KR20170044464A - Compositions for treating, improving or preventing for diseases derived from helicobacter pylori - Google Patents

Abstract

The present invention relates to a composition for preventing, ameliorating or treating Helicobacter pylori-derived diseases, and more particularly to a composition for preventing, ameliorating or treating Helicobacter pylori-derived diseases, which comprises a complex extract of broccoli, cabbage, carrot, burdock, squash, It is derived from Helicobacter pylori which shows antioxidant activity and inhibits the growth inhibition activity of pylori and Helicobacter pylori Urease and is safe to human body by using safe natural materials and can suppress the generation of resistant bacteria by abuse of antibiotics and low toxicity To a composition for preventing, ameliorating or treating diseases.

Description

[0001] The present invention relates to a composition for preventing, ameliorating or treating Helicobacter pylori-derived diseases,

The present invention relates to a composition for preventing, ameliorating or treating Helicobacter pylori-derived diseases, and more particularly, to a composition for preventing, ameliorating or treating Helicobacter pylori-derived diseases. More particularly, the present invention relates to a Helicobacter pylori composition comprising a complex extract of broccoli, cabbage, carrot, burdock, And to a composition for preventing, ameliorating or treating a disease caused by pyrethroids.

Modern people who live in a complicated competitive society are very easy to get gastrointestinal diseases by irregular eating habits and excessive stress. Moreover, because Koreans often eat foods with high irritation, gastrointestinal diseases such as gastric ulcers, gastritis, and stomach cancer occur more frequently than in developed countries such as the US, Europe, and Japan.

Helicobacter pylori has been successfully isolated and cultivated by Dr. Marshall and Warren in Australia in 1983 and its name has been named. In many articles, it has been reported that this bacterium has a high rate of hygiene in gastritis, gastric ulcer and duodenal ulcer patients Is now known to be one of the etiologic factors of gastric ulcer, gastritis, gastric cancer and duodenal ulcer.

The Helicobacter pylori (H. pylori) is a gram-negative bacillus as the optimum growth pH inhabiting the junction between the upper epithelial cells from 7.0 to 7.4 and the temperature is in the growth Miho aerobic condition of 30 ~ 37 ℃. In addition, when the growth condition is not satisfied or when the environment changes, the bacillary form is changed to a cocoid form. Infections caused by Helicobacter pylori are caused by the bacteria that have entered the stomach and duodenal mucosa together with the food. When we look at the infection factors of Helicobacter pylori, we can see that the urease secreted to survive in the stomach juice, , Monocotyledon for maintaining mobility, and outer membrane protein on the cell surface that facilitate adhesion to gastric epithelial cells. The double urease has the property of neutralizing the gastric juice of the strong acid secreted from the stomach by changing the elements around gastric mucosal tissues into ammonia and carbon dioxide and changing the acidic to the alkali around the cells.

To date, the treatment of Helicobacter pylori infection has relied on the use of antibiotics or vaccines and has been used to treat Helicobacter pylori using a variety of natural materials such as garlic (Antimicrob. Agents and Chemother., 1999, 43, 1811-1812) . ≪ / RTI > In recent years, efforts have been made to find an active ingredient capable of inhibiting H. pylori from various natural materials. Ohta et al. Reported various active substances from garlic extract (Antimicrob. Agents and Chemother., 1999, 43, 1811-1812). Mabe et al. Reported that catechin- (Antimicrob. Agents and Chemother., 1999, 43, 1788-1791) in vitro as well as in vivo. In addition, strong activity has also been reported from thyroid (J. Appl. Bacteriol., 1996, 80, 667-672) and various flavonoids (Arzneim.-Forsch./Drug Res., 1995, 45, 697-700).

Korean Patent No. 10-0844577

It is another object of the present invention to provide a composition for preventing, ameliorating or treating Helicobacter pylori-derived diseases, which exhibits an activity of inhibiting the growth of Helicobacter pylori and an activity of inhibiting the growth of Helicobacter pylori.

It is another object of the present invention to provide a composition for preventing, improving or treating Helicobacter pylori-derived diseases which is excellent in antioxidative effect and safe for human body by using natural materials and capable of inhibiting the generation of resistant bacteria by low toxicity and abuse of antibiotics .

To achieve the above object, the present invention provides a food composition for preventing or ameliorating a disease caused by Helicobacter pylori comprising a combined extract of broccoli, cabbage, carrot, burdock, squash, oats, milky oil, milky white, cloves and broiler .

Preferably, the complex extract comprises 20 to 30 parts by weight of broccoli, 10 to 15 parts by weight of cabbage, 1 to 10 parts by weight of carrot, 1 to 10 parts by weight of burdock, 1 to 10 parts by weight of pumpkin, 1 to 5 parts by weight of oats, 0.01 to 1 part by weight of whey, 0.01 to 1 part by weight of whey, 0.01 to 1 part by weight of clove and 0.01 to 1 part by weight of broiler.

The complex extract may be prepared by dissolving the compound of the present invention in water such as broth, cabbage, carrot, burdock, squash, oats, milky oil, 1,3-butylene glycol, and a mixed solvent thereof.

The present invention also provides a pharmaceutical composition for preventing or treating Helicobacter pylori-derived diseases, which comprises a combined extract of broccoli, cabbage, carrot, burdock, squash, amber, oil, milk, clove and broiler.

According to the present invention, it is possible to prevent, ameliorate, or treat Helicobacter pylori-derived diseases by exhibiting the activity of inhibiting the growth of Helicobacter pylori and inhibiting the activity of Helicobacter pylori effect and exhibiting excellent antioxidative effect and being safe for human body using natural materials, It is possible to inhibit the generation of resistant bacteria by the abuse of antibiotics.

FIG. 1 is a graph showing the H. pylor inhibiting area (concentration: 133 mg / L, dose: 150 μg) of 30 kinds of vegetables, fruits and medicines according to one embodiment of the present invention by disc diffusiong method.
FIG. 2 is a graph showing the H. pylor inhibitory area (concentration: 133) by the disc diffusiong method for a combined extract of broccoli, cabbage, carrot, burdock, squash, oak, papaya, milfoil , clove and broiler according to an embodiment of the present invention. Mg / mu l, dose of 150 mu g).
FIG. 3 is a graph showing the results of Urease test on H. pylor infection + compound extract administration group, H. pylor infection group, and normal group according to an embodiment of the present invention.
FIG. 4 is a graph showing the results of gastric mucosal tissue examination for H.pylor infected + complex extract administered group, H.pylor infected group, and normal group according to an embodiment of the present invention.
5 to 7 are graphs showing DPPH levels of a single extract of broccoli, carrot, cabbage, squash, burdock, oats, tallow, wheat bark, clove and broiler chickens and a complex extract obtained by mixing 10 kinds of ingredients according to an embodiment of the present invention. Fig. 3 shows the measurement result of free radical scavenging action.
FIGS. 8 to 10 are graphs showing the effect of the extracts of broccoli, carrot, cabbage, squash, burdock, oats, milky oil, Fig. 7 is a graph showing the results of the erasing ability measurement.
FIG. 11 is a graph showing the results of MTS (Cells) analysis of a single extract of broccoli, carrot, cabbage, squash, burdock, oats, viability) measurement results.

Hereinafter, the present invention will be described in detail.

The present invention verifies Helicobactor's inhibitory power against dozens of fruit and vegetable fruits with inhibition and killing effect of Helicobacter. In consideration of the nutritional aspect among the raw materials excellent in the Helicobacter-suppressing ability, organic raw materials and intimacy of the people to the raw materials, The present invention has been accomplished by confirming the antibacterial activity of Helicobacter against the combined extract.

The present invention provides a food composition for preventing or ameliorating a disease caused by Helicobacter pylori comprising a combined extract of broccoli, cabbage, carrot, burdock, squash, oats, tallow, milky white, cloves and broiler chickens.

The combined extract can be obtained by extracting, isolating and fractionating from the natural using methods known in the art for extracting, isolating and fractionating broccoli, cabbage, carrot, burdock, squash, oats, Can be used. The 'extract' defined in the present invention is obtained by extraction treatment from each raw material using an appropriate solvent and includes, for example, a crude extract, a polar solvent-soluble extract or a non-polar solvent-soluble extract.

The complex extract can be extracted according to various extraction solvents and extraction methods and pharmaceutically acceptable as a suitable solvent for extracting broccoli, cabbage, carrot, burdock, pumpkin, oats, milky oil, milky white pepper, Any organic solvent may be used, and water or an organic solvent may be used, but the present invention is not limited thereto. Examples of the solvent include water, alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propanol, isopropanol, and butanol, acetone, ethyl acetate, butyl acetate , Chloroform, hexane, diethyl ether, 1,3-butylene glycol, and the like, or a mixture of two or more thereof. Particularly, as the solvent, methanol or ethanol (alcohol) is preferably used, and ethanol (alcohol) is particularly preferably used.

The extraction temperature is preferably 50 to 100 ° C. As the extraction method, methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method and compression method can be used and more preferred is the use of hot water extraction method or reflux cooling extraction method Do.

After the extract is obtained using water or an organic solvent as described above, a liquid material can be obtained by freezing, heating and filtering at room temperature by a conventional method known in the art, or a liquid material can be obtained by further evaporating, spray drying or freeze- You may.

In addition, the complex extract of the present invention may be prepared in powder form by an additional process such as vacuum distillation, freeze-drying, or spray drying. The extract can also be obtained as an additional purified fraction using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like .

Therefore, the complex extract used in the present invention is a concept that includes all the extracts, fractions and tablets obtained in each step of extraction, fractionation or purification, their diluted solutions, concentrates or dried products.

Specifically, the combined extract of the present invention can be prepared as follows.

First, broccoli, carrots, cabbage, squash, burdock, and oats are washed with each raw material, and then the water is removed and cut to a predetermined size. Purified water is added to each raw material, and then infiltrated and aged. The infiltrated and aged raw material is heated with steam using a steam extractor to first extract the juice at an internal temperature of 95 to 96 ° C. Subsequently, the juice of the primary extract is refluxed while maintaining the internal temperature of the steam extractor at 94 to 96 캜, followed by further secondary extraction. The secondary juice extracted from the juice is firstly sterilized for 10 to 20 minutes by increasing the internal temperature of the steam extractor to 110 ~ 115 ° C and then subjected to primary filtration. The first filtered juice of the extracted juice can be secondarily filtered and then sterilized in a high-temperature sterilizer at 95 ° C or higher to prepare broccoli, carrot, cabbage, squash, burdock and oat extract.

At this time, it is preferable that the infiltration is aged at 40 ° C for 40 to 60 minutes, preferably about 40 minutes in summer and 60 minutes in winter. Also, it is preferable that the temperature of the extracted juice after the primary filtration step, the temperature of the extracted juice through the secondary filtration step, and the temperature of the extracted juice after the secondary sterilizing step are maintained at 95 ° C or higher. It should be careful not to come into contact with air when each step is carried out in order to prevent other bacteria or yeast from multiplying and changing product quality or decaying.

In addition, the crude oil, milky blood, cloves and broiler chick were thoroughly washed and then extracted in a constant temperature water bath at 50 ° C for 72 hours in 30 to 35% ethanol (alcohol), and then they were completely concentrated in a vacuum rotary condenser to obtain an extract .

The complex extract of the present invention comprises 20 to 30 parts by weight of broccoli, 10 to 15 parts by weight of cabbage, 1 to 10 parts by weight of carrot, 1 to 10 parts by weight of burdock, 1 to 10 parts by weight of pumpkin, 1 to 5 parts by weight of oats, 0.01 1 to 1 part by weight of whey protein, 0.01 to 1 part by weight of whey protein, 0.01 to 1 part by weight of clove and 0.01 to 1 part by weight of broiler chick. When the contents of the respective raw materials are mixed within the above-mentioned range, the synergistic effect of the respective ingredients can exert a greater effect on the inhibitory activity of H. pylori, and the taste, Can be increased.

In addition, the compound extract of the present invention may further contain other ingredients such as broccoli, cabbage, carrots, burdock, squash, amber, milk powder, milfoil, clove and broiler, rhubarb, cinnabar, frankincense, myrrh, Extracts such as Chinese cabbage, Chinese cabbage, Chinese cabbage, Chinese cabbage, potato, radish, waku, green tea, red beet, shiitake, shiitake mushroom, kaisan, kale, turmeric, mulberry leaf can be further mixed.

The complex extract of the present invention exhibits an inhibitory activity against Helicobacter pylori Urease and is also excellent in antioxidative activity and can be used as a food or medicine useful for preventing, treating and improving Helicobacter pylori -derived diseases or complications.

The Helicobacter pylori-derived diseases or complications include, but are not limited to, gastritis, gastric ulcer, gastroduodenal ulcer, peptic ulcer, gastric cancer and the like.

The present invention provides a food composition comprising a combined extract of broccoli, cabbage, carrot, burdock, squash, oats, jute, milky white, clove and broiler. When the complex extract of the present invention is used as a food additive, it can be suitably used according to a conventional method such as adding the complex extract as it is or mixing it with another food or food ingredient.

The amount of the complex extract may be suitably changed according to the intended use (prevention, health or therapeutic treatment), and it is preferable that the complex extract is contained in an amount of 0.01 to 95% by weight based on the total weight of the food composition By weight, more preferably 1 to 80% by weight. If the content is less than 0.01% by weight, the efficiency of taking may be lowered. If the content is more than 95% by weight, formulation may be difficult.

As a specific example, the compound extract of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight, based on the raw material. However, when it is intended for health and hygiene purposes or for the purpose of controlling health, it can be added in an amount below the above range, and there is no problem in terms of safety. Therefore, the active ingredient can be used in an amount exceeding the above range have.

There is no particular limitation on the kind of the food. Examples of the foods to which the compound extract of the present invention can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, , A drink, an alcoholic beverage, a vitamin complex, and the like.

When the complex extract of the present invention is prepared as a beverage, it may contain additional ingredients such as various flavors or natural carbohydrates such as ordinary beverages. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame, and the like. The natural carbohydrate is contained in an amount of 0.01 to 10% by weight, preferably 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

In addition to the above, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , Carbonating agents used in carbonated beverages, and the like. In addition, the compositions of the present invention may include flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of the above additives is not limited to a great extent, but may be in the range of 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

The present invention also provides a pharmaceutical composition comprising a combined extract of broccoli, cabbage, carrot, burdock, squash, oats, tallow, whey, clove and broiler.

The complex extract is preferably contained in an amount of 0.01 to 95% by weight, more preferably 1 to 80% by weight, based on the total weight of the pharmaceutical composition. If the content is less than 0.01% by weight, the efficiency of taking may be lowered. If the content is more than 95% by weight, formulation may be difficult.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral preparations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents which may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The present invention relates to pharmaceutical compositions comprising an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, physician or other clinician, RTI ID = 0.0 > effective < / RTI > amount. It will be apparent to those skilled in the art that the therapeutically effective dose and frequency of administration for the pharmaceutical compositions of the present invention will vary with the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. For a desired effect, the pharmaceutical composition of the present invention may be administered in an amount of 1 to 10,000 mg / kg / day, preferably 1 to 200 mg / kg / day, or may be administered once a day, It may be administered separately.

The pharmaceutical compositions of the present invention can be administered to a subject in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

In addition, the composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy or biological response modifiers for the prevention or treatment of obesity, obesity-related diseases or complications.

Hereinafter, the present invention will be described in more detail with reference to examples. These embodiments are for purposes of illustration only and are not intended to limit the scope of protection of the present invention.

Example 1: Helicobacter-suppressing ability of 30 kinds of vegetables, fruits and medicinal foods was confirmed.

Organic raw materials were purchased from the market, and broccoli, cabbage, carrots, burdock, squash, potato, radish, green tea, red beet, shiitake mushroom, , Rhubarb, Rhodiola, Fermented myrrh, Myrrhin, Injinhwa, Goryeonpyeol, Lilium, Rhododendron, Rhododendron, Chungho,

Each of the prepared raw materials was subjected to hot air drying at 40 ° C, 100% ethyl alcohol corresponding to 3 times (v / w) of the mass of the dry raw material (1 kg) was added and extracted at 50 ° C for 48 hours. The mixture was filtered and concentrated under reduced pressure Followed by lyophilization and used as an experimental sample. 2 g of each test sample was dissolved in 15 mL of 60% ethanol to give a concentration of 133 mg / mL. In the control group, the commercial injections of metronidazole (MTZ, 5 mg / ml, CJ), amoxicillin sodium (AMX, 25 mg / ml, Ildong Pharm) and Isepamicin sulfate (ICP, 100 mg / .

Each of the prepared test samples was applied to a paper disk, and the H. pylori inhibitory activity was measured as follows. A strain of H. pylori Sydney strain (SS1-Passed-4 ') purchased from Gyeongsang National University Helicobacter strain bank was streaked with 1.0 × 10 ⁶ CFU on a Brucella agar plate supplemented with 10% horse serum, (Diameter 0.8 mm) three times by 50 μL each, and then dried and placed on a disc. After culturing in a thermostat maintained at 10% CO ₂ and 100% humidity for 48 hours, the clear zone area and the paper disc area were measured using the ImagJ program, and the area excluding the paper disc area was compared to calculate the area And H. pylori was more effective than H. pylori .

The results are shown in Fig. 1 and Table 1 below.

Example 2. Preparation of complex extract

From the results of the above example 1, it was confirmed that the synergistic effect of H. pylor inhibition among the raw materials showing excellent H. pylor inhibiting ability, nutritional aspect, organic raw materials and intimacy of the public were considered, and broccoli, cabbage, carrot, burdock, , Olive oil, ground oil, milky white pepper, cloves and broiler chickens were selected.

Broccoli, carrots, cabbage, squash, burdock, and oats are washed with water and then cut into a uniform size. After adding 2 to 4 times of purified water to each raw material, 40 ~ 60 Min for aging. The infiltrated and aged raw material was put into a steam extractor and heated with steam to first extract the juice at an internal temperature of 95 to 96 ° C. Then, the above-mentioned primary juice was further refluxed while maintaining the internal temperature of the steam extractor at 94 to 96 ° C and further subjected to secondary extraction. The secondary juice extracted from the juice was firstly sterilized for 10 to 20 minutes by increasing the internal temperature of the steam extractor to 110 ~ 115 ° C and then subjected to primary filtration. The primary filtered juice was secondarily filtered and then sterilized in a high-temperature sterilizer at 95 ° C or higher to prepare broccoli, carrot, cabbage, squash, burdock and oat extract.

Each raw material was thoroughly cleaned, and then it was extracted in a constant temperature water bath at 50 ° C for 72 hours in 30 ~ 35% ethanol (alcohol), and it was completely concentrated in a vacuum rotary condenser to produce papermaking, milky white, Respectively.

The combined extracts were prepared by mixing the broccoli, carrot, cabbage, squash, burdock, oats, tallow, milky white, cloves and broiler chick extracts as described in Table 2 below.

The units in Table 2 are parts by weight.

Example 3 In Vitro Helicobacter Antibacterial Activity Test

The in vitro helicobacter antimicrobial activity test was carried out using the compound extract prepared in Example 2 above. The H. pylori strain (SS1-Passed-4 ') strain used in the experiment was inoculated into Brucella Broth (BD, USA) agar medium containing 10% horse serum and purchased from Glycotechnology, After incubation at 37 ° C in a humidified incubator at ₂ and 100% humidity, H. pylori cultured for 24 h for each experiment was used as a viable cell count of 1.0 × 10 ⁶ colony-forming unit (CFU) And examined for contamination.

After the compound extract prepared in Example 2 was applied to a paper disk, the H. pylori inhibitory activity was measured as follows. H. pylori strains were streaked on a Brucella agar plate supplemented with 10% horse serum at a concentration of 1.0 × 10 ⁶ CFU, and the cells were absorbed three times in 50 μL of sterilized paper disks (diameter 0.8 mm) Lt; / RTI > After culturing in a thermostat maintained at 10% CO ₂ and 100% humidity for 48 hours, the clear zone area and the paper disc area were measured using the ImagJ program, and the area excluding the paper disc area was compared to calculate the area And H. pylori was more effective than H. pylori . (AMX, 25 mg / ㎖, Ildong Pharmaceuticals) and Isepamicin sulfate (ICP, 100 mg / ㎖, Yuhan) were used as a control group without dilution or concentration control Respectively. The results are shown in Fig. 2 and Table 3 below.

Raw material AREA (㎟) MTZ 50.329 AMX 16.279 Complex extract 140.55

As shown in Fig. 2 and Table 3, according to the present invention, the combined extract of Example 2, which includes broccoli, carrot, cabbage, squash, burdock, oats, milky oil, It was confirmed that the synergistic effect shows a marked inhibition of H. pylori against commercially available antibiotic components.

Example 3 In vivo Helicobacter Antibacterial Efficacy Test (Urease test)

The in vivo Helicobacter antimicrobial activity test was carried out using the compound extract prepared in Example 2 above. Seven-week-old female C57BL / 6N, maintained in a specific-pathogenfree (SPF) state, was supplied from Orient Bio and used for experiments for one week in the experimental animal breeding room. Breeding in an environment of temperature 22 ± 1 ℃, humidity 50 ± 5%, noises 60phone or less, illumination time 08: 00 ~ 20: 00 (12 hours a day), illumination 150 ~ 300 Lux and 10-12 times per hour during rearing . The feed was sterilized with 2.0 Mrad of feed for experimental animal feed (Samtaco, Korea) and fed free. The filtered water was filtered and filtered with ultraviolet sterilized water.

For the mouse infection, H. pylori strain (SS1-Passed-4 ') strain was cultured at 37 ° C for 24 hours, and the final concentration was adjusted to an absorbance of 10 at a wavelength of 600 nm. 10 ⁹ cfu / mouse) were infused into the stomach using an oral sonde needle. All mice were inoculated for 7 days through a purifying period and used repeatedly for 3 days in the same volume for 3 times. Mice were fasted for at least 3 hours before infection.

Five mice were divided into two groups: H. pylori infection + compound extract (group 1, group 2), H. pylori infection (group 3), and normal group (group 4). Groups 1 and 2 of the mice administered with the compound extract were injected into the stomach with an oral sonde needle to a mouse which had been inoculated with 0.5 ml of the compound extract prepared in Example 2 for 3 hours or more, On a regular basis for 4 weeks.

After 4 weeks, mice were sacrificed through cervical dislocation and aseptically stomach was removed and used for urine test. The fore-stomach portion was removed from the antrum and the corpus of the stomach, and then incised vertically along the lesser curvature from the knee to the knee. The stomach contents of the developed gastric mucosal surface were carefully removed and then symmetrically cut along the great curvature to balance the stomach in two parts. The area of the duodenum adjacent to the duodenum was homogenized to a size of about 0.5 × 0.5 cm or less. Urease test kit (Asan Pharmaceutical Co., Ltd.) was used to determine whether the urease was positive or not. Respectively.

In Table 4, P indicates positive and N indicates negative.

Category (n) One 2 3 4 5 1 group (5) H. pylori + sample P P N N N 2 groups (5) H. pylori + sample N P N N N 3 groups (5) H. pylori P P P P P 4 groups (5) Normal N N N N N

As shown in Fig. 3 and Table 4, in a mouse group administered with Helicobacter infections and combined extracts, 3 out of 10 mice were positive in a Urease test KIT (Asan Pharmacopoeia) by a total of 10 gastric tissue extracts , And 7 were negative. Thus, anti - helicobacter activity was recognized by administration of the combined extract. Urease test was positive in all three groups of Helicobacter infections, and all of them were negative in all four groups.

Example 4. Antibacterial efficacy test of in vivo helicobacter (gastric mucosal tissue examination)

Using the mouse as in Example 3, the gastric mucosal portions of the tissues were fixed with a needle without overlapping and observed with a dissecting microscope (Nikon) to evaluate the helicobacter inhibiting ability by observing the presence or absence of gastric ulcer lesions in the gastric mucosa.

As a result, as shown in Fig. 4, inflammation such as gastric mucosal hemorrhage and hyperemia were observed in 3 groups in which only Helicobacter Infection was performed, but in the groups 1 and 2 administered with the complex extract of the present invention, The results of this study are as follows.

Example 5. Antioxidative effect of the combined extract (DPPH free radical scavenging method)

The combined extracts of Example 2, in which a sole extract of broccoli, carrot, cabbage, squash, burdock, oats, milfoil, milky white, cloves and broiler, and the above 10 components were mixed in a 96 well plate was diluted with EtOH to 1, 10, 100 (DPPH) (EtOH) was added to the solution prepared at a concentration of 500 μg / ml, 0.2 mM of 1,1-diphenyl-2-picrylhydrazyl After shaking for 10 seconds, it was left at room temperature for 30 minutes and absorbance was measured at 520 nm using a microplate reader. L-ascorbic acid was used as the reference drug. The antioxidant effect is shown in a graph in comparison with the absorbance of the control group to which no sample is added. The DPPH free radical scavenging activity of each sample was measured three times.

The results are shown in Figs. 5 to 7 and Table 5 below.

500 100 10 One Vit.C 84.3349 84.3349 6.15281 6.27954 BHA 81.6835 80.9845 46.5855 11.6971 Broiler 78.5861 76.2314 22.643 9.78173 Milky blood 76.5907 78.348 32.9573 10.0888 Oil 77.5019 80.5934 50.1211 12.5303 cloves 81.3953 74.6978 36.269 12.8329 carrot 63.1206 35.8323 21.2903 5.94315 broccoli 44.5993 26.109 19.8198 4.90323 cabbage 56.7036 28.0769 19.0968 5.81395 Sweet pumpkin 27.7066 8.20707 5.27671 4.77419 burdock 81.4141 77.0115 11.9675 7.37982 Oh 50.5003 27.4074 7.0898 8.44024 Total 81.0721 57.6534 9.80392 6.29655

As shown in Table 5 and FIGS. 5 to 7, the results of DPPH free radical scavenging activity showed Vit. C 84.3349, BHA was 81.6835, and the compound extract of Example 2 showed excellent antioxidative activity as 81.0721 there was. Compared to the case of using broccoli, carrot, cabbage, squash, burdock, oats, milky oil, milky white, clove, clove and broiler extract alone, 10 kinds of ingredients are mixed to show DPPH free radical scavenging effect And it was found.

Example 6. Antioxidative Effect of Compound Extract (Superoxide scavenging ability)

The superoxide scavenging ability of the extract of Example 2 in which the extracts of broccoli, carrot, cabbage, squash, burdock, oats, milky oil, (NBT) were used to measure photochemical behavior. The reaction mixtures consisted of 2.6 μM riboflavin, 13 mM methionine, 75 μM NBT, 0.1 mM EDTA, PBS (pH 7.4) and samples at concentrations of 1, 10, 100 and 500 μg / ml. The mixture was placed in a light box and allowed to react for 15 minutes at random intervals every 5 minutes. The temperature inside the light box was 20 ± 1 ° C and the brightness of the light was maintained at 5500 lux. NBT is reduced from light to blue formazan, which is measured at 560 nm. Suppression of formation of blue formazan is superoxide scavenging ability. The results are shown in Figures 8-10 and Table 6 below.

500 100 10 One Vit.C 88.6364 89.5928 8.76404 -2.0316 BHA 84.7534 81.8594 32.1918 -3.8636 Broiler 79.2715 69.7202 22.2542 -1.709 Milky blood 77.5155 85.0489 34.0634 -3.2884 Oil 77.3297 85.8042 36.9515 -2.6057 cloves 76.3571 80.3091 26.2854 -1.6715 carrot 48.8335 40.9396 5.29355 -0.081 broccoli 46.8691 34.598 4.87106 -3.431 cabbage 65.4364 62.7843 17.7482 -2.0781 Sweet pumpkin 38.1961 23.0666 16.0083 3.362 burdock 88.7356 84.0547 30.5239 -6.3636 Oh 80.1354 40.3189 -2.9545 -3.8855 Total 86.2222 85.3273 29.2517 -2.0833

As shown in Table 6 and FIG. 8 to FIG. 10, the superoxide inhibition activity of Vit.C 88.6364 and BHA was 84.7534, and that of the compound extract of Example 2 was 86.2222, indicating excellent antioxidative activity. In contrast to the use of broccoli, carrots, cabbage, squash, burdock, oats, myrtle, milky, clove, clove and broiler extracts alone, 10 compounds were found to be superior to Superoxide inhibitory activity I could.

Example 7. Measurement of Human Stability of Compound Extract

The cell suspension was counted using a haemacytometer, and then 180 μl of the cell suspension was added to each well of a 96-well plate. At this time, blank was prepared with 180ul of medium and 20ul of PBS, and control was performed with 180ul of cell suspension and 20ul of PBS.

The extracts of broccoli, carrot, cabbage, squash, burdock, oats, milfoil, milfoil, clove and broiler and the combined extract of Example 2 were dissolved in PBS, ) Were added to each well. After incubation, samples were removed and 100 μl of MTT solution was added to each well. After incubation for 4 hours, the MTT dilution was carefully removed. 100 μl of DMSO was added to each well, and the plate was shaken with a plate shaker for 15 to 20 minutes. Then, the absorbance was measured using an ELISA reader at a wave length of 540 nm. This absorbance represents the amount of MTT reduced by cells and is proportional to the number of viable cells in each well.

As a result, as shown in FIG. 11, the cell viability test for the safety test showed that a single extract of broccoli, carrot, cabbage, squash, burdock, oats, milky oil, And the compound extract of Example 2 showed a control value of over 100 at a concentration of 0.25 占 퐂 / ml, confirming the safety of the human body.

Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.

Claims (7)

A food composition for preventing or ameliorating a Helicobacter pylori-derived disease comprising a combination extract of broccoli, cabbage, carrot, burdock, squash, oats, milky oil, milky white, clove, The method according to claim 1,
The complex extract contains 20 to 30 parts by weight of broccoli, 10 to 15 parts by weight of cabbage, 1 to 10 parts by weight of carrot, 1 to 10 parts by weight of burdock, 1 to 10 parts by weight of pumpkin, 1 to 5 parts by weight of oats, 0.01 to 1 part by weight of clove, 0.01 to 1 part by weight of broiler, 0.01 to 1 part by weight of whey protein, 0.01 to 1 part by weight of clove and 0.01 to 1 part by weight of broiler chick. The method according to claim 1,
The complex extract may be prepared by dissolving the compound of the present invention in water such as broth, cabbage, carrot, burdock, squash, oats, milky oil, 1,3-butylene glycol, and a mixed solvent thereof. The composition for preventing or improving Helicobacter pylori-derived diseases according to claim 1, The method according to claim 1,
The food composition may be selected from the group consisting of a rhubarb, a cinnabar, a frankincense, a myrrh, an angina, a lavender, a lobule, a rhododendron, a rhubarb, a rhubarb, Wherein the composition further comprises at least one selected from the group consisting of kale, turmeric, and mulberry leaves. The method according to claim 1,
Wherein said Helicobacter pylori-derived disease is gastritis, gastric ulcer, gastroduodenal ulcer, peptic ulcer or gastric cancer. The method according to claim 1,
The composition for preventing or improving Helicobacter pylori-derived diseases is characterized in that the complex composition exhibits a Helicobacter pylori oil-restraining activity. A pharmaceutical composition for preventing or treating Helicobacter pylori-derived diseases, which comprises a combination extract of broccoli, cabbage, carrot, burdock, squash, oats, milfoil, milky white, clove, and broiler.

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