Freeze-dried powder and capsule for treating intractable helicobacter pylori infection as well as preparation method and application of freeze-dried powder and capsule

Abstract

The invention discloses freeze-dried powder and capsules for treating intractable helicobacter pylori infection as well as a preparation method and application of the freeze-dried powder and capsules, and belongs to the technical field of biological medicines. The invention provides a preparation method of freeze-dried powder for treating intractable helicobacter pylori infection, which comprises the following steps: acquiring gastric mucous layer flushing fluid which contains gastric microorganisms; filling the obtained gastric mucous layer flushing fluid into an eggplant-shaped bottle, connecting a low-temperature rotator, and rotating for a certain time under a low-temperature condition until the bacterial liquid adheres to the wall; and after the bacterial liquid adheres to the wall, taking down the eggplant-shaped bottle, mounting the eggplant-shaped bottle on a freeze-drying machine, and freeze-drying at low temperature for a certain time to prepare freeze-dried powder. The capsule prepared from the freeze-dried powder prepared by using the preparation method disclosed by the invention can overcome gastrointestinal flora disorder caused by conventional anti-helicobacter pylori infection treatment, repair unbalanced gastric microbiota of an infected patient, and avoid complex operations or invasive operations possibly involved in traditional flora transplantation, such as gastroscopy or nasogastric tubes and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a freeze-dried powder, a capsule for treating refractory Helicobacter pylori infection, a preparation method thereof, and an application thereof. Background Art

[0002] Helicobacter pylori (Hp) infection is a global health problem. According to epidemiological surveys, approximately 50% of the global population is infected with Hp. Hp infection can cause varying degrees of gastric flora disorders. The research results of Schulz et al. show that in the gastric juice and gastric mucosa of people infected with Helicobacter pylori, at the phylum level, the relative abundance of Proteobacteria significantly increases, while the relative abundances of Firmicutes, Bacteroidetes, Actinobacteria, and Fusobacteria decrease. The gastric microecological imbalance caused by Hp infection is also a potential mechanism leading to gastric diseases. Eradicating Hp can significantly improve the prognosis of diseases such as gastritis and peptic ulcers, and reduce the incidence of gastric cancer.

[0003] Currently, the main regimens for eradicating Hp are based on the combination of antibiotics and proton pump inhibitors. However, these regimens have many limitations: on the one hand, with the widespread use of antibiotics, the resistance of Hp to various antibiotics gradually increases, resulting in a decrease in the eradication rate. On the other hand, the use of antibiotics and proton pump inhibitors can disrupt the balance of the gastrointestinal microbiota, leading to dysbiosis, and further causing adverse reactions such as bad breath, abdominal pain, diarrhea, abdominal distension, and constipation, reducing patient compliance and affecting the eradication efficacy. In addition, it takes a long time for the gastrointestinal microbiota to return to its original state after antibiotic treatment, and some patients may not be able to fully recover.

[0004] In recent years, the concept of "bacteria inhibiting bacteria" has been proposed, and the growth of Hp is inhibited and the gastrointestinal microecology is improved through the action of probiotics. However, the existing probiotic preparations (such as lactic acid bacteria, bifidobacteria, etc.) have a single strain structure and insufficient diversity, and cannot effectively restore the imbalanced gastrointestinal microbiota. In addition, the survival rate of probiotics in the gastric acid environment is relatively low, and it is difficult to colonize in the stomach and play a continuous role. Therefore, it is of great clinical significance to find safer and more effective antibacterial therapeutic drugs or regimens for treating refractory Helicobacter pylori infection. Summary of the Invention

[0005] Aiming at the above existing problems, the present invention aims at refractory Helicobacter pylori infection, and proposes a freeze-dried powder, a capsule for treating refractory Helicobacter pylori infection, a preparation method thereof, and an application thereof. By obtaining natural gastric microorganisms from the human gastric mucus layer, and through low-temperature treatment and freeze-drying technology to make a freeze-dried powder and prepare it into a capsule, it can colonize in the stomach and restore the imbalanced gastrointestinal microbiota, thereby effectively eradicating Hp infection and reducing antibiotic resistance and side effects.

[0006] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0007] A preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection, comprising the following steps:

[0008] S1. Obtain gastric mucus layer rinsing fluid, which contains gastric microorganisms;

[0009] S2. Load the obtained gastric mucus layer rinsing fluid into an eggplant-shaped flask, connect a low-temperature rotator, and rotate it under low-temperature conditions until the bacterial solution adheres to the wall;

[0010] S3. Remove the eggplant-shaped flask and install it on a freeze-dryer for low-temperature freeze-drying to obtain a freeze-dried powder for treating refractory Helicobacter pylori infection.

[0011] Through the above technical solution, further, in step S1, the gastric mucus layer rinsing fluid is obtained by thoroughly rinsing the gastric mucus layer of a donor with an eluent.

[0012] Through the above technical solution, further, the eluent is prepared by mixing 20000 u pronase, 1 g sodium bicarbonate with warm water at 40 °C to 50 ml.

[0013] Through the above technical solution, further, in step S2, the temperature of low-temperature rotation is -40 to -20 °C, and the time is 30 - 40 minutes.

[0014] Through the above technical solution, further, in step S3, the low-temperature freeze-drying temperature on the freeze-dryer is -40 to -20 °C.

[0015] Through the above technical solution, further, in step S3, the low-temperature freeze-drying time on the freeze-dryer is 16 - 20 h.

[0016] Through the above technical solution, further, the finished product in step S3 is vacuum-packed.

[0017] Through the above technical solution, further, the capsule for treating refractory Helicobacter pylori infection is prepared from the freeze-dried powder prepared by a preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection.

[0018] Through the above technical solution, further, the capsule for treating refractory Helicobacter pylori infection is prepared by adding pharmaceutically acceptable excipients to the freeze-dried powder prepared above.

[0019] The technical effects achieved by the invention are as follows:

[0020] The present invention provides a freeze-dried powder, a capsule, a preparation method and an application thereof for safely and effectively treating refractory Helicobacter pylori infection. By utilizing natural gastric microorganisms and low-temperature freeze-drying technology, the present invention overcomes the deficiencies of traditional antibiotic treatments, can significantly improve the gastrointestinal microecology of patients, increase the Hp eradication rate, and reduce the occurrence of long-term adverse reactions. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is a schematic diagram of a low-temperature rotator used in step S2 of the preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection in an embodiment of the present application;

[0022] Figure 2 It is a schematic diagram of a freeze dryer used for making freeze-dried powder in an eggplant-shaped flask in step S3 of the preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection in an embodiment of the application;

[0023] Figure 3 It is a result display diagram after filling the freeze-dried powder into a capsule in an embodiment of the present application;

[0024] Figure 4 It is an endoscopic and pathological examination diagram before treatment of a refractory Helicobacter pylori infection patient using the capsule for treating refractory Helicobacter pylori infection prepared in the present application in an embodiment of the present application;

[0025] Figure 5 It is an endoscopic and pathological examination diagram after treatment of a refractory Helicobacter pylori infection patient using the capsule for treating refractory Helicobacter pylori infection prepared in the present application in an embodiment of the present application;

[0026] Figure 6 It is a treatment effect diagram of a capsule for treating refractory Helicobacter pylori infection on refractory Helicobacter pylori infection in an embodiment of the present application;

[0027] Figure 7A It is a change diagram of the gastric microecology of a refractory Helicobacter pylori patient before and after treatment in an embodiment of the present application;

[0028] Figure 7B It is a change diagram of the gastric microecology of a refractory Helicobacter pylori patient before and after treatment in an embodiment of the present application;

[0029] Figure 7C It is a change diagram of the gastric microecology of a refractory Helicobacter pylori patient before and after treatment in an embodiment of the present application. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0030] To enable those skilled in the art to better understand the solution of the present invention, the following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part, rather than all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention. It should be noted that the terms "first", "second", etc. in the specification and claims of the present invention and the above-mentioned drawings are used to distinguish similar objects, and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged under appropriate circumstances so that the embodiments of the present invention described herein can be implemented in an order different from those illustrated or described herein. In addition, the terms "comprising" and "having" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, system, product or device comprising a series of steps or units is not necessarily limited to those steps or units clearly listed, but may include other steps or units not clearly listed or inherent to these processes, methods, products or devices.

[0031] The present invention and its implementation manners are described below, and this description is not restrictive. The actual implementation manners are not limited thereto. Generally speaking, if those of ordinary skill in the art are inspired by it and, without departing from the purpose of the present invention, design similar structural manners and embodiments without creative efforts, they shall fall within the protection scope of the present invention. The test methods described in the following embodiments are conventional methods unless otherwise specified; the instruments and materials are commercially available unless otherwise specified.

[0032] Hp infection is a global health problem. Through epidemiological investigations, it has been found that approximately 50% of the global population is infected with Hp. Hp infection can cause varying degrees of gastric flora disorders. Eradicating Hp can significantly improve the prognosis of diseases such as gastritis and peptic ulcers, and reduce the incidence of gastric cancer. Currently, the main regimens for eradicating Hp are based on antibiotics combined with PPI, but they also have limitations: on the one hand, with the widespread use of antibiotics, the resistance of Hp to various antibiotics has gradually increased, resulting in a decline in the eradication rate. On the other hand, the use of antibiotics and proton pump inhibitors can disrupt the balance of the gastrointestinal microbiota, leading to dysbiosis, and further causing adverse reactions such as bad breath, abdominal pain, diarrhea, abdominal distension, and constipation, reducing patient compliance and affecting the eradication efficacy. In addition, it takes a long time for the gastrointestinal microbiota to return to its original state after antibiotic treatment, and some patients may not be able to fully recover.

[0033] In recent years, the concept of "controlling bacteria with bacteria" has been proposed, aiming to inhibit the growth of Hp and improve the gastrointestinal microecology through the action of probiotics. However, the existing probiotic preparations (such as lactic acid bacteria, bifidobacteria, etc.) have a single strain structure and insufficient diversity, and are unable to effectively restore the imbalanced gastrointestinal microbiota. In addition, probiotics have a low survival rate in the gastric acid environment and are difficult to colonize in the stomach and play a continuous role.

[0034] Currently, fecal microbiota transplantation has been applied to the treatment of diseases with intestinal microbiota dysregulation such as antibiotic-associated diarrhea, Clostridium difficile infection, and inflammatory bowel disease, and has shown good ability to reconstruct the imbalanced intestinal microbiota in the treatment of these diseases. At the same time, there are also a large number of microorganisms in the stomach. Moreover, there is a certain degree of gastric microbiota disorder in patients with Helicobacter pylori infection before and after traditional eradication treatment.

[0035] Based on the above findings, this application proposes a treatment plan for refractory Helicobacter pylori infection by gastric microbiota transplantation (GMT). Gastric microbiota transplantation is similar to fecal microbiota transplantation. By transplanting the microbiota extracted from the donor's stomach into the patient's stomach, the disease is treated by repairing the imbalanced gastric microbiota of the patient. However, for this treatment plan of gastric microbiota transplantation, the microbiota extracted from the donor cannot be directly transplanted into the patient's stomach for capsule treatment of refractory Helicobacter pylori infection. It is necessary to prepare the gastric microbiota of the donor into gastric bacteria capsules for use by patients with indications.

[0036] Based on the above, the technical solution of this application aims to provide a preparation method, capsule and application of a freeze-dried powder for treating refractory Helicobacter pylori infection, which can effectively eliminate drug-resistant Helicobacter pylori strains, maintain the stability of the gastrointestinal microbiota at the same time, and thus improve the long-term health prognosis of patients. It can overcome problems faced by existing treatment plans such as increased drug resistance and disruption of microecological balance, effectively treat and eradicate refractory Helicobacter pylori infection, and the patients are convenient to take the medicine, have good compliance, reduce the incidence of adverse reactions of antibiotics, and also reduce the antibiotic resistance rate.

[0037] The following combines specific embodiments to introduce and describe the technical solution of this application in detail.

[0038] Example 1:

[0039] The present invention provides a preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection, which is prepared from gastric microorganisms extracted from the stomach of a donor not infected with Helicobacter pylori, and is used to repair the imbalanced gastric microbiota in patients infected with Helicobacter pylori, and overcome the gastrointestinal flora dysbiosis caused by conventional anti-Helicobacter pylori infection treatment, can overcome problems such as increased drug resistance and disruption of microecological balance faced by existing treatment regimens, effectively treat and eradicate refractory Helicobacter pylori infection, and the patients are convenient to take the medicine, have good compliance, reduce the incidence rate of adverse reactions of antibiotics, and also reduce the antibiotic resistance rate.

[0040] A preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection in an embodiment of the present invention includes:

[0041] S1. Obtain gastric mucus layer rinsing fluid, and the gastric mucus layer rinsing fluid includes gastric microorganisms;

[0042] S2. Load the obtained gastric mucus layer rinsing fluid into an eggplant-shaped flask, connect a low-temperature rotary instrument, and rotate at a low temperature until the bacterial solution adheres to the wall;

[0043] S3. After rotating at a low temperature until the bacterial solution adheres to the wall, remove the eggplant-shaped flask and install it on a freeze dryer, and perform low-temperature freeze drying to obtain a freeze-dried powder for treating refractory Helicobacter pylori infection.

[0044] In an embodiment of the present application, the donor refers to a donor not infected with Helicobacter pylori, and the gastric mucus layer rinsing fluid in step S1 is obtained by sufficiently rinsing the gastric mucus layer of the donor with an eluent.

[0045] It should be noted that in the process of obtaining the gastric mucus layer rinsing fluid from the donor using the eluent, it is necessary to thoroughly rinse the donor's stomach first to remove impurities and suck out the rinsing fluid. After that, the gastric mucus layer is treated with the eluent to obtain the required rinsing fluid.

[0046] Specifically, in an embodiment of the present application, for the treatment of the donor, different operations are performed according to the presence or absence of a mucus lake: if there is a mucus lake, the gastric juice is removed first and then rinsed; if not, rinsing is directly performed. In this process, safe cleaning fluids are used, and strict ethical norms are followed to ensure the health and privacy of the donor.

[0047] Specifically, the eluent in an embodiment of the present application is prepared by adding 20000u of pronase and 1 g of sodium bicarbonate to 50 ml of warm water at 40°C.

[0048] In an embodiment of the present application, after obtaining the gastric mucus layer rinsing fluid, in step S2, the obtained gastric mucus layer rinsing fluid is loaded into an eggplant-shaped flask, connected to a low-temperature rotary instrument, and the temperature is controlled at -40°C to -20°C, and rotated at a low temperature for 30 min - 40 min until the bacterial solution adheres to the wall.

[0049] In the embodiments of the present application, the time for the gastric mucus layer flushing solution to be swirled at low temperature is 30 - 40 minutes. This range helps to ensure the activity and stability of microorganisms while inhibiting other unwanted biological activities. The low-temperature freeze-drying temperature of the eggplant-shaped flask on the freeze dryer is controlled at -40 to -20 °C. This range helps to effectively remove moisture while protecting the structure and activity of microorganisms.

[0050] As shown in the Figure 1 appendix, Figure 1 Figure 1 is a schematic diagram of the low-temperature swirl apparatus used in step S2 of the embodiments of the application. After the gastric mucus layer flushing solution is filled into the eggplant-shaped flask in step S2, it is connected to the low-temperature swirl apparatus and subjected to swirl treatment at a controlled temperature. After 30 minutes of rotation operation, the bacterial liquid in the liquid is effectively adhered to the wall. While ensuring the effective removal of moisture from the gastric mucus layer flushing solution, it helps to protect the structure and activity of microorganisms and can inhibit the activities of other unnecessary microorganisms.

[0051] Specifically, in step S3, the time for the low-temperature freeze-drying of the eggplant-shaped flask on the freeze dryer is 16 - 20 h.

[0052] As shown in the Figure 2 appendix, Figure 2 Figure 2 is a schematic diagram of the freeze dryer used to produce the freeze-dried powder of the eggplant-shaped flask in step S3 of the embodiments of the present application. In step S3, after swirling at low temperature until the bacterial liquid adheres to the wall, the eggplant-shaped flask is removed and installed on the freeze dryer. The low-temperature freeze-drying process is carried out for 16 to 20 hours through the freeze dryer, and the liquid in the eggplant-shaped flask is converted into a freeze-dried powder, which can ensure that moisture can be effectively removed during the freeze-drying process while maintaining the quality and activity of the product.

[0053] In some embodiments of the present application, the finished product in step S3, that is, the finally obtained freeze-dried powder for treating refractory Helicobacter pylori infection, is vacuum-packed. By using vacuum packaging technology for the prepared freeze-dried powder for treating refractory Helicobacter pylori infection, its activity and stability can be effectively maintained, the shelf life can be extended, contamination can be reduced, and it is convenient to use. By selecting appropriate packaging materials, strict process flows, and reasonable storage conditions, the quality and safety of the freeze-dried powder can be ensured.

[0054] It should be noted that the packaging materials used in the present application for the freeze-dried powder for treating refractory Helicobacter pylori infection can be packaging materials with good barrier properties to ensure minimal penetration of oxygen, moisture, and light. The storage temperature can be selected to be stored in a low-temperature environment of -80 °C to avoid a decrease in activity caused by high temperature.

[0055] In the embodiment of the present application, a preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection includes obtaining gastric mucus layer rinsing fluid; filling the obtained gastric mucus layer rinsing fluid into an eggplant-shaped flask, connecting a low-temperature rotary instrument, and rotating at low temperature until the bacterial solution adheres to the wall; after rotating at low temperature until the bacterial solution adheres to the wall, removing the eggplant-shaped flask and installing it on a freeze-dryer, and freeze-drying at low temperature to obtain the freeze-dried powder for treating refractory Helicobacter pylori infection. After the freeze-dried powder is made, the obtained freeze-dried powder can be made into capsules for the treatment of patients with refractory Helicobacter pylori infection, which can effectively eliminate drug-resistant Helicobacter pylori strains and maintain the stability of the gastrointestinal microbiota, thereby improving the long-term health prognosis of patients. It can overcome problems faced by existing treatment regimens such as increased drug resistance and disruption of the microecological balance, effectively treat and eradicate refractory Helicobacter pylori infection, and the patients can take the medicine conveniently with good compliance, reducing the incidence of adverse reactions of antibiotics and also reducing the drug resistance rate of antibiotics.

[0056] The freeze-dried powder in the present application can be used in the preparation of drugs for treating refractory Helicobacter pylori infection. In some embodiments of the present application, after the freeze-dried powder for treating refractory Helicobacter pylori infection is prepared, the obtained freeze-dried powder can be made into capsules for the treatment of patients with refractory Helicobacter pylori infection.

[0057] It should be noted that in some implementations, after the freeze-dried powder for treating refractory Helicobacter pylori infection is prepared, it can also be made into capsules for the treatment of patients with refractory Helicobacter pylori infection by adding pharmaceutically acceptable excipients.

[0058] It should be noted that the excipients used to prepare capsules for the treatment of patients with refractory Helicobacter pylori infection by adding pharmaceutically acceptable excipients to the freeze-dried powder in the embodiment of the present application can be selected from microcrystalline cellulose, lactose, etc. By using these excipients, the stability of the drug can be improved, the solubility and absorbability of the drug can be improved, the compliance of the patients can be improved, production and quality control can be facilitated, and the release time of the drug can be extended.

[0059] As Figure 3 shown, attached Figure 3This is the result display after filling the freeze-dried powder into the capsules in the embodiments of this application. The powder obtained after pretreatment and low-temperature freeze-drying is carefully transferred into the capsule shell to facilitate further storage, transportation, and application. The preparation of the capsules ensures the stability of the ingredients and is convenient for quantitative use. In the embodiments of this application, by using the freeze-dried powder for treating refractory Helicobacter pylori infection to prepare capsules for patients to take, it can overcome the gastrointestinal flora imbalance caused by conventional anti-Helicobacter pylori infection treatment and repair the imbalanced gastric microbiota in infected patients. By orally taking this capsule, the capsule gradually dissolves in the stomach, and the released live bacteria can directly colonize and play a role in the stomach, avoiding the complex surgeries or invasive operations that may be involved in traditional flora transplantation, such as gastroscopy or nasogastric tube.

[0060] Application Case

[0061] In view of the technical problems that there is a certain degree of gastric flora disorder in the existing treatment plan for Helicobacter pylori infection and it cannot be completely eradicated, the applicant proposes a capsule, a preparation method and an application for treating refractory Helicobacter pylori infection, which can overcome the gastrointestinal flora imbalance caused by conventional anti-Helicobacter pylori infection treatment and repair the imbalanced gastric microbiota in infected patients. Specifically, the present invention relates to a preparation method of a capsule for treating refractory Helicobacter pylori infection, which includes the following steps:

[0062] (1) Preparation before preparation:

[0063] 1) Establish a standard laboratory and obtain corresponding qualifications;

[0064] 2) Form a professional team, implement standardized management, and strengthen the supervision and management of GMT technology access, ethics, the preparation and quality control of bacterial liquid and capsules, clinical application, evaluation, and information materials;

[0065] 3) Strictly screen and manage donors;

[0066] (2) Capsule preparation

[0067] 1) After inserting the BL-7000 magnifying glass into the stomach, for donors with a mucus lake, insert an irrigation tube into the biopsy channel of the gastroscope, connect one end to a 50 ml sterile syringe, and the other end is guided by the gastroscope to penetrate deep into the mucus lake to aspirate the gastric juice in it; if there is no mucus lake, directly proceed to step 2);

[0068] 2) Thoroughly rinse the entire stomach with an appropriate amount of clear water once, and aspirate the rinsing fluid with a negative pressure aspirator;

[0069] 3) Replace the irrigation tube and syringe and use an eluent (20000 u of pronase + 1 g of sodium bicarbonate added to 40 °C warm water to 50 ml) to thoroughly rinse the gastric mucus layer, and aspirate the air in the stomach repeatedly 3 times, lasting about 3 - 5 min to achieve the purpose of sufficient elution;

[0070] 4) Take 50 ml of the mucus layer rinse solution and pour it into an eggplant-shaped flask. Connect a low-temperature rotary instrument and control the temperature at -40 to

[0071] -20 °C, rotate at low temperature for 30 min - 40 min until the bacterial solution adheres to the wall;

[0072] 5) Remove the eggplant-shaped flask from the rotary instrument, then install it on a freeze dryer and freeze-dry it into powder at low temperature for about 16 - 20 h to obtain the freeze-dried powder for treating refractory Helicobacter pylori infection;

[0073] 6) Make the obtained freeze-dried powder into capsules.

[0074] Specifically, in step 6), capsules can be made by adding pharmaceutically acceptable excipients such as microcrystalline cellulose and lactose to the obtained freeze-dried powder.

[0075] (3) Capsule storage

[0076] Store the capsules in a refrigerator at -80 °C for patients to use.

[0077] The working principle of a capsule for treating refractory Helicobacter pylori infection according to the present invention: (1) Oral administration: By orally taking the capsules made from the freeze-dried powder for treating refractory Helicobacter pylori infection, it avoids the complex surgeries or invasive operations that may be involved in traditional microbiota transplantation, such as gastroscopy or nasogastric tube. (2) Gastric release: The capsules gradually dissolve in the stomach, and the active microorganisms can colonize in the stomach and continuously play their roles without complex surgeries or invasive operations.

[0078] The structural characteristics of a capsule for treating refractory Helicobacter pylori infection according to the present invention: (1) High activity: The live bacteria in the gastric bacteria capsules (corresponding to the capsules made from the freeze-dried powder for treating refractory Helicobacter pylori infection mentioned above) maintain a high activity state. Protected by the capsules, the live bacteria can play their full roles when reaching the stomach. (2) Easy to store: The gastric bacteria capsules are made in the form of freeze-dried powder, which is convenient for storage and transportation. They can be stably stored at -80 °C for a long time, maintaining the activity of microorganisms, reducing the risk of contamination, extending the shelf life, and maintaining the physical properties of the capsules.

[0079] The following is a specific application example of the present invention regarding a capsule for treating refractory Helicobacter pylori infection:

[0080] Application case 1:

[0081] Such as Figures 4 - 5 , Figure 4 are the endoscopic and pathological examination images of a patient with refractory Helicobacter pylori infection before using the capsules for treating refractory Helicobacter pylori infection prepared in this application in the examples of this application, Figure 5This is the endoscopic and pathological examination images of patients with refractory Helicobacter pylori infection after treatment with the capsules prepared in this application for treating refractory Helicobacter pylori infection. Combined with Figure 4 It can be seen that in the gastroscopy images of patients with refractory Helicobacter pylori infection before treatment, the mucosa of the corpus in the corpus part shows congestion and erythema, the mucosal folds in the antrum part are thickened and widened, and there are scattered erosions on the surface of the mucosa in the angulus part, with congestion and accompanied by edema, making the mucosa look swollen and thickened, losing its usual smoothness and color. The histological examination results of the antrum part and the corpus part show that the nuclear morphology is irregular, the staining is deepened, the cytoplasmic staining is uneven, the glandular structure is disordered, the glandular arrangement is disordered, and the interstitial structure is significantly thickened. The histological examination results of the antrum part and the corpus part show the characteristic manifestations of Helicobacter pylori infection. Combined with Figure 5 It can be seen that in the gastroscopy images of patients with refractory Helicobacter pylori infection after treatment, the mucosa colors of the corpus, antrum and angulus parts are light red or pink, with uniform color, no obvious congestion or erythema, the mucosal surface is smooth, with regular folds, no obvious erosions, ulcers or nodules, and no manifestations of Helicobacter pylori infection are seen. The histological examination results of the antrum part and the corpus part show the normal gastric mucosal structure, the nuclei are evenly distributed in the cells, without abnormal aggregation or dispersion, there are no obvious abnormal organelles in the cytoplasm, the cell boundaries are clear, and there are no manifestations of Helicobacter pylori infection.

[0082] From the above Figures 4 - 5 comparison, it can be seen that after the patients use the capsules prepared in this application for treating refractory Helicobacter pylori infection, the indicators of Helicobacter pylori infection show normal manifestations. Thus, it can be seen that the capsules prepared in this application can effectively treat and eradicate refractory Helicobacter pylori infection, and it is convenient for patients to take the medicine.

[0083] Application case 2: The effect of patients taking the capsules of this application for treating refractory Helicobacter pylori infection.

[0084] Patient 1:

[0085] The patient had symptoms of abdominal distension and belching. The carbon breath test showed Hp(+). After evaluation, the patient had the indication to take the gastric bacteria capsules. The patient was given oral gastric bacteria capsules, 6 capsules each time, once a day. Antibiotics were avoided during the medication period. After taking the medicine for 5 days, it was stopped.

[0086] Specific effect: The patient's carbon breath test was rechecked 2 months after treatment, and the patient's Helicobacter pylori turned negative, and there were no adverse reactions.

[0087] Patient 2:

[0088] The patient had symptoms of abdominal distension and abdominal burning sensation. The carbon breath test indicated Hp(+). After evaluation, the patient had the indication to take Weijun Capsules. The patient was given oral administration of Weijun Capsules, 6 capsules each time, once a day. During the medication period, avoid taking antibiotics. After taking the medicine for 5 days, stop using it.

[0089] Specific effect: One month after the treatment, the patient had a reexamination of the carbon breath test. The patient's Helicobacter pylori turned negative and there were no adverse reactions.

[0090] Patient 3:

[0091] The patient had symptoms of abdominal distension and abdominal burning sensation. The carbon breath test indicated Hp(+). After evaluation, the patient had the indication to take Weijun Capsules. The patient was given oral administration of Weijun Capsules, 6 capsules each time, once a day. During the medication period, avoid taking antibiotics. After taking the medicine for 5 days, stop using it.

[0092] Specific effect: Three months after the treatment, the patient had a reexamination of the carbon breath test. The patient's Helicobacter pylori turned negative and there were no adverse reactions.

[0093] The present invention relates to a capsule for treating refractory Helicobacter pylori infection, which can overcome the gastrointestinal flora imbalance caused by conventional anti-Helicobacter pylori infection treatment. For the quadruple anti-Helicobacter pylori infection treatment regimen, the patient's medication experience is poor. Some patients fail to take the medicine regularly, especially elderly patients who cannot correctly follow the doctor's advice to take the medicine, etc., and achieve the purpose of eradicating Helicobacter pylori infection. It is simple to operate, has good effects and few side effects.

[0094] The capsule for treating refractory Helicobacter pylori infection of the present invention has the following advantages:

[0095] 1. High flora diversity: The capsule of the present invention is derived from natural gastric microorganisms of the human body, with a rich flora structure, and can more effectively restore the imbalanced gastrointestinal microbiota.

[0096] 2. Strong colonization ability: After the capsule dissolves in the stomach, the released active microorganisms can colonize in the stomach and continuously play a role, without complex surgeries or invasive operations.

[0097] 3. Reducing antibiotic resistance: Different from traditional antibiotic treatment, the present invention inhibits the growth of Hp by restoring the gastrointestinal microecological balance, reducing the dependence on antibiotics and the generation of drug resistance.

[0098] 4. High patient compliance: The capsule is convenient to take, reducing the adverse reactions of antibiotic treatment (such as abdominal pain, diarrhea, etc.), and improving the patient's treatment compliance.

[0099] As Figure 6 shown, Figure 6This is a diagram showing the therapeutic effect of a capsule for treating refractory Helicobacter pylori infection in the embodiments of the present application. Combining Figure 6 with the Prior-GMT row in Figure 6 , it can be seen that the Helicobacter pylori infection indicators of patients were all positive before treatment. Using the commonly used treatment regimen for eradicating Helicobacter pylori infection (such as the regimen based on the combination of antibiotics and PPI) in the Post-GMT row in Figure 6 , it can be seen that the Helicobacter pylori infection indicators of most patients were still positive after treatment. It can be seen that the existing treatment regimen has poor efficacy in eradicating refractory Helicobacter pylori infection; the third row, Second-linetherapy, shows the effect of using the capsule for treating refractory Helicobacter pylori infection in the present application. It can be seen that the Helicobacter pylori infection indicators of patients became negative after using the capsule for treating refractory Helicobacter pylori infection in the present application. Thus, it can be known that the capsule of the present application can effectively treat and eradicate refractory Helicobacter pylori infection, and the patients are convenient to take the medicine, with good compliance, reducing the incidence of adverse reactions of antibiotics, and at the same time reducing the antibiotic resistance rate.

[0100] Application case 3: Changes in the microecology before and after treatment.

[0101] As Figures 7A - 7C shown, Figures 7A - 7C This is a diagram showing the changes in the gastric microecology of patients with refractory Helicobacter pylori before and after treatment in the embodiments of the present application.

[0102] As Figure 7A , Figure 7B and Figure 7C shown, before and after GMT treatment, the dominant phyla of the microbial community were mainly Bacteroidetes, Bacillota, Pseudomonadota, Actinomycetota, and Fusobacteriota, although there were significant differences in species abundance. Patients with refractory Helicobacter pylori showed multiple independent microbial species before GMT treatment, while several specific species from the donor were obtained after GMT treatment. In addition, the richness of bacterial species at the phylum and genus levels in patients increased after GMT treatment, and the abundance gradually approached that of the bacterial species in the donor group. The changes in the composition of viruses and fungi were relatively small. These results indicate that microbiota transplantation can effectively change the structure of the recipient's gastric microbiota.

[0103] From the above, combining with the diagram showing the changes in the gastric microecology of patients with refractory Helicobacter pylori before and after treatment in Figure 7, it can be seen that the capsule prepared in the embodiments of the present application can effectively eliminate drug-resistant Helicobacter pylori strains, while maintaining the stability of the gastrointestinal microbiota, thereby improving the long-term health prognosis of patients, overcoming problems such as increased drug resistance and disruption of the microecological balance faced by the existing treatment regimen, and effectively treating and eradicating refractory Helicobacter pylori infection.

Claims

1. A preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection, characterized in that, It includes the following steps: S1. Obtain gastric mucus layer rinsing fluid, which contains gastric microorganisms; S2. Load the obtained gastric mucus layer rinsing fluid into an eggplant-shaped flask, connect a low-temperature rotary instrument, and rotate it under low-temperature conditions until the bacterial solution adheres to the wall; S3. Remove the eggplant-shaped flask and install it on a freeze dryer, and perform low-temperature freeze-drying to obtain freeze-dried powder for treating refractory Helicobacter pylori infection.

2. The preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection as described in claim 1, wherein, In step S1, the gastric mucus layer rinsing fluid is obtained by sufficiently rinsing the gastric mucus layer of the donor with an eluent.

3. The preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection according to claim 2, characterized in that, The eluent is prepared by mixing 20000u pronase, 1 g sodium bicarbonate with warm water at 40 °C to 50 ml.

4. The preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection according to claim 1, characterized in that, In step S2, the temperature of low-temperature rotation is -40 °C to -20 °C, and the time is 30 - 40 minutes.

5. The preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection according to claim 1, characterized in that, In step S3, the freeze-drying temperature of the freeze dryer is -40 to -20 °C.

6. The preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection according to claim 1, wherein, In step S3, the freeze-drying time of the freeze dryer is 16 - 20 hours.

7. The preparation method of a freeze-dried powder for treating refractory Helicobacter pylori infection according to claim 1, characterized in that, The freeze-dried powder prepared in step S3 is vacuum-packed.

8. A capsule for treating refractory Helicobacter pylori infection, characterized in that, The capsule is made of freeze-dried powder prepared by the preparation method according to any one of claims 1 - 7.

9. A capsule for treating refractory Helicobacter pylori infection according to claim 8, characterized in that, The capsule is prepared by adding pharmaceutically acceptable excipients to the freeze-dried powder prepared by the preparation method according to any one of claims 1 - 7.

10. Use of a freeze-dried powder for treating refractory Helicobacter pylori infection, characterized in that, The freeze-dried powder is used for preparing a drug for treating refractory Helicobacter pylori infection.

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