks it into pieces for easy digestion, and regulates the delivery of food to the duodenum so that it can be efficiently digested and absorbed in harmony with the secretion of pancreatic enzymes. When food comes in, the stomach secretes gastric acid, a strong acid to digest it. At this time, the protective layer of the gastric mucosa acts to prevent damage to the gastric mucosa by gastric acid.

Factors that adversely affect the functioning of a person's gastrointestinal system are extremely diverse in nature. Such disorders can occur in the upper gastrointestinal tract, lower gastrointestinal tract, or both, and there are a wide range of gastrointestinal disorders, including genetic, physiological, environmental and mental factors. Chronic diseases of the upper gastrointestinal tract include gastritis and gastric ulcer, and inflammation occurs when the secretion of gastric acid is promoted or the protective layer of the gastric mucosa is damaged due to various causes. When this inflammatory lesion is confined to the gastric mucosa, it is called gastric ulcer, when it penetrates the gastric mucosa and forms an ulcer to the submucosal tissue and muscle layer. In addition, ulcers in the duodenum are called duodenal ulcers, and gastric ulcers and duodenal ulcers are collectively referred to as peptic ulcers.

The main symptoms of gastritis and peptic ulcer include abdominal pain, heartburn, bloating, indigestion, belching, nausea, vomiting, and bleeding. Among these, the most painful of these is abdominal pain, the area of which is the myeongchi area, the heart and the right upper abdomen, and in the case of gastric ulcer, it appears between 30 to 60 minutes after meals, and in the case of duodenal ulcer, it is very painful on an empty stomach, that is, 2-3 hours after meals. It is known to appear.

Gastritis and peptic ulcer are caused by excessive secretion of gastric juice or a decrease in the resistance of the lining of the stomach to digestive acids and pepsin. Therefore, as drugs used to treat gastritis and peptic ulcer, drugs to reduce gastric acid, pepsin, stress, etc., which are factors that promote and recur the ulcer, are mainly used, and among them, drugs that inhibit excessive secretion of gastric acid are used. It is used the most.

Conventional treatments for gastritis and peptic ulcer include antacids that neutralize excessive gastric secretion, histamine H ₂ receptor antagonists and proton pump inhibitors that inhibit gastric acid secretion, and increase the resistance of the gastric lining to digestive juices. And prostaglandin, or gastric mucosa protective agent, which can inhibit acid secretion.

Among the above drugs, antacids are not for fundamental treatment, but for temporary fast-acting, and show medicinal effects by lowering the activity of pepsin by raising the pH in the stomach. However, side effects such as constipation, diarrhea, metabolic alkalosis, and urolithiasis have been reported by the administration of inorganic substances, and recently, acid rebound caused by antacids has been a problem.

Cimetidine, the most widely used histamine H ₂ receptor antagonist among the above drugs, blocks histamine receptors in the gastric mucosa and blocks histamine molecules by gastric cells, thereby preventing the histamine molecules from signaling the secretion of acid to the gastric cells. do. The full-fledged treatment of peptic ulcer began when cimetidine was released in 1977, and several types of derivatives were developed. Among them, ranitidine, famotidine, roxatidine, nizatidine, etc., exhibit excellent anti-ulcer effects in clinical practice due to the shortening of the treatment period. There is a disadvantage of high recurrence rate. In addition, cimetidine exhibits side effects such as decreased libido in men, gynecomastia, and neutropenia, as well as drug interactions, so the use of cimetidine is avoided, and ranitidine and famotidine, the most recent drugs, are effective in long-term use. Significant decline is being observed.

In addition, proton pump inhibitors include omeprazole and lansoprazol, and are known to exhibit a strong acid secretion inhibitory effect by inhibiting acid secretion in gastric parietal cells in the final stage. However, the recurrence rate is not decreased even in patients with ulcer healing by these drugs, and side effects such as soft stool, diarrhea, fever, headache, and fatigue have been reported.

As described above, with the development of antisecretory agents such as histamine H ₂ receptor antagonists and proton pump inhibitors, gastritis and peptic ulcers requiring surgery have been cured with drug therapy. However, most of the healed gastritis and peptic ulcers recur or recur and require long-term maintenance therapy even after they are completely healed, and a phenomenon of recurrence or recurrence is frequently observed during maintenance therapy. Therefore, when only histamine H ₂ receptor antagonist and proton pump inhibitor are used for the purpose of inhibiting gastric acid secretion, the most important recurrence rate inhibitory effect in the treatment of gastritis and peptic ulcer cannot be expected.

In addition, gastric mucosa protective agents generally have a short-acting effect, a large dose, and have a disadvantage of having to be taken for a relatively long period of time, but it is known that the regenerated mucosa recovers similar to normal. Therefore, in recent years, with gastric acid inhibitors such as antacids, histamine H ₂ receptor antagonists or proton pump inhibitors, such as sucralfat and bismuth chelate, which protect mucous membranes, promote tissue repair, and increase mucosal blood flow. A gastric mucosa protectant is being used at the same time.

In general, in order to relieve pain, which is a major symptom of gastritis and peptic ulcer, there are cases of fast-acting antacids or eating, but this only makes temporary pain disappear, and steroidal and nonsteroidal anti-inflammatory analgesics are not used due to side effects. It is limited. In particular, nonsteroidal anti-inflammatory analgesics such as aspirin and ibuprofen are severely restricted in their use, such as intensifying peptic ulcer, so that patients with ulcers are extremely refrained from using it for other diseases or taken together with antacids. Accordingly, there is a need to develop a drug that can relieve pain and protect the stomach, which is a major symptom of gastritis and peptic ulcer.

Therefore, the present inventors experimented with a number of natural substances to protect gastritis and gastric mucosa. As a result, Eoseongcho extract increases the occurrence of prostagladin E2, which protects the gastric mucosa, while rapidly recovering damaged epithelial cells. It was found that there is, and the present invention was completed.

Korean Patent Registration No. 10-1961028 Korean Patent Registration No. 10-1329575 Korean Patent Registration No. 10-1045368 Korean Patent Registration No. 10-1615929 Korean Patent Registration No. 10-0921006

One object of the present invention is to provide a composition for acute gastritis and gastric mucosa protection comprising Eoseongcho extract.

Another object of the present invention is to provide the composition as a pharmaceutical composition for the prevention and treatment of acute gastritis and protection of the gastric mucosa, and a food composition for the prevention and improvement of acute gastritis and protection of the gastric mucosa.

Another object of the present invention is to provide a method of preventing, treating, or improving acute gastritis in an individual and protecting the gastric mucosa by administering the composition to an individual.

In one aspect, the present invention provides a composition for acute gastritis and gastric mucosa protection comprising Eoseongcho extract.

In the present invention, Houttuynia cordata thunb.) is an outpost of Yakmomil, a plant of the Saururaceae family, and inhabits southeastern Asia and especially Japan and Korea. Eoseongcho is also called Sip-Yak because it has ten medicinal effects, and the stem is like a sweet potato leaf, and it is also called Eoseongcho because it originates from the fact that when you touch the raw leaves, fish smells badly. Eoseongcho is also available for medicinal and edible use. It is classified as animals and plants that can only be used in a minimum amount as an auxiliary ingredient in the classification of food raw materials in the food code. Pharmacologically, it is widely known for its strong heart, diuretic, antibacterial, detoxifying, and anticancer effects. Is used as a cosmetic and health functional food that helps detoxification and beauty.

In the present invention, “eoseongcho extract” is an extract obtained by extracting Eoseongcho by a conventional method, and includes not only the extract extracted from Eoseongcho, but also dry powder thereof or any form formulated using the same.

The Eoseongcho extract may be extracted using water or an organic solvent, and the extracted liquid may be used in a liquid form or concentrated and/or dried. The organic solvent is methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1 ,3-butylene glycol, propylene glycol, or a mixed solvent thereof, and can be extracted by heating or at room temperature under conditions that do not destroy or minimize the active ingredient of the extract.

The extraction method is not limited, for example, cold sediment extraction, ultrasonic extraction, reflux cooling extraction, and the like.

According to a specific embodiment of the present invention, the Eoseongcho extract is a hot water extract obtained by adding 5 to 15 times the volume of Eoseongcho outpost and extracting it at 100 to 110°C for 3 to 10 hours.

Eoseongcho extract of the present invention may be all extracts, fractions, purified products, diluted solutions, concentrates, or dried products obtained in each step of extraction, fractionation, or purification (separation, fractionation).

The composition of the present invention may contain 0.0001 to 80% by weight, more preferably 0.001 to 10% by weight, and most preferably 0.1 to 5% by weight, based on the total weight of the Eoseongcho extract. If the Eoseongcho extract is less than 0.0001% by weight of the total composition, the prevention, treatment or improvement effect of acute gastritis and the protective effect of the gastric mucosa are too weak, and if it exceeds 80% by weight, the increase in the effect due to the increase in content In addition to being insufficient, there is a problem in that the stability of the formulation is not secured.

In the present invention, "acute gastritis" is induced by non-steroidal anti-inflammatory drugs (NSAIDs), steroidal anti-inflammatory drugs, drugs, alcohol, food poisoning, food hypersensitivity, smoking, or stress. Refers to the acute inflammatory disease of the stomach.

In the present invention, the acute gastritis includes paroxysmal gastritis, erosive gastritis, hemorrhagic gastritis, bile reflux gastritis, chronic superficial gastritis, atrophic gastritis, metabolic gastritis, hypertrophic gastritis, and the like.

In the present invention, "gastric mucosa damage" refers to a non-steroidal anti-inflammatory drug (NSAID), a steroidal anti-inflammatory drug, a drug, alcohol, food poisoning, food hypersensitivity, smoking, or stress due to single-layer columnar epithelium. It refers to the destruction of the mucous membrane that covers the inner surface of the gastric wall. The above-described acute gastritis may occur due to the gastric mucosa injury.

In the present invention, "protecting the gastric mucosa" refers to preventing or preventing damage to the gastric mucosa or restoring the damaged gastric mucosa.

According to a specific embodiment of the present invention, the Eoseongcho extract may reduce the wound area by forming a new tissue in the wound area. In the wound healing process, blood vessel constriction and platelet coagulation occur along with the inflammatory phase at the wound site after the wound, followed by complex pathological processes such as synthesis of collagen and the formation of new blood vessels, after re-epithelialization, in which granulation tissue fills the wound area. It goes through the process of contracting the wound area.

Therefore, the extract of Eoseongcho of the present invention can heal wounds by forming a new tissue in the wound area and contracting the wound area.

According to a specific embodiment of the present invention, Eoseongcho extract reduces severe hemorrhagic mucosal damage that occurs in gastric mucosa in an HCl/Ethanol-induced gastritis animal model, increases physiologically active substance (PGE2), and swells and collapses of gastric epithelial cells. It has an excellent effect in restoring the boundaries of the liver.

Therefore, the Eoseongcho extract of the present invention can protect the gastric mucosa and prevent, improve, and treat acute gastritis.

According to one specific aspect of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of acute gastritis, comprising an Eoseongcho extract as an active ingredient.

According to another specific aspect of the present invention, the present invention provides a pharmaceutical composition for gastric mucosa protection comprising an Eoseongcho extract as an active ingredient.

In the present invention, the "active ingredient" refers to a component that exhibits a desired activity by itself or can exhibit activity together with a carrier that is not active by itself.

When the composition of the present invention is a pharmaceutical composition, it may further contain a pharmaceutically acceptable carrier in addition to the Eoseongcho extract as an active ingredient. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and carbohydrate compounds (eg, lactose, amylose, dextrose, sucrose, sorbitol, mannitol, starch, cellulose, etc.) , Gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, salt solution, alcohol, gum arabic, vegetable oil (e.g. corn oil, cotton seed oil , Soy milk, olive oil, coconut oil), polyethylene glycol, methyl cellulose, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, mineral oil, etc., but are not limited thereto. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (latest edition), Mach Publishing Company, Easton, PA.

A suitable dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the severity of the disease, the form of the drug, and the time, but can be appropriately selected by a person skilled in the art. It is 0.001 mg/kg to 50 mg/kg, and it can be administered once to several times a day as needed.

Formulations of the pharmaceutical composition of the present invention include oral formulations such as powders, granules, tablets, capsules, emulsions, syrups, aerosols, etc., and sterile injectable solutions in any form suitable for pharmaceutical formulations according to conventional methods. It can be used in a form.

According to another specific aspect of the present invention, the present invention provides a food composition for preventing or improving acute gastritis, comprising an extract of Eoseongcho as an active ingredient.

According to another specific aspect of the present invention, the present invention provides a food composition for gastric mucosa protection comprising an extract of Eoseongcho as an active ingredient.

When the composition of the present invention is a food composition, as an active ingredient, various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening 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, carbonates used in carbonated beverages, and the like may be contained. In addition, it may contain pulp for the manufacture of natural fruit juices, synthetic fruit juices and vegetable beverages. These components may be used independently or in combination. In addition, the food composition may be in the form of any one of meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, drink, alcohol and vitamin complex. have.

In addition, the food composition may additionally contain food additives, and whether it is suitable as a food additive is determined according to the general rules of the Food Additive Code approved by the Ministry of Food and Drug Safety and general test methods, etc. And the criteria.

For example, ketones, glycine, potassium citrate, nicotinic acid, chemical synthetic products such as cinnamic acid, dark pigment, licorice extract, crystalline cellulose, high cooling pigment, natural additives such as guar gum, L-glutamic acid And mixed preparations such as sodium preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.

At this time, the content of the composition according to the present invention added to food in the process of manufacturing the food composition may be appropriately added or subtracted as needed.

In another specific aspect, the present invention provides a method of preventing, treating or improving acute gastritis in an individual and protecting the gastric mucosa by administering a composition containing Eoseongcho extract as an active ingredient to an individual, and a use thereof.

In the present invention, the term "subject" refers to monkeys, cattle, horses, pigs, sheep, dogs, cats, rats, mice, including humans with conditions or diseases in which symptoms can be improved by administering the composition of the present invention, It means a mammal such as a chimpanzee.

The administration is oral or parenteral administration, preferably oral administration, and its dosage is based on factors such as formulation method, administration method, age, weight, sex, and pathological condition of the patient. It can be easily changed and used by the possessor.

On the other hand, the Eoseongcho extract of the present invention has no cytotoxicity and is excellent in human stability, so it has almost no side effects to the human body, so it can be safely used even when used for a long time, so it can be safely used in medicines and foods.

Eoseongcho extract of the present invention reduces severe hemorrhagic mucosal damage in the gastric mucosa in an HCl/Ethanol-induced gastritis animal model, increases the physiologically active substance (PGE2), and restores the swelling of epithelial cells and the boundary between broken cells. It can be usefully used as a pharmaceutical composition for the prevention and treatment of gastritis and protection of the gastric mucosa, as a food composition for prevention and improvement of acute gastritis and protection of the gastric mucosa. In addition, the Eoseongcho extract of the present invention has no cytotoxicity and has excellent human stability, so there are almost no side effects to the human body, so it can be safely used even when used for a long time, so it can be safely used in medicines and foods.

1 is a diagram illustrating the effect of Eoseongcho water extract on gastric lesions in an HCl/Ethanol-induced gastritis animal model.
FIG. 2 is a diagram illustrating the effect of Eoseongcho water extract on PGE2 concentration in an HCl/Ethanol-induced gastritis animal model.
3 is a diagram illustrating the effect of Eoseongcho water extract on gastric tissue morphology change in an HCl/Ethanol-induced gastritis animal model.

Hereinafter, the present invention will be described in more detail with reference to examples. These examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention.

<Example 1> Preparation and experimental method of extract

1-1. Preparation of Eoseongcho extract

Houttuynia used in this experiment cordata thunb.) The outpost was washed and dried for a week in a hot air dryer at 50℃.

After 3,080 g of the dried sample was pulverized, water equivalent to 10 times the weight of the sample was added, and extraction was repeated at 105° C. for 5 hours using a reflux cooling extraction method. The extracted sample was concentrated at 55°C using a large rotary vacuum concentrator (Eyela, NVC-2200, ToKyo, Japan), and then filtered using filter paper (Thermo, Waltham, MA, USA), and the filtered extract was subjected to a freeze dryer. And dried.

The experimental material was 908 g to obtain a yield of about 29%, and the dried powder was used after dissolving in 0.5% carboxymethyl cellulose (CMC, Sigma, St. Louis, MO, USA).

1-2. Laboratory animals and treatment

To measure the gastritis prevention effect, male 6-week-old SD rats were purchased from Orient Bio (Jeongeup, Korea), and were divided into five groups, and the number of digits was as follows (normal control; n = 7, negative control; n = 7, positive control; n = 7, 100 mg/kg Eoseongcho water extract administration group; n = 7, 500 mg/kg Eoseongcho water extract administration group; n = 7).

The negative control group was orally administered 1 ml of water for 3 days. The positive control group was orally administered omeprazole at a concentration of 20 mg/kg for 3 days, and the Eoseongcho water extract group was orally administered at a concentration of 100 or 500 mg/kg for 3 days, respectively. On the last day of the experiment 30 minutes after oral administration, gastritis was induced for 30 minutes by oral administration of 150 mM HCl (in 60% EtOH, 1 ml) per animal, and gastritis induction concentration was selected by referring to several experiments (Choi et al., 2012; Hwang et al., 2017; Lee et al., 2016), after the experiment, all experimental animals induce euthanasia by overbreathing CO ₂ in accordance with the IACUC laboratory animal management criteria.

1-3. Gastritis lesion and inhibition rate measurement

In order to measure the degree of inhibition by gastritis lesions and extract administration, the extracted stomach was incised and fixed, and the length of damage to the stomach wall was measured through a microscope, and the area and inhibition rate of the lesion were obtained.

1-4. Hematoxylin and eosin staining

The extracted stomach was fixed in 10% NBF and embedded in paraffin through tissue processing. Embedded stomach tissue was sectioned to a thickness of 5 µm so that the mucosal layer of the stomach wall could be identified, and Dako Mayer's Hematoxylin (Agilent, Santa clara, California, USA) and Eosin Y (Sigma, St. Louis, Missouri, USA) were used. After staining using, it was observed under a microscope.

1-5. PGE ₂ measurement

Experiments were conducted using a PGE ₂ ELISA kit (Enzo, Farmingdale, USA). Part of the extracted stomach was cut to a constant weight, dissolved in RIPA buffer (Cell signaling technology, Danvers, Massachusetts, USA) to separate the protein, and then centrifuged for 10 minutes at 4℃ and 13,000×g with a centrifuge. The experiment was performed according to the protocol suggested by the manufacturer using the protein lysate. In addition, the PGE2 concentration per weight of the stomach tissue was expressed as a graph.

1-6. Statistical processing

Statistical processing was performed using SPSS (Chicago, IL, USA), and the mean value and standard deviation were calculated. To verify the mean difference between the experimental groups, one-way ANOVA was performed, and then statistical difference was verified at the significance level p <0.05 using the Tukey-Kramer Multiple Comparison Test.

< Experimental example 1> Eoseongcho water extract in HCl/Ethanol-induced gastritis animal model Stomach injury Analysis of the effect on the lesion

The inhibitory effect on gastric damage by HCl-ethanol was observed. An incision was made in the upper Taiwan area, and the degree of damage to the gastric mucosa was observed with the naked eye. In the control group in which gastritis was induced, hemorrhage of the dark red line due to bleeding occurred in the upper body and the lower part of the stomach, and serious hemorrhagic mucosal damage was observed in the entire stomach. On the other hand, the positive control drug omeprazole administration group (POS) significantly reduced gastric damage compared to the HCl/Ethanol control group.

The drug group administered orally at concentrations of 100 and 500 mg/kg of Eoseongcho water extract showed a decreasing tendency compared to the HCl/Ethanol control group, and suppressed the lesions of gastritis by 31% and 73.3%, respectively (see FIG. 1).

< Experimental example 2> Eoseongcho water extract in HCl/Ethanol-induced gastritis animal model PGE ₂ Analysis of the effect on concentration

General inflammatory response, the PGE ₂ enzyme is known to be increased, but the inhibition of the gastric tissue PGE2 enzyme gastric mucosal barrier preserving the above, the role of promoting cell proliferation occurs when the gastritis or gastric ulcer is PGE2 enzyme is reduced. Gastric tissue was analyzed by ELISA method to observe the PGE ₂ change in the gastritis animal model of Eoseongcho water extract.

As a result, as can be seen in Fig. 2, 42.1±1.6 ng/mg in the normal control group, 11.1±0.2 ng/mg in the HCl/Ethanol control group (CON), and 36.9±3.2 ng in the group administered omeprazole (POS). /Mg also showed a significant increase compared to the control group. The PGE ₂ was significantly increased in the Eoseongcho water extract-administered group, 16.3±4 ng/mg and 23.3±2.4 ng/mg, respectively, compared to the control group.

< Experimental example 3> Analysis of the effect of Eoseongcho water extract on gastric tissue morphology in an HCl/Ethanol-induced gastritis animal model

In order to investigate the effect of Eoseongcho water extract on gastric epithelial cell injury in gastritis-induced mice, gastric tissue was H&E stained. First, as a result of observing the change in the morphology of the gastric tissue of the mouse, it was observed that in the control group that induced gastritis, the swelling of the gastric epithelial cells and the boundary between the cells were significantly broken. In the group administered with the positive control omeprazole (POS), the preservation of epithelial cells increased compared to the control group. In the group to which Eoseongcho water extract was administered 100 mg/kg and 500 mg/kg, it was confirmed that it was suppressed compared to the control group (see FIG. 3).

Claims (4)

A pharmaceutical composition for the prevention or treatment of gastric mucosa protection and acute gastritis, comprising Eoseongcho extract as an active ingredient.
The pharmaceutical composition according to claim 1, wherein the Eoseongcho extract is Eoseongcho water extract.
Food composition for preventing or improving gastric mucosa and preventing or improving acute gastritis, comprising Eoseongcho extract as an active ingredient.
The food composition according to claim 3, wherein the Eoseongcho extract is Eoseongcho water extract.

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