Lonicerae flos water extract-containing pharmaceutical composition for preventing or treating helicobacter pylori infection

Abstract

To provide a pharmaceutical composition for preventing or treating Helicobacter pylori infection, or a health functional food, containing Lonicerae flos water extract.SOLUTION: The present invention provides a composition for preventing or treating Helicobacter pylori infection, using Lonicerae flos water extract containing secoxyloganin as an active ingredient.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a pharmaceutical composition for preventing or treating Helicobacter pylori infection containing an aqueous extract of honeysuckle flower.

Background Art

[0002] Helicobacter pylori (H. pylori) was first discovered in 1983 by Marshall and Warren in Australia. It is a spiral-shaped microaerophilic, Gram-negative bacterium with 4 to 6 flagella, and it is reported that more than half of the world's population is infected. In the case of Korea, an infection rate of more than 70% is reported among adults over 40 years old. Helicobacter pylori is mainly closely related to upper gastrointestinal diseases and is known as a causative bacterium that causes chronic gastritis, gastric and duodenal ulcers, gastric cancer, etc. In particular, in 1994, the IARC (International Agency for Research on Cancer) and the World Health Organization defined Helicobacter pylori as a carcinogenic factor (Class I carcinogen). In relation to the prevention and diagnosis of gastric cancer, the infectivity of Helicobacter pylori is accepted as a very important issue, and correcting the infection factors of Helicobacter pylori is accepted as an important issue for national health.

[0003] Helicobacter pylori has 5 to 6 flagella, which are locomotor organs similar to the tails of animals, and uses these to stick to the surface of the gastric mucosa by passing through the viscous mucus directly above the gastrointestinal wall. Urease is a common feature found in most strains of Helicobacter pylori and is well conserved genetically among individuals. Urease decomposes urea into ammonia (NH 3 ) and carbon dioxide (CO 2) It decomposes into [substances] to increase the surrounding pH, neutralize gastric acid, and survive. Due to the influence of the ammonia thus generated, cell destruction and mutations of the gastric mucosa are induced, resulting in the loss of the gastric acid defense function. A vicious cycle occurs where gastric acid attacks the gastric mucosa, leading to gastric tissue deformation and inflammatory reactions. The gastric mucosal inflammation caused by various cytotoxic factors secreted by Helicobacter pylori induces the activation of various inflammatory cells such as lymphocytes and macrophages. Such inflammatory cells have been reported to show adverse effects in vivo such as tissue damage and cytotoxicity, and to induce inflammatory mediators to exacerbate the inflammation.

[0004] Helicobacter pylori is a very dangerous microorganism in gastric diseases, but the development of appropriate antibacterial substances and the like is insufficient. Currently, in the treatment of Helicobacter pylori, there are common triple therapies that combine Bismuth preparations, Metronidazole, and Tetracycline or Amoxicillin, and quadruple therapies that mix Bismuth preparations, Omeprazole, Tetracycline, and Metronidazole. The above-mentioned treatment methods have been reported to have excellent therapeutic effects against Helicobacter pylori. However, repeated use of such drugs has been reported to cause an increase in resistance to antibiotics and various side effects. Currently, efforts are continuing to find extracts and active ingredients that can suppress Helicobacter pylori using various natural product materials.

[0005] Flos Lonicerae is the flower of Lonicera japonica Thunb. belonging to the family Caprifoliaceae, and has been used in traditional Chinese medicine and folk medicine for diuresis, stomach strengthening, arthritis, suppurative dermatitis, and bronchitis.

[0006] Among the components of honeysuckle, tannin, inositol, sterol, chlorogenic acid, isochlorogenic acid, etc. have been reported. Among the flavonoid components, luteolin, apigenin, luteolin-7-O-rhamnoglucoside, quercetin, etc. have been reported. The flavonoid components of honeysuckle are known to have anti-inflammatory and antimutagenic effects, etc.

[0007] As conventional prior literature related to a composition for preventing or treating Helicobacter pylori infection containing an aqueous extract of honeysuckle, the prior papers [Ma, F. et al., World J Gastroenterol., 16(44), 5629-5634, 2010], [Xiao, Z.P. et al., J Agric Food Chem., 60(42), 10572-10577, 2012] and [Yi, Hona, etc., Korean J Vet Res., 53(2), 117-123, 2013] disclose the anti-Helicobacter pylori effects on various types of natural product extracts containing honeysuckle extract. The prior literature [Oku, H. et al., Biol.Pharm.Bull., 34(8), 1330-1333, 2011] discloses a honeysuckle-derived compound containing secoxyloganin and its anti-allergic effect. The prior literature [Xiong, J. et al., Food Chemistry, 138, 327-333, 2013] has disclosed the antibacterial effects of secoxyloganin against Escherichia coli and Staphylococcus aureus.

[0008] In the process of researching the aqueous extract of Lonicera japonica Thunb., the inventors confirmed that among the active ingredients of the aqueous extract of Lonicera japonica Thunb., secoxyloganin shows a selective antibacterial effect against Helicobacter pylori, which is more than 20 times that against Escherichia coli and Staphylococcus aureus under the same conditions. The inventors also confirmed that the aqueous extract of Lonicera japonica Thunb. containing secoxyloganin at a specific content or more has excellent effects in suppressing and treating Helicobacter pylori infection, and thus completed the present invention.

Summary of the Invention

Problems to be Solved by the Invention

[0009] An object of the present invention is to provide a pharmaceutical composition or a health functional food for preventing or treating Helicobacter pylori infection containing the aqueous extract of Lonicera japonica Thunb.

Means for Solving the Problems

[0010] The present invention relates to a pharmaceutical composition for preventing or treating Helicobacter pylori infection, which is an aqueous extract of Lonicerae Flos and contains Secoxyloganin as an active ingredient.

[0011] The above aqueous extract of Lonicera japonica Thunb. can be obtained by mixing Lonicera japonica Thunb. with 1500 - 2500 parts by weight of water based on 100 parts by weight of Lonicera japonica Thunb. and then extracting at a temperature of 90°C or higher for 1 hour or more. The extraction equipment can be ordinary extraction equipment, ultrasonic crushing extraction equipment or fractionation equipment. Preferably, it is extracted at a temperature of 90°C or higher for 1 to 5 hours so that it contains 0.1 to 10% by weight of the active ingredient secoxyloganin.

[0012] Secoxyloganin, which is contained as an active ingredient in the above honeysuckle water extract, is contained in the honeysuckle water extract at 0.1 to 10% by weight. If it is less than 0.1% by weight or exceeds 10% by weight after removing the above weight%, the effect of inhibiting the infection of Helicobacter pylori is low and not preferable, or the increase range of the effect of inhibiting the infection of Helicobacter pylori is not large, so it is not economical and not preferable. Most preferably, for the effective treatment of Helicobacter pylori infection, secoxyloganin, which is contained as an active ingredient in the above honeysuckle water extract, is contained in the honeysuckle water extract at 0.5 to 5% by weight.

[0013] The above Helicobacter pylori infection is a disease caused by being infected with Helicobacter pylori, and may be selected from the group consisting of gastritis, gastric ulcer, duodenal ulcer, non-ulcer dyspepsia syndrome, gastric MALT lymphoma, gastric hyperplastic polyp, gastric cancer, digestive tract cancer, pancreatitis, inflammatory bowel disease and functional digestive disorders (such as upper abdominal fullness, upper abdominal pain, belching, abdominal distension, early satiety, nausea, vomiting, reflux, stomachache, loss of appetite, etc.). The honeysuckle water extract of the present invention shows a particularly selective therapeutic effect on Helicobacter pylori infectious diseases compared to Helicobacter pylori non-infectious diseases.

[0014] The pharmaceutical composition according to the present invention may be formulated into a suitable form together with a commonly used pharmaceutically acceptable carrier. "Pharmaceutically acceptable" refers to a composition that is physiologically acceptable and does not cause allergic reactions or similar reactions such as gastrointestinal disorders and dizziness when administered to humans. In addition, the above compositions may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories and sterile injection solutions by ordinary methods respectively.

[0015] Carriers, excipients, and diluents that may be included in the above composition can include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginic acid, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulating, it is prepared using diluents or excipients such as commonly used fillers, stabilizers, binders, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. Such solid preparations are prepared by mixing at least one or more excipients, such as starch, microcrystalline cellulose, sucrose or lactose, low-substituted hydroxypropyl cellulose, hypromellose, etc., with the extract of the present invention. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, internal liquids, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients, such as wetting agents, sweeteners, flavoring agents, preservatives, etc., may be included. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. In non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As the base of suppositories, witepsol, macrogol, tween 61, cocoa butter, laurin oil, glycerol, gelatin, etc. may be used. To formulate into a parenteral dosage form, the above extract or its pharmaceutically acceptable salt is mixed with water together with adjuvants such as sterilizing / sterilized agents, preservatives, stabilizers, hydrating agents or emulsification promoters, salts for osmotic pressure adjustment and / or buffers, and other therapeutically useful substances to produce a solution or suspension, which can be manufactured in ampoule or vial unit dosage forms.

[0016] The pharmaceutical composition containing the extract disclosed in the present invention as an active ingredient can be administered to mammals such as mice, livestock, and humans by various routes. All modes of administration are contemplated, and for example, it can be administered orally, rectally, or by intravenous, intramuscular, subcutaneous, intrauterine dural, or intracerebrovascular injection. The dosage will vary depending on the age, sex, weight of the subject being treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the time of administration, the route of administration, the absorption, distribution, and excretion rates of the drug, the types of other drugs used, and the judgment of the prescriber, etc. Determination of the dosage based on such factors is within the level of those skilled in the art, and generally the dosage ranges from 0.01 mg / kg / day to about 2000 mg / kg / day. A more preferred dosage is from 1 mg / kg / day to 500 mg / kg / day. The administration can be carried out once a day or divided into several times. The above dosage does not limit the scope of the present invention.

[0017] On the other hand, the present invention also relates to a health functional food for preventing or improving Helicobacter pylori infection, which is an aqueous extract of Lonicerae Flos and contains Secoxyloganin as an active ingredient.

[0018] The above health functional food refers to foods manufactured or processed using raw materials and components with useful functions, and includes, for example, health supplements, functional foods, nutritional agents, adjuvants, etc.

[0019] The above extract can be added preferably in an amount of 0.001% by weight to 50% by weight, more preferably 0.001% by weight to 30% by weight, and most preferably 0.001% by weight to 10% by weight based on the total weight of the health functional food. The health functional food of the present invention includes forms such as tablets, capsules, pills, or liquids, and examples of foods to which the extract of the present invention can be added include various foods, drinks, gums, teas, vitamin complexes, etc.

[0020] The present invention also relates to a pharmaceutical composition for preventing or treating Helicobacter pylori infection, which contains secoxyloganin as an active ingredient.

Effects of the Invention

[0021] The present invention relates to a composition for preventing or treating Helicobacter pylori infection, which contains an aqueous extract of Lonicera japonica Thunb. containing secoxyloganin as an active ingredient. By containing a specific content of secoxyloganin, the above extract has excellent antibacterial effects when treating Helicobacter pylori bacteria, and when treating mice infected with Helicobacter pylori, it has excellent effects of reducing the expression of Helicobacter pylori IgG antibodies in the blood, improving histopathological lesions, and reducing cytokine expression. Therefore, the aqueous extract of Lonicera japonica Thunb. of the present invention can be usefully used as a composition for preventing or treating Helicobacter pylori infection.

Brief Description of the Drawings

[0022]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Best Mode for Carrying Out the Invention

[0023] Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Instead, the content introduced here is provided to be thorough and complete and to fully convey the idea of the present invention to those skilled in the art.

[0024] <Example 1. Production of Honeysuckle Water Extract (1)> 2000 ml of water was added to 100 g of honeysuckle and hot water extraction was carried out at 90 °C for 3 hours to obtain 1400 ml of honeysuckle extract. After filtering the above extract with a 5 μm filter paper, it was concentrated under reduced pressure at 50 °C for 3 hours. Thereafter, the above concentrate was dried under reduced pressure at 50 °C overnight to obtain 25 g of the honeysuckle water extract of Example 1 of the present invention.

[0025] <Example 2. Production of Honeysuckle Water Extract (2)> 2000 ml of water was added to 100 g of honeysuckle and hot water extraction was carried out at 90 °C for 1 hour to obtain 1400 ml of honeysuckle extract. After filtering the above extract with a 5 μm filter paper, it was concentrated under reduced pressure at 50 °C for 3 hours. Thereafter, the above concentrate was dried under reduced pressure at 50 °C overnight to obtain 25 g of the honeysuckle water extract of Example 2 of the present invention.

[0026] <Example 3. Production of Honeysuckle Water Extract (3)> 2000 ml of water was added to 100 g of honeysuckle and hot water extraction was carried out at 90 °C for 5 hours to obtain 1400 ml of honeysuckle extract. After filtering the above extract with a 5 μm filter paper, it was concentrated under reduced pressure at 50 °C for 3 hours. Thereafter, the above concentrate was dried under reduced pressure at 50 °C overnight to obtain 25 g of the honeysuckle water extract of Example 3 of the present invention.

[0027] <Comparative Example 1. Production of Comparative Target Honeysuckle Ethanol Extract> It was produced in the same manner as in Example 1 above, except that a 70% ethanol aqueous solution was used as the extraction solvent instead of water to produce the honeysuckle ethanol extract of Comparative Example 1.

[0028] <Comparative Example 2. Preparation of the honeysuckle water extract for comparison (1)> It was produced in the same manner as in Example 1 above, except that it was extracted at 60 °C for 3 hours instead of hot water extraction at 90 °C for 3 hours to produce the honeysuckle water extract of Comparative Example 2.

[0029] <Comparative Example 3. Preparation of the honeysuckle water extract for comparison (2)> It was produced in the same manner as in Example 1 above, except that it was extracted at 30 °C for 3 hours instead of hot water extraction at 90 °C for 3 hours to produce the honeysuckle water extract of Comparative Example 3.

[0030] <Comparative Example 4. Preparation of the honeysuckle water extract for comparison (3)> It was produced in the same manner as in Example 1 above, except that it was extracted at 90 °C for 30 minutes instead of hot water extraction at 90 °C for 3 hours to produce the honeysuckle water extract of Comparative Example 4.

[0031] <Experimental Example 1. Confirmation of the content of the index components contained in the honeysuckle extract> To confirm the content of the index components contained in the honeysuckle extract, a standard solution was first prepared. The standard solution was prepared by putting about 5 mg of secoxyloganin standard into a 50 ml flask, adding water, sonicating to completely dissolve it, and then cooling the flask to make the liquid level coincide with the calibration mark. The resulting liquid was used as a high-concentration (0.1 mg / ml) secoxyloganin solution.

[0032] Next, about 200 mg each of the examples and comparative examples were taken and put into a 50 ml flask, water was added, and they were completely dissolved by sonication. After cooling the flask to make the liquid level coincide with the calibration mark, the remaining liquid filtered through a membrane filter with a pore size of 0.22 μm was used as the test solution (4 mg / ml).

[0033] The HPLC running conditions for the above standard product, examples, and comparative examples are as shown in Table 1 below, and the content of secoxyloganin among the index components contained in the examples and comparative examples is as shown in Table 2 below.

[0034]

Table 1

[0035]

Table 2

[0036] From the above Table 2, it was found that the aqueous extract of Flos Lonicerae in the examples of the present invention had a high content of secoxyloganin when the extraction temperature was 90 °C or higher.

[0037] <Experimental Example 2. Confirmation of antibacterial activity against Helicobacter pylori> Helicobacter pylori (ATCC43504) strain was smeared on Brucella agar medium containing 10% horse serum and cultured in an incubator at 37 °C and 10% CO 2 for 3 days. Thereafter, the cultured Helicobacter pylori cells were collected and suspended in sterilized Brucella liquid medium to prepare a cell suspension with an absorbance of 1.0 at 600 nm.

[0038] After dissolving 14 g of Brucella medium in 450 ml of purified water, 6 g of agar was added and suspended, and sterilized at 121 °C for 15 minutes. 50 ml of horse serum was mixed with the medium sterilized at about 40 °C, and 25 ml each was subcultured on a plate with a diameter of 90 mm to solidify the agar medium, and then 0.2 ml of the above Helicobacter pylori cell suspension was smeared.

[0039] The above examples and comparative examples were dissolved in water according to the concentration, sterilized by filtration through a 0.2 μm filter, and then 20 μl each was treated on a sterilized paper disc (Paper disc, 6 mm in diameter) and placed on the plate smeared with the strain. After culturing in an incubator at 37 °C and 10% CO 2 for 72 hours, the diameter of the generated clear zone was measured, and the diameter of the pure ring excluding the perforation diameter is shown in Table 3.

[0040]

Table 3

[0041] As can be seen from Table 3 above, when treating Helicobacter pylori with the honeysuckle water extract of Examples 1 to 3 containing 0.5% by weight or more of the index component secoxyloganin, it was confirmed that a clear zone of 10 mm or more was shown and the antibacterial activity was excellently exhibited.

[0042] In particular, the present inventors confirmed the Helicobacter pylori infection inhibitory effect in mice by oral administration of 100 mg / kg each using the honeysuckle water extract according to the content of secoxyloganin. As a result, different from Comparative Examples 1 to 4, it was confirmed that the Helicobacter pylori infection inhibitory effect was shown only in the honeysuckle water extracts of Examples 1 to 3 containing 0.5% by weight or more of secoxyloganin. Hereinafter, in the animal experiment, the composition of Example 1 was orally administered to mice infected with Helicobacter pylori at 100 mg / kg, 200 mg / kg, and 400 mg / kg, and the Helicobacter pylori infection inhibitory effect depending on the dose was confirmed.

[0043] <Experimental Example 3. Confirmation of Helicobacter pylori Infection Inhibitory Effect> Experimental Example 3-1. Production of Helicobacter pylori-Infected Mice First, after inoculating the Helicobacter pylori strain (H. pylori SS1, Korean Helicobacter Bank) into a trypticase soy agar medium supplemented with 5% sheep blood, it was cultured at 10% CO 2 , 37 °C and micro-aerobic conditions for 2 to 3 days.

[0044] To increase the Helicobacter pylori infection rate, antacids were administered to the mice 2 days before and on the day of Helicobacter pylori infection. All groups orally administered 5% NaHCO 3 (sodium bicarbonate) at 0.2 ml per mouse once a day for a total of 3 days using a mouse zonde.

[0045] Mice before Helicobacter pylori infection were fasted for 12 hours, and all groups except the negative control group (G1) were orally administered 0.2 ml each of Helicobacter pylori culture solution at a bacterial count of 5.0×10 9 / ml CFU (colony-forming unit) three times at two-day intervals using a mouse sonde to infect them.

[0046] After inducing Helicobacter pylori infection, to confirm the maintenance of infection, blood was collected from the facial vein of all mice one week after Helicobacter pylori infection, and plasma was separated. For the measurement of Helicobacter pylori antibodies, only individuals in whom an increase in antibodies was confirmed by infection with the Mouse H.pylori antibody (IgG) ELISA Kit (Cusabio Biotech Co., USA) were selected and used in the test.

[0047] All test groups were suspended in distilled water and orally administered 5 ml per kg of mouse at the same time every day, once a day for 28 days (positive control group 1 was administered once a day for 14 days on the first and third weeks).

[0048]

Table 4

[0049] Experimental Example 3-2. Helicobacter pylori PCR test of gastric mucosa Genomic DNA was collected from aseptically collected gastric mucosa tissue, and a Helicobacter pylori PCR test was performed under the conditions shown in Table 5 below. The target gene used in the experiment is CagA, a toxin-type gene that exists specifically only in Helicobacter pylori and is a gene that does not exist in humans or mice. Therefore, specific bands generated by treating each test group were confirmed from mice infected with Helicobacter pylori, and positive individuals were determined and shown in Figure 2.

[0050]

Table 5

[0051] Figure 2 shows the result of measuring the presence or absence of Helicobacter pylori in gastric mucosal tissue by PCR and calculating the cure rates of each test group based on this. The honeysuckle extract (G5 - G7) of Example 1 of the present invention showed cure rates of 40 - 60% for each concentration compared to the infected group (G2). Therefore, it was found that the honeysuckle extract of the present invention is a composition that reduces the expression of specific genes in gastric mucosa increased by Helicobacter pylori infection.

[0052] Experimental Example 3-3. Comparison of the titers of Helicobacter pylori antibody IgG in blood In the above Experimental Example 3-1, one week after inducing Helicobacter pylori infection, the plasma separated from the facial vein of the mice and the plasma separated after blood sampling from the abdominal vena cava at the end of the experiment were used to measure the antibody titers of Helicobacter pylori in each plasma with a Mouse H.pylori antibody (IgG) ELISA Kit, and the results are shown in Figure 3.

[0053] Looking at Figure 3, as a result of measuring the Helicobacter pylori antibody in the blood at the time of autopsy, it was found that the Helicobacter pylori antibody in Example 1 (G5 - G7) of the present invention decreased in a concentration-dependent manner compared to the infected group (G2).

[0054] Through this, it was found that the aqueous extract of honeysuckle of the present invention containing secoxyloganin as an active ingredient is a composition having an excellent effect of suppressing Helicobacter pylori infection.

[0055] Experimental Example 3-4. Macroscopic lesion observation and histological pathological analysis comparison of gastric tissue The gastric tissue excised on the autopsy day was longitudinally incised and spread from the esophagus along the great curvature towards the duodenum to observe specific lesions of the inner mucosa. After observing the lesions macroscopically, the spread gastric tissue was fixed in 10% neutral formalin, paraffin-embedded using the usual method for pathological tissue examination, sectioned at a thickness of 4 μm, stained with H&E (hematoxylin and eosin), and then subjected to pathological tissue examination. The histological score represented the degree of infiltration of inflammatory cells (neutrophils & mononuclear cells) (indicated by yellow arrows) and the degree of atrophic gastritis with atrophic deformation comprehensively in the general parts of the Corpus and Antrum per individual, and the grade of each tissue was converted into a score and shown in Figure 4 according to the criteria in Table 6.

[0056]

Table 6

[0057] Referring to Figure 4, for the histological pathological results, the findings observed in the gastric tissue were scored for inflammatory cell infiltration and atrophic change according to the criteria of the prior literature [Lee, J.Y. et al., J Cancer Prev., 19(2), 144-151, 2014]. In the infection group (G2), compared with the normal group, inflammation and atrophic deformation increased in the gastric tissue due to Helicobacter pylori infection, while in Example 1 (G5 - G7), inflammation and atrophic deformation in the gastric tissue decreased in a concentration-dependent manner.

[0058] Through this, it was found that the honeysuckle extract of the present invention can reduce the symptoms of gastritis caused by infiltration of inflammatory cells and atrophic deformation during Helicobacter pylori infection.

[0059] Experimental Examples 3 - 5. Rapid urease test (CLO test) When Helicobacter pylori is present in the gastric mucosa, the bacteria secrete urease while growing in the test reagent medium, hydrolyzing the urea present in the test reagent to produce ammonia. As a result, the overall pH of the test reagent increases. Based on the color change (red) of such a pH indicator, a rapid urease test is performed, and the test results are indicated by the cure rate and the CLO score.

[0060] For the rapid urease test, gastric mucosa tissue excised on the day of autopsy was aseptically collected and tested using the CLO (campylobacter‐like organism, Asan Pharm Co., Ltd., Korea) of the test reagent. After culturing at 37°C for 2 hours in an incubator, if the color of the reagent changed from yellow to red, it was judged as positive. The positive rate was obtained by calculating the percentage of the number of individuals judged as positive, and the cure rate for Helicobacter pylori eradication by sample treatment was calculated by the following formula and shown in Fig. 5. [Formula] {(Number of specimens - Number of positive specimens) / Number of specimens} × 100

[0061] Also, after the CLO test, when there was no change in the color of the medium, it was measured as 0 points, when it showed a little red as 1 point, when it showed a light purple as 2 points, and when it showed a dark purple as 3 points. The average and standard deviation of each group were obtained and the difference in inter-group values was compared, and the CLO score is shown in Fig. 6.

[0062] Looking at the cure rate and CLO score results from the rapid urease test in Figs. 5 and 6, the honeysuckle extract (G5 - G7) of Example 1 of the present invention showed a cure rate of 40 - 60% compared to the infected group (G2), and also showed a CLO score reduction effect of 60 - 90%.

[0063] As a result, it was found that the honeysuckle extract of the present invention is a composition that reduces the expression of rapid urease in the gastric mucosa increased by Helicobacter pylori infection.

[0064] Experimental Example 3-6. Cytokine analysis in gastric mucosa tissue To measure the pro-inflammatory cytokines in the gastric mucosa tissue, the aseptically collected gastric tissue was finely crushed in liquid nitrogen and proteins were extracted using cell lysis buffer for analysis. The separated proteins were analyzed for TNF-α (tumor necrosis factor-α) and IL-1β (Interleukin-1β) using an ELISA kit (R&D system, Minneapolis, MN, USA), and the results are shown in Figure 7.

[0065] As shown in Figure 7, the honeysuckle extract of Example 1 of the present invention dose-dependently decreased the expression of TNF-α and IL-1β in the gastric mucosa tissue increased by Helicobacter pylori infection.

[0066] <Formulation Example 1. Production of Tablets> 20 g of the honeysuckle extract of Example 1 of the present invention was mixed with 175.9 g of lactose, 180 g of potato starch, and 32 g of colloidal silica. After adding a 10% gelatin solution to this mixture, it was pulverized and passed through a 14-mesh sieve. This was dried, and 160 g of potato starch, 50 g of talc, and 5 g of magnesium stearate were added thereto, and the resulting mixture was made into tablets.

[0067] <Formulation Example 2. Production of Capsules> 100 mg of the honeysuckle extract of Example 1 of the present invention, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate were mixed, and then the above components were mixed according to the usual method for producing capsules and filled into gelatin capsules to produce capsules.

[0068] <Formulation Example 3. Production of Injections> 1 g of the honeysuckle extract of Example 1 of the present invention, 0.6 g of sodium chloride, and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. This solution was placed in a bottle and sterilized by heating at 20°C for 30 minutes.

[0069] <Formulation Example 4. Production of Health Functional Food> 20 g of the honeysuckle extract of Example 1 of the present invention, an appropriate amount of vitamin mixture, 70 μg of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.5 mg of vitamin B6, 0.2 μg of vitamin B12, 10 mg of vitamin C, 10 μg of biotin, 1.7 mg of nicotinamide, 50 μg of folic acid, 0.5 mg of calcium pantothenate, an appropriate amount of inorganic substance mixture, 1.75 mg of ferrous sulfate, 0.82 mg of zinc oxide, 25.3 mg of magnesium carbonate, 15 mg of potassium phosphate monobasic, 55 mg of calcium phosphate dibasic, 90 mg of potassium citrate, 100 mg of calcium carbonate, 24.8 mg of magnesium chloride were mixed and made into granules, but it can be produced by being deformed into various dosage forms depending on the use. Further, the composition ratio of the above vitamin and mineral mixture may be arbitrarily deformed and implemented, or the above components can be mixed and produced according to the production method of ordinary health functional foods.

[0070] <Formulation Example 5. Production of Health Functional Beverage> 1 g of the honeysuckle extract of Example 1 of the present invention, 0.1 g of citric acid, 100 g of fructooligosaccharide, and 900 g of purified water were mixed and stirred, heated, filtered, sterilized, and refrigerated according to the ordinary beverage production method to produce a beverage.

Claims

1. A pharmaceutical composition for preventing or treating Helicobacter pylori infections, comprising an aqueous extract of Lonicera flos and secoxyloganin as an active ingredient, The secoxyloganin is contained in the aqueous extract of the rhododendron amplexicaule in an amount of 0.5 to 5% by weight. The aqueous extract of rhododendron amplexicaule is obtained by mixing rhododendron amplexicaule with 1,500 to 2,500 parts by weight of water per 100 parts by weight of rhododendron amplexicaule, and extracting the mixture at a temperature of 90° C. for 1 to 5 hours; The pharmaceutical composition has antibacterial activity against Helicobacter pylori and is used for preventing or treating Helicobacter pylori infection.

2. The pharmaceutical composition for preventing or treating Helicobacter pylori infection according to claim 1, wherein the composition is formulated into a pharmaceutical dosage form by adding a pharma-ceutical acceptable carrier, excipient or diluent.

3. A health functional food for preventing or improving Helicobacter pylori infection, comprising secoxyloganin as an active ingredient in the form of a water extract of Lonicera flos, The secoxyloganin is contained in the aqueous extract of the rhododendron amplexicaule in an amount of 0.5 to 5% by weight. The aqueous extract of rhododendron amplexicaule is obtained by mixing rhododendron amplexicaule with 1,500 to 2,500 parts by weight of water per 100 parts by weight of rhododendron amplexicaule, and extracting the mixture at a temperature of 90° C. for 1 to 5 hours; The above-mentioned health functional food has antibacterial activity against Helicobacter pylori and is a health functional food for preventing or ameliorating Helicobacter pylori infection.

4. 4. The health functional food for preventing or improving Helicobacter pylori infection according to claim 3, characterized in that the dosage form of the health functional food is selected from the group consisting of tablets, capsules, pills and liquid preparations.

5. A pharmaceutical composition for preventing or treating Helicobacter pylori infections, comprising secoxyloganin as an active ingredient, The water extract of rhododendron amplexicaule is obtained by mixing rhododendron amplexicaule with 1,500 to 2,500 parts by weight of water per 100 parts by weight of rhododendron amplexicaule, and extracting the mixture at a temperature of 90°C for 1 to 5 hours. A pharmaceutical composition for preventing or treating Helicobacter pylori infection, which has antibacterial activity against Helicobacter pylori.

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