KR102588240B1 - Composition for Improving, Preventing or Treating Helicobacter pylori Infection - Google Patents

Abstract

The present invention relates to a composition for improving, preventing or treating Helicobacter pylori infection, and specifically, to a composition for improving, preventing or treating Helicobacter pylori infection containing estafiatin as an active ingredient. It's about.

Description

Composition for improving, preventing or treating Helicobacter pylori infection {Composition for Improving, Preventing or Treating Helicobacter pylori Infection}

The present invention relates to a composition for improving, preventing or treating Helicobacter pylori infection, and specifically, to a composition for improving, preventing or treating Helicobacter pylori infection containing estafiatin as an active ingredient. It's about.

Helicobacter is a Gram-negative bacterium that lives between the gastric mucosa and mucus and is the cause of gastritis and peptic ulcer disease around the world. The World Health Organization (WHO) defined Helicobacter pylori infection as a carcinogen in 1994. Helicobacter pylori is so common that more than half of the world's population is infected, and Korea is no exception. The infection rate is 47% of the entire population and approximately 70% among adults. Additionally, Helicobacter pylori is found in 90-95% of duodenal ulcer patients and 60-80% of gastric ulcer patients.

Symptoms of Helicobacter pylori infection appear differently depending on the host's susceptibility and the diversity of the strain. Even if infected with bacteria, there are often no symptoms, and only in some cases symptoms such as indigestion, acute gastritis, chronic gastritis, gastric ulcer, duodenal ulcer, and stomach cancer appear. Helicobacter pylori infection can be diagnosed through gastroscopy, blood test, and urea breath test.

Treatment is necessary after endoscopic resection for patients with gastric ulcer, duodenal ulcer, gastric mucosa-associated lymphoid tissue lymphoma, and early gastric cancer, and for patients with chronic idiopathic thrombocytopenic purpura. In addition, treatment is required if there is a direct family history of stomach cancer or iron deficiency anemia with no clear cause.

Generally, amoxicillin, clarithromycin, and gastric acid secretion suppressants are taken for bacterial eradication treatment, and about 70-80% of people who receive treatment succeed in eradicating bacteria. However, there may be cases where sterilization fails, and antibiotic-resistant bacteria may develop. Additionally, side effects such as diarrhea, loss of taste, and skin abnormalities may occur during eradication treatment. The reinfection rate in Korea after treatment for Helicobacter pylori is approximately 2-4%.

Meanwhile, research on bioactive substances that can prevent or suppress diseases from natural products is increasing. In particular, research on the search for marine plants known to contain bioactive substances and materials that can suppress or improve diseases from seaweed are increasing. Research to find out is actively underway.

Artemisia scoparia is a perennial plant belonging to the Asteraceae family and is an edible and medicinal halophyte that grows in the sandy soil of the seaside. Estafiatin, isolated from the ethanol extract of Mugwort, is known as a representative sesquiterpene lactone, and sesquiterpene lactones are mostly found in Asteraceae plants. Although it has been scientifically proven that various compounds of the sesquiterpene lactone series have anti-inflammatory, anti-obesity, anti-cancer, and antioxidant effects, there is no known research on the antibacterial effect of estaphiatin against Helicobacter pylori.

The present inventors have made extensive research efforts to develop a composition for improving, preventing, or treating Helicobacter pylori infection. As a result, the present invention was completed by confirming that estafiatin is effective against Helicobacter pylori infection.

Accordingly, the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating Helicobacter pylori infection containing estapiatin as an active ingredient.

Another object of the present invention is to provide a food composition for preventing or improving Helicobacter pylori infection containing estapiatin as an active ingredient.

Another object of the present invention is to provide a use of estapiatin to improve, prevent, or treat Helicobacter pylori infection.

Another object of the present invention relates to a method of treating Helicobacter pylori infection using estapiatin.

The present invention relates to a pharmaceutical composition for preventing or treating Helicobacter pylori infection containing estafiatin as an active ingredient, and a food composition for preventing or improving Helicobacter pylori infection.

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

Estapiatin is a compound with the molecular formula of C ₁₅ H ₁₈ O ₃ and has a molar mass of 246.3 g/mol, and its structural formula is as shown in Structural Formula I below.

[Structural Formula I]

According to one aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating Helicobacter pylori infection, containing mugwort extract as an active ingredient.

In this specification, the term “Artemisia” refers to a perennial plant belonging to the Artemisia genus of the Compositae family. It is a perennial plant that grows on sandy beach soil and resembles mugwort, but has a different smell. Mugwort has tree-like stems, so it does not die even in winter, but mugwort has a characteristic that its stems completely dry out and die in winter. Mugwort is a rare herb that is rarely used in oriental medicine.

In the present invention, mugwort may be one or more species selected from the group consisting of leaves, stems, fruits, roots and flowers, for example, it may be leaves, but is not limited thereto.

The term “extract” used herein has the meaning commonly used in the art as a crude extract, but in a broad sense also includes fractions obtained by additional fractionation of the extract. In other words, mugwort extracts include not only those obtained using extraction solvents, but also those obtained by additionally applying a purification process. For example, fractions obtained by passing the extract through an ultrafiltration membrane with a certain molecular weight cut-off value, separation by various chromatographs (designed for separation according to size, charge, hydrophobicity, or affinity), etc. Fractions obtained through purification methods are also included in the mugwort extract of the present invention. Additionally, mugwort extract may be in solution, concentrate, or powder form.

In the present invention, the polar solvent may be water, alcohol, acetic acid, dimethyl formamide (DMFO), dimethyl sulfoxide (DMSO), or a mixture thereof, but is not limited thereto.

In the present invention, the alcohol is a straight-chain or branched alcohol having 1 to 4 carbon atoms, for example, methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, 2-butoxy. It may be ethanol, ethylene glycol, or a mixture thereof, but is not limited thereto.

In the present invention, the solvent has an alcohol content of 10 or more to less than 100% (v/v), 20 or more to less than 100% (v/v), 30 or more to less than 100% (v/v), or 40 or more to 100%. (v/v), 50 or more to 100% (v/v), 50 or more to less than 95% (v/v), 50 or more to less than 92% (v/v), 60 or more to 100% (v/v) /v), 60 or more to less than 95% (v/v), 60 or more to less than 92% (v/v), 70 or more to less than 100% (v/v), 70 or more to 95% (v/v) ) less than, 70 or more to less than 92% (v/v), 80 or more to less than 100% (v/v), 80 or more to less than 95% (v/v), 80 or more to less than 92%, for example, It may be a 90% (v/v) alcohol aqueous solution, but is not limited thereto.

In the present invention, the fractions are the normal hexane fraction obtained by solvent fractionating the crude extract extracted with a solvent with normal hexane, the chloroform fraction obtained by solvent fractionating the crude extract extracted with a solvent with normal hexane, and then solvent fractionating the remaining aqueous solution fraction with chloroform. After solvent fractionation of the extracted crude extract with normal hexane, the aqueous solution remaining after solvent fractionation with chloroform, the ethyl acetate fraction obtained by solvent fractionation with ethyl acetate, and the solvent-extracted crude extract into the aqueous solution remaining after solvent fractionation with normal hexane. It may be one or more fractions selected from the group consisting of a saturated butanol fraction obtained by solvent fractionating the aqueous solution fraction remaining after solvent fractionation with chloroform and solvent fractionation with ethyl acetate, and a mixture thereof. It is not limited.

In the present invention, the Helicobacter pylori infection may be one or more types selected from the group consisting of gastritis, gastroduodenal ulcer, gastric cancer, gastric burr lymphoma, anemia, purpura, arteriosclerosis, migraine, infertility, and chronic urticaria, but is limited thereto. That is not the case.

As used herein, the term “gastritis” refers to a condition that causes indigestion and discomfort in the upper abdomen. It is used not only in cases of organic lesions such as peptic ulcers and narrow-sense gastritis, but also in functional gastrointestinal disorders such as irritable bowel and non-ulcer dyspepsia. Includes all. Symptoms range from mild symptoms of indigestion, heartburn, sores, and abdominal discomfort to severe symptoms of severe abdominal pain, vomiting, headache, and fever. The causes of gastritis are very diverse, including overeating, uncleanly prepared foods, spicy or irritating foods, and irritation of the mucous membrane from various medications. In particular, acute gastritis is often caused by infection with bacteria, viruses, and Helicobacter pylori.

As used herein, the term “gastroduodenal ulcer” refers to a depression in the stomach or duodenal mucosa as if digging into the ground with a shovel. In this case, if the depression goes beyond the mucosal muscle plate, it is called an ulcer, and if it does not go beyond the mucosal muscle plate, it is called an erosion. The most important causes of peptic ulcers are known to be Helicobacter pylori bacteria and the use of non-steroidal analgesics and anti-inflammatory drugs. Symptoms vary greatly from patient to patient, and may complain of non-specific abdominal discomfort or indigestion without typical ulcer symptoms. Many cases are even observed without any symptoms at all.

As used herein, the term “gastric cancer” refers to all cancers that occur in the stomach, but mainly refers to gastric adenocarcinoma that arises from glandular cells of the gastric mucosa. Stomach cancer is caused by a combination of several factors rather than by any single factor. Currently known, it is related to Helicobacter pylori, stomach cancer-related diseases, diet, smoking, drinking, and family history. It is known that genetic and environmental factors act simultaneously in family history.

As used herein, the term “gastric lymphoma” refers to gastric malt lymphoma, which is the second most common malignant tumor of the stomach after gastric adenocarcinoma. It is a low-grade lymphoma derived from B cells of mucosa associated lymphoid tissue (MALT). Occurs in the stomach. In the human body, Helicobacter pylori infection and autoimmune conditions produce reactive MALT in the gastric mucosa, and it is known that eradication of Helicobacter pylori causes this MALT lymphoma to disappear. There are no clinically specific symptoms, but if the cancer has progressed, abdominal pain, bleeding, lumps, weight loss, and loss of appetite may appear.

As used herein, the term “anemia” refers to a case where the blood does not sufficiently supply oxygen necessary for metabolism of human tissues, resulting in hypoxia of the tissues, and anemia is diagnosed based on the hemoglobin (hemoglobin) in red blood cells. According to recent studies, Helicobacter pylori is known to increase the risk of anemia and glaucoma.

As used herein, the term “purpura” is a disease in which small purple spots occur due to bleeding in the skin or mucous membranes of the kidney. It is closely related to a decrease in platelets, and the blood coagulation ability is reduced, and the capillaries of the skin and mucous membranes are affected. Bleeding occurs. Symptoms of low platelet levels include increased bleeding after injury, gum or nose bleeding, increased bleeding when urinating or defecating, nausea, vomiting, diarrhea, and proteinuria. Helicobacter pylori is known to be one of the pathogens.

As used herein, the term “arteriosclerosis” refers to a phenomenon in which fat settles and sticks in blood vessels, causing the arteries to narrow and lose elasticity. It can be seen as a type of aging phenomenon, mainly due to the deposition of cholesterol or neutral fat, which reduces the elasticity of blood vessels and narrows arteries, including the formation of blood clots. The causes are known to be abnormalities in lipid metabolism, abnormal hormone metabolism, genetic predisposition, dietary problems, and lack of exercise. Symptoms include narrowing of the arteries, which prevents blood from reaching the end of the tissue sufficiently, leading to tissue necrosis and pain during exercise. , various symptoms such as numbness and paralysis appear. In particular, Helicobacter pylori is known to produce inflammatory mediators that travel through the blood and damage blood vessel walls, creating atherosclerosis, which is the early stage of arteriosclerosis.

As used herein, the term “migraine” is a type of headache that occurs paroxysmal and periodically due to dysfunction of blood vessels in the head, and the pain mainly occurs on only one side of the head. Although the mechanism by which migraine occurs is not yet clearly known, functional changes in the brain, changes in neurotransmitter concentration, and inflammatory responses in the trigeminal nerve and surrounding blood vessels are known to be important migraine pathogenesis mechanisms. Symptoms include pain in the head, indigestion and nausea, sensitivity to sound or light, and frequent complaints of dizziness. Recently, research has shown that Helicobacter pylori causes migraines.

In this specification, the term “infertility” is defined as a case where a couple who did not use contraception fails to achieve pregnancy within one year despite normal marital relations, and the cause is male factor (male factor), decreased ovarian function (decreased ovarian function) Reserve), ovulatory disorder, fallopian tube damage, ligation (knot), or paratubal adhesions (tubal injury, blockage, or paratubal adhesions), cervical or immunologic factors, etc. Recently, research has shown that Helicobacter pylori causes infertility.

As used herein, the term “chronic urticaria” refers to an increase in the permeability of blood vessels of the skin or mucous membrane (a property that allows the passage or invasion of substance molecules), causing blood plasma components to temporarily accumulate in the tissue, causing the skin to swell red or white. It is a skin disease accompanied by severe itching. In many cases, the exact cause cannot be found, and the skin swells rapidly, accompanied by severe itching, and the skin symptoms often disappear within 3 to 4 hours and then reappear in another location. The U.S. Centers for Disease Control and Prevention (CDC) states that infection with Helicobacter pylori can cause hives.

As used herein, the term “treatment” refers to any action in which symptoms of Helicobacter pylori infection are improved or completely cured by administration of the composition according to the present invention.

As used herein, the term “prevention” refers to all actions that suppress or delay the symptoms of Helicobacter pylori infection by administering the composition according to the present invention.

As used herein, the term “improvement” refers to any action in which symptoms of Helicobacter pylori infection are improved or completely cured by administration of the composition according to the present invention.

The mugwort extract contained in the composition of the present invention is a natural plant material and has no cytotoxicity. The upper quantitative limit of the mugwort extract contained in the composition of the present invention can be selected and implemented by a person skilled in the art within an appropriate range.

Another aspect of the present invention relates to a method for producing mugwort extract.

The extract of Mugwort includes a solvent extract of Mugwort, as described above.

The manufacturing process of the mugwort extract according to the present invention is described in more detail as follows: extraction is performed at room temperature with an extraction solvent of about 8 to 50 times the weight of the mugwort powder. After extraction, filter and collect the filtrate. The extraction temperature is not particularly limited, but is preferably 15 to 110°C, preferably 20 to 90°C.

The extraction process can be repeated once or several times, and in a preferred example of the present invention, a method of re-extraction after the first extraction can be adopted, which means that even if the extract is effectively filtered when mass-producing the extract, its moisture content is reduced. This is to prevent loss as it is high and the extraction efficiency decreases with primary extraction alone. In addition, as a result of verifying the extraction efficiency at each stage, it was found that about 80 to 90% of the total extraction amount was extracted through secondary extraction.

In one example of the present invention, the extraction process is performed twice, preferably at 8 to 12 volumes by reflux extraction. When filtering after extraction and repeating the previously obtained residue, the obtained residue is again combined with the extraction solvent, about 5 to 15 times by volume, and the filtrate, and then concentrated under reduced pressure to prepare a mugwort extract. In this way, extraction efficiency can be increased by performing two extractions and mixing the filtrate obtained after each extraction, but the extract of the present invention is not limited to extraction recovery.

If the amount of solvent used in preparing the Mugwort extract is too small, stirring becomes difficult and the solubility of the extract decreases, resulting in lower extraction efficiency. If it is too large, the amount of solvent used in the next purification step increases, making it uneconomical. Since handling problems may occur, it is recommended that the amount of solvent used be within the above range.

The filtered extract obtained in this way is about 10 to 30 times, preferably 15 to 25 times, more preferably about 20 times the total amount of the concentrate in order to control the content of remaining lower alcohol to be suitable for use as a pharmaceutical raw material. It can be prepared as a powdered mugwort extract by azeotropically concentrating it with pear water 1 to 5 times, preferably 2 to 3 times, adding the same amount of water again to homogeneously suspend it, and then freeze-drying it.

The extraction method used in the present invention may be any commonly used method, for example, cold immersion, hot water extraction, ultrasonic extraction, or reflux cooling extraction, but is not limited thereto.

The pharmaceutical composition of the present invention can be used as a pharmaceutical composition containing a pharmaceutically effective amount of mugwort extract and/or a pharmaceutically acceptable carrier.

The term “pharmaceutically effective amount” as used herein refers to an amount sufficient to achieve the efficacy or activity of the above-described mugwort extract.

As used herein, the term “including as an active ingredient” means containing a sufficient amount to achieve the efficacy or activity of the mugwort extract.

Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are those commonly used in preparation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, Includes, but is limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. It doesn't work. In addition to the above ingredients, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc.

The pharmaceutical composition according to the present invention can be administered to mammals, including humans, through various routes. The administration method may be any commonly used method, for example, oral, dermal, intravenous, intramuscular, subcutaneous, etc. administration, and is preferably administered orally.

The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, administration method, patient's age, body weight, gender, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity. Usually, a skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prevention.

The composition of the present invention may further contain pharmaceutically suitable and physiologically acceptable auxiliaries such as carriers, excipients, and diluents in addition to the mixed extract.

Carriers, excipients and diluents that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate. , cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

When formulating, commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants can be used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the extract with at least one excipient, such as starch, calcium carbonate, sucrose ( It can be prepared by mixing sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate talc are also used.

Preparations for oral use include suspensions, oral solutions, emulsions, syrups, ointments, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is.

Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, transdermal preparations, etc. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.

Preparations for suppositories include witepsol, macrogol, tween 61, cacao, laurin, glycerogenatin, etc. can be used.

In an embodiment of applying the composition of the present invention to humans, the herbal extract composition of the present invention may be administered alone, but is generally administered with a pharmaceutical carrier selected in consideration of the administration method and standard pharmaceutical practice (standard phamaceutical practice). It can be administered by mixing.

For example, the herbal extract-containing composition of the present invention may be in the form of tablets containing starch or lactose, or in the form of capsules alone or containing excipients, or as elixirs or suspensions containing flavoring or coloring chemicals. It can be administered orally, intraorally, or sublingually in dosage form. These liquid preparations may contain suspending agents (e.g. methylcellulose, semi-synthetic glycerides such as witepsol or a mixture of apricot kernel oil and PEG-6 esters or PEG-8 and caprylic/capric It can be formulated with pharmaceutically acceptable excipients such as mixtures of glycerides).

The administered dose of the extract-containing composition of the present invention may vary depending on the patient's age, weight, gender, dosage form, health condition, and disease level, and may be administered once or several times a day at regular time intervals according to the judgment of a doctor or pharmacist. It may also be administered in divided doses.

In the present invention, the pharmaceutical composition has a content of mugwort extract of 5 to 500 mg/day, 5 to 400 mg/day, 5 to 300 mg/day, 5 to 200 mg/day, 5 to 100 mg/kg of body weight. day, 10 to 500 mg/day, 10 to 400 mg/day, 10 to 300 mg/day, 10 to 200 mg/day, 10 to 100 mg/day, 20 to 500 mg/day, 20 to 400 mg/day , 20 to 300 mg/day, 20 to 200 mg/day, 20 to 100 mg/day, 50 to 500 mg/day, 50 to 400 mg/day, 50 to 300 mg/day or 50 to 200 mg/day, For example, it may be administered in the range of 50 to 100 mg/day.

If the daily dosage of the composition containing the mugwort extract of the present invention is less than the above dosage, no significant effect can be obtained, and if it exceeds the dosage, it is not only uneconomical but also outside the range of the usual dosage, so there is a risk of undesirable side effects occurring. Since this may occur, it is better to keep it within the above range.

Another aspect of the present invention relates to a food composition for preventing or improving Helicobacter pylori infection, comprising mugwort extract as an active ingredient.

In the present invention, mugwort may be one or more species selected from the group consisting of leaves, stems, fruits, roots and flowers, for example, it may be leaves, but is not limited thereto.

In the present invention, the mugwort extract may be a crude extract obtained by extracting mugwort with a polar solvent, a non-polar solvent, or a mixed solvent thereof.

In the present invention, the polar solvent may be water, alcohol, acetic acid, dimethyl formamide (DMFO), dimethyl sulfoxide (DMSO), or a mixture thereof, but is not limited thereto.

In the present invention, the mugwort extract may be an extract obtained by extraction with one or more solvents selected from the group consisting of water and straight-chain or branched alcohols having 1 to 4 carbon atoms, but is not limited thereto.

In the present invention, the solvent has an alcohol content of 10 or more to less than 100% (v/v), 20 or more to less than 100% (v/v), 30 or more to less than 100% (v/v), or 40 or more to 100%. (v/v), 50 or more to 100% (v/v), 50 or more to less than 95% (v/v), 50 or more to less than 92% (v/v), 60 or more to 100% (v/v) /v), 60 or more to less than 95% (v/v), 60 or more to less than 92% (v/v), 70 or more to less than 100% (v/v), 70 or more to 95% (v/v) ) less than, 70 or more to less than 92% (v/v), 80 or more to less than 100% (v/v), 80 or more to less than 95% (v/v), 80 or more to less than 92%, for example, It may be a 90% (v/v) alcohol aqueous solution, but is not limited thereto.

In the present invention, the solvent may be anhydrous alcohol, but is not limited thereto.

In the present invention, the solvent may be one or more selected from the group consisting of ethanol, butanol, ethyl ether, and ethyl acetate, but is not limited thereto.

In the present invention, the fractions are the normal hexane fraction obtained by solvent fractionating the crude extract extracted with a solvent with normal hexane, the chloroform fraction obtained by solvent fractionating the crude extract extracted with a solvent with normal hexane, and then solvent fractionating the remaining aqueous solution fraction with chloroform. After solvent fractionation of the extracted crude extract with normal hexane, the aqueous solution remaining after solvent fractionation with chloroform, the ethyl acetate fraction obtained by solvent fractionation with ethyl acetate, and the solvent-extracted crude extract into the aqueous solution remaining after solvent fractionation with normal hexane. It may be one or more fractions selected from the group consisting of a saturated butanol fraction obtained by solvent fractionating the aqueous solution fraction remaining after solvent fractionation with chloroform and solvent fractionating the aqueous solution fraction remaining after solvent fractionation with ethyl acetate with hydrated butanol, and mixtures thereof. It is not limited.

In the present invention, the Helicobacter pylori infection may be one or more types selected from the group consisting of gastritis, gastroduodenal ulcer, gastric cancer, gastric burr lymphoma, anemia, purpura, arteriosclerosis, migraine, infertility, and chronic urticaria, but is limited thereto. That is not the case.

When using the food composition of the present invention as a food additive, the food composition can be added as is or used together with other foods or food ingredients, and can be used appropriately according to conventional methods. In general, when manufacturing food or beverages, the food composition of the present invention may be added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw materials.

There are no special restrictions on the types of foods above. Examples of foods to which the above substances can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, etc. These include alcoholic beverages and vitamin complexes, and include all foods in the conventional sense.

The beverage may contain various flavors or natural carbohydrates as additional ingredients. The above-mentioned natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, and synthetic sweeteners such as saccharin and aspartame. . The ratio of the natural carbohydrates can be appropriately determined by the selection of a person skilled in the art.

In addition to the above, the food composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, and alcohol. , may contain carbonating agents used in carbonated drinks, etc. Additionally, the food composition of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages. These ingredients can be used independently or in combination. The proportions of these additives can also be appropriately selected by those skilled in the art.

In the food composition, content that overlaps with the pharmaceutical composition is omitted in consideration of the complexity of the present specification.

The present invention relates to a composition for improving, preventing or treating Helicobacter pylori infection, and specifically, to a composition for improving, preventing or treating Helicobacter pylori infection containing estafiatin as an active ingredient. It's about.

Figure 1 is the structural formula of estafiatin.
Figure 2 is a graph showing the OD value of the bacterial growth inhibitory effect of estapiatin when culturing Helicobacter pylori P1WT strain.
Figure 3 is a graph showing the OD value of the effect of estapiatin on inhibiting bacterial growth when culturing Helicobacter pylori SS1 strain.

Hereinafter, the present invention will be described in more detail through the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Throughout this specification, “%” used to indicate the concentration of a specific substance means (weight/weight) % for solid/solid, (weight/volume) % for solid/liquid, and Liquid/liquid is (volume/volume) %.

Preparation Example 1. Preparation of Mugwort ethanol aqueous solution extract

140 g of mugwort aerial part powder was ground with a homogenizer by adding 2.2 L of 95% (V/V) ethanol, then extracted at room temperature for 24 hours, and filtered through filter paper (No. 2, Whatman) to obtain an ethanol aqueous solution and extract filtrate. got it Additionally, 1 L of 95% (V/V) ethanol was added to the residue, and it was extracted and filtered again in the same manner as above. The obtained ethanol aqueous solution extraction filtrate was concentrated under reduced pressure at 45°C using a vacuum concentrator equipped with an aspirator (A-3S, Eyela, Tokyo, Japan) to prepare an aqueous mugwort ethanol solution extract.

Preparation Example 2. Preparation of estapiatin

The ethanol aqueous solution extract of Mugwort obtained in Preparation Example 1 was suspended in double distilled water (1.2 L) and solvent fractionated three times with normal hexane (1.2 L) to obtain a normal hexane fraction, and then the aqueous solution fraction was mixed with chloroform (CHCl ₃ , 1.2 L) repeated solvent fractionation three times to obtain a chloroform fraction. Then, the aqueous solution fraction was subjected to solvent fractionation three times with ethyl acetate (EtOAc, 1.2 L) to obtain an ethyl acetate fraction, and the aqueous solution fraction was subjected to repeated solvent fractionation three times with hydrated butanol (BuOH, 0.9 L) to obtain a water-saturated butanol fraction. . Each obtained fraction was concentrated under reduced pressure at 45°C using a vacuum concentrator to prepare normal hexane fraction, chloroform fraction, ethyl acetate fraction, and saturated butanol fraction.

The ethyl acetate fraction (13.74 g) was purified using MPLC equipped with a silica gel column. That is, the ethyl acetate fraction (13.74 g) was adsorbed onto silica gel, filled into a column, and then connected to a silica gel column (SNAP KP-Sil 340 g, Biotage, Seongnam, Korea). Afterwards, it was eluted and fractionated using a step-wise elution method (3,400 mL, 100 mL/min for each step) using chloroform/methanol = 9:1, 4:6, and 0:10 (v/v) solvent system.

Fraction A (6.37 g, chloroform/methanol = 9:1, v/v) obtained from the silica gel column chromatography was repurified using MPLC equipped with a silica gel column. That is, fraction A was adsorbed onto silica gel in the same manner as above, filled into a column, and then connected to a silica gel column for compound separation (ZIP KP-Sil 120 g, Biotage, Seongnam, Korea). Then, the gradient elution method (50 mL) was performed with normal hexane/ethyl acetate = 90:10 (v/v) for the first 10 minutes, and normal hexane/ethyl acetate = 0:100 (v/v) from 10 to 40 minutes. /min) to obtain fraction A7 (1.14 g, retention value 23-25 min) containing estaphiatin.

Experimental Example 1. Confirmation of the structure of estaphiatin isolated from ethanol aqueous solution extract of Mugwort

The structure of estapiatin, the compound isolated in Preparation Example 2, was confirmed through MS and NMR analysis. From the HR-ESI-MS (positive) spectrum of estapiatin, a molecular weight ion peak of m/z 269.1254 [M+Na] ⁺ was observed, and it was found that the molecular weight of this compound was 246 (C ₁₅ H ₁₈ O ₃ ). .

¹ H- and ¹³ C-NMR spectra suggested that this compound was a guaiadienolide-based compound. That is, in the ¹ H-NMR (500 MHz, CD ₃ OD) spectrum, four types of proton signals attributed to two types of sp ² methylene [δ 6.21 (1H, d, J = 3.6 Hz, H-13a), 5.48 (1H, d, J = 3.6 Hz, H-13b), 4.95 (1H, br. s, H-14a), 4.86 (1H, br. s, H-14b)] and five types of methine proton signals [ δ 2.97 (1H, dt, J = 11.0, 8.0 Hz, H-1), 3.37 (1H, s, H-3), 2.31 (1H, dd, J = 11.0, 8.5 Hz, H-5), 4.08 ( 1H, dd, J = 11.0, 8.5 Hz, H-6), 2.87 (1H, m, H-7)] were observed. In addition, five types of proton signals attributed to three types of methylene groups [δ 2.06 (1H, dt, J = 11.0, 8.0 Hz, H-2a), 2.06 (1H, dd, J = 14.0, 7.0 Hz, H -2b), 2.26 (2H, m, H-8), 2.21 (1H, m, H-9a), 1.53 (1H, m, H-9b)] and one methine proton signal [δ 1.62 (3H) , s, H-15)] was observed. ^{13 In} the C-NMR (125 MHz) spectrum, one carbonyl carbon signal [δ 169.7 (C-12)] and three quaternary signals [δ 65.8 (C-4), 146.1 (C-10), A total of 15 types of carbon signals, including [139.5 (C-11)], were observed, which was consistent with the analysis results of ESI-MS and ¹ H-NMR spectrum. From the above results, the structure of this compound was analyzed as estapiatin, a guaianolide-based compound.

Table 1 below shows ¹ H- (500 MHz) and ¹³ C-NMR (125 MHz) data (CD ₃ OD) of estapiatin, and its structure is as shown in FIG. 1.

location δH (rel. Int., Multi., J in Hz) δC One 2.97 (1H, td, 11.0, 8.0) 44.8 2a 2.06 (1H, dd, 14.0, 7.0) 33.1 2b 1.81 (1H, dd, 3.5, 1.0) 3 3.37 (1H, s) 63.3 4 - 65.8 5 2.31 (1H, dd, 11.0, 8.5) 50.8 6 4.08 (1H, dd, 11.0, 8.5) 80.5 7 2.87 (1H, m) 44.1 8 2.26 (2H, m) 28.6 9a 2.21 (1H, m) 29.2 9b 1.53 (1H, m) 10 - 146.1 11 - 139.5 12 - 169.7 13a 6.21 (1H, d, 3.5) 120.2 13b 5.48 (1H, d, 3.5) 14a 4.95 (1H, br. s) 115.3 14b 4.86 (1H, br. d, 2) 15 1.62 (3H, s) 18.6

Experimental Example 2: Evaluation of Helicobacter pylori P1WT growth inhibition using estapiatin isolated from mugwort ethanol aqueous extract (Optical Density; OD)

Helicobacter pylori P1WT strain was dispensed at a concentration of 1x10 ⁶ CFU/ml at 2ml/well in a 6-well plate, and then estaphiatin isolated from the mugwort ethanol extract was added at 0, 12.5, 25, 50, or 100. After treatment at a concentration of μM, the cells were incubated at 37°C and 5% CO ₂ for 24 or 48 hours. Inhibition of Helicobacter pylori growth was confirmed through the OD value before culture and the OD value 24 or 48 hours after culture.

The growth inhibitory effect (OD) of Helicobacter pylori P1WT strain in each experimental group treated with 0, 12.5, 25, 50, or 100 μM of estaphiatin isolated from mugwort ethanol extract is classified as follows and is shown in Table 2. , the results are shown in Figure 2.

Ethanol aqueous solution extract 24 hours 48 hours 0 (μM) 0.087 (OD) 0.570 12.5 0.045 0.219 25 0.042 0.109 50 0.042 0.044 100 0.044 0.043

To check the growth rate of bacteria, the growth rate was measured at OD 600 nm, and as a result, there was no significant difference between each sample treatment 24 hours after treatment. However, after 48 hours, it was found that in the experimental group treated with 0, 12.5, 25, 50, or 100 μM of estapiatin, the inhibitory effect on the growth of Helicobacter pylori P1WT increased as the treatment amount increased. However, in the experimental group treated with 50 or 100 μM, the effect of inhibiting the growth of Helicobacter pylori P1WT was found to be similar. As a result, it was confirmed that the growth inhibitory effect of Helicobacter pylori P1WT became significant when treated with 50 μM or more of the ethanol aqueous solution extract.

Experimental Example 3: Evaluation of Helicobacter pylori SS1 growth inhibition using estapiatin isolated from mugwort ethanol aqueous extract (Optical Density; OD)

Helicobacter pylori SS1 strain was dispensed at a concentration of 1x10 ⁶ CFU/ml into a 6-well plate at 2ml/well, and then treated with estaphiatin isolated from the Mugwort ethanol extract at a concentration of 0, 12.5, 25, 50, or 100 μM. Cultured at 37°C and 5% CO ₂ for 24 or 48 hours. Inhibition of Helicobacter pylori growth was confirmed through the OD value before culture and the OD value 24 or 48 hours after culture.

The growth inhibitory effect (OD) of Helicobacter pylori SS1 strain in each experimental group treated with 0, 12.5, 25, 50, or 100 μM of estaphiatin isolated from the ethanol extract of Mugwort is classified as follows and is shown in Table 3, and the results are presented. Figure 3 shows.

Ethanol aqueous solution extract 24 hours 48 hours 0 (μM) 0.245 (OD) 0.551 12.5 0.118 0.590 25 0.060 0.570 50 0.044 0.046 100 0.043 0.043

To check the growth rate of bacteria, the growth rate was measured at OD 600nm. As a result, 24 hours after treatment, in the experimental group treated with 12.5, 25, or 50 μM of ethanol aqueous solution extract, the effect of inhibiting the growth of Helicobacter pylori SS1 increased as the treatment amount increased. In the experimental group treated with 50 or 100 μM, the Helicobacter pylori SS1 growth inhibition effect was found to be similar. However, after 48 hours, it was confirmed that the experimental group treated with 12.5 or 25 μM of the ethanol aqueous solution extract showed no difference in the inhibitory effect of Helicobacter pylori SS1 growth compared to the control group. As a result, it was confirmed that the inhibitory effect on Helicobacter pylori SS1 growth was significant when treated with 50 μM or more of estapiatin.

Claims (12)

Pharmaceutical composition for preventing or treating Helicobacter pylori bacterial infection containing estaphiatin represented by the following structural formula I as an active ingredient:

[Structural Formula I]
. The method of claim 1, wherein the Helicobacter pylori infection is one or more types selected from the group consisting of gastritis, gastroduodenal ulcer, gastric cancer, gastric burr lymphoma, anemia, purpura, arteriosclerosis, migraine, infertility, and chronic urticaria. Pharmaceutical composition for preventing or treating bacterial infections. The pharmaceutical composition for preventing or treating Helicobacter pylori infection according to claim 1, wherein the estaphiatin is derived from an extract of Artemisia scoparia . The pharmaceutical composition for preventing or treating Helicobacter pylori infection according to claim 3, wherein the mugwort extract is a crude extract extracted with a solvent consisting of water, methanol, ethanol, or a combination thereof. The method of claim 4, wherein the mugwort extract is a pharmaceutical for preventing or treating Helicobacter pylori infection, which is obtained by fractionating the crude extract with one or more solvents selected from the group consisting of normal hexane, chloroform, ethyl acetate, and saturated butanol. Composition. The pharmaceutical composition for preventing or treating Helicobacter pylori infection according to claim 3, wherein the mugwort is at least one selected from the group consisting of leaves, stems, fruits, roots, and flowers of mugwort. Food composition for preventing or improving Helicobacter pylori bacterial infection containing estaphiatin represented by the following structural formula I as an active ingredient:

[Structural Formula I]
. The method of claim 7, wherein the Helicobacter pylori infection is at least one selected from the group consisting of gastritis, gastroduodenal ulcer, gastric cancer, gastric burr lymphoma, anemia, purpura, arteriosclerosis, migraine, infertility, and chronic urticaria. Food composition for preventing or improving bacterial infections. The food composition for preventing or improving Helicobacter pylori infection according to claim 7, wherein the estaphiatin is derived from an extract of Artemisia scoparia . The food composition for preventing or improving Helicobacter pylori infection according to claim 9, wherein the mugwort extract is a crude extract extracted with a solvent consisting of water, methanol, ethanol, or a combination thereof. The method of claim 10, wherein the mugwort extract is a food composition for preventing or improving Helicobacter pylori infection, wherein the crude extract is fractionated with one or more solvents selected from the group consisting of normal hexane, chloroform, ethyl acetate, and saturated butanol. . The food composition for preventing or improving Helicobacter pylori infection according to claim 9, wherein the mugwort is at least one selected from the group consisting of leaves, stems, fruits, roots, and flowers of mugwort.