Application of cyclocarya paliurus leaves or cyclocarya paliurus leaf extract in preparation of helicobacter pylori bacteriostatic agent

Abstract

The invention relates to the technical field of biological medicines, in particular to application of cyclocarya paliurus leaves or an extract thereof in preparation of a helicobacter pylori bacteriostatic agent. The helicobacter pylori bacteriostatic agent is used for inhibiting the helicobacter pylori with the preservation number of GDMCC 1.1820, the helicobacter pylori with the preservation number of GDMCC 1.1823 and the helicobacter pylori B5-3 with the preservation number of GDMCC 1.1823. The preparation method of the extract comprises the following steps: mixing crushed cyclocarya paliurus leaves and water, heating and extracting to prepare the extract. The invention provides a new scheme for inhibiting helicobacter pylori.

Description

Technical Field

[0001] The present application relates to the field of biomedicine technology, and in particular to an application of Cyclocarya paliurus leaves or an extract thereof in the preparation of a Helicobacter pylori inhibitor. Background Art

[0002] Helicobacter pylori (Hp) is a Gram-negative bacterium that mainly lives in the human stomach and duodenum. The morphological characteristics of Helicobacter pylori include: spiral, S-shaped, arc-shaped, with flagella, and the ability to move, which makes it have strong mobility in the mucus layer of the gastric mucosa. The length is generally 2.5 to 4.0 μm, and the diameter is about 0.5 to 1.0 μm.

[0003] Helicobacter pylori mainly causes disease by destroying the gastric mucosal barrier. Its flagella enable it to penetrate the gastric mucus layer and reach the surface of gastric epithelial cells. Toxins secreted by the bacteria, such as vacuolating toxin (VacA) and cytotoxin-associated protein (CagA), can damage gastric epithelial cells and cause inflammatory reactions. Long-term infection with Helicobacter pylori may lead to chronic inflammation of the gastric mucosa, and then cause diseases such as atrophic gastritis, gastric ulcer, and duodenal ulcer. In addition, it is also closely related to the occurrence and development of gastric cancer and gastric mucosa-associated lymphoid tissue lymphoma.

[0004] At present, conventional drugs for controlling Helicobacter pylori mainly include proton pump inhibitors (omeprazole, lansoprazole, rabeprazole, etc.), bismuth agents (bismuth potassium citrate, etc.) and antibiotics (amoxicillin, clarithromycin, metronidazole, levofloxacin, etc.). It is necessary to provide new Helicobacter pylori inhibitors and new solutions for Helicobacter pylori control. Summary of the invention

[0005] Based on this, one or more embodiments of the present application provide the use of Cyclocarya paliurus leaves or extracts thereof in the preparation of Helicobacter pylori antibacterial agents, including the following technical solutions:

[0006] One or more embodiments of the present application provide a use of Cyclocarya paliurus leaves or an extract thereof in the preparation of a Helicobacter pylori inhibitor.

[0007] In some embodiments of the present application, the Helicobacter pylori inhibitor inhibits Helicobacter pylori with a deposit number of GDMCC1.1820.

[0008] In some embodiments of the present application, the Helicobacter pylori inhibitor inhibits Helicobacter pylori with a deposit number of GDMCC1.1823.

[0009] In some embodiments of the present application, the Helicobacter pylori inhibitor inhibits Helicobacter pylori B5-3.

[0010] In some embodiments of the present application, the method for preparing the extract comprises the following steps: mixing the pulverized Cyclocarya paliurus leaves and water, and heating and extracting to prepare the extract.

[0011] In some embodiments of the present application, the heating extraction satisfies one or more of the conditions shown in items (1) to (3):

[0012] (1) The weight of the water is 5 to 25 times the weight of the crushed Cyclocarya paliurus leaves;

[0013] (2) The temperature of the heating extraction is 70°C to 110°C;

[0014] (3) The number of heating extractions is 1 to 5 times, each time for 1 to 3 hours.

[0015] In some embodiments of the present application, the heating extraction satisfies the following conditions:

[0016] The weight of the water is 5 to 15 times the weight of the crushed Cyclocarya paliurus leaves;

[0017] The temperature of heating extraction is 80℃~95℃;

[0018] The heating extraction is performed once to three times, each time for 1.5 to 2.5 hours.

[0019] In some embodiments of the present application, the heating extraction satisfies the following conditions:

[0020] The weight of the water is 5 to 10 times the weight of the crushed Cyclocarya paliurus leaves;

[0021] The temperature of heating extraction is 80℃~91℃;

[0022] The heating extraction is performed once to three times, each time for 1.8 hours to 2.2 hours.

[0023] In some embodiments of the present application, the Helicobacter pylori inhibitor includes drugs for preventing and treating gastric diseases caused by Helicobacter pylori.

[0024] In some embodiments of the present application, the drug includes the Cyclocarya paliurus leaf or an extract thereof and a pharmaceutically acceptable excipient.

[0025] The details of one or more embodiments of the present application are set forth in the description which follows, and other features, objects, and advantages of the present application will be apparent from the description and its claims. DETAILED DESCRIPTION

[0026] Below in combination, embodiment and example, the application is further described in detail.It should be understood that these embodiments and examples are only used to illustrate the application and are not used to limit the scope of the application. The purpose of providing these embodiments and examples is to make the understanding of the disclosure of the application more thorough and comprehensive.It should also be understood that the application can be implemented in many different forms, is not limited to the embodiment and example described herein, and those skilled in the art can make various changes or modifications without violating the connotation of the application, and the equivalent form obtained also falls within the protection scope of the application.In addition, in the description below, a large number of specific details are given in order to provide a more comprehensive understanding of the application, and it should be understood that the application can be implemented without one or more of these details.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing implementation modes and embodiments and are not intended to limit this application.

[0028] the term

[0029] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0030] The terms "and / or", "or / and", and "and / or" used in this article include any one of two or more related listed items, and also include any and all combinations of related listed items, and the arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions that are all connected by "logical and", and undoubtedly includes technical solutions that are all connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution that is all connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the combination of four of A, B, C, and D (that is, the technical solution that is all connected by "logical AND").

[0031] In the present application, "plurality", "multiple", "multiple times", "multiples", etc., unless otherwise specified, refer to a number greater than 2 or equal to 2. For example, "one or more" means one or greater than or equal to two.

[0032] As used herein, "combination thereof", "any combination thereof", "any combination thereof" etc. include all suitable combinations of any two or more of the listed items.

[0033] Herein, the “suitable” mentioned in “suitable combination”, “suitable method”, “any suitable method”, etc., shall be based on the ability to implement the technical solution of this application, solve the technical problems of this application, and achieve the expected technical effects of this application.

[0034] Herein, “preferred”, “better”, “more preferred” and “suitable” are merely used to describe implementation methods or examples with better effects, and it should be understood that they do not constitute limitations on the scope of protection of this application.

[0035] In the present application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0036] In this application, "optionally", "optional", and "optional" mean optional or dispensable, that is, any one of the two parallel schemes of "yes" or "no". If multiple "options" appear in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" is independent.

[0037] In the present application, the terms "first", "second", "third", "fourth", etc. in "the first aspect", "the second aspect", "the third aspect", "the fourth aspect", etc. are used only for descriptive purposes and cannot be understood as indicating or implying relative importance or quantity, nor can they be understood as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first", "second", "third", "fourth", etc. only serve the purpose of non-exhaustive enumeration and description, and it should be understood that they do not constitute a closed limitation on quantity.

[0038] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0039] In the present application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to the integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In this article, it is equivalent to directly listing each integer, such as t is an integer selected from 1 to 10, indicating that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0040] The temperature parameters in this application, unless otherwise specified, are allowed to be either constant temperature treatment or to vary within a certain temperature range. It should be understood that the constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. Fluctuations within the range of ±5°C, ±4°C, ±3°C, ±2°C, and ±1°C are allowed.

[0041] In the present application, % (w / w) and wt% both represent weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass volume percentage.

[0042] The present application embodiment provides a use of Cyclocarya paliurus leaves or an extract thereof in the preparation of a Helicobacter pylori antibacterial agent.

[0043] The present application does not specifically limit the Helicobacter pylori inhibited by the Helicobacter pylori inhibitor, including but not limited to Helicobacter pylori with a deposit number of GDMCC 1.1820, Helicobacter pylori with a deposit number of GDMCC 1.1823, and Helicobacter pylori B5-3.

[0044] In some examples, the method for preparing the extract includes the following steps: mixing crushed Cyclocarya paliurus leaves and water, heating and extracting, to prepare the extract.

[0045] Optionally, the heating extraction satisfies one or more of the conditions shown in items (1) to (3):

[0046] (1) The weight of the water is 5 to 25 times (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 times) the weight of the crushed Cyclocarya paliurus leaves;

[0047] (2) The temperature of the heating extraction is 70° C. to 110° C. (e.g., 70, 75, 80, 85, 90, 95, 100, 105, 110° C.);

[0048] (3) The number of times of heating extraction is 1 to 5 times (for example, 1, 2, 3, 4, 5 times), and each time is 1 to 3 hours (for example, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, 3 hours).

[0049] It is understood that, if the number of times of heating extraction is multiple, the filter residue of the previous heating extraction is added with water and then heated and extracted again, and so on, and the extracts of multiple heating extractions are mixed. It is understood that after obtaining the extract, the solvent therein can be removed by heating and concentration.

[0050] Further optionally, the heating extraction satisfies the following conditions:

[0051] The weight of the water is 5 to 15 times the weight of the crushed Cyclocarya paliurus leaves;

[0052] The temperature of heating extraction is 80℃~95℃;

[0053] The heating extraction is performed once to three times, each time for 1.5 to 2.5 hours.

[0054] Further optionally, the heating extraction satisfies the following conditions:

[0055] The weight of the water is 5 to 10 times the weight of the crushed Cyclocarya paliurus leaves;

[0056] The temperature of heating extraction is 80℃~91℃;

[0057] The heating extraction is performed once to three times, each time for 1.8 hours to 2.2 hours.

[0058] In some examples, the Helicobacter pylori inhibitor includes a drug for preventing and treating gastric diseases caused by Helicobacter pylori. It is understood that the drug can be used to prevent and treat gastritis, gastric cancer, gastric ulcer, etc. caused by Helicobacter pylori.

[0059] In the present application, the "subject" of the drug is an animal, preferably a mammal, more preferably a human, and the subject includes but is not limited to a patient with a disease, illness and / or symptom. The subject in the present application is preferably a mammal. The term "mammal" mainly refers to warm-blooded vertebrate mammals, including but not limited to: such as cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice (such as rats, mice), pigs, cattle, sheep, horses, humans, etc., preferably primates, more preferably humans.

[0060] In some examples, the drug includes the Cyclocarya paliurus leaf or an extract thereof and a pharmaceutically acceptable excipient.

[0061] In the present application, the administration mode of the drug includes but is not limited to: oral, rectal, parenteral (intravenous, intramuscular or subcutaneous) injection, and topical administration, inhalation. Solid dosage forms for oral administration may include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain a buffer. Solid dosage forms such as tablets, sugar pills, capsules, pills and granules may be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active ingredient in such compositions may be delayed in a certain part of the digestive tract. Examples of embedding components that may be used are polymeric substances and waxes. If necessary, the active ingredient may also be formed into microcapsules with one or more of the above-mentioned excipients.

[0062] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, specifically, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances. In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and spices. For example, the suspension may contain a suspending agent, specifically, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitan esters, microcrystalline cellulose, aluminum methylate and agar or mixtures of these substances.

[0063] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous or non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0064] Dosage forms for topical administration include ointments, powders, patches, sprays and inhalants, which are prepared by mixing the active ingredient with pharmaceutically acceptable excipients and any preservatives, buffers, or propellants that may be required under sterile conditions.

[0065] The embodiments of the present application will be described in detail below in conjunction with examples. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manual or normal conditions in this area, can also be based on the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0066] In the following specific embodiments, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0067] Example 1

[0068] This example demonstrates the inhibitory effect of Cyclocarya paliurus leaf extract on Helicobacter pylori.

[0069] 1. Preparation of Cyclocarya paliurus leaf extract

[0070] Weigh 500g of room temperature dried Cyclocarya paliurus leaves, crush them, add 10 times the weight of pure water, and heat them for extraction. The extraction temperature range is 70-110°C, the number of extractions is 1-5 times, and the extraction time is 1-3 hours each time. Determine the inhibitory effect of each extract on the growth of Helicobacter pylori.

[0071] Referring to the antibacterial test results, the extraction method was optimized and the Cyclocarya paliurus leaf extract was prepared for testing. The preferred extraction method was: extract at 80°C for 2 hours, filter, add 5 times the weight of pure water to the filter residue, and extract again at 80°C for 2 hours. The filtrates after the two extractions were combined, concentrated, and freeze-dried to obtain 42.6g of Cyclocarya paliurus leaf extract powder.

[0072] According to the General Chapter 0401 of the 2020 edition of the Chinese Pharmacopoeia, the polysaccharide content of the Cyclocarya paliurus leaf extract powder prepared in the present application was measured by UV-visible spectrophotometry, with D-glucose, rutin, diosgenin and ursolic acid as controls, respectively, and the content was 10.26wt% for polysaccharide, 1.36wt% for flavonoids, 5.02wt% for saponin, and 0.68t% for triterpenoids. The Cyclocarya paliurus extract was prepared with pure water to form a 128mg / mL Cyclocarya paliurus leaf extract solution, which was subsequently used for antibacterial experiments and stored at 4°C.

[0073] 2. Strains and drugs

[0074] Strains: Helicobacter pylori GDMCC 1.1820 and GDMCC 1.1823 (purchased from Institute of Microbiology, Guangdong Academy of Sciences / Guangdong Microbiological Analysis and Testing Center); Helicobacter pylori B5-3 (isolated, identified and preserved in West China Hospital, Sichuan University; accession number: HX-B5-3-IVR).

[0075] Drugs: the above-mentioned Cyclocarya paliurus leaf extract solution; the negative control loquat leaf and the positive control antibiotics for antibacterial effect are amoxicillin trihydrate and clarithromycin, both of which are commercially available products.

[0076] Preparation of negative control extract solution for antibacterial effect: Weigh 2.5g loquat leaves and place them in a 50mL centrifuge tube, crush them with a glass rod, add 45mL of pure water, and turn them upside down to completely soak the tea leaves. Place the centrifuge tube in a 40℃ water bath for 1h, and turn the centrifuge tube upside down several times every 20min during the water bath. After the water bath, filter and collect the liquid with a 4-layer 300-mesh filter, continue to add pure water to the remaining loquat leaves, extract them again according to the previous method, and collect the extract. Centrifuge the tea extracts collected twice at 4000rpm for 10min, filter the supernatant with a 0.22μm filter membrane, and freeze-dry to obtain loquat leaf extract powder. The above powder is prepared into a 128mg / mL loquat leaf extract solution with pure water and stored at 4℃.

[0077] Preparation of antibiotics: Amoxicillin trihydrate and clarithromycin were prepared into solutions with a concentration of 100 mg / mL using distilled water, and stored at -20°C after aliquoting.

[0078] Culture medium: Every 5 mL of Hp complete liquid culture medium is prepared with 4.25 mL of Skirrow medium, 0.25 mL of 10% glucose solution, and 0.5 mL of calf serum.

[0079] Preparation of bacterial solution: Inoculate Helicobacter pylori strains stored at -80℃ into Hp complete liquid culture medium at 10%

[0080] (v / v)CO2, 5% (v / v)O2, 85% (v / v)N2, 37°C, 220rpm shaking culture for 24h. Determine the OD of the bacterial solution 600 Value, spare.

[0081] 3. Test on the inhibitory effect of Cyclocarya paliurus leaf extract on Helicobacter pylori

[0082] Take the prepared bacterial solution and dilute it to 5×10 5 CFU / mL of bacterial solution, take 5mL and inoculate it into a culture bottle, and then add drugs to the bacterial solution.

[0083] The final concentrations of Cyclocarya paliurus leaf extract in the bacterial solution were 0.25, 0.5, 1, 2, and 4 mg / mL, respectively;

[0084] The final concentrations of the negative control extract in the bacterial solution were 0.25, 0.5, 1, 2, and 4 mg / mL;

[0085] The final concentration of antibiotics in the bacterial solution was 0.5 μg / mL.

[0086] The culture medium was placed in 10% (v / v) CO2, 5% (v / v) O2, 85% (v / v) N2, 37°C, and shaken at 220 rpm for 24 h.

[0087] After the culture is completed, take a sample of the bacterial solution and measure the OD of the sample. 600 Value, to judge the growth of Helicobacter pylori.

[0088] The effect of Cyclocarya paliurus leaf extract on the growth of Helicobacter pylori is shown in Table 1. The effect of negative control extract on the growth of Helicobacter pylori is shown in Table 2. The effect of positive control antibiotics on the growth of Helicobacter pylori is shown in Table 3.

[0089] Table 1. Effects of Cyclocarya paliurus leaf extract on the growth of Helicobacter pylori

[0090]

[0091]

[0092] Table 2. Effect of negative control extracts on the growth of Helicobacter pylori

[0093]

[0094] Table 3. Effects of positive control antibiotics on the growth of Helicobacter pylori

[0095]

[0096] Note: OD 600The lower the value, the smaller the amount of bacteria and the better the antibacterial effect of the drug.

[0097] The experimental results show that the Cyclocarya paliurus leaf extract provided in the present application has a significant inhibitory effect on the growth of Helicobacter pylori. Under the condition of a concentration of 0.5 to 4 mg / mL, it can achieve the inhibitory effect of conventional dosages of classical antibiotics on Helicobacter pylori. However, the negative control extract solution had no inhibitory effect on Helicobacter pylori at a concentration of 4 mg / mL. The Cyclocarya paliurus leaf extract provided in the present application can be prepared into an antibacterial drug or antibacterial agent for Helicobacter pylori, and has excellent application prospects in solving the global health and sanitation problem of Helicobacter pylori infection.

[0098] The technical features of the above-mentioned implementation modes and examples can be combined in any appropriate manner. To make the description concise, not all possible combinations of the technical features in the above-mentioned implementation modes and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0099] The above-described embodiments only express several implementation methods of the present application, which is convenient for understanding the technical solution of the present application in detail, but it cannot be understood as a limitation on the scope of protection of the patent application. It should be pointed out that for ordinary technicians in this field, without departing from the concept of the present application, several deformations and improvements can be made, which all belong to the protection scope of the present application. In addition, it should be understood that after reading the above-mentioned teaching content of the present application, the technicians in this field can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the protection scope of the present application. It should also be understood that the technical solutions obtained by the technicians in this field through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided in the present application are all within the protection scope of the claims attached to the present application. Therefore, the protection scope of the patent of the present application shall be based on the content of the attached claims, and the description can be used to interpret the content of the claims.

Claims

1. Application of Cyclocarya paliurus leaves or extracts thereof in the preparation of Helicobacter pylori inhibitors.

2. The use according to claim 1, characterized in that: The Helicobacter pylori inhibitor inhibits Helicobacter pylori with a deposit number of GDMCC 1.1820.

3. The use according to claim 1, characterized in that: The Helicobacter pylori inhibitor inhibits Helicobacter pylori with a deposit number of GDMCC 1.1823.

4. The use according to claim 1, characterized in that: The Helicobacter pylori inhibitor inhibits Helicobacter pylori B5-3.

5. The use according to any one of claims 1 to 4, characterized in that The preparation method of the extract comprises the following steps: mixing the pulverized Cyclocarya paliurus leaves and water, and heating and extracting to prepare the extract.

6. The use according to claim 5, characterized in that: Heating extraction satisfies one or more of the conditions shown in items (1) to (3): (1) The weight of the water is 5 to 25 times the weight of the crushed Cyclocarya paliurus leaves; (2) The temperature of the heating extraction is 70°C to 110°C; (3) The number of heating extractions is 1 to 5 times, each time for 1 to 3 hours.

7. The use according to claim 6, characterized in that: Heating extraction meets the following conditions: The weight of the water is 5 to 15 times the weight of the crushed Cyclocarya paliurus leaves; The temperature of heating extraction is 80℃~95℃; The heating extraction is performed once to three times, each time for 1.5 to 2.5 hours.

8. The use according to claim 7, characterized in that: Heating extraction meets the following conditions: The weight of the water is 5 to 10 times the weight of the crushed Cyclocarya paliurus leaves; The temperature of heating extraction is 80℃~91℃; The heating extraction is performed once to three times, each time for 1.8 hours to 2.2 hours.

9. The use according to any one of claims 1 to 4 and 6 to 8, characterized in that The Helicobacter pylori inhibitors include drugs for preventing and treating gastric diseases caused by Helicobacter pylori.

10. The use according to claim 9, characterized in that: The medicine comprises the Cyclocarya paliurus leaf or its extract and pharmaceutically acceptable auxiliary materials.

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