Anti-helicobacter pylori pharmaceutical composition containing lactobacillus reuteri as well as preparation method and application of anti-helicobacter pylori pharmaceutical composition

Abstract

The invention discloses an anti-helicobacter pylori pharmaceutical composition containing lactobacillus reuteri as well as a preparation method and application of the anti-helicobacter pylori pharmaceutical composition, and relates to the technical field of medicines. The anti-helicobacter pylori pharmaceutical composition comprises a traditional Chinese medicine composition and a lactobacillus reuteri liquid, the traditional Chinese medicine composition comprises the following components: uncinata spikemoss herb, dianthus chinensis root, airpotato yam rhizome, rabdosia rubescens and rabdosia amethystoides. The mass volume ratio of the traditional Chinese medicine composition to the lactobacillus reuteri liquid is (30-50) g: (7-15) mL. The traditional Chinese medicine composition and the lactobacillus reuteri liquid are combined for use, helicobacter pylori in the oral cavity and digestive tract of the human body is efficiently removed, and other side effects are not generated.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an anti-Helicobacter pylori pharmaceutical composition containing Lactobacillus reuteri, a preparation method and application thereof. Background Art

[0002] Helicobacter pylori (Hp) is a Gram-negative, microaerophilic bacillus that colonizes the human gastric mucosa. Studies have shown that H. pylori infection affects approximately 40% of the global population, primarily through oral-oral and fecal-oral transmission, water sources, and within households. H. pylori infection is a major risk factor for chronic gastritis, peptic ulcers, gastric mucosa-associated lymphoid tissue lymphoma, and gastric cancer. Data show that my country is one of the countries with the highest incidence of H. pylori infection and gastric cancer, with a concentrated H. pylori infection rate of 49.6% and a gastric cancer incidence rate of 28.68 per 100,000 people. The H. pylori infection rate increases with age, ranging from 28.0% in children and adolescents to 46.1% in adults. This is accompanied by an increased risk of gastric cancer, which can lead to adverse outcomes for patients. Therefore, early screening for H. pylori infection and eradication treatment are essential.

[0003] However, the resistance to commonly used antibiotics currently used for eradication treatment has generally increased from 2013 to 2023. With the increase in the number of medications used, the course of treatment has been prolonged and related adverse reactions have also increased, with diarrhea, abdominal distension and other digestive tract adverse reactions being the most common. Due to the increase in H. pylori resistance and adverse drug reactions, the eradication rate of H. pylori has declined year by year. The main reason for this is related to the increase in H. pylori resistance to antimicrobial drugs caused by factors such as strain variation, secondary resistance, cross-resistance, and cross-infection between different strains. Therefore, providing a safe, side-effect-free, and highly effective anti-Helicobacter pylori pharmaceutical composition has become an urgent problem to be solved. Summary of the Invention

[0004] In view of the above-mentioned defects of the prior art, the present invention proposes an anti-Helicobacter pylori pharmaceutical composition containing Lactobacillus reuteri, a preparation method and application thereof, so as to solve the problems raised by the above-mentioned background technology.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] A Lactobacillus reuteri-containing anti-Helicobacter pylori pharmaceutical composition comprises a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 15 to 30 parts of Selaginella uncinata (Pteridophyta, Lycopods, Selaginel laceae, Selaginella), 12 to 25 parts of Dianthus root (the root or whole herb of Dianthus chinensis L., a plant of the Caryophyllaceae family), 10 to 18 parts of Dioscorea bulbifera (Dioscorea bulbifera, a plant of the Dioscoreaceae family), and 10 to 18 parts of Dioscorea bulbifera (Dioscorea bulbifera, a plant of the Dioscoreaceae family). L.), 7-14 parts of Rubescens rubescens (the dried aerial parts of Rabdosia rubescens (Hemsl.) Hara, a plant of the Lamiaceae family), and 5-10 parts of Isodon macrocalyx (the dried tubers or aerial parts of Rabdosia amethystoides (Benth.) H. Hara or Isodon macrocalyx (Dunn) Kudo, a plant of the Lamiaceae family).

[0007] Preferably, the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri liquid is (30-50) g: (7-15) mL.

[0008] Preferably, the number of viable bacteria in the Lactobacillus reuteri liquid is 2.0×10 8 CFU / mL~2.5×10 10 CFU / mL.

[0009] Preferably, the preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0010] (1) Wash and dry Herba Coptidis, Dianthus root, Dioscorea bulbifera, Rabdosia rubescens, and Herba Citri Reticulatae, crush them to obtain a mixture, and set aside;

[0011] (2) soaking the mixture obtained in step (1) in an ethanol solution with a volume fraction of 70% to 80% for 50 hours to 80 hours to obtain a mixture;

[0012] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, ultrasonically extracting 3 to 5 times, and filtering to obtain a Chinese medicine composition;

[0013] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 30° C. to 35° C. for a fermentation time of 24 h to 48 h to obtain a fermentation product;

[0014] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0015] Preferably, the mass volume ratio of the mixture to the ethanol solution in step (2) is 1 g: (8-16) mL.

[0016] Preferably, the temperature of ultrasonic extraction in step (3) is 30° C. to 45° C., the power is 200W to 300W, and the time is 15 min to 30 min.

[0017] Preferably, the anti-Helicobacter pylori pharmaceutical composition further comprises pharmaceutically acceptable excipients and is prepared into any dosage form including powder, decoction, pill, capsule, tablet, granule, and oral liquid.

[0018] Preferably, the anti-Helicobacter pylori pharmaceutical composition of the present application can be used in the preparation of anti-Helicobacter pylori drugs.

[0019] Preferably, the anti-Helicobacter pylori pharmaceutical composition can be used in combination with an antibiotic preparation. After Helicobacter pylori patients are treated with antibiotic preparations for one week, they are then treated with the anti-Helicobacter pylori pharmaceutical composition, or Helicobacter pylori patients are treated with antibiotic preparations and the anti-Helicobacter pylori pharmaceutical composition at the same time.

[0020] Preferably, the antibiotic preparation comprises a proton pump inhibitor PPI, the antibiotics are amoxicillin and clarithromycin, and the bismuth preparation is potassium bismuth citrate.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] (1) The present invention found that the Chinese medicine composition of Herba Coptidis, Root of Dianthus, Rhizoma Dioscoreae Bulbiferae, Rhizoma Rubescens and Rhizoma Cibotii has a combined antibacterial effect and can effectively eliminate Helicobacter pylori in the oral cavity and digestive tract. The anti-Helicobacter pylori pharmaceutical composition provides a new direction for the subsequent development of drugs for the prevention and treatment of Helicobacter pylori.

[0023] (2) This application not only discovers for the first time that Herba Coptidis, Root of Dianthus, Dioscorea Bulbifera, Rabdosia Rubescens, and Herba Citri Reticulatae have anti-Helicobacter pylori effects, which has never been proposed in the prior art, but also broadens the scope of application of these five Chinese herbs, greatly facilitating the development of these five Chinese herbs. These five Chinese herbs are all easy to harvest and inexpensive, providing new ideas for the subsequent development of cost-effective anti-Helicobacter pylori drugs. DETAILED DESCRIPTION

[0024] The technical solution of the present invention will be further described in detail below in conjunction with specific implementation methods.

[0025] Example 1

[0026] A pharmaceutical composition for resisting Helicobacter pylori containing Lactobacillus reuteri, comprising a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 15 parts of Herba Coptidis, 12 parts of Dianthus root, 10 parts of Dioscorea bulbifera, 7 parts of Rubescens herba, and 5 parts of Camellia sinensis; the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri bacterial solution is 30 g:7 mL; the number of viable bacteria in the Lactobacillus reuteri bacterial solution is 2.0×10 8 CFU / mL.

[0027] The preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0028] (1) Wash and dry Herba Coptidis, Dianthus root, Dioscorea bulbifera, Rabdosia rubescens, and Herba Citri Reticulatae, crush them to obtain a mixture, and set aside;

[0029] (2) soaking the mixture obtained in step (1) in a 70% by volume ethanol solution for 50 hours to obtain a mixture; wherein the mass volume ratio of the mixture to the ethanol solution is 1 g:8 mL;

[0030] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, performing ultrasonic extraction three times at a temperature of 30° C. and a power of 200 W, each extraction time being 15 min, and filtering to obtain a Chinese medicine composition;

[0031] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 30° C. for 24 h to obtain a fermentation product;

[0032] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0033] Example 2

[0034] A pharmaceutical composition for resisting Helicobacter pylori containing Lactobacillus reuteri, comprising a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 30 parts of Herba Coptidis, 25 parts of Dianthus root, 18 parts of Dioscorea bulbifera, 14 parts of Rubescens herba, and 10 parts of Camellia sinensis; the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri bacterial solution is 50 g:15 mL; the number of viable bacteria in the Lactobacillus reuteri bacterial solution is 2.5×10 10 CFU / mL.

[0035] The preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0036] (1) Wash and dry Herba Coptidis, Dianthus root, Dioscorea bulbifera, Rabdosia rubescens, and Herba Citri Reticulatae, crush them to obtain a mixture, and set aside;

[0037] (2) soaking the mixture obtained in step (1) in an 80% by volume ethanol solution for 80 hours to obtain a mixture; wherein the mass volume ratio of the mixture to the ethanol solution is 1 g:16 mL;

[0038] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, performing ultrasonic extraction 5 times at a temperature of 45° C. and a power of 300 W, each time for 30 min, and filtering to obtain a Chinese medicine composition;

[0039] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 35° C. for 48 h to obtain a fermentation product;

[0040] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0041] Example 3

[0042] A pharmaceutical composition for resisting Helicobacter pylori containing Lactobacillus reuteri, comprising a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 25 parts of Herba Coptidis, 20 parts of Dianthus root, 15 parts of Dioscorea bulbifera, 10 parts of Rubescens rubescens, and 7 parts of Herba Citriodorae; the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri bacterial solution is 40 g:13 mL; the number of viable bacteria in the Lactobacillus reuteri bacterial solution is 2.0×10 10 CFU / mL.

[0043] The preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0044] (1) Wash and dry Herba Coptidis, Dianthus root, Dioscorea bulbifera, Rabdosia rubescens, and Herba Citri Reticulatae, crush them to obtain a mixture, and set aside;

[0045] (2) soaking the mixture obtained in step (1) in a 75% by volume ethanol solution for 60 hours to obtain a mixture; wherein the mass volume ratio of the mixture to the ethanol solution is 1 g:10 mL;

[0046] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, performing ultrasonic extraction 4 times at a temperature of 40° C. and a power of 250 W, each extraction time being 25 min, and filtering to obtain a Chinese medicine composition;

[0047] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 32° C. for 40 h to obtain a fermentation product;

[0048] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0049] Example 4

[0050] A pharmaceutical composition for resisting Helicobacter pylori containing Lactobacillus reuteri, comprising a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 25 parts of Herba Coptidis, 20 parts of Dianthus root, 15 parts of Dioscorea bulbifera, 10 parts of Rubescens rubescens, and 7 parts of Herba Citriodorae; the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri bacterial solution is 40 g:13 mL; the number of viable bacteria in the Lactobacillus reuteri bacterial solution is 2.0×10 10 CFU / mL.

[0051] The preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0052] (1) Wash and dry Herba Coptidis, Dianthus root, Dioscorea bulbifera, Rabdosia rubescens, and Herba Citri Reticulatae, crush them to obtain a mixture, and set aside;

[0053] (2) soaking the mixture obtained in step (1) in a 75% by volume ethanol solution for 60 hours to obtain a mixture; wherein the mass volume ratio of the mixture to the ethanol solution is 1 g:10 mL;

[0054] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, performing ultrasonic extraction 4 times at a temperature of 40° C. and a power of 250 W, each extraction time being 25 min, and filtering to obtain a Chinese medicine composition;

[0055] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 32° C. for 40 h to obtain a fermentation product;

[0056] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0057] When treating Helicobacter pylori patients, antibiotic preparations and the anti-Helicobacter pylori pharmaceutical composition prepared in the present application are used simultaneously for treatment. The antibiotic preparations include a proton pump inhibitor PPI, antibiotics such as amoxicillin and clarithromycin, and a bismuth preparation such as potassium bismuth citrate.

[0058] Comparative Example 1

[0059] This comparative example differs from Example 3 in that no Herba Cynanchifolia is added to the Chinese medicine composition, and the other ingredients and steps remain unchanged.

[0060] A pharmaceutical composition for resisting Helicobacter pylori containing Lactobacillus reuteri, comprising a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 20 parts of Dianthus chinensis roots, 15 parts of Dioscorea bulbifera seeds, 10 parts of Rubescens rubescens, and 7 parts of Camellia sinensis; the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri bacterial solution is 40 g:13 mL; the number of viable bacteria in the Lactobacillus reuteri bacterial solution is 2.0×10 10 CFU / mL.

[0061] The preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0062] (1) Wash and dry dianthus root, Dioscorea bulbifera, Rubescens herb and Camellia sinensis, crush them to obtain a mixture, and set aside;

[0063] (2) soaking the mixture obtained in step (1) in a 75% by volume ethanol solution for 60 hours to obtain a mixture; wherein the mass volume ratio of the mixture to the ethanol solution is 1 g:10 mL;

[0064] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, performing ultrasonic extraction 4 times at a temperature of 40° C. and a power of 250 W, each extraction time being 25 min, and filtering to obtain a Chinese medicine composition;

[0065] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 32° C. for 40 h to obtain a fermentation product;

[0066] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0067] Comparative Example 2

[0068] This comparative example differs from Example 3 in that no Dianthus root is added to the Chinese medicine composition, and the other ingredients and steps remain unchanged.

[0069] A pharmaceutical composition for resisting Helicobacter pylori containing Lactobacillus reuteri, comprising a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 25 parts of Herba Coptidis, 15 parts of Dioscorea bulbifera, 10 parts of Rabdosia rubescens, and 7 parts of Herba Citri Reticulatae; the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri bacterial solution is 40 g:13 mL; the number of viable bacteria in the Lactobacillus reuteri bacterial solution is 2.0×10 10CFU / mL.

[0070] The preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0071] (1) Wash and dry Herba Coptidis, Dioscorea bulbifera, Rabdosia rubescens, and Herba Cibotii, crush them to obtain a mixture, and set aside;

[0072] (2) soaking the mixture obtained in step (1) in a 75% by volume ethanol solution for 60 hours to obtain a mixture; wherein the mass volume ratio of the mixture to the ethanol solution is 1 g:10 mL;

[0073] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, performing ultrasonic extraction 4 times at a temperature of 40° C. and a power of 250 W, each extraction time being 25 min, and filtering to obtain a Chinese medicine composition;

[0074] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 32° C. for 40 h to obtain a fermentation product;

[0075] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0076] Comparative Example 3

[0077] This comparative example differs from Example 3 in that the rhizome of Dioscorea bulbifera is replaced with Sanguisorba officinalis in the Chinese medicine composition, and the other ingredients and steps remain unchanged.

[0078] A pharmaceutical composition for resisting Helicobacter pylori containing Lactobacillus reuteri, comprising a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 25 parts of Herba Coptidis, 20 parts of Dianthus root, 15 parts of Sanguisorba officinalis, 10 parts of Rubescens rubescens, and 7 parts of Camellia sinensis; the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri bacterial solution is 40 g:13 mL; the number of viable bacteria in the Lactobacillus reuteri bacterial solution is 2.0×10 10 CFU / mL.

[0079] The preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0080] (1) Wash and dry Herba Coptidis, Dianthus root, Sanguisorba officinalis, Rubescens rubescens and Herba Citri Reticulatae, crush them to obtain a mixture, and set aside;

[0081] (2) soaking the mixture obtained in step (1) in a 75% by volume ethanol solution for 60 hours to obtain a mixture; wherein the mass volume ratio of the mixture to the ethanol solution is 1 g:10 mL;

[0082] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, performing ultrasonic extraction 4 times at a temperature of 40° C. and a power of 250 W, each extraction time being 25 min, and filtering to obtain a Chinese medicine composition;

[0083] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 32° C. for 40 h to obtain a fermentation product;

[0084] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0085] Comparative Example 4

[0086] This comparative example differs from Example 3 in that Rabdosia rubescens in the Chinese medicine composition is replaced with Scutellaria baicalensis, and the other ingredients and steps remain unchanged.

[0087] A pharmaceutical composition for resisting Helicobacter pylori containing Lactobacillus reuteri, comprising a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 25 parts of Herba Coptidis, 20 parts of Dianthus Root, 15 parts of Dioscorea Bulbifera, 10 parts of Scutellaria Baicalensis, and 7 parts of Camellia sinensis; the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri bacterial solution is 40 g:13 mL; the number of viable bacteria in the Lactobacillus reuteri bacterial solution is 2.0×10 10 CFU / mL.

[0088] The preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0089] (1) Wash and dry Herba Coptidis, Dianthus root, Dioscorea bulbifera, Scutellaria baicalensis and Herba Citri Reticulatae, crush them to obtain a mixture, and set aside;

[0090] (2) soaking the mixture obtained in step (1) in a 75% by volume ethanol solution for 60 hours to obtain a mixture; wherein the mass volume ratio of the mixture to the ethanol solution is 1 g:10 mL;

[0091] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, performing ultrasonic extraction 4 times at a temperature of 40° C. and a power of 250 W, each extraction time being 25 min, and filtering to obtain a Chinese medicine composition;

[0092] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 32° C. for 40 h to obtain a fermentation product;

[0093] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0094] Comparative Example 5

[0095] This comparative example differs from Example 3 in that the herb is replaced with radix schoenoprasum in the Chinese medicinal composition, and the other ingredients and steps remain unchanged.

[0096] A pharmaceutical composition for resisting Helicobacter pylori containing Lactobacillus reuteri, comprising a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial solution; the traditional Chinese medicine composition comprises the following components in parts by weight: 25 parts of Herba Coptidis, 20 parts of Dianthus Root, 15 parts of Dioscorea Bulbifera, 10 parts of Rubescens Herba, and 7 parts of Scutellariae; the mass-to-volume ratio of the traditional Chinese medicine composition to the Lactobacillus reuteri bacterial solution is 40 g:13 mL; the number of viable bacteria in the Lactobacillus reuteri bacterial solution is 2.0×10 10 CFU / mL.

[0097] The preparation method of the anti-Helicobacter pylori pharmaceutical composition comprises the following steps:

[0098] (1) Wash and dry Herba Coptidis, Dianthus root, Dioscorea bulbifera, Rabdosia rubescens, and Scutellaria baicalensis, crush them to obtain a mixture, and set aside;

[0099] (2) soaking the mixture obtained in step (1) in a 75% by volume ethanol solution for 60 hours to obtain a mixture; wherein the mass volume ratio of the mixture to the ethanol solution is 1 g:10 mL;

[0100] (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, performing ultrasonic extraction 4 times at a temperature of 40° C. and a power of 250 W, each extraction time being 25 min, and filtering to obtain a Chinese medicine composition;

[0101] (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 32° C. for 40 h to obtain a fermentation product;

[0102] (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

[0103] Experimental Example 1 Pharmacological test against Helicobacter pylori

[0104] 1.1 Materials and test methods

[0105] 1.1.1 Experimental strains: Helicobacter pylori SS1 was purchased from Chengdu Rongshengke Biotechnology Co., Ltd.; Lactobacillus reuteri was purchased from Lanzhou Waterless Biotechnology Co., Ltd.

[0106] 1.1.2 Anti-Helicobacter pylori Pharmaceutical Composition: The pharmaceutical composition prepared in Example 3 was used in the experiment and sterilized under UV for 2 hours. 3.0 g of the pharmaceutical composition powder was weighed into a centrifuge tube and filled to 10 mL with sterile water to prepare a suspension with a concentration of 300 mg / mL. The suspension was sonicated at room temperature for 2 hours and centrifuged at 10,000 rpm for 15 minutes. The supernatant was aspirated and sterilized using a 0.45 μm filter. At this point, 1 mL of the pharmaceutical solution is equivalent to 300 mg of the crude drug. The suspension was set aside.

[0107] 1.1.3 Helicobacter pylori SS1 culture method: Take 100 μL of glycerol culture and spread it into Helicobacter pylori culture medium. Incubate at 37°C with microaerophilicity (5% O2) for 72 hours. Pick a single colony and inoculate it into Helicobacter pylori liquid culture medium. Incubate at 37°C with microaerophilicity (5% O2) and shaking at 150 rpm for 48 hours. Determine the bacterial concentration by McFarland turbidimetry. Adjust the bacterial concentration to 1×10 8 CFU / mL, bacterial solution is reserved.

[0108] 1.1.4 Determination of minimum inhibitory concentration and minimum bactericidal concentration: The inhibitory effect of the pharmaceutical composition on Helicobacter pylori was determined by the broth microdilution method.

[0109] In row A of a 96-well plate, add 100 μL of sterile Helicobacter pylori liquid culture medium to each well, then add 100 μL of the drug composition supernatant to each well, A1 and A2. Mix well in well A2, then add 100 μL of the mixture from well A2 to well A3. Repeat this process until well A9 is reached. Mix well in well A9, then aspirate and discard 100 μL. Add 100 μL of bacterial solution to each well A1 to A9. Separately, establish a sterile water control group, a culture medium negative control group, and a positive control group. Repeat the same steps as in row A for rows B and C, three times. The drug composition concentrations in columns 1-12 of the 96-well plate are 150 mg / mL, 75 mg / mL, 37.5 mg / mL, 18.75 mg / mL, 9.38 mg / mL, 4.69 mg / mL, 2.34 mg / mL, 1.17 mg / mL, 0.59 mg / mL, and a sterile water control, respectively. The cells were cultured at 37°C under microaerophilic conditions (5% O2) for 72 h, 100 μL of 2% TTC staining solution was added, and the cells were incubated in the dark for 30 min. The lowest drug concentration that did not turn red under naked eye observation was the MIC (Minimum Inhibitory Concentration).

[0110] Based on the broth dilution method for determining the MIC of drugs against bacteria, 50 μL of the test bacterial solution was taken from the wells without bacterial growth and spread onto Helicobacter pylori culture medium without antibiotics. The culture was microaerophilic (5% O2) at 37°C for 72 hours. The minimum bacterial concentration (MBC) at which no bacteria grew was defined as the minimum bactericidal concentration.

[0111] 1.2 Test Results

[0112] The minimum inhibitory concentration (MIC) of the pharmaceutical composition of Example 3 against Helicobacter pylori was measured to be 9.38 mg / mL. Spreading the plate revealed that no colonies grew on the solid culture plate at concentrations of 150 mg / mL, 75 mg / mL, 37.5 mg / mL, and 18.75 mg / mL, indicating a minimum bactericidal concentration (MBC) of 18.75 mg / mL.

[0113] Experimental Example 2 Antibacterial Rate

[0114] 2.1 Under the same MIC concentration of 9.38 mg / mL, the effects of the anti-Helicobacter pylori pharmaceutical compositions prepared in Examples 1 to 4 and Comparative Examples 1 to 5 on the growth of Helicobacter pylori were tested, and the inhibition rates were calculated. 8 The CFU / mL Helicobacter pylori culture was inoculated into each experimental group at a ratio of 1:100. Meanwhile, a culture medium containing bacteria but no drug composition was used as a negative control group. The OD (optical density, absorbance) value of the drug was deducted. The culture was shaken at 150 rpm at 37°C and microaerophilic (5% O2) conditions. The OD was measured at 0 and 48 h. 600 , repeated three times. Calculate the antibacterial rate of the pharmaceutical composition. The calculation formula of the antibacterial rate is as follows:

[0115]

[0116] 2.2 Test Results

[0117] As shown in Table 1:

[0118] Table 1 Inhibition rate of Helicobacter pylori

[0119]

[0120]

[0121] As shown in Table 1, the pharmaceutical compositions prepared in Examples 1 to 4 exhibited inhibition rates against Helicobacter pylori ranging from 91.36% to 94.12%. This indicates that the pharmaceutical compositions of the present invention exhibit a highly potent inhibitory effect against Helicobacter pylori, and that, at the same concentration, the pharmaceutical compositions prepared in Examples 1 to 4 exhibited superior inhibitory effects against Helicobacter pylori compared to the pharmaceutical compositions prepared in Comparative Examples 1 to 5. Comparison of the inhibition rates in Examples 3 and 4 demonstrates that the pharmaceutical compositions of the present invention can be used in combination with antibiotic preparations, achieving superior technical benefits compared to using the pharmaceutical compositions alone. Comparison of Example 3 with Comparative Examples 1 and 2 shows that the complete Chinese medicine composition components of Herba Cinnamomi, Dianthus Root, Dioscorea Bulbifera, Rabdosia Rubescens and Camellia Sinensis have a greater impact on the inhibition rate of Helicobacter pylori. If a certain component is missing, the synergistic effect of multiple components will be destroyed to exert the efficacy, resulting in a decrease in the overall efficacy. Comparison of Example 3 with Comparative Examples 3 to 5 shows that replacing Dioscorea Bulbifera, Rabdosia Rubescens and Camellia Sinensis with similar efficacy of Radix Sanguisorbae, Radix Scutellariae and Radix Scutellariae, even if the efficacy of the replaced Chinese medicines is similar, may result in a decrease in the inhibition rate of Helicobacter pylori due to differences in specific active ingredients or the lack of certain key antibacterial components. Therefore, only by combining the Chinese medicine compositions of the present application can an anti-Helicobacter pylori pharmaceutical composition with the best antibacterial effect be prepared.

[0122] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. An anti-Helicobacter pylori pharmaceutical composition containing Lactobacillus reuteri, characterized in that: The anti-Helicobacter pylori pharmaceutical composition comprises a traditional Chinese medicine composition and a Lactobacillus reuteri bacterial liquid; the traditional Chinese medicine composition comprises the following components in parts by weight: 15 to 30 parts of Herba Coptidis, 12 to 25 parts of Dianthus root, 10 to 18 parts of Dioscorea bulbifera, 7 to 14 parts of Rabdosia rubescens, and 5 to 10 parts of Camellia sinensis.

2. The anti-Helicobacter pylori pharmaceutical composition according to claim 1, characterized in that The mass-to-volume ratio of the traditional Chinese medicine composition and the Lactobacillus reuteri liquid is (30-50) g: (7-15) mL.

3. The anti-Helicobacter pylori pharmaceutical composition according to claim 1, characterized in that The number of viable bacteria in the Lactobacillus reuteri liquid is 2.0×10 8 CFU / mL~2.5×10 10 CFU / mL.

4. The method for preparing the anti-Helicobacter pylori pharmaceutical composition according to claim 1, characterized in that: The following steps are involved: (1) Wash and dry Herba Coptidis, Dianthus root, Dioscorea bulbifera, Rabdosia rubescens, and Herba Citri Reticulatae, crush them to obtain a mixture, and set aside; (2) soaking the mixture obtained in step (1) in an ethanol solution with a volume fraction of 70% to 80% for 50 hours to 80 hours to obtain a mixture; (3) placing the mixture obtained in step (2) in an ultrasonic extraction tank, ultrasonically extracting 3 to 5 times, and filtering to obtain a Chinese medicine composition; (4) mixing the Chinese medicine composition obtained in step (3) with the Lactobacillus reuteri bacterial solution, and performing sealed anaerobic fermentation at a fermentation temperature of 30° C. to 35° C. for a fermentation time of 24 h to 48 h to obtain a fermentation product; (5) freeze-drying the fermentation product obtained in step (4) to finally obtain an anti-Helicobacter pylori pharmaceutical composition.

5. The method for preparing the anti-Helicobacter pylori pharmaceutical composition according to claim 4, characterized in that: The mass volume ratio of the mixed material and the ethanol solution in step (2) is 1g:(8-16)mL.

6. The method for preparing the anti-Helicobacter pylori pharmaceutical composition according to claim 4, characterized in that: The temperature of ultrasonic extraction in step (3) is 30° C. to 45° C., the power is 200W to 300W, and the time is 15min to 30min.

7. The method for preparing the anti-Helicobacter pylori pharmaceutical composition according to claim 4, characterized in that: The anti-Helicobacter pylori pharmaceutical composition further comprises pharmaceutically acceptable excipients and is prepared into any dosage form including powder, decoction, pill, capsule, tablet, granule and oral liquid.

8. Use of the anti-Helicobacter pylori pharmaceutical composition according to any one of claims 1 to 3 or the anti-Helicobacter pylori pharmaceutical composition prepared by the preparation method according to any one of claims 4 to 7 in the preparation of anti-Helicobacter pylori drugs.

9. The use according to claim 8, characterized in that The anti-Helicobacter pylori pharmaceutical composition is used in combination with an antibiotic preparation.

10. The use according to claim 9, characterized in that The antibiotic preparation includes omeprazole, amoxicillin, clarithromycin, and bismuth potassium citrate.

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