A pharmaceutical composition for treating inflammatory bowel disease and use thereof

Abstract

The application belongs to the field of biological medicine, and particularly relates to a pharmaceutical composition for treating inflammatory bowel disease and application of the pharmaceutical composition. The disclosed pharmaceutical composition comprises anti-TNF monoclonal antibody, phosphoketolase-producing bacteria or a pharmaceutically acceptable viable bacterial preparation or extract thereof. The application comprehensively utilizes multi-dimensional data analysis and experimental verification of'microbe-immune', and finds that the phosphoketolase-producing bacteria can regulate host immune metabolic microenvironment, actively stabilize blood drug concentration of the anti-TNF monoclonal antibody and reduce immunogenicity, so as to achieve the purposes of stabilizing blood drug concentration of the anti-TNF monoclonal antibody and delaying clearance, significantly improve clinical response rate and remission rate, and is particularly suitable for a patient population with primary non-response or secondary non-response to the anti-TNF monoclonal antibody, thereby providing an effective solution for solving this clinical difficulty.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to a pharmaceutical composition for treating inflammatory bowel disease and the application of said pharmaceutical composition. Background Technology

[0002] Inflammatory bowel disease (IBD), primarily comprising two subtypes—Crohn's disease (CD) and ulcerative colitis (UC)—is a group of refractory chronic inflammatory bowel diseases that severely impair patients' quality of life. Currently, treatment options for IBD are increasingly diverse, including aminosalicylic acid derivatives, corticosteroids, immunomodulators, small molecule inhibitors, and various biologics, such as monoclonal antibodies targeting tumor necrosis factor (TNF), interleukin-12 / 23 (IL-12 / 23), or integrins.

[0003] Anti-TNF antibodies are one of the first-line treatment strategies for moderate to severe active IBD. The first anti-TNF antibody, infliximab (IFX), was approved by the US FDA in 1998 for the treatment of Crohn's disease, followed by approval for ulcerative colitis in 2005, and approval by the China National Medical Products Administration (NMPA) for the treatment of Crohn's disease and ulcerative colitis in 2007 and 2013, respectively. Another anti-TNF monoclonal antibody, adalimumab, was approved by the US FDA in 2007 and 2012 for the treatment of Crohn's disease and ulcerative colitis, and by the NMPA in 2010 and 2020, respectively. Despite the significant efficacy of anti-TNF therapy in IBD treatment, approximately 10%–45% of patients experience primary non-response, and approximately 23%–55% experience secondary non-response within 12 months of treatment. Given that a significant proportion of IBD patients fail to achieve clinical remission with existing therapies, exploring the underlying mechanisms and finding strategies to improve the efficacy of anti-TNF therapy is particularly urgent and important.

[0004] The efficacy of anti-TNF monoclonal antibodies is influenced by a complex interplay of multiple factors. Pharmacokineticly, adequate serum drug concentration is fundamental to efficacy, while accelerated drug clearance can lead to insufficient concentration. Immunogenicity-wise, the production of anti-drug antibodies neutralizes drug activity and accelerates its clearance. Host factors include genetic background, obesity, and baseline inflammatory burden; disease characteristics such as IBD subtype, disease duration, severity, and early intervention also significantly affect response. Adequate serum drug concentration is a crucial cornerstone for maintaining drug efficacy. A clinical study published in *The Lancet Gastroenterology & Hepatology* showed that in Crohn's disease patients, low drug concentration at week 14 was the only factor independently associated with primary non-response. This study, through a systematic analysis of patients receiving anti-TNF monoclonal antibody induction therapy, found that even after excluding confounding variables such as anti-drug antibody interference, differences in disease activity, and concomitant medications, insufficient serum drug trough concentration remained the strongest predictor of early treatment failure. This finding further emphasizes the importance of therapeutic drug monitoring (TDM) during the induction phase, suggesting that in clinical practice, dynamic monitoring of blood drug concentrations and timely adjustment of dosage are necessary to optimize initial treatment strategies and improve early response rates.

[0005] Currently, although therapeutic drug monitoring is widely used in clinical practice to assess the blood concentration of anti-TNF monoclonal antibodies and to adjust the dosage accordingly (e.g., increasing the dose or shortening the dosing interval), a fundamental intervention method that can actively, stably, and persistently maintain the target blood drug concentration is still lacking. Existing strategies are mostly reactive adjustments to address low concentrations rather than preventative homeostasis. Whether optimizing dosing regimens, combining immunosuppressants to reduce anti-drug antibody production, or attempting predictive dosing based on pharmacokinetic models, none can completely overcome the significant individual differences in drug metabolism and the fluctuations in clearance rates over time. Therefore, developing novel formulations or adjuvant therapies that can precisely regulate drug exposure and counteract accelerated clearance remains a key unmet clinical need. Summary of the Invention

[0006] The purpose of this invention is to overcome the problems existing in the prior art and provide a pharmaceutical composition that can stabilize the blood concentration of anti-TNF monoclonal antibodies, delay their clearance, and enhance the clinical efficacy of anti-TNF monoclonal antibodies, thereby improving the response rate and remission rate of IBD patients. Simultaneously, this invention also provides the application of the pharmaceutical composition in the preparation of medicaments for treating inflammatory bowel disease.

[0007] To achieve the above objectives, in a first aspect, the present invention provides a pharmaceutical composition for treating inflammatory bowel disease, said pharmaceutical composition comprising an anti-TNF monoclonal antibody, phosphotransketase-producing bacteria, or a pharmaceutically acceptable live bacterial preparation or extract thereof.

[0008] Currently, anti-TNF monoclonal antibody therapy for inflammatory bowel disease (IBD) faces two major clinical challenges. First, maintaining stable blood drug concentrations is difficult. Although therapeutic drug monitoring can guide dose adjustments, existing methods are all passive responses, lacking proactive and sustained intervention methods to maintain effective blood drug concentrations. This leads to primary non-response or secondary loss of response in some patients due to insufficient concentrations. Second, existing efficacy enhancement strategies have limitations. Traditional methods such as combining with immunosuppressants and dose optimization cannot completely overcome individual pharmacokinetic differences and immunogenicity effects, and fail to address the gut microenvironment, a key factor influencing efficacy. Based on this, the inventors of this application discovered in their research that regulating the host immune metabolic microenvironment through specific gut microbiota intervention can stabilize anti-TNF monoclonal antibody blood concentrations, delay their clearance, and ultimately enhance the clinical efficacy of anti-TNF monoclonal antibodies, improving response and remission rates in IBD patients.

[0009] The inventors of this application, by selecting phosphotransketolase-producing bacteria to combine with anti-TNF monoclonal antibodies, discovered that these bacteria can regulate the host's immune metabolic microenvironment, actively stabilize the blood concentration of anti-TNF monoclonal antibodies, and reduce their immunogenicity. This achieves the goal of stabilizing the blood concentration of anti-TNF monoclonal antibodies and delaying their clearance, significantly improving clinical response and remission rates. It is particularly suitable for patients with primary or secondary non-response to anti-TNF monoclonal antibodies, providing an effective solution to this clinical challenge. Moreover, compared to traditional synergistic regimens combining immunosuppressants (such as azathioprine and methotrexate), the proposed regimen uses microbial intervention, which does not cause systemic immunosuppression and significantly reduces the risk of serious adverse reactions such as infection and liver damage.

[0010] In this application, the anti-TNF monoclonal antibody is a monoclonal antibody or its biosimilar capable of inhibiting the biological activity of tumor necrosis factor (TNF). The anti-TNF monoclonal antibody includes, but is not limited to, at least one of infliximab, adalimumab, golimumab, and sertozumab. Preferably, the anti-TNF monoclonal antibody is at least one of infliximab and adalimumab.

[0011] In this application, the bacteria producing phosphotransketase are probiotics, symbiotics, or engineered strains that possess phosphotransketase activity and can regulate the intestinal immune microenvironment through phosphotransketase production. They are naturally occurring strains capable of synthesizing and secreting phosphotransketase. Pharmaceutically acceptable live bacterial preparations of the phosphotransketase-producing bacteria refer to live bacterial products that comply with pharmaceutical manufacturing standards, possess phosphotransketase activity, and can regulate the intestinal immune microenvironment through phosphotransketase production. For example, live bacteria can be protected using techniques such as encapsulation or freeze-drying to resist damage from gastric acid and bile, maintaining their activity after reaching the intestines. The aforementioned phosphotransketase-producing bacterial extract refers to phosphotransketase proteins, metabolites, or bacterial components extracted from probiotics, symbiotics, or engineered strains that possess phosphotransketase activity and can regulate the intestinal immune microenvironment through phosphotransketase production. It does not rely on live bacterial activity, is more stable, and has higher safety, making it suitable for people who are intolerant to live bacteria. Its core activity is phosphotransketase itself or its metabolites (such as short-chain fatty acids), which can directly exert enzymatic regulatory effects.

[0012] Preferably, the bacteria producing phosphotransketase are selected from at least one strain of Bifidobacterium, Roselle, or other bacteria that secrete phosphotransketase and have phosphotransketase activity.

[0013] The *Bifidobacterium* genus includes, but is not limited to, *Bifidobacterium longum*, *Bifidobacterium bifidum*, *Bifidobacterium adolescentis*, and *Bifidobacterium animalis*. The *Roseobacterium* genus includes, but is not limited to, *Roseobacterium faecalis* and *Roseobacterium enterica*. Other bacteria that secrete phosphotransketolase include, but are not limited to, *Lactobacillus* and *Leuconostoc*.

[0014] Specifically, the bacteria producing phosphotransketolase may be, for example, but not limited to, *Bifidobacterium longum*. Specific strains of *Bifidobacterium longum* may include, for example, BB536, etc.

[0015] Addressing the issue of unsatisfactory clinical remission rates in some patients undergoing anti-TNF monoclonal antibody treatment for IBD due to unstable blood drug concentrations and primary or secondary loss of response, the inventors of this application conducted an integrated analysis of the "microbe-host immunity" mechanism using multi-omics high-throughput sequencing technologies (including metagenomics and single-cell transcriptomics). The analysis revealed that phosphotransketase-producing bacteria can delay the clearance of anti-TNF monoclonal antibodies and improve their blood drug concentration stability by regulating host metabolic homeostasis and modulating immune cell function. Further experimental verification showed that, compared to using anti-TNF monoclonal antibodies alone to treat IBD, the combination of anti-TNF monoclonal antibodies and phosphotransketase-producing bacteria significantly improves the clinical remission rate and is expected to reduce the potential adverse reaction risks associated with dose escalation or combination with immunosuppressants.

[0016] Preferably, the number of viable bacteria producing phosphotransketolase in the pharmaceutical composition is not less than 2 × 10⁸.

[0017] The ratio of the anti-TNF monoclonal antibody to phosphotransketolase-producing bacteria is determined based on the clinically effective dose. In one specific embodiment, the anti-TNF monoclonal antibody is administered intravenously or subcutaneously at a standard body weight dose, simultaneously with an oral dose containing 2×10⁻⁶ mg / L of the anti-TNF monoclonal antibody daily. 8 A preparation of live phosphate transketolase-producing bacteria was used for continuous intervention for 14 weeks.

[0018] Secondly, this invention also provides the use of the pharmaceutical composition described above in the preparation of a medicament for treating inflammatory bowel disease (IBD). Addressing the key technical problem of the lack of a targeted method in the prior art for actively maintaining anti-TNF drug blood concentrations by regulating the gut microbiota, thereby achieving synergistic effects, this application provides a synergistic treatment strategy that can maintain anti-TNF monoclonal antibody blood concentrations and significantly improve the clinical response rate of anti-TNF monoclonal antibody treatment for IBD. Specifically, combining anti-TNF monoclonal antibodies with phosphotransferase-producing bacteria or their pharmaceutically acceptable live bacterial preparations or extracts for the treatment of IBD has shown that the combination can exert synergistic anti-inflammatory and immunomodulatory effects, effectively improving the clinical response rate and mucosal healing rate of single-agent anti-TNF monoclonal antibody treatment for IBD.

[0019] Preferably, the inflammatory bowel disease includes Crohn's disease and ulcerative colitis.

[0020] Preferably, the anti-TNF monoclonal antibody in the pharmaceutical composition is an injectable form. The anti-TNF monoclonal antibody in the pharmaceutical composition is preferably an injectable form, administered intravenously or subcutaneously at a standard body weight dose.

[0021] Preferably, the phosphotransketase-producing bacteria or a pharmaceutically acceptable live bacterial preparation or extract thereof in the pharmaceutical composition is an oral formulation. The phosphotransketase-producing bacteria or a pharmaceutically acceptable live bacterial preparation or extract thereof in the pharmaceutical composition is preferably an oral formulation, containing 2 × 10⁻⁶ bacteria orally daily. 8 Preparations of live phosphate transketolase-producing bacteria, when used in conjunction with injection regimens of anti-TNF monoclonal antibodies, do not increase the treatment burden on patients.

[0022] In specific treatment applications, a standard regimen can be used to administer anti-TNF monoclonal antibodies, along with daily oral administration of phosphotransketase-producing bacterial preparations for at least 14 weeks.

[0023] Thirdly, the present invention also provides the use of phosphotransketase-producing bacteria or pharmaceutically acceptable live bacterial preparations or extracts thereof in the preparation of medicaments that enhance the efficacy of anti-TNF monoclonal antibodies, prolong their duration of action, or reduce their loss of response rate.

[0024] The inventors of this application discovered in their research that phosphotransketolase-producing bacteria can regulate the local and systemic immune status of the intestine and stabilize the blood concentration of anti-TNF monoclonal antibodies, thereby synergistically enhancing the efficacy of anti-TNF monoclonal antibodies, prolonging their duration of action, or reducing their loss of response rate. This provides a new drug combination strategy for improving the efficacy of biological agents.

[0025] The inventors of this application, through comprehensive analysis of multidimensional "microbiome-immunity" data and experimental verification, have for the first time elucidated the synergistic mechanism and clinical potential of the combined application of phosphate transketolase-producing bacteria and anti-TNF monoclonal antibodies. This combined strategy not only effectively addresses the issue of insufficient response in some patients when anti-TNF monoclonal antibodies are used to treat IBD, but also provides a new therapeutic paradigm for enhancing the efficacy of biologics by regulating the gut microenvironment, possessing significant clinical application value and development prospects. Specifically, this invention has the following beneficial effects: (1) Traditional strategies to address the inadequacy of anti-TNF monoclonal antibody efficacy mainly rely on passive monitoring and dose adjustment. However, this invention is the first to actively stabilize the blood concentration of anti-TNF monoclonal antibody and reduce its immunogenicity by supplementing specific intestinal bacteria that produce phosphotransketonease, starting from the fundamental approach of regulating the host immune metabolic microenvironment. This provides a new scientific path to enhance the efficacy of biological agents and achieves important innovation in the treatment mechanism.

[0026] (2) Clinical efficacy has been significantly improved. Verification has shown that the combination therapy regimen of this application can effectively delay the clearance of anti-TNF monoclonal antibodies, significantly improving clinical response rate and remission rate. More importantly, the regimen of this application is particularly suitable for patients with spontaneous or secondary non-response to anti-TNF monoclonal antibodies, providing an effective solution to this clinical challenge.

[0027] (3) Compared with traditional synergistic regimens combining immunosuppressants (such as azathioprine and methotrexate), the microbial intervention strategy adopted in this invention does not cause systemic immunosuppression and significantly reduces the risk of serious adverse reactions such as infection and liver damage. Based on the development path of human symbiotic bacteria or recognized safe strains, the biosafety of the treatment regimen is further ensured, and the safety and tolerability are excellent.

[0028] (4) In the pharmaceutical composition of this application, the bacterial preparation can be administered orally and combined with existing injection treatment regimens of anti-TNF monoclonal antibodies without increasing the treatment burden on patients. At the same time, the bacterial strains can be precisely matched according to the individual gut microbiota characteristics, providing an innovative tool for personalized precision medicine of inflammatory bowel disease, which has important clinical value and market development potential, and has a good prospect for clinical translation. Attached Figure Description

[0029] Figure 1This is a comparison chart of changes in body weight and disease activity index in different groups of mice in the DSS-induced enteritis model.

[0030] Figure 2 This is a schematic diagram showing the colon length and histological changes in mice in different groups of the DSS-induced enteritis model.

[0031] Figure 3 This is a comparison of the changes in anti-TNF antibody concentration in the serum of mice in different groups of a DSS-induced enteritis model.

[0032] Figure 4 This is a comparison chart of changes in body weight and disease activity index in different groups of mice in a TNBS-induced enteritis model.

[0033] Figure 5 This is a schematic diagram showing the colon length and histological changes in mice in different groups of a TNBS-induced enteritis model.

[0034] Figure 6 This is a comparison of the changes in anti-TNF antibody concentration in the serum of mice in different groups of a TNBS-induced enteritis model.

[0035] Figure 7 A statistical chart comparing the endoscopic scores of patients in each group.

[0036] Figure 1-3 In the study, NC was the control group, DSS was the model group, and DSS+anti-TNF was the DSS+anti-TNF monoclonal antibody group. B.longum The DSS+ Bifidobacterium longum group, DSS+ anti-TNF+ B.longum The group consisted of DSS, anti-TNF monoclonal antibody, and Bifidobacterium longum.

[0037] Figure 4-6 In the study, NC served as the control group, TNBS as the model group, and TNBS+anti-TNF as the TNBS+anti-TNF monoclonal antibody group. B.longum The TNBS+ Bifidobacterium longum group, TNBS+ anti-TNF+ B.longum The group consisted of TNBS + anti-TNF monoclonal antibody + Bifidobacterium longum.

[0038] Figure 7 The left image shows a comparison of endoscopic scores before and after treatment in patients with ulcerative colitis, where IFX is the control group (infliximab + placebo) and IFX+LBC is the combination therapy group (infliximab + bacterial agent); the right image shows a comparison of endoscopic scores before and after treatment in patients with Crohn's disease, where ADA is the control group (adalimumab + placebo) and ADA+LBC is the combination therapy group (adalimumab + bacterial agent). Detailed Implementation

[0039] The technical solution and the technical effects achieved by the present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0040] The *Bifidobacterium longum* used in the following examples is *Bifidobacterium longum* (ATCC 15707), obtained from the U.S. Type Culture Collection. Unless otherwise specified, all other materials and equipment used are common in the art and can be prepared using conventional methods or purchased directly.

[0041] Example 1 Animal experiments on the treatment of inflammatory bowel disease with the pharmaceutical composition of the present invention. This embodiment uses an animal model experiment to examine the efficacy of combined treatment of phosphotransketolase-producing bacteria and anti-TNF monoclonal antibody in the treatment of inflammatory bowel disease, specifically including the following steps; 1. Animal model making Sixty 5-week-old male C57BL / 6 mice were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd. All mice were housed in an SPF-grade environment at the Animal Center of Renji Hospital, affiliated with Shanghai Jiao Tong University School of Medicine. A chronic colitis model induced by sodium dextran sulfate (DSS) was used in the experiment. The modeling method was as follows: mice were allowed free access to drinking water containing 3% DSS (molecular weight 36,000-50,000 Da) for 7 days, followed by 2 days of normal drinking water.

[0042] Mice in the anti-TNF monoclonal antibody treatment group were administered the antibody on day 0. The anti-TNF monoclonal antibody was administered intraperitoneally at a dose of 5 mg / kg, given only once. Mice in the phosphotransferase-producing bacterial intervention group were administered the antibody daily by gavage containing 1×10⁻⁶ mg / kg starting 7 days prior to the start of the experiment. 9 A 200 μL suspension of a specific CFU strain (Bifidobacterium longum was used in this example) was administered. Colonic histology was examined on day 9 after the start of DSS intervention. The animal experiment was approved by the Ethics Committee of Renji Hospital, with protocol number 21-E-05.

[0043] 2. Experimental Grouping Sixty mice were randomly divided into the following 6 groups, with 10 mice in each group: (1) Control group: fed with normal drinking water, no modeling was performed; (2) Model group (DSS): DSS induced colitis, and an equal volume of physiological saline was administered; (3) DSS + Bifidobacterium longum group: DSS induced colitis, and Bifidobacterium longum was administered by gavage daily; (4) DSS + anti-TNF monoclonal antibody group: DSS induces colitis, and anti-TNF monoclonal antibody is injected intraperitoneally on day 0; (5) DSS + anti-TNF monoclonal antibody + Bifidobacterium longum group: DSS induced colitis, combined with anti-TNF monoclonal antibody treatment and Bifidobacterium longum gavage.

[0044] 3. Experimental detection indicators (1) Disease Activity Index (DAI) score: Record daily changes in weight, stool characteristics and blood in stool, and calculate the DAI score.

[0045] (2) Colon length measurement: At the end of the experiment, the mice were anesthetized and euthanized. The colon was dissected and removed. It was gently straightened along the mesenteric axis, rinsed with physiological saline and excess tissue was removed. The total length of the colon from the cecum to the rectum was measured using a ruler and the data was recorded to assess the degree of colon damage.

[0046] (3) Histopathological evaluation of colon tissue: Colon tissue was taken at the end of the experiment, fixed in formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE). The degree of inflammation, crypt structure destruction, and ulcer extent were assessed using a histological damage scoring system.

[0047] (4) Serum drug concentration detection: Serum of mice in each group was collected at 1, 3, 5, 7 and 9 days after administration of anti-TNF monoclonal antibody, and the serum concentration of anti-TNF monoclonal antibody was determined by ELISA.

[0048] 4. Experimental Results (1) Changes in body weight and disease activity index of mice in each group are shown in the attached figure. Figure 1 As shown, by appendix Figure 1 It can be seen that the combination therapy significantly improved disease activity: the DSS + anti-TNF monoclonal antibody + Bifidobacterium longum group had lower DAI scores and less weight loss.

[0049] (2) The colon length and histological changes of mice in each group are shown in the attached figure. Figure 2 As shown, by appendix Figure 2 It can be seen that the combined treatment group showed significant pathological improvement: the combined treatment group had a longer colon, a significantly lower colon tissue pathological score, a basically restored mucosal structure, and a significant reduction in inflammatory cell infiltration.

[0050] (3) The changes in the concentration of anti-TNF antibody in the serum of mice in each group are shown in the attached figure. Figure 3 As shown, by appendix Figure 3 It can be seen that the blood drug concentration in the combined treatment group remained more stable: the concentration of anti-TNF monoclonal antibody in the serum of mice in the combined treatment group was higher.

[0051] Example 2 Another animal experiment on the treatment of inflammatory bowel disease with the pharmaceutical composition of the present invention. This embodiment uses another animal model experiment to examine the efficacy of combined treatment of phosphotransketase-producing bacteria and anti-TNF monoclonal antibody for inflammatory bowel disease, specifically including the following steps: 1. Animal model making Sixty 5-week-old male C57BL / 6 mice were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.

[0052] All mice were housed in an SPF-grade environment at the Animal Center of Renji Hospital, affiliated with Shanghai Jiao Tong University School of Medicine. An acute colitis model induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS) was used in the experiment.

[0053] The modeling method was as follows: 150 μL of TNBS sensitization solution was applied to the back of shaved mice for immunization. Seven days later, 100 μL of 2.5% (w / v) TNBS containing 50% ethanol was administered rectally via a catheter at a depth of 4 cm.

[0054] Mice in the anti-TNF monoclonal antibody treatment group were administered the antibody on day 0. The anti-TNF monoclonal antibody was administered via intraperitoneal injection at a dose of 5 mg / kg, once only.

[0055] Mice in the TNBS-producing bacterial intervention group were administered 200 μL of a bacterial suspension containing 1 × 10^9 CFU of a specific strain (Bifidobacterium longum was used in this example) daily by gavage for 7 days prior to the start of the experiment. Colonic histological examination was performed on day 4 after the start of the TNBS intervention. The animal experiment was approved by the Ethics Committee of Renji Hospital, and the experimental protocol number is 21-E-05.

[0056] 2. Experimental Grouping Sixty mice were randomly divided into the following 6 groups, with 10 mice in each group: (1) Control group: fed with normal drinking water, no modeling was performed; (2) Model group (TNBS): TNBS induced colitis, and an equal volume of physiological saline was administered; (3) TNBS + Bifidobacterium longum group: TNBS induced colitis, and Bifidobacterium longum was administered by gavage daily; (4) TNBS + anti-TNF monoclonal antibody group: TNBS induced colitis, and anti-TNF monoclonal antibody was injected intraperitoneally on day 0; (5) TNBS + anti-TNF monoclonal antibody + Bifidobacterium longum group: TNBS-induced colitis, combined with anti-TNF monoclonal antibody treatment and Bifidobacterium longum gavage; 3. Experimental detection indicators (1) Colon length measurement: At the end of the experiment, the mice were anesthetized and euthanized, and the colon was dissected and removed. The colon was gently straightened along the mesenteric axis, rinsed with physiological saline and excess tissue was removed. The total length of the colon from the cecum to the rectum was measured using a ruler and the data was recorded to assess the degree of colon damage.

[0057] (2) Histopathological evaluation of colon tissue: Colon tissue was taken at the end of the experiment, fixed in formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE). The degree of inflammation, crypt structure destruction, and ulcer extent were assessed using a histological damage scoring system.

[0058] (3) Serum drug concentration detection: Serum of mice in each group was collected 1 and 4 days after administration of anti-TNF monoclonal antibody, and the serum concentration of anti-TNF monoclonal antibody was determined by ELISA.

[0059] 4. Experimental Results (1) Changes in body weight and disease activity index of mice in each group are shown in the attached figure. Figure 4 As shown, by appendix Figure 4 It can be seen that the combination therapy significantly improved disease activity: the TNBS + anti-TNF monoclonal antibody + Bifidobacterium longum group experienced less weight loss.

[0060] (2) The colon length and histological changes of mice in each group are shown in the attached figure. Figure 5 As shown, by appendix Figure 5 It can be seen that the combined treatment group showed significant pathological improvement: the combined treatment group had a longer colon, a significantly lower colon tissue pathological score, a basically restored mucosal structure, and a significant reduction in inflammatory cell infiltration.

[0061] (3) The changes in the concentration of anti-TNF antibody in the serum of mice in each group are shown in the attached figure. Figure 6 As shown, by appendix Figure 6 It can be seen that the blood drug concentration in the combined treatment group remained more stable: the concentration of anti-TNF monoclonal antibody in the serum of mice in the combined treatment group was higher.

[0062] Example 3 Clinical trials of the pharmaceutical composition of the present invention for the treatment of inflammatory bowel disease In this embodiment, a randomized controlled clinical trial was conducted using the pharmaceutical composition described in this invention, specifically including the following steps: 1. Study Design and Patient Grouping Based on the Department of Gastroenterology at Renji Hospital affiliated with Shanghai Jiao Tong University School of Medicine, the inventors of this application conducted a single-center, randomized, double-blind, placebo-controlled clinical study on the effect of phosphate transketolase-producing bacterial preparations on the efficacy of anti-TNF monoclonal antibodies in patients with inflammatory bowel disease (Chinese Clinical Trial Registry No.: ChiCTR2500108246).

[0063] This study recruited 40 patients with moderate to severe active inflammatory bowel disease, including 20 patients with ulcerative colitis and 20 patients with Crohn's disease, who were randomly assigned to two groups in a 1:1 ratio: Ulcerative colitis: Combination therapy group (infliximab + bacterial agent): 10 patients received standardized infliximab induction therapy (5 mg / kg intravenously at weeks 0, 2, and 6), along with daily oral administration of phosphate transketolase-producing bacterial capsules (5 × 10 mg / kg per capsule). 8 Live bacteria), the phosphate transketolase-producing bacteria being Bifidobacterium longum, 2 tablets twice daily for 14 consecutive weeks.

[0064] Control group (IFX + placebo): 10 patients received the same regimen of infliximab treatment without oral phosphotransketase-producing bacterial capsules.

[0065] Crohn's disease: Combination therapy group (adalimumab + bacterial agent): 10 patients received standard adalimumab induction therapy (initial 160 mg, week 2 80 mg, then 40 mg subcutaneously every 2 weeks), and daily oral administration of phosphate transketolase-producing bacterial capsules (5 × 10 capsules per capsule). 8 Live bacteria), the phosphate transketolase-producing bacteria being Bifidobacterium longum, 2 tablets twice daily for 14 consecutive weeks.

[0066] Control group (ADA + placebo): 10 patients received the same regimen of adalimumab treatment but did not take oral phosphotransketase-producing bacterial capsules.

[0067] 2. Efficacy evaluation and experimental results The primary endpoint for patients with ulcerative colitis was: modified Mayo score ≤2, bowel frequency score ≤1 and not higher than baseline, and rectal bleeding score =0.

[0068] The primary endpoint for Crohn's disease patients was a CDAI score below 150.

[0069] The endoscopic scores of patients in each group were calculated, as shown in the attached figure. Figure 7 As shown. (From the appendix) Figure 7 It can be seen that the endoscopic score of the combined treatment group at week 14 was significantly lower than that of the control group.

[0070] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A pharmaceutical composition for treating inflammatory bowel disease, characterized in that, The pharmaceutical composition comprises an anti-TNF monoclonal antibody, a phosphotransketase-producing bacterium, or a pharmaceutically acceptable live bacterial preparation or extract thereof.

2. The pharmaceutical composition for treating inflammatory bowel disease as described in claim 1, characterized in that, The anti-TNF monoclonal antibody includes at least one of infliximab, adalimumab, golimumab, and sertozumab.

3. The pharmaceutical composition for treating inflammatory bowel disease as described in claim 1, characterized in that, The bacteria that produce phosphotransketonease are probiotics, symbiotics, or engineered strains that have phosphotransketonease activity and can regulate the intestinal immune microenvironment by producing phosphotransketonease.

4. The pharmaceutical composition for treating inflammatory bowel disease as described in claim 1 or 3, characterized in that, The bacteria that produce phosphotransketase are selected from at least one strain of Bifidobacterium, Roselle, or other bacteria that secrete phosphotransketase and have phosphotransketase activity.

5. The pharmaceutical composition for treating inflammatory bowel disease as described in claim 4, characterized in that, The bacteria that produce phosphotransketolase are Bifidobacterium longum.

6. The pharmaceutical composition for treating inflammatory bowel disease as described in claim 1, characterized in that, The number of viable bacteria producing phosphotransketonease in the pharmaceutical composition is not less than 2 × 10⁻⁶. 8 indivual.

7. Use of the pharmaceutical composition according to any one of claims 1-6 in the preparation of a medicament for treating inflammatory bowel disease.

8. The use of the pharmaceutical composition of claim 7 in the preparation of a medicament for treating inflammatory bowel disease, characterized in that, The inflammatory bowel disease mentioned includes Crohn's disease and ulcerative colitis.

9. The use of the pharmaceutical composition of claim 7 in the preparation of a medicament for treating inflammatory bowel disease, characterized in that, The anti-TNF monoclonal antibody in the pharmaceutical composition is an injectable formulation. And / or, the phosphotransketase-producing bacteria in the pharmaceutical composition or a pharmaceutically acceptable live bacterial preparation or extract thereof is an oral preparation.

10. Use of phosphotransketolase-producing bacteria or pharmaceutically acceptable live bacterial preparations or extracts thereof in the preparation of drugs that enhance the efficacy of anti-TNF monoclonal antibodies, prolong their duration of action, or reduce their loss of response rate.

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