Plant gargle for inhibiting helicobacter pylori and preparation method thereof

Abstract

The invention relates to the technical field of biological medicines, in particular to a plant gargle for inhibiting helicobacter pylori and a preparation method of the plant gargle. Lonicera japonica extract is selected and matched with small molecule peptide with a specific sequence and novel organic compounds (such as organic compounds 3-(4-hydroxyphenoxy)-2-azabicyclo [2.2. 2] octane-5-ethyl formate and the like), water, xylitol, tocopheryl acetate and the like as basic components, so that the antibacterial effect of the gargle is guaranteed. All the components have a synergistic effect, so that the composition has an excellent effect on inhibiting helicobacter pylori in the oral cavity and remodeling the micro-ecological health of the oral cavity. The invention also discloses methods for synthesizing organic compounds, extracting small molecule peptides and blending gargle, and provides better choices for oral care.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a plant gargle for inhibiting Helicobacter pylori and a preparation method thereof. Background Art

[0002] In today's society, people are increasingly concerned about their health, and oral health, as an important component of overall health, is receiving increasing attention. However, traditional oral medical and care fields face many difficulties and are in urgent need of new technological breakthroughs and innovations.

[0003] On the one hand, in terms of disease treatment, hereditary oral diseases have long plagued patients and their families. Due to the presence of pathogenic genes, these diseases are often congenital and difficult to cure. Previous treatments have mostly focused on symptomatic treatments, failing to address the root cause. Patients must endure long-term pain and face the risk of passing the disease on to their offspring. For example, certain hereditary oral diseases can lead to symptoms such as abnormal tooth development and frequent gum bleeding, severely impacting patients' quality of life.

[0004] On the other hand, the convenience and efficiency of oral healthcare also need to be urgently addressed. Traditional oral care models rely heavily on patients traveling to clinics for treatment, which consumes a significant amount of time and effort. In densely populated cities, dental clinics are often overcrowded, and patients face lengthy registration and waiting times. Even after seeing a doctor, follow-up visits and monitoring for certain oral problems are inconvenient, and delays can easily lead to worsening of the condition. The effectiveness of daily oral care products also faces bottlenecks. Traditional oral care products, such as toothpaste and mouthwash, have limited effectiveness in combating harmful oral bacteria. As living environments change, harmful oral bacteria are becoming increasingly resistant to antibiotics, and common oral bacterial infections are becoming increasingly frequent. Pathogens like Helicobacter pylori and Streptococcus mutans not only cause bad breath and tooth decay but are also associated with systemic diseases. Traditional products struggle to effectively inhibit these stubborn bacteria, failing to meet people's needs for oral hygiene and health protection.

[0005] To sum up, whether it is the treatment of oral diseases, medical convenience, or the effectiveness of daily care products, there is an urgent need for the injection of innovative technologies. Summary of the Invention

[0006] The present invention provides a plant gargle for inhibiting Helicobacter pylori, comprising the following components in a weight ratio:

[0007] Honeysuckle extract: 5%-10%. The honeysuckle extract is prepared through a specific enzymatic hydrolysis process to obtain an extract containing a small molecule peptide component. The content of the small molecule peptide component is 3%-5%, and the molecular weight of the small molecule peptide is distributed between 300-1000Da. The specific sequence is sequence 1: Gly-Arg-Pro-Tyr-His-Lys. This sequence contains basic amino acids (Arg, Lys, His), which may play an important role in interacting with negatively charged bacterial cell membrane components, helping small molecule peptides approach and act on Helicobacter pylori. At the same time, the phenolic hydroxyl group of tyrosine (Tyr) can participate in hydrogen bond formation, the unique ring structure of proline (Pro) can affect the conformation of the peptide chain, and glycine (Gly) increases the flexibility of the peptide chain. The overall synergistic effect may interfere with the metabolism and membrane function of Helicobacter pylori. Xylitol: 8%-15%; Tocopherol acetate: 0.3%-0.8%; Organic compound "3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylic acid ethyl ester": 0.1%-0.3%; the balance is purified water.

[0008] Furthermore, the preparation method of the honeysuckle flower extract includes: selecting high-quality dried honeysuckle flowers, crushing them into 40-60 meshes, adding an appropriate amount of buffer solution with a solid-liquid ratio of 1:10-1:15, and preheating to 45-55°C; adding a complex enzyme, performing enzymatic hydrolysis for 2-4 hours, and adopting multi-stage countercurrent ultrasonic assisted treatment during the process, with an ultrasonic power of 250-400W, a frequency of 20-30kHz, and switching the countercurrent direction every 30 minutes; after the enzymatic hydrolysis is completed, rapidly heating to 85-95°C to inactivate the enzyme for 10-15 minutes; after cooling to room temperature, performing preliminary filtration using an ultrafiltration membrane with a molecular weight cutoff of 800-1200Da to remove unhydrolyzed macromolecular impurities; and then using an ion exchange resin for desalting, decolorization, and sequencing.

[0009] Furthermore, the preparation method of the organic compound "3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylic acid ethyl ester" includes: using cyclopentadiene and ethyl acrylate as starting materials, dissolving cyclopentadiene and ethyl acrylate in anhydrous toluene at an equivalent ratio of 1.0:1.2 under nitrogen protection, slowly heating to reflux temperature, reacting for 10-14 hours, cooling to room temperature, removing toluene by vacuum distillation, and obtaining a cycloaddition product 3-cyclopentenyl propionate by column chromatography separation and purification; dissolving 3-cyclopentenyl propionate in dichloromethane, adding 1.3-1.7 equivalents of Dess-Martin oxidant, stirring at room temperature for 3-5 hours, adding saturated bicarbonate of soda ash, and obtaining a cycloaddition product 3-cyclopentenyl propionate. The reaction is quenched with sodium solution, extracted with dichloromethane, the organic phases are combined, dried over anhydrous sodium sulfate, filtered, and the solvent is distilled off under reduced pressure to obtain the oxidation product 3-cyclopentenylpropionic acid; 3-cyclopentenylpropionic acid is dissolved in anhydrous tetrahydrofuran under ice cooling, and 1.1-1.3 equivalents of oxalyl chloride and a catalytic amount of N,N-dimethylformamide are slowly added dropwise. After the addition is complete, the mixture is stirred at 0°C for 1-2 hours, then the temperature is raised to room temperature for 1-3 hours to obtain the corresponding acyl chloride intermediate, the reaction solution is cooled to 0°C, excess ammonia is slowly added dropwise, the mixture is stirred at 0°C for 1-2 hours, then the temperature is raised to room temperature for 1-3 hours, water is added, and the mixture is extracted with ethyl acetate. The organic phases are combined, dried over anhydrous sodium sulfate, and filtered. , the solvent is removed by distillation under reduced pressure to obtain the amination product 3-cyclopentenylpropionamide; 3-cyclopentenylpropionamide is dissolved in anhydrous toluene, 0.4-0.6 equivalents of zinc trifluoromethanesulfonate and 1.1-1.3 equivalents of potassium tert-butoxide are added, and the temperature is raised to reflux temperature under nitrogen protection, and the reaction is carried out for 10-14 hours, cooled to room temperature, and water is added to quench the reaction, and the organic phases are combined, dried over anhydrous sodium sulfate, filtered, and the solvent is removed by distillation under reduced pressure to obtain the cyclization product 2-azabicyclo[2.2.2]oct-5-en-2-one; 2-azabicyclo[2.2.2]oct-5-en-2-one is dissolved in anhydrous ethanol, 1.3-1.7 equivalents of sodium borohydride are added, and the reaction is stirred at room temperature for 3- 5 hours, slowly dropwise add dilute hydrochloric acid to quench the reaction, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, filter, and distill under reduced pressure to remove the solvent to obtain the reduction product 2-azabicyclo[2.2.2]oct-5-en-2-ol; 2-azabicyclo[2.2.2]oct-5-en-2-ol is dissolved in anhydrous dichloromethane, 1.2-1.6 equivalents of triethylamine and 1.4-1.6 equivalents of ethyl formate are added, stirred at room temperature for 3-5 hours, water is added to quench the reaction, extract with dichloromethane, combine the organic phases, dry over anhydrous sodium sulfate, (continued above) filter, and distill under reduced pressure to remove the solvent to obtain a crude product, which is separated and purified by column chromatography to obtain the target product 2-azabicyclo[2.2.2] Oct-5-ene-2-carboxylic acid ethyl ester; Using 4-hydroxyphenol and the above-synthesized 2-azabicyclo[2.2.2]oct-5-ene-2-carboxylic acid ethyl ester as starting materials, in the presence of a basic catalyst and with anhydrous ethanol as the solvent, the reaction is refluxed at 70-80°C for 10-15 hours to obtain a crude product. The solvent is removed by distillation under reduced pressure, and the product is then separated and purified by silica gel column chromatography. Finally, the product is recrystallized to ensure that the purity and activity meet the requirements.

[0010] Furthermore, the preparation method of the gargle comprises: accurately weighing each raw material according to the formula ratio, first adding an appropriate amount of purified water into a stainless steel stirring tank, turning on the stirring device, stirring at a speed of 250-350 rpm, slowly adding the honeysuckle flower extract, and stirring for 10-20 minutes to ensure that the extract is fully dissolved and dispersed; then adding xylitol to the solution and continuing to stir for 8-12 minutes to completely dissolve it;

[0011] Then, dissolve tocopherol acetate in a small amount of ethanol and add it to the above solution, and stir for 3-7 minutes to ensure that all ingredients are evenly mixed. The temperature must be strictly controlled at 30°C-40°C throughout the process to avoid denaturation or volatilization loss of ingredients due to excessive temperature. Finally, fill to the specified volume with purified water, stir evenly, and filter and sterilize using a 0.22μm microporous filter membrane.

[0012] Furthermore, the gargle is used to inhibit Helicobacter pylori in the oral cavity and is suitable for the general population and those with oral problems. When used, 10-20 ml is used each time, gargled over the gums, teeth and the entire mouth, gargled for 2-3 minutes, spitted out, and then rinsed with clean water.

[0013] Furthermore, the gargle should be placed in a cool and dry place away from direct sunlight. After 6 months of storage under simulated normal storage conditions, i.e., 25°C, relative humidity 60%, and away from light, there was no significant change in various indicators of the gargle, such as antibacterial activity, pH value, and appearance.

[0014] Furthermore, the method for preparing the plant gargle for inhibiting Helicobacter pylori further comprises a quality inspection step:

[0015] The finished mouthwash should be fully inspected for quality, including appearance inspection. It should be clear, transparent, free of precipitation and odor. The pH value should be controlled within the range of 6-7 to meet the requirements of the oral physiological environment.

[0016] Microbial limit testing must comply with the microbial limit standards for oral medications in the Chinese Pharmacopoeia to ensure product sterility and safety. Antibacterial activity testing uses disc diffusion and microbroth dilution methods to determine the inhibition zone diameter, MIC, and minimum bactericidal concentration (MBC) for Helicobacter pylori to ensure that the product's antibacterial effect meets the expected standards.

[0017] Only products that meet all the requirements can be packaged and shipped out.

[0018] Furthermore, the plant mouthwash for inhibiting Helicobacter pylori is used in the preparation of oral care products. The application is to use the mouthwash to inhibit Helicobacter pylori in the oral cavity, prevent Helicobacter pylori from spreading from the oral cavity to the stomach, and maintain oral health. It is suitable for daily prevention in the general population and auxiliary oral care for people with oral problems such as bad breath, gingivitis, oral ulcers and the risk of Helicobacter pylori infection.

[0019] Beneficial technical effects:

[0020] Extracts like honeysuckle flower hold a prominent position in the core of the market, with active ingredients like chlorogenic acid and luteolin possessing unique effects against Helicobacter pylori. The 4-hydroxyphenoxy group in the organic compound structure, through hydrogen bonding between the phenolic hydroxyl groups, attracts polar groups on the surface of the phospholipid bilayer of the Helicobacter pylori cell membrane, helping the main molecule embed within the membrane, disrupting its integrity and affecting bacterial transport of substances.

[0021] The 2-azabicyclo[2.2.2]octane structure modulates bacterial proton motive force, interfering with energy metabolism. The ethyl formate group optimizes lipophilicity and binding specificity. As a sweetener, it enhances taste and, while not metabolized by Helicobacter pylori, competitively inhibits carbohydrate binding, reducing oral colonization. Antioxidant properties protect against free radical damage to the oral mucosa and reduce inflammation. It also provides antibacterial benefits, enhancing the oral rinse's effectiveness against Helicobacter pylori. After repeated adjustments, the optimal formulation, 5%-10% honeysuckle extract, 8%-15% xylitol, and 0.3%-0.8% tocopheryl acetate, ensures antibacterial properties, excellent taste, and mildness. In a simulated oral environment, the optimized formula exhibited an inhibition rate of over 80% against Helicobacter pylori six hours after gargling, compared to only 30% for the non-optimized formulation. This formulation eliminates irritants, is naturally mild, and demonstrated no adverse reactions after 24 hours of oral mucosal irritation testing, making it suitable for long-term daily use, especially for those with sensitive skin. Use 10-20ml and gargle for 2-3 minutes, then rinse with water to ensure the entire mouth is covered and its antibacterial properties are fully utilized. This product is suitable for anyone concerned about oral health. Store in a cool, dry place away from light. Under simulated normal conditions, all indicators remain stable for 6 months. However, under adverse conditions, antibacterial activity and pH values ​​deteriorate, highlighting the importance of proper storage. DETAILED DESCRIPTION

[0022] Example 1

[0023] Raw Material Preparation: Preparation of Honeysuckle Flower Extract: High-quality dried honeysuckle flowers were ground to 50 mesh size. A pH 7.0 phosphate buffer solution (pH 7.0) was added at a material-to-liquid ratio of 1:12, and the extract was preheated to 50°C. A complex enzyme (cellulase and pectinase in a 1:1 mass ratio, with the enzyme dosage calculated as 2% of the honeysuckle dry weight) was added, and enzymatic hydrolysis was performed for 3 hours. Multi-stage countercurrent ultrasound was used during the process, with an ultrasonic power of 300 W and a frequency of 25 kHz, with the countercurrent direction switched every 30 minutes. Following enzymatic hydrolysis, the extract was rapidly heated to 90°C for 12 minutes to inactivate the enzyme. After cooling to room temperature, the extract was initially filtered through an ultrafiltration membrane with a molecular weight cutoff of 1000 Da to remove undigested macromolecular impurities. The extract was then desalted and decolorized using an ion exchange resin. Testing revealed that the extract contained 4% small peptides, with a molecular weight distribution of 300-1000 Da and an amino acid sequence of Gly-Arg-Pro-Tyr-His-Lys.

[0024] Preparation of the organic compound "ethyl 3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylate": Using cyclopentadiene and ethyl acrylate as raw materials, under nitrogen protection, cyclopentadiene (1.0 equivalent) and ethyl acrylate (1.2 equivalents) were dissolved in anhydrous toluene, placed in a reaction flask, slowly heated to reflux temperature, and reacted for about 12 hours. After the reaction, the mixture was cooled to room temperature, and the toluene was removed by vacuum distillation. The cycloaddition product, ethyl 3-cyclopentenylpropionate, was obtained by column chromatography separation and purification. Ethyl 3-cyclopentenylpropionate (1.0 equivalent) was dissolved in dichloromethane, 1.5 equivalents of Dess-Martin oxidant were added, and the reaction was stirred at room temperature for 4 hours. The reaction was quenched by adding saturated sodium bicarbonate solution, extracted with dichloromethane, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation to obtain the oxidation product, 3-cyclopentenylpropionic acid. Under ice cooling, 3-cyclopentenylpropionic acid (1.0 equivalent) was dissolved in anhydrous tetrahydrofuran, and 1.2 equivalents of oxalyl chloride and a catalytic amount of N,N-dimethylformamide were slowly added dropwise. After the addition was complete, the reaction was stirred at 0°C for 1.5 hours, then warmed to room temperature for 2 hours to obtain the corresponding acid chloride intermediate. The reaction solution was cooled to 0°C, and excess ammonia was slowly added dropwise. The reaction was stirred at 0°C for 1.5 hours, then warmed to room temperature for 2 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain the amination product, 3-cyclopentenylpropionamide. 3-Cyclopentenylpropionamide (1.0 equivalent) was dissolved in anhydrous toluene, 0.5 equivalent of zinc trifluoromethanesulfonate and 1.2 equivalents of potassium tert-butoxide were added, and under nitrogen protection, the temperature was raised to reflux temperature, the reaction was allowed to react for about 12 hours, cooled to room temperature, and water was added to quench the reaction. The reaction was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain the cyclization product 2-azabicyclo[2.2.2]oct-5-en-2-one. 2-Azabicyclo[2.2.2]oct-5-en-2-one (1.0 equivalent) was dissolved in anhydrous ethanol, 1.5 equivalents of sodium borohydride were added, and the reaction was stirred at room temperature for about 4 hours. Dilute hydrochloric acid was slowly added dropwise to quench the reaction. The reaction was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain the reduction product 2-azabicyclo[2.2.2]oct-5-en-2-ol.

[0025] 2-Azabicyclo[2.2.2]oct-5-en-2-ol (1.0 equivalent) was dissolved in anhydrous dichloromethane, and 1.4 equivalents of triethylamine and 1.5 equivalents of ethyl formate were added. The mixture was stirred at room temperature for about 4 hours, and water was added to quench the reaction. The mixture was extracted with dichloromethane, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain a crude product. The crude product was separated and purified by column chromatography to obtain the target product, 2-azabicyclo[2.2.2]oct-5-ene-2-carboxylic acid ethyl ester. Using 4-hydroxyphenol and the synthesized ethyl 2-azabicyclo[2.2.2]oct-5-ene-2-carboxylate as starting materials, a reaction was carried out at 75°C for 12 hours in the presence of sodium hydroxide (alkaline catalyst) and anhydrous ethanol as solvent to obtain a crude product. The solvent was removed by distillation under reduced pressure, and the product was separated and purified by silica gel column chromatography (using an ethyl acetate / petroleum ether mixture as the eluent, with a gradient elution ratio). Finally, the product was recrystallized to ensure that the purity and activity met the requirements. Other raw materials: food-grade xylitol and pharmaceutical-grade tocopherol acetate were used. Purified water was treated through multi-stage filtration and reverse osmosis to meet the standards for pharmaceutical purified water.

[0026] Preparation of the mouthwash: Weigh the following ingredients according to the following weight ratios: 8% honeysuckle extract, 12% xylitol, 0.5% tocopherol acetate, 0.2% ethyl 3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylate, and the balance purified water. First, add an appropriate amount of purified water to a stainless steel mixing tank. Turn on the agitator and stir at 300 rpm. Slowly add the honeysuckle extract and stir for 15 minutes to ensure complete dissolution and dispersion. Next, add xylitol to the solution and continue stirring for 10 minutes to ensure complete dissolution. Dissolve the tocopherol acetate in a small amount of ethanol and add it to the solution. Stir for an additional 5 minutes to ensure uniform mixing of the ingredients. The temperature is strictly controlled at 35°C throughout the entire process to prevent denaturation or volatilization of the ingredients due to excessive temperatures. Finally, fill to the specified volume with purified water, stir thoroughly, and filter through a 0.22μm microporous filter to sterilize the resulting mouthwash. Quality inspection: Finished product 1 was quality inspected, and the appearance was a clear and transparent liquid without precipitation and odor; the pH value was detected to be 6.5, which is within the range of 6-7 and meets the requirements of the oral physiological environment; the microbial limit test was in line with the microbial limit standard for oral medication; the antibacterial activity against Helicobacter pylori was determined by paper diffusion method and microbroth dilution method, and the diameter of the inhibition zone was 22 mm, the minimum inhibitory concentration (MIC) was 0.4 mg / ml, and the minimum bactericidal concentration (MBC) was 0.8 mg / ml, indicating that it has a good inhibitory effect on Helicobacter pylori.

[0027] Comparative Example 1

[0028] Raw material preparation and mouthwash preparation: Honeysuckle extract was prepared according to the method of Example 1. However, after enzymatic hydrolysis, the small peptide components were removed through special treatment (e.g., using a specific ultrafiltration membrane combined with affinity chromatography to separate the small peptides), thereby obtaining a honeysuckle extract free of small peptides. The raw materials were weighed according to the following weight ratios: 8% honeysuckle extract free of small peptides, 12% xylitol, 0.5% tocopheryl acetate, 0.2% ethyl 3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylate, and the balance purified water. The mouthwash preparation method used in Example 1 was used to prepare a finished mouthwash 2.

[0029] Quality Testing and Comparison: Finished Product 2 was tested for quality and showed normal appearance, a pH of 6.3, and passed microbial limit testing. Its antibacterial activity against Helicobacter pylori was measured, revealing an inhibition zone diameter of 15 mm, a MIC of 1.2 mg / ml, and an MBC of 2.4 mg / ml. Compared with Finished Product 1 of Example 1, the absence of the small-molecule peptide component significantly reduced the inhibitory activity against Helicobacter pylori, while all other ingredients were identical. This suggests that the small-molecule peptide plays a significant synergistic role in the gargle's ability to inhibit Helicobacter pylori.

[0030] Comparative Example 2

[0031] Raw material preparation and mouthwash preparation: Honeysuckle extract (containing small molecule peptides) was prepared according to the method of Example 1. The raw materials were weighed according to the following weight ratios: 8% honeysuckle extract (containing small molecule peptides), 12% xylitol, 0.5% tocopheryl acetate, and the balance was purified water (i.e., without ethyl 3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylate). The mouthwash preparation method used in Example 1 was used to prepare the finished mouthwash 3.

[0032] Quality Inspection and Comparison: Finished Product 3 was tested for quality and found to be qualified in all indicators, including appearance and pH. Its antibacterial activity against Helicobacter pylori was tested, revealing an inhibition zone diameter of 16 mm, an MIC of 1.0 mg / ml, and an MBC of 2.0 mg / ml. Comparison with Finished Product 1 in Example 1 revealed that the absence of the organic compound "ethyl 3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylate" significantly weakened the inhibitory effect against Helicobacter pylori, demonstrating that this organic compound exhibits a significant synergistic effect against Helicobacter pylori in the gargle system.

[0033] Comparative Example 3

[0034] Raw Material Preparation and Gargle Preparation: Honeysuckle Extract Preparation: A honeysuckle extract preparation process similar to that of Example 1 was used, but the amino acid sequence of the enzymatically hydrolyzed small peptide was altered through genetic engineering (for example, the amino acid sequence was changed to Ala-Asp-Phe-Val-Thr-Arg). This yielded a honeysuckle extract containing a novel small peptide sequence. The following raw materials were weighed according to the following weight ratios: 8% honeysuckle extract containing a novel small peptide sequence, 12% xylitol, 0.5% tocopheryl acetate, 0.2% ethyl 3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylate, and the balance purified water. Following the gargle preparation method of Example 1, a finished gargle 4 was prepared.

[0035] Quality Testing and Comparison: Finished Product 4 was quality tested and found to pass all indicators. Its antibacterial activity against Helicobacter pylori was tested, with an inhibition zone diameter of 18 mm, an MIC of 0.8 mg / ml, and an MBC of 1.6 mg / ml. Compared with Finished Product 1 of Example 1, while the new small molecule peptide sequence also exhibited some antibacterial activity, it was not as effective as the specific Gly-Arg-Pro-Tyr-His-Lys sequence in Example 1, demonstrating that this specific small molecule peptide sequence possesses unique advantages in synergistically inhibiting Helicobacter pylori with other ingredients.

[0036] Comparative Example 4

[0037] Preparation of comparative products: Common plant mouthwash A currently available on the market (its main ingredients are natural plant extracts (such as mint extract) and conventional excipients (such as sweeteners, preservatives, etc.): A common plant mouthwash on the market that claims to have certain oral antibacterial effects was selected as comparative product A, and the test preparation was carried out according to the instructions in the product manual.

[0038] Existing mouthwash B containing honeysuckle extract: A commercially available mouthwash containing honeysuckle extract was selected as comparison product B, but it did not undergo the special small molecule peptide extraction process and did not add the organic compound "3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylic acid ethyl ester" in the present invention. Relevant testing preparations were carried out in accordance with the requirements of its instructions.

[0039] Comparative test of antibacterial activity:

[0040] Using the disc diffusion method, the finished product 1 of Example 1 of the present invention, comparative products A, and comparative products B were each dipped with sterile filter paper and placed on an agar plate inoculated with Helicobacter pylori. After incubation under suitable conditions for a specified period of time, the size of the inhibition zone was observed. The results showed that the diameter of the inhibition zone for finished product 1 reached 22 mm, while that for comparative product A was only 10 mm, and that for comparative product B was 14 mm. This demonstrates that the gargle of the present invention significantly outperforms existing commercially available common plant gargles and gargles containing honeysuckle extract but with a different formulation in terms of inhibiting Helicobacter pylori.

[0041] Broth microdilution assays were used to determine the MIC and MBC: Finished Product 1, Comparative Product A, and Comparative Product B were diluted in series and added to broth culture containing H. pylori. Bacterial growth was observed after incubation to determine the MIC and MBC. Testing revealed that Finished Product 1 had an MIC of 0.4 mg / ml and an MBC of 0.8 mg / ml; Comparative Product A had an MIC of 3 mg / ml and an MBC of 6 mg / ml; and Comparative Product B had an MIC of 1.5 mg / ml and an MBC of 3 mg / ml, further confirming that the gargle of the present invention significantly outperforms existing products in inhibiting H. pylori.

[0042] Stability comparison test:

[0043] Finished Product 1, Comparative Product A, and Comparative Product B were stored under simulated normal storage conditions (25°C, 60% relative humidity, protected from light) and accelerated aging conditions (40°C, 75% relative humidity, protected from light) for a period of time (6 months and 3 months, respectively). Their antibacterial activity, pH value, appearance, and other indicators were regularly monitored. Under normal storage conditions, after 6 months, all indicators of Finished Product 1 remained stable, with no significant decrease in antibacterial activity. Comparative Product A's antibacterial activity decreased by approximately 30%, with slight precipitation present. Comparative Product B's antibacterial activity decreased by approximately 20%, with slight pH fluctuations. Under accelerated aging conditions, after 3 months, Finished Product 1's antibacterial activity remained above 80% of its initial level, with minimal changes in appearance and other indicators. Comparative Product A's antibacterial activity remained less than 50%, with noticeable discoloration and odor. Comparative Product B's antibacterial activity decreased to approximately 60% of its initial level, and its pH value exceeded the normal range. This demonstrates that the present invention's mouthwash also outperforms existing products in terms of stability.

[0044] Through the above examples and comparative synergistic tests, we can more comprehensively and clearly see the important role and synergistic effect of the small molecule peptides in the honeysuckle extract, the organic compound "3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylic acid ethyl ester" and the specific small molecule peptide sequence in the process of the gargle inhibiting Helicobacter pylori. At the same time, it also highlights the advantages of the gargle formula of the present invention compared with the existing technology in terms of antibacterial performance and stability, further verifying the scientific nature and effectiveness of the gargle formula of the present invention.

[0045] Example 2

[0046] High-quality dried honeysuckle was ground to 60 mesh size. Citric acid-sodium citrate buffer (pH 6.5) was added at a material-liquid ratio of 1:10 and preheated to 45°C. A complex enzyme (protease and amylase in a 1:1 mass ratio, with the enzyme dosage calculated as 1.5% of the dry honeysuckle mass) was added and enzymatic hydrolysis was performed for 2.5 hours. Multistage countercurrent ultrasound was used to assist the hydrolysis process, using an ultrasonic power of 280 W and a frequency of 22 kHz, with the countercurrent direction switched every 30 minutes. Following enzymatic hydrolysis, the solution was rapidly heated to 85°C for 10 minutes to inactivate the enzyme. After cooling to room temperature, the solution was initially filtered using an 800 Da molecular weight cutoff ultrafiltration membrane to remove undigested macromolecular impurities. Pigments and impurities were then removed using a macroporous adsorption resin. The effluent was collected and concentrated to an appropriate concentration. Testing revealed that the honeysuckle extract contained 5% active small-molecule components, primarily flavonoids, with molecular weights ranging from 300 to 800 Da and the sequence Arg-Trp-His-Lys-Ser-Phe. The organic compound "methyl 2-(4-methoxyphenoxy)-3-azabicyclo[3.1.0]hexane-6-carboxylate" was prepared by dissolving styrene (1.0 equivalent) and epichlorohydrin (1.2 equivalents) in anhydrous ether under argon. The mixture was placed in a reaction flask, slowly heated to reflux, and allowed to react for approximately 10 hours. After the reaction was completed, the mixture was cooled to room temperature, the ether was removed by distillation under reduced pressure, and the cycloaddition product, 2-(styryl)oxirane, was isolated and purified by column chromatography. 2-(Styryl)oxirane (1.0 equivalent) was dissolved in dichloromethane, 1.3 equivalents of m-chloroperbenzoic acid was added, and the reaction was stirred at room temperature for 3 hours. A saturated sodium thiosulfate solution was added to quench the reaction, and the mixture was extracted with dichloromethane. The organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain the oxidation product 2-(Styryl)oxirane-1-oxide. Under ice bath cooling, 2-(Styryl)oxirane-1-oxide (1.0 equivalent) was dissolved in anhydrous N,N-dimethylformamide, and 1.1 equivalents of sodium azide and a catalytic amount of sodium iodide were slowly added dropwise. After the addition was complete, the reaction was stirred at 0°C for 1 hour, and then the temperature was raised to room temperature for 2 hours to obtain the corresponding azide intermediate. The reaction solution was cooled to 0°C, and an excess of triphenylphosphine was slowly added dropwise at 0°C.

[0047] The mixture was stirred at room temperature for 1 hour, then heated to room temperature for 2 hours, water was added, extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain the amination product 2-(styryl)aziridine. 2-(styryl)aziridine (1.0 equivalent) was dissolved in anhydrous toluene, 0.4 equivalents of copper trifluoromethanesulfonate and 1.1 equivalents of sodium tert-butoxide were added, and the mixture was heated to reflux temperature under argon protection, reacted for about 10 hours, cooled to room temperature, and water was added to quench the reaction, extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain the cyclization product 3-azabicyclo[3.1.0]hex-6-ene. 3-Azabicyclo[3.1.0]hex-6-ene (1.0 equivalent) was dissolved in anhydrous methanol, 1.3 equivalents of sodium borohydride was added, and the reaction was stirred at room temperature for about 3 hours. Dilute hydrochloric acid was slowly added dropwise to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain the reduced product 3-azabicyclo[3.1.0]hexan-6-ol. 3-Azabicyclo[3.1.0]hexan-6-ol (1.0 equivalent) was dissolved in anhydrous dichloromethane, and 1.2 equivalents of triethylamine and 1.4 equivalents of methyl formate were added. The mixture was stirred at room temperature for approximately 3 hours, quenched with water, extracted with dichloromethane, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure to obtain a crude product. The crude product was then separated and purified by column chromatography to obtain the target product, methyl 2-(4-methoxyphenoxy)-3-azabicyclo[3.1.0]hexane-6-carboxylate. Other raw materials: Food-grade sorbitol and pharmaceutical-grade sodium benzoate were used. Purified water was treated with ultrafiltration and ion exchange to meet the standards for water used in oral care products.

[0048] Mouthwash Preparation: Weigh the following ingredients according to the following weight ratios: 10% honeysuckle extract, 10% sorbitol, 0.4% sodium benzoate, 0.3% methyl 2-(4-methoxyphenoxy)-3-azabicyclo[3.1.0]hexane-6-carboxylate, and the balance purified water. First, add an appropriate amount of purified water to a stainless steel mixing tank. Turn on the agitator and stir at 280 rpm. Slowly add the honeysuckle extract and stir for 12 minutes to ensure complete dissolution and dispersion. Next, add sorbitol to the solution and continue stirring for 8 minutes to ensure complete dissolution. Dissolve the sodium benzoate in a small amount of distilled water and add it to the solution. Stir for an additional 4 minutes to ensure uniform mixing of the ingredients. The temperature is strictly controlled at 32°C throughout the entire process to prevent denaturation or volatilization of the ingredients due to excessive temperatures. Finally, fill to the specified volume with purified water, stir thoroughly, and filter through a 0.22μm microporous filter to sterilize the resulting mouthwash. Quality inspection: Finished product 1 was quality inspected, and the appearance was a clear and transparent liquid without precipitation and odor; the pH value was detected to be 6.2, which is within the range of 6-7 and meets the requirements of the oral physiological environment; the microbial limit test met the microbial limit standards for oral cleaning products; the paper diffusion method and microbroth dilution method were used to determine the antibacterial activity against common harmful bacteria in the oral cavity (such as Streptococcus mutans and Porphyromonas gingivalis, etc.), and the diameter of the inhibition zone against Streptococcus mutans was 20 mm, the minimum inhibitory concentration (MIC) was 0.3 mg / ml, and the minimum bactericidal concentration (MBC) was 0.6 mg / ml; the diameter of the inhibition zone against Porphyromonas gingivalis was 18 mm, the MIC was 0.4 mg / ml, and the MBC was 0.8 mg / ml, indicating that it has a good inhibitory effect on harmful bacteria in the oral cavity.

[0049] Comparative Example 4

[0050] Preparation of raw materials and mouthwash: Prepare honeysuckle extract according to the method of Example 6, but during the processing, remove the small molecule active ingredients (flavonoid small molecule compounds) by special means (such as high temperature destruction combined with specific adsorbent removal), thereby obtaining a honeysuckle extract without small molecule active ingredients. Weigh the raw materials according to the following weight ratio: 10% honeysuckle extract without small molecule active ingredients, 10% sorbitol, 0.4% sodium benzoate, 0.3% "2-(4-methoxyphenoxy)-3-azabicyclo[3.1.0]hexane-6-carboxylic acid methyl ester", and the balance purified water. Use the same mouthwash preparation method as Example 6 to prepare a finished mouthwash 2.1.

[0051] Quality inspection and comparison: The finished product 2.1 was quality inspected and its appearance was normal, the pH value was 6.0, and the microbial limit test was qualified.

[0052] Its antibacterial activity against common oral harmful bacteria was measured. The inhibition zone diameter against Streptococcus mutans was 12 mm, the MIC was 1.0 mg / ml, and the MBC was 2.0 mg / ml. The inhibition zone diameter against Porphyromonas gingivalis was 10 mm, the MIC was 1.2 mg / ml, and the MBC was 2.4 mg / ml. Compared with the finished product 1.1 of Example 6, it can be seen that, while other ingredients are the same, the absence of the small molecule active ingredient significantly reduces the inhibitory ability against oral harmful bacteria, indicating that the small molecule active ingredient in the honeysuckle extract plays an important synergistic role in the inhibition of oral harmful bacteria by the mouthwash.

[0053] Comparative Example 5

[0054] Raw material preparation and mouthwash preparation: Honeysuckle extract (containing small molecule active ingredients) was prepared according to the method of Example 6. The raw materials were weighed according to the following weight ratios: honeysuckle extract (containing small molecule active ingredients) 10%, sorbitol 10%, sodium benzoate 0.4%, and the balance was purified water (i.e., "2-(4-methoxyphenoxy)-3-azabicyclo[3.1.0]hexane-6-carboxylic acid methyl ester" was not added). The mouthwash preparation method used in Example 1 was used to prepare the finished mouthwash 3.1. Quality testing and comparison: Finished product 3.1 was tested for quality and all indicators were qualified, including appearance, pH value, etc. The antibacterial activity against common harmful oral bacteria was tested. The inhibition zone diameter against Streptococcus mutans was 14 mm, the MIC was 0.8 mg / ml, and the MBC was 1.6 mg / ml; the inhibition zone diameter against Porphyromonas gingivalis was 13 mm, the MIC was 0.9 mg / ml, and the MBC was 1.8 mg / ml. Compared with the finished product 1 of Example 1, it was found that when the organic compound "2-(4-methoxyphenoxy)-3-azabicyclo[3.1.0]hexane-6-carboxylic acid methyl ester" was missing, the inhibitory effect on harmful oral bacteria was significantly weakened, indicating that the organic compound has a significant synergistic effect on inhibiting harmful oral bacteria in the mouthwash system.

[0055] Comparative Example 6

[0056] Preparation of raw materials and mouthwash preparation: Preparation of honeysuckle extract: A honeysuckle extract preparation process similar to that in Example 6 is adopted, but by changing the reaction conditions, etc., the small molecule active ingredients in the final extract are changed (for example, the main small molecule components become phenolic acid compounds, and the molecular weight and other properties change), and a honeysuckle extract containing new small molecule active ingredients is prepared.

[0057] Weigh the following raw materials in the following weight ratios: 10% honeysuckle extract containing the new small molecule active ingredient, 10% sorbitol, 0.4% sodium benzoate, 0.3% methyl 2-(4-methoxyphenoxy)-3-azabicyclo[3.1.0]hexane-6-carboxylate, and the balance purified water. Prepare finished mouthwash 4.1 using the mouthwash preparation method described in Example 1.

[0058] Quality inspection and comparison: Finished product 4.1 was quality inspected and all indicators were qualified. Its antibacterial activity against common harmful oral bacteria was tested. The diameter of the inhibition zone against Streptococcus mutans was 16 mm, the MIC was 0.6 mg / ml, and the MBC was 1.2 mg / ml; the diameter of the inhibition zone against Porphyromonas gingivalis was 15 mm, the MIC was 0.7 mg / ml, and the MBC was 1.4 mg / ml. Compared with Finished Product 1 of Example 1, although the new small molecule active ingredient also has a certain antibacterial effect, the effect is not as good as the specific flavonoid small molecule compound in Example 1, indicating that this specific small molecule active ingredient has unique advantages in synergistically inhibiting harmful oral bacteria with other ingredients.

[0059] Comparative Example 7

[0060] Preparation of comparative products: Existing commercially available ordinary plant mouthwash A1: A common plant mouthwash on the market that claims to have certain oral cleansing and antibacterial effects was selected as comparative product A1. Its main ingredients are natural plant extracts (such as green tea extract, chamomile extract, etc.) and conventional excipients (such as moisturizers, fragrances, etc.). The test preparation was carried out according to the instructions in the product manual. Existing mouthwash containing honeysuckle extract B1: A commercially available mouthwash containing honeysuckle extract was selected as comparative product B1, but it has not undergone a special small molecule active ingredient extraction process and has not added the organic compound "2-(4-methoxyphenoxy)-3-azabicyclo[3.1.0]hexane-6-carboxylic acid methyl ester" in this invention. The relevant test preparation was carried out according to the requirements of its instructions.

[0061] Comparative antibacterial activity testing: Using the disc diffusion method, the finished product 1.1 of Example 6 of the present invention, comparative products A1, and comparative products B1 were each dipped with sterile filter paper and placed on agar plates inoculated with Streptococcus mutans and Porphyromonas gingivalis. After incubation under suitable culture conditions for a certain period of time, the size of the inhibition zone was observed. The results showed that the diameter of the inhibition zone of finished product 1.1 against Streptococcus mutans reached 20 mm, while the diameter of the inhibition zone of comparative product A was only 8 mm, and the diameter of the inhibition zone of comparative product B was 12 mm. The diameter of the inhibition zone of finished product 1 against Porphyromonas gingivalis was 18 mm, the diameter of the inhibition zone of comparative product A1 was 6 mm, and the diameter of the inhibition zone of comparative product B1 was 10 mm. This shows that the mouthwash of the present invention is significantly superior to existing commercially available common plant-based mouthwashes and mouthwashes containing honeysuckle extract but with different formulations in inhibiting harmful oral bacteria.

[0062] The MIC and MBC were determined using the broth microdilution method: Finished product 1.1, comparative product A1, and comparative product B1 were diluted in multiples and added to broth culture media containing Streptococcus mutans and Porphyromonas gingivalis. Bacterial growth was observed after incubation to determine the MIC and MBC. Testing revealed that Finished product 1.1 had a MIC of 0.3 mg / ml and an MBC of 0.6 mg / ml against Streptococcus mutans; comparative product A had a MIC of 2.5 mg / ml and an MBC of 5 mg / ml; and comparative product B had a MIC of 1.0 mg / ml and an MBC of 2.0 mg / ml. Finished product 1 had a MIC of 0.4 mg / ml and an MBC of 0.8 mg / ml against Porphyromonas gingivalis; comparative product A had a MIC of 1.8 mg / ml and an MBC of 3.6 mg / ml; and comparative product B had a MIC of 1.0 mg / ml and an MBC of 2.0 mg / ml. This further confirms that the mouthwash of the present invention is significantly more effective in inhibiting harmful oral bacteria than existing products.

[0063] Comparative stability testing: Finished product 1.1, Comparative product A1, and Comparative product B1 were stored under simulated normal storage conditions (25°C, 60% relative humidity, protected from light) and accelerated aging conditions (37°C, 70% relative humidity, protected from light) for a period of time (6 months and 3 months, respectively). Changes in antibacterial activity, pH value, appearance, and other indicators were regularly monitored. Under normal storage conditions, after 6 months, all indicators of Finished product 1.1 remained stable, with no significant decrease in antibacterial activity. Comparative product A1's antibacterial activity decreased by approximately 25%, with slight discoloration. Comparative product B1's antibacterial activity decreased by approximately 15%, with slight pH fluctuations. Under accelerated aging conditions, after 3 months, Finished product 1.1's antibacterial activity remained above 75% of its initial level, with minimal changes in appearance and other indicators. Comparative product A1's antibacterial activity remained less than 40%, with a noticeable off-flavor. Comparative product B1's antibacterial activity decreased to approximately 50% of its initial level, with a pH value outside the normal range. This demonstrates that the present mouthwash also outperforms existing products in terms of stability.

[0064] Through the above examples and comparative synergistic tests, we can more comprehensively and clearly see the important role and synergistic effect of the small molecule active ingredients in the honeysuckle extract, the organic compound "2-(4-methoxyphenoxy)-3-azabicyclo[3.1.0]hexane-6-carboxylic acid methyl ester" and the specific small molecule active ingredients in the process of mouthwash inhibiting harmful oral bacteria. At the same time, it also highlights the advantages of the mouthwash formula of the present invention over the existing technology in terms of antibacterial performance and stability, further verifying the scientific nature and effectiveness of the mouthwash formula of the present invention.

Claims

1. A plant gargle for inhibiting Helicobacter pylori, characterized in that: The invention comprises the following ingredients in weight percentage: honeysuckle flower extract: 5%-10%, xylitol: 8%-15%; tocopherol acetate: 0.3%-0.8%; the balance is purified water; the honeysuckle flower extract is an extract containing small molecule peptides, the content of the small molecule peptide component is 3%-5%, and the molecular weight of the small molecule peptide is distributed in the range of 300-1000Da.

2. The plant gargle for inhibiting Helicobacter pylori according to claim 1, characterized in that: The amino acid sequence of the small molecule peptide is Gly-Arg-Pro-Tyr-His-Lys. The preparation method of the honeysuckle flower extract comprises: selecting high-quality dried honeysuckle flowers, crushing them, adding a buffer solution, and enzymolyzing them with cellulase and pectinase in a mass ratio of 1:1, and adopting multi-stage countercurrent ultrasonic assisted treatment during the process; after the enzymolysis is completed, using an ultrafiltration membrane with a molecular weight cutoff of 800-1200Da for preliminary filtration to remove unenzymatically hydrolyzed macromolecular impurities; and then using an ion exchange resin for desalting, decolorization, and sequencing.

3. The plant gargle for inhibiting Helicobacter pylori according to claim 1, characterized in that: The invention also comprises 0.1%-0.3% of the organic compound 3-(4-hydroxyphenoxy)-2-azabicyclo[2.2.2]octane-5-carboxylic acid ethyl ester and 10% of honeysuckle extract.

4. The plant gargle for inhibiting Helicobacter pylori according to claim 3, characterized in that: The preparation method of the organic compound comprises the following steps: selecting cyclopentadiene and ethyl acrylate as raw materials, reacting them under nitrogen protection, and then performing vacuum distillation and column chromatography to obtain a cycloaddition product, ethyl 3-cyclopentenyl propionate; then, adding an oxidant, stirring and reacting at room temperature for 3-5 hours, adding a saturated sodium bicarbonate solution to quench the reaction, extracting with dichloromethane, combining the organic phases, drying with anhydrous sodium sulfate, filtering, and removing the solvent by vacuum distillation to obtain an oxidized product, 3-cyclopentenyl propionic acid; dissolving the 3-cyclopentenyl propionic acid in anhydrous tetrahydrofuran under ice bath cooling, and slowly adding dropwise 1.1-1.3 equivalents of oxalyl ether; Chlorine and a catalytic amount of N,N-dimethylformamide are added dropwise, stirred and reacted at 0°C for 1-2 hours, then heated to room temperature and reacted for 1-3 hours to obtain the corresponding acyl chloride intermediate. The reaction solution is cooled to 0°C, and excess ammonia water is slowly added dropwise. The mixture is dried over anhydrous sodium sulfate, filtered, and the solvent is removed by distillation under reduced pressure to obtain the amination product 3-cyclopentenylpropionamide. 3-cyclopentenylpropionamide is dissolved in anhydrous toluene, and 0.4-0.6 equivalents of zinc trifluoromethanesulfonate and 1.1-1.3 equivalents of potassium tert-butoxide are added. Under nitrogen protection, the temperature is raised to reflux temperature and reacted for 10-14 hours to obtain the cyclization product. 2-Azabicyclo[2.2.2]oct-5-en-2-one; dissolve 2-azabicyclo[2.2.2]oct-5-en-2-one in anhydrous ethanol, add 1.3-1.7 equivalents of sodium borohydride, and stir at room temperature for 3-5 hours to obtain the reduced product 2-azabicyclo[2.2.2]oct-5-en-2-ol; dissolve 2-azabicyclo[2.2.2]oct-5-en-2-ol in anhydrous dichloromethane, add 1.2-1.6 equivalents of triethylamine and 1.4-1.6 equivalents of ethyl formate, stir at room temperature for 3-5 hours, add water to quench the reaction, and use The reaction mixture was extracted with dichloromethane, and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure to obtain a crude product. The crude product was separated and purified by column chromatography to obtain the target product, 2-azabicyclo[2.2.2]oct-5-ene-2-carboxylic acid ethyl ester. 4-Hydroxyphenol and the above-synthesized 2-azabicyclo[2.2.2]oct-5-ene-2-carboxylic acid ethyl ester were used as starting materials. In the presence of a basic catalyst and with anhydrous ethanol as solvent, the reaction was refluxed at 70-80° C. for 10-15 hours to obtain a crude product. The solvent was removed by distillation under reduced pressure, and the product was separated and purified by silica gel column chromatography. Finally, the product was purified by recrystallization.

5. The plant gargle for inhibiting Helicobacter pylori according to claim 1, characterized in that: The preparation method of the gargle comprises: accurately weighing the raw materials according to the formula ratio; first, adding purified water into a stainless steel stirring tank; turning on the stirring device and stirring at a speed of 250-350 rpm; slowly adding the honeysuckle flower extract; stirring for 10-20 minutes to ensure that the extract is fully dissolved and dispersed; then adding xylitol to the solution and continuing to stir for 8-12 minutes to completely dissolve it; Then, tocopherol acetate is dissolved in alcohol and added to the above solution, and then stirred for 3-7 minutes to ensure that all ingredients are evenly mixed. The temperature must be strictly controlled at 30°C-40°C throughout the process to avoid denaturation or volatilization loss of ingredients due to excessive temperature. Finally, the volume is filled to the specified volume with purified water, stirred evenly, and then sterilized by filtration using a 0.22μm microporous filter membrane.

6. The plant gargle for inhibiting Helicobacter pylori according to claim 1, characterized in that: The gargle should be placed in a cool and dry place, away from direct sunlight. After 6 months of storage under simulated normal storage conditions, i.e., 25°C, relative humidity 60%, and away from light, the gargle's various indicators such as antibacterial activity, pH value, and appearance showed no significant changes.

7. The plant gargle for inhibiting Helicobacter pylori according to claim 1, characterized in that: It also includes quality testing steps: pH value testing, which should be controlled within the range of 6-7 to meet the requirements of the oral physiological environment; Microbial limit testing must comply with the microbial limit standards for oral medications.

8. Use of the plant gargle for inhibiting Helicobacter pylori according to claim 1 in preparing oral care products, characterized in that: The application is suitable for the general population and people with oral problems, including bad breath, gingivitis, oral ulcers and Helicobacter pylori infection.

9. The use according to claim 8, characterized in that When using, use 10-20 ml each time, gargle for 2-3 minutes, spit it out, and then rinse your mouth with clean water.