Bifidobacterium animalis subsp. lactis BX-186, a strain that alleviates the adverse effects of semaglutide

Abstract

The present invention belongs to the field of biotechnology, and specifically relates to a strain of animal Bifidobacterium lactis subsp. lactis BX‑186 that alleviates the adverse effects of semaglutide. The classification name of the animal Bifidobacterium lactis subsp. lactis provided by the present invention is: Bifidobacterium animalis subsp. lactis, the deposit unit is: General Microbiology Center of China Microbiological Culture Collection Administration Committee, the deposit number is: CGMCC No. 27837, and the deposit time is July 7, 2023. The animal Bifidobacterium lactis subsp. BX‑186 provided by the present invention directly interacts with the intestinal flora, thereby affecting the metabolic activities and physiological functions of the intestinal microorganisms, restoring the balance of the intestinal flora, and can alleviate the gastrointestinal discomfort symptoms caused by semaglutide, including nausea, vomiting and diarrhea, etc., and can also reduce body weight and the content of glycosylated hemoglobin HbA1c in the blood, and has broad application prospects.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and particularly relates to a strain of animal Bifidobacterium lactis subspecies BX-186 capable of alleviating the adverse effects of semaglutide. Background Art

[0002] Semaglutide is a human glucagon-like peptide-1 (GLP-1) analog produced by Saccharomyces cerevisiae cells through genetic recombination technology, which inhibits the release of glucagon, which can increase blood sugar. Studies have found that in addition to regulating blood sugar, the drug can also help the body increase its sensitivity to insulin, increase satiety after meals, slow down gastric emptying, and suppress appetite, thereby reducing weight. In addition, in June 2021, the U.S. FDA officially approved semaglutide for the treatment of simple obesity. Studies have shown that semaglutide 2.4 mg once a week can reduce the weight of non-diabetic simple fat friends by about 15%, and it has good safety. Therefore, semaglutide plays an important role in reducing weight. However, semaglutide can have some side effects on the body, mainly including gastrointestinal reactions: such as nausea and vomiting, and some are also accompanied by diarrhea, constipation and other symptoms.

[0003] Bifidobacterium is a genus of Gram-positive, non-motile, rod-shaped cells that are sometimes forked at one end and are strictly anaerobic. It is widely found in the digestive tract, vagina, oral cavity and other habitats of humans and animals. Bifidobacterium is an important member of the intestinal flora of humans and animals. Animal experiments have found that multiple species of Bifidobacterium (such as Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium animalis) can improve the host's glucose tolerance.

[0004] Chinese invention patent CN116376740A discloses a strain of Bifidobacterium longum with hypoglycemic effect and its application. The Bifidobacterium longum is named WHH2270 and is classified as Bifidobacterium longum subsp Longum in microbial classification. It was deposited in Guangdong Microbiological Culture Collection Center on October 18, 2022, with the preservation address at No. 100, Xianlie Middle Road, Guangzhou City, and the preservation number is GDMCC No: 62900. The Bifidobacterium longum subsp Longum of the invention can increase the abundance of beneficial bacteria in the intestines, increase the content of short-chain fatty acids in the intestines, and has excellent hypoglycemic effect. The strain is isolated from healthy infant feces, has no virulence genes, has no toxic side effects in animal evaluation, and is sensitive to multiple antibiotics. It has certain advantages over traditional drugs currently used to treat lipid metabolism disorders.

[0005] Another Chinese invention patent CN115804454A discloses a probiotic composition, a nutritional composition and its application, which contains Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium animalis, Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus helveticus, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus acidophilus and Streptococcus thermophilus. The probiotic composition of the invention promotes the metabolism of probiotics in the human intestine through the effective compounding of multiple probiotics, prebiotics such as complex polysaccharides, specific types of proteins and other ingredients, which can help inhibit weight gain or even reduce weight, regulate blood sugar and blood lipids, and have the effects of anti-oxidation, anti-aging and improving skin condition.

[0006] Probiotics have been approved for clinical treatment of many diseases. They are generally defined as a class of active microorganisms that can regulate and improve intestinal flora and produce clear benefits to human health when the body consumes sufficient amounts from the diet. However, the prior art does not disclose strains that can alleviate the side effects of semaglutide. Since probiotics themselves have the function of regulating intestinal flora, the effect of probiotics on alleviating the adverse reactions of semaglutide is a possible breakthrough. Summary of the invention

[0007] The present invention aims at the problems existing in the prior art and provides a strain of animal Bifidobacterium lactis subspecies BX-186 which can alleviate the adverse effects of semaglutide and can reduce body weight and the content of glycosylated hemoglobin HbA1c in blood.

[0008] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0009] A strain of Bifidobacterium animalis subsp. lactis BX-186 that alleviates the adverse effects of semaglutide. The Bifidobacterium animalis subsp. lactis BX-186 is classified and named: Bifidobacterium animalis subsp. lactis, and the preservation unit is the General Microbiology Center of China Microbiological Culture Collection Administration, the preservation number is CGMCC No. 27837, the preservation time is July 7, 2023, and the preservation address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences.

[0010] The present invention also provides a product for preventing, alleviating and / or treating the adverse effects of semaglutide, wherein the product comprises animal Bifidobacterium lactis subspecies BX-186 and / or a fermentation metabolite of animal Bifidobacterium lactis subspecies BX-186.

[0011] Preferably, the adverse effects are gastrointestinal discomfort symptoms, including diarrhea, vomiting and nausea.

[0012] The present invention also provides a product for reducing the content of glycosylated hemoglobin in blood, wherein the product comprises animal Bifidobacterium lactis subspecies BX-186 and / or a fermentation metabolite of animal Bifidobacterium lactis subspecies BX-186.

[0013] The present invention also provides a product for reducing body weight, which comprises Bifidobacterium animalis subspecies lactis BX-186 and / or a fermentation metabolite of Bifidobacterium animalis subspecies lactis BX-186.

[0014] Preferably, the daily intake of Bifidobacterium lactis subspecies BX-186 is not less than 1.0×10 6 CFU / d.

[0015] Preferably, the product is a pharmaceutical product.

[0016] Preferably, the medicine comprises pharmaceutical excipients.

[0017] Preferably, the pharmaceutical excipients include at least one of fillers, binders, wetting agents, disintegrants, lubricants and flavoring agents commonly used in pharmacy.

[0018] Preferably, the drug is in the form of tablets, powders, capsules or granules.

[0019] The present invention also provides the use of the above-mentioned animal Bifidobacterium lactis subspecies BX-186 in the preparation of any one of the following products (1)-(3):

[0020] (1) Products that prevent, alleviate and / or treat the adverse effects of semaglutide, wherein the adverse effects are gastrointestinal discomfort symptoms, including diarrhea, vomiting and nausea;

[0021] (2) Products for weight loss;

[0022] (3) Products that reduce the level of glycated hemoglobin in the blood.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The animal Bifidobacterium lactis subspecies BX-186 provided by the present invention directly interacts with the intestinal flora, thereby affecting the metabolic activities and physiological functions of the intestinal microorganisms and restoring the balance of the intestinal flora. It can alleviate the gastrointestinal discomfort symptoms (including nausea, vomiting and diarrhea, etc.) caused by semaglutide, and can also reduce body weight and the content of glycosylated hemoglobin HbA1c in the blood, and has broad application prospects.

[0025] Biological Deposit Description

[0026] A strain of Bifidobacterium animalis subsp. lactis BX-186 that alleviates the adverse effects of semaglutide is classified and named: Bifidobacterium animalis subsp. lactis; the preservation unit: China General Microbiology Center of Microbiological Culture Collection, abbreviated as CGMCC; the preservation number: CGMCC No.27837; the preservation time: July 7, 2023; the preservation address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences; the test result of the strain is survival. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Figure 2 shows the levels of glycated hemoglobin HbA1c in the blood of the four groups of subjects. DETAILED DESCRIPTION

[0028] The present invention is further described below in conjunction with specific examples, and the advantages and features of the present invention will become clearer as the description progresses. However, these examples are exemplary only and do not constitute any limitation to the scope of the present invention. The methods, ingredients and dosages involved in the following embodiments, unless otherwise specified, are conventional methods known to those skilled in the art. It is worth noting that the raw materials used in the present invention are all common commercial products, among which Bifidobacterium animalis subsp. lactis Bb-12 was purchased from Chr. Hansen Co., Ltd.; soy peptone was purchased from Angel Yeast Co., Ltd.; yeast peptone was purchased from Angel Yeast Co., Ltd.; semaglutide was purchased from Shanghai Zaifan Biotechnology Co., Ltd.

[0029] Example 1

[0030] A bacterial powder of animal Bifidobacterium lactis subspecies BX-186 is prepared according to a conventional bacterial powder preparation process, and the preparation process comprises the following steps:

[0031] 1. Activation of strains: The animal Bifidobacterium lactis subsp. BX-186 strain stored at -80°C was inoculated into the corresponding liquid culture medium sterilized at 121°C for 15 minutes, and cultured anaerobically at 37°C for 18-24 hours. The activated strains were obtained by subculturing 1-2 times; animal Bifidobacterium lactis subsp. BX-186 was cultivated using a lactic acid bacteria optimized culture medium.

[0032] 2. Preparation of bacterial solution: Take each bacterial strain activated in step (1) and inoculate it into the optimized lactic acid bacteria culture medium, and culture it anaerobically at 37°C until the pH value reaches 4.5-4.8 to obtain seed solution.

[0033] The composition of the optimized culture medium for lactic acid bacteria is as follows: sucrose 23.5Kg, lactose 12.0Kg, soy peptone 15.0Kg, yeast powder 5.0Kg, yeast peptone 12Kg, Na2HPO3 21.0Kg, citric acid 2.0Kg, MgSO4·7H2O 0.6Kg, MnSO4·5H2O0.3Kg, Tween-80 1.0Kg, L-cysteine ​​hydrochloride 0.3Kg, and distilled water to make up to 1000L.

[0034] 3. Inoculation and fermentation: The seed liquid of step (2) is inoculated into the lactic acid bacteria optimized culture medium prepared in step (2) at 1‰, and fermented for 18 hours under controlled fermentation conditions.

[0035] The fermentation conditions are controlled as follows: in the early stage of fermentation, the temperature is kept constant at 30°C, and the fermentation is carried out naturally until the pH reaches 5.0; then the fermentation temperature is adjusted to 37°C, and the pH is kept constant at 6.0 to maintain anaerobic fermentation. In this step, the pH is kept constant at 6.0 by adding a neutralizer, and the neutralizer is NaOH.

[0036] The anaerobic fermentation conditions are maintained by passing nitrogen through the chamber every two hours.

[0037] 4. Termination of fermentation: When the bacteria stop producing acid, the fermentation is terminated to obtain a high-density fermentation liquid of the strain, and the number of live bacteria in the fermentation liquid reaches 2×10 10 CFU / mL or more. The cessation of acid production is determined by the cessation of pH decrease and the cessation of neutralizing agent addition.

[0038] 5. Freeze drying: a. Bacteria concentration: The high-density fermentation broth is concentrated by centrifugation at 12000g;

[0039] b. Add protective agent: add 5 times the weight of protective agent solution to the bacterial concentrate; the protective agent solution is composed of: skim milk 15Kg, lactose 12Kg, vitamin C 1Kg, sodium glutamate 1Kg, distilled water 1000L;

[0040] c. Drying: The above-mentioned bacterial suspension after adding the protective agent is freeze-dried to obtain freeze-dried bacterial powder, and the bacterial powders of each strain are mixed according to the ratio (ratio of the number of colony forming units) to control the total number of viable bacteria in the mixed bacterial powder to reach 2.0×10 11 CFU / g and above.

[0041] The number of live bacteria in the animal Bifidobacterium lactis subspecies BX-186 bacterial powder prepared in this example reaches 200 billion / g-500 billion / g.

[0042] Comparative Example 1

[0043] A kind of animal Bifidobacterium lactis subspecies Bb-12 bacterial powder, the preparation process of which is the same as that of Example 1, and the number of viable bacteria in the prepared animal Bifidobacterium lactis subspecies Bb-12 bacterial powder reaches 200 billion / g-500 billion / g.

[0044] Experimental example

[0045] 1. Experimental subjects

[0046] 120 adult volunteers were recruited, including 60 males and 60 females, each with a BMI ≥ 27 kg / m 2 or ≥30kg / m 2 All subjects first underwent a 20-week semaglutide run-in period, and only those who tolerated the 2.4 mg dose at week 20 could undergo the randomized trial. Due to intolerance to 2.4 mg semaglutide and some personal reasons, 20 subjects were excluded, and finally 100 subjects participated in the trial.

[0047] 2. Experimental Methods

[0048] 2.1 Grouping

[0049] One hundred volunteers were randomly divided into four groups, each with 25 people. The four groups were as follows: probiotic group: animal Bifidobacterium lactis subsp. BX-186 bacterial powder + semaglutide; comparative group: animal Bifidobacterium lactis subsp. Bb-12 bacterial powder + semaglutide; control group: semaglutide; placebo group: placebo (normal saline).

[0050] 2.2 Administration

[0051] The intake dose of animal Bifidobacterium lactis subspecies BX-186 powder and animal Bifidobacterium lactis subspecies Bb-12 powder was 30 billion CFU / d per person, the injection dose of semaglutide was 32.4 mg / d per person, and the injection dose of placebo was 32.4 mg / d per person.

[0052] 2.3 Index detection

[0053] At baseline and 6 months after the intervention, the subjects in each group were evaluated by questionnaire on body weight and gastrointestinal adverse reactions (nausea, diarrhea, vomiting), and blood samples were collected for determination of glycated hemoglobin to evaluate the regulatory effect of animal Bifidobacterium lactis subsp. lactis BX-186 on the adverse reactions caused by semaglutide.

[0054] 3. Experimental results

[0055] 3.1 Analysis of subject weight

[0056] The weight of the subjects was evaluated at baseline and after 6 months of intervention. The results are shown in Table 1. Compared with the placebo group, the weight of the subjects in the probiotic group, the comparison group and the control group decreased significantly after 6 months of intervention, and the reduction in the probiotic group was higher than that in the other groups. This shows that the intervention of animal Bifidobacterium lactis subspecies BX-186 can reduce weight more effectively.

[0057] Table 1 Weight indicators of the four groups of subjects

[0058]

[0059] 3.2 Analysis of nausea in subjects

[0060] The subjects were asked to conduct a questionnaire analysis on nausea at baseline and after 6 months of intervention. The results are shown in Table 2. After 6 months of intervention, the nausea of ​​the subjects in the probiotic group was significantly improved compared with the control group and the control group, and there was no significant difference with the placebo group. This shows that the intervention of animal Bifidobacterium lactis subspecies BX-186 can alleviate the nausea symptoms caused by the injection of semaglutide.

[0061] Table 2 Nausea indicators of the four groups of subjects

[0062]

[0063]

[0064] 3.3 Analysis of diarrhea in subjects

[0065] The subjects were asked to conduct diarrhea questionnaire analysis at baseline and after 6 months of intervention. The results are shown in Table 3. After 6 months of intervention, the probiotic group had significantly improved diarrhea compared with the control group and the control group, and there was no significant difference with the placebo group. This shows that the intervention of animal Bifidobacterium lactis subspecies BX-186 can alleviate the diarrhea symptoms caused by semaglutide injection.

[0066] Table 3 Diarrhea indicators of the four groups of subjects

[0067]

[0068] 3.4 Analysis of vomiting in subjects

[0069] The subjects were asked to conduct a questionnaire analysis on vomiting at baseline and after 6 months of intervention. The results are shown in Table 4. After 6 months of intervention, the vomiting of the subjects in the probiotic group was significantly improved compared with the control group and the control group, and there was no significant difference with the placebo group. This shows that the intervention of animal Bifidobacterium lactis subspecies BX-186 can effectively alleviate the vomiting symptoms caused by the injection of semaglutide.

[0070] Table 4 Vomiting indicators of the four groups of subjects

[0071]

[0072] 3.5 Analysis of HbA1c Index of Subjects

[0073] Blood samples were collected from the subjects at baseline and after 6 months of intervention. Figure 1 As shown in the figure, after 6 months of intervention, the HbA1c levels in the blood of the subjects in the probiotic group, the comparison group and the control group were significantly lower than those in the placebo group, and the decrease in the probiotic group was the most obvious. This shows that the intervention of animal Bifidobacterium lactis subspecies BX-186 can effectively reduce the HbA1c level in the blood of the subjects.

[0074] In summary, intervention with animal Bifidobacterium lactis subspecies BX-186 can effectively alleviate the gastrointestinal discomfort symptoms (including nausea, vomiting and diarrhea) of subjects caused by semaglutide, and can achieve the effect of reducing body weight and blood glycosylated hemoglobin HbA1c content.

[0075] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A strain of Bifidobacterium animalis subsp. lactis BX-186 for alleviating the adverse effects of semaglutide, characterized in that: The classification name of the animal Bifidobacterium lactis subspecies BX-186 is: Bifidobacterium animalis subsp. lactis, the preservation unit is: General Microbiology Center of China Microbiological Culture Collection Administration, the preservation number is: CGMCC No. 27837, and the preservation time is: July 7, 2023.

2. A product for preventing, alleviating and / or treating the adverse effects of semaglutide, characterized in that: The product comprises the animal Bifidobacterium lactis subspecies BX-186 as described in claim 1.

3. The product according to claim 2, characterized in that The adverse effects are gastrointestinal upset symptoms including diarrhea, vomiting and nausea.

4. The product according to any one of claims 2-3, characterized in that: The product described is a pharmaceutical product.

5. The product according to claim 4, characterized in that The medicine includes pharmaceutical excipients, and the medicine is in the form of tablets, powders, capsules or granules.

6. The product according to claim 5, characterized in that The pharmaceutical excipients include at least one of fillers, binders, wetting agents, disintegrants, lubricants and flavoring agents commonly used in pharmacy.

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