Application of bendamustine combined with rituximab in the preparation of drugs for the treatment of DLBCL

Abstract

The present invention provides the use of bendamustine combined with rituximab in preparing a drug for regulating the body's immune ability, in particular, in preparing an anti-tumor drug for treating DLBCL combined with symptoms of heart failure.

Description

[0001] Application technology

[0002] The present invention relates to the technical field of anti-tumor compositions, and in particular to a composition for treating patients with high-tumor-loaded DLBCL combined with symptoms of heart failure. Background Art

[0003] Diffuse large B cell lymphoma (DLBCL) is the most common aggressive non-Hodgkin's B cell lymphoma, accounting for approximately 30%-40% of all non-Hodgkin's lymphomas. Lymphoma cells can originate from different differentiation and maturation stages of B lymphocytes. According to the source stage of tumor cells, they can be divided into germinal center (GCB) and non-germinal center (non-GCB) sources. Before the application of CD20 monoclonal antibody, CHOP regimen (cyclophosphamide + doxorubicin + vincristine + prednisone) was the classic treatment for DLBCL. With the application of CD20 monoclonal antibody rituximab (R), a large number of large-scale clinical studies have confirmed that R-CHOP regimen significantly improves the survival of patients compared with CHOP regimen alone. R-CHOP has become the standard treatment for various molecular subtypes. However, in actual clinical work, some patients cannot tolerate the CHOP regimen due to heart failure, myocardial damage, etc. In this case, medication should be used with caution. Secondly, some DLBCL patients have a high tumor burden at the onset of the disease, and the risk of tumor lysis syndrome caused by the use of the R-CHOP regimen in the first course of treatment is relatively high. Therefore, there is an urgent need to find an effective treatment option to replace the classic R-CHOP therapy.

[0004] At present, bendamustine is widely used in CLL and indolent lymphoma at home and abroad. The literature has clearly reported that in the treatment of CLL patients, the BR regimen (bendamustine + rituximab) is not inferior to the FCR regimen (fludarabine + cyclophosphamide + rituximab), and has lower toxicity. The BR regimen is more effective for patients over 65 years old. In addition, actual clinical work often faces patients with high tumor load, such as large abdominal masses, multiple tumor lesions throughout the body, etc. The R-CHOP regimen for the treatment of such patients leads to a high incidence of tumor syndrome and high treatment-related mortality. Some patients also have symptoms of heart failure. At present, there is still room for optimization and improvement in the treatment of patients with high tumor load DLBCL with symptoms of heart failure. Based on the characteristics of low toxicity and side effects of BR chemotherapy, it can be used as the first course of treatment for patients with high tumor load DLBCL with symptoms of heart failure, providing a guarantee for subsequent treatment. Summary of the invention

[0005] In view of the problems existing in the prior art, the present invention first provides a composition for regulating the body's immune ability, characterized in that the composition comprises the following components: rituximab and bendamustine.

[0006] Furthermore, the so-called regulation of the body's immune ability is to induce immunogenic death of lymphoma cells, upregulate pro-inflammatory factors, form an immune-hot tumor microenvironment, and promote the recruitment of immune cells.

[0007] Furthermore, the improvement of the body's immune capacity is manifested as the treatment of tumor symptoms.

[0008] Furthermore, the reported tumor symptoms are in a high tumor burden state.

[0009] Furthermore, the reported tumor symptoms are also combined with symptoms of heart failure or myocardial damage.

[0010] Furthermore, the tumor symptoms reported are a high tumor burden state combined with symptoms of heart failure or myocardial damage.

[0011] Furthermore, the composition also includes other cytotoxic agents.

[0012] In a preferred embodiment, the cytotoxic agent comprises gefitinib, erlotinib, afatinib, osimertinib, carboplatin, paclitaxel, gemcitabine or an anti-tumor antibody.

[0013] Furthermore, the composition also includes the addition of human serum containing complement.

[0014] Furthermore, the composition also includes a tumor stem cell differentiation agent.

[0015] In a preferred embodiment, the tumor stem cell differentiation agent is selected from: retinoic acid, BRD7552, noggin, niacinamide, dexamethasone, and cytarabine.

[0016] Furthermore, the composition also contains a pharmaceutically acceptable carrier or excipient.

[0017] In a preferred embodiment, the pharmaceutically acceptable carrier or excipient includes one or more of starch, dextrin, microcrystalline cellulose, inorganic salts and powdered sugar.

[0018] On the other hand, the present invention also provides the use of rituximab and bendamustine in the combined preparation of a drug for regulating the body's immune ability.

[0019] Furthermore, the so-called regulation of the body's immune ability is to induce immunogenic death of lymphoma cells, upregulate pro-inflammatory factors, form an immune-hot tumor microenvironment, and promote the recruitment of immune cells.

[0020] Furthermore, the regulation of the body's immune ability is manifested as the treatment of tumor symptoms.

[0021] Furthermore, the reported tumor symptoms are in a high tumor burden state.

[0022] Furthermore, the reported tumor symptoms are also combined with symptoms of heart failure or myocardial damage.

[0023] Furthermore, the tumor symptoms reported are a high tumor burden state combined with symptoms of heart failure or myocardial damage.

[0024] Furthermore, the tumor symptoms reported are non-Hodgkin's lymphoma.

[0025] Furthermore, the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.

[0026] Furthermore, the drug also includes other cytotoxic agents.

[0027] In a preferred embodiment, the cytotoxic agent comprises gefitinib, erlotinib, afatinib, osimertinib, carboplatin, paclitaxel, gemcitabine or an anti-tumor antibody or complement.

[0028] When human serum containing complement is added, the tumor killing rate of the drug is increased by about 30%.

[0029] Furthermore, the drug also includes a tumor stem cell differentiation agent.

[0030] In a preferred embodiment, the tumor stem cell differentiation agent is selected from: retinoic acid, BRD7552, noggin, niacinamide, dexamethasone, and cytarabine.

[0031] Furthermore, the drug also contains a pharmaceutically acceptable carrier or excipient.

[0032] In a preferred embodiment, the pharmaceutically acceptable carrier or excipient includes one or more of starch, dextrin, microcrystalline cellulose, inorganic salts and powdered sugar.

[0033] The compounds, compositions and applications provided by the embodiments of the present invention have the following beneficial effects:

[0034] 1. A new composition is provided, which can improve the body's immune ability and treat diffuse large B-cell lymphoma in a state of high tumor load and combined with symptoms of heart failure or myocardial damage.

[0035] 2. After adding human serum containing complement, the killing rate of the combination on tumors can be increased by about 30%, with a significant synergistic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The method of the present invention and its beneficial effects are described in detail below in conjunction with the accompanying drawings and specific implementation methods.

[0037] Figure 1 PET-CT evaluated the efficacy of the BR regimen. After 4 courses of BR treatment, PET-CT images of the three patients showed that the high-metabolic tumor lesions were inactivated, and the clinical evaluation showed that the disease was completely relieved.

[0038] Figure 2 :In vitro experiments verified that the BR regimen has a synergistic tumor killing effectA:Database sequencing analysis found that OCI-LY1, SU-DHL2 and KARPAS-422 cell lines expressed CD20.B,C:Rituximab and bendamustine were treated with three lymphoma cell lines for 12 hours respectively, and the proportion of PI and AnnexinV double-negative cells was detected by flow cytometry. Rituximab 5uM achieved the maximum tumor cell killing effect; bendamustine had a significant concentration dependence;D:Rituximab (10μM), bendamustine (200μM) and the combination of the two drugs were used to treat OCI-LY1 cell lines, and the proportion of PI and AnnexinV double-negative cells in each group was detected by flow cytometry. The cell survival rate of the combination of the two drugs was the lowest, with statistical significance.

[0039] Figure 3 : BR regimen induces tumor cell apoptosis and inhibits cell proliferation. A: Western-Blot detected that after 36 hours of BR treatment, the anti-apoptotic protein Bcl-2 decreased, and pro-apoptotic proteins such as Caspase3, Caspase8 and cleaved-Caspase8 increased significantly; B: GSEA analysis showed that the BR regimen blocked cell differentiation from G1 phase to S phase; C: Flow cytometry detected that the number of S phase cells decreased significantly after BR treatment; D, E: PCR and WB detected that the cell cycle-related CDKN1A gene encoding P21 protein increased significantly after BR treatment; F: The volcano plot results showed that compared with the control group, the expression of genes such as CDKN1A, STING, CXCL10, and IFNB1 were upregulated after BR treatment.

[0040] Figure 4 : BR regimen induces late pyroptosis in lymphoma cells A: Flow cytometry showed that PI+ cells increased significantly after 12h and 36h of drug treatment, especially after 36h of treatment, and the most obvious was in the BR combined treatment group; B: WB detected the protein levels of pyroptosis-specific genes GSDMD and CASP1, and the expression levels in the BR group were significantly increased; C: RNA sequencing showed that the expression of pyroptosis-related genes increased significantly after BR treatment for 36h. D: PCR detected the expression levels of mRNA of pyroptosis-specific genes GSDMD, CASP1, and NLRP1, and the BR group was significantly higher than the control group.

[0041] Figure 5 : BR regimen activates cGAS-STING signaling pathwayA: RNAseq analysis showed that the expression of genes related to the cGAS-STING signaling pathway was upregulated after BR treatment; B: GSEA analysis showed that dsDNA and type I interferon response pathways were significantly enhanced after BR treatment; C: Cluster analysis showed that the mechanism of action of the BR regimen may focus on the cGAS-STING signaling pathway; D: Analysis of the DLBCL patient database found that patients with high expression of STING had significantly better long-term production than those with low STING expression; E, F: PCR and WB verified that the mRNA and protein levels of cGAS, STING and IFNB1 were significantly increased after BR treatment.

[0042] Figure 6 :cGAS-STING is closely related to cell pyroptosisA: Analysis of the GDC-DLBCL-2018 database found that pyroptosis-related genes: GSDMD, NLRP1, and CASP1 were positively correlated with STING; B: Single-cell sequencing showed that the STING gene was mainly expressed in tumor cells and B lymphocytes, and was highly consistent with the expression distribution of pyroptosis genes; C, D: The ability of the BR regimen to kill tumor cells was significantly reduced by knocking out the STING gene using shRNA technology; E: Flow cytometry detection of PI+ cells found that the STING knockout group showed a significant decrease, indicating that cell pyroptosis depends on the STING gene; F: WB detection found that after the cells were treated with the BR regimen, the STING genome was knocked out, and the expression of pyroptosis- and apoptosis-related proteins was significantly reduced.

[0043] Figure 7 :cGAS-STING is closely related to cell pyroptosisA: Normal human serum (NHS) promotes BR to kill tumor cells, while serum with inactivated complement (ICNHS) has no synergistic effect;B: RNAseq analysis shows that after BR treatment, tumor cells produce more inflammatory chemokines and receptors;C, GDC-DLBCL-2018 database cohort weighted, WGCNA analysis results;D, pathway enrichment analysis of two co-expression modules related to cGAS-STING;E: The results of two co-expression modules related to cGAS-STING enriched in the TNF pathway;F: DLBCL single-cell sequencing results show that TNF mainly comes from tumor cells;G, H: Flow cytometry detection found that the expression level of TNFα was significantly increased after BR treatment;I: PCR detection detected that the expression level of TNF mRNA was significantly increased after BR treatment.

[0044] Figure 8:cGAS-STING signaling pathway promotes tumor immunogenic deathA: TCGA database analysis of DLBCL patients showed that patients with high STING expression had more abundant T cell infiltration in tumor tissues than patients with low STING expression;B: Transwell chamber experiments showed that BR regimen treated lymphoma cells, which could induce T lymphocytes in the upper chamber to shuttle from the lower chamber, indicating that BR regimen participated in tumor killing by recruiting T lymphocytes;C: IFN-γ detection of tumor supernatant, tumor cells and T lymphocytes co-culture system was established respectively, and the results showed that T cells could be more activated to secrete INF-γ in the co-culture system of tumor cells and T lymphocytes;D: IFN-γ detection of tumor supernatant, tumor cells and T lymphocytes co-culture system was established respectively, and the results showed that T cells could be more activated to secrete INF-γ in the co-culture system of tumor cells and T lymphocytes;E: RNAseq analysis showed that tumor cells highly expressed MHC class I and class II antigens after BR treatment;F, G, H, I: Flow cytometry detection of tumor cells expressing MHC after BR treatment Class I and class II molecules were significantly elevated. DETAILED DESCRIPTION

[0045] The technical scheme in the embodiment of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiment of the present invention. Obviously, the described embodiment is only a part of the embodiment of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention. Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention in this specification are only for the purpose of describing specific embodiments and are not used to limit the present invention. The experimental methods in the following embodiments are conventional methods unless otherwise specified. The experimental materials in the following embodiments are purchased from conventional biochemical reagent stores unless otherwise specified.

[0046] Example 1 Bendamustine combined with rituximab (BR) regimen directly kills lymphoma cells

[0047] 1.1BR regimen can directly induce tumor cell apoptosis and inhibit tumor cell cycle, thereby directly killing DLBCL cells

[0048] Experimental steps: According to the previous experiments, we selected rituximab 10μM and bendamustine 200μM as the drug concentrations for subsequent experiments. The experimental groups were: normal saline control group (C), rituximab group (R), bendamustine group (B), BR combined drug group (BR).

[0049] (1) The above-mentioned groups were treated with drugs for 12 h and 36 h respectively, and the cell cycle changes were detected by flow cytometry; CCK8 kit was used to detect cell proliferation changes;

[0050] (2) Collect cells from each group, and use qRT-PCR and Western Blot to detect the expression of related proteins such as Bcl-2, Caspase-3, Caspase-8, and GSDMD;

[0051] We selected three cell lines, OCI-LY1, KARPAS-422 and SU-DHL2, which highly expressed CD20 ( Figure 2 A) Human DLBCL cell lines were used in our preliminary experiments. Rituximab has a certain direct effect of inducing cell apoptosis, but it is not concentration-dependent, while bendamustine kills tumor cells in a concentration-dependent manner. The BR regimen of the two drugs combined has a significant synergistic effect of killing tumor cells ( Figure 2 BD), accompanied by a decrease in the anti-apoptotic protein Bcl-2 and a significant increase in pro-apoptotic proteins such as Caspase3, Caspase8 and cleaved-Caspase8 ( Figure 3 A), indicating that the BR regimen has the effect of inducing tumor cell apoptosis. This is because the BR regimen inhibits the progression of lymphoma cells from the G1 phase to the S phase ( Figure 3 B, 3C), and the expression of the cell cycle-related CDKN1A gene encoding the P21 protein was significantly increased, further confirming this hypothesis ( Figure 3 DF). The above experimental results clearly show that the BR regimen can directly induce tumor cell apoptosis and inhibit tumor cell cycle, thereby directly killing DLBCL cells.

[0052] 1.2BR regimen induces cell pyroptosis and promotes tumor killing

[0053] Experimental steps: We selected rituximab 10μM and bendamustine 200μM as the drug concentrations for subsequent experiments. The experimental groups were: saline control group (C), rituximab group (R), bendamustine group (B), and BR combined drug group (BR).

[0054] (1) After drug treatment for 12 h and 36 h, flow cytometry was used to detect the changes in the proportion of early and late apoptosis of cells; CCK8 method was also used to detect changes in cell proliferation;

[0055] (2) Collect cells from each group, and use qRT-PCR and Western-Blot to detect the expression of related proteins such as Bcl-2, Caspase-3, Caspase-8, and GSDMD;

[0056] Pyroptosis, also known as inflammatory cell death, is different from apoptosis and necrosis. It is a programmed cell death that depends on GSDMD family proteins and Caspase, and is usually triggered by changes in internal and external environmental factors of the cell. It manifests as continuous cell swelling until the cell membrane ruptures, leading to the release of cell contents and activating a strong inflammatory response. It is an important natural immune response of the body. By comparing BR treatment at different times, it was found that long-term BR treatment led to PI + A large number of cells appear, which is also an important feature of cell pyroptosis ( Figure 4 A). Our further experiments found that after BR treatment of lymphoma cells, the RNA and protein expression levels of pyroptosis-specific genes, such as GSDMD and CASP1, were significantly increased (4B-D). It was confirmed that the BR regimen induced cell apoptosis in the early stage and caused cell pyroptosis in the later stage.

[0057] 1.3BR regimen activates cGAS-STING signaling pathway to kill tumor cells

[0058] Experimental steps: (1) Taking advantage of the fact that mRNA of higher organisms all have a Poly(A) tail, magnetic beads with Poly(T) probes are used to hybridize with total RNA. (2) The Poly(T) probe binds to the mRNA with a Poly(A) tail. Next, the magnetic beads are recovered, and the mRNA with Poly(A) is eluted from the magnetic beads. The eluted mRNA is then treated with a magnesium ion solution. (3) The magnesium ion solution will break the mRNA, and the broken mRNA fragments are reverse transcribed using random primers. (4) After reverse transcription into (first-strand) cDNA, the second-strand (cDNA) is synthesized. This creates a double-stranded cDNA. (5) A "Y"-shaped adapter is then added to both ends of the double-stranded cDNA to form a standard sequencing library, which is then sequenced on the HiSeq sequencer.

[0059] In order to explore the molecular mechanism of BR regimen in killing lymphoma, we performed RNAseq sequencing. The results showed that after BR treatment, the expression of genes related to the cGAS-STING signaling pathway was significantly upregulated ( Figure 5 A). Signaling pathway cluster analysis showed that the BR regimen significantly enhanced dsDNA and type I interferon response pathways as well as pattern recognition receptor pathways ( Figure 5B, 5C). Currently, a large number of literatures have reported that the cGAS-STING signaling pathway is closely related to tumors. The intracellular DNA receptor cyclic GMP-AMP synthase (cGAS) is an important indicator for detecting abnormal DNA fragments in the cytoplasm. Once DNA abnormalities are detected, cGAS transduces signals to STING, thereby initiating downstream signal transduction, ultimately leading to the production of IFN-I and the expression of other immune-related genes [15,16]. Our preliminary study analyzed the database information and found that the long-term survival (OS) of DLBCL patients with high STING expression was significantly better than that of patients with low STING expression ( Figure 5 D). Therefore, we speculate that the killing of lymphoma cells by the BR regimen may be related to the cGAS-STING signaling pathway. In addition, RNA sequencing results showed that genes such as cGAS, STING, and INFB1 were significantly upregulated after BR treatment, and this was verified at the mRNA and protein levels ( Figure 5 EF), further suggesting BR may activate cGAS-STING signaling The mitochondrial pathway kills DLBCL cells.

[0060] To determine the possibility that BR regimen kills lymphoma cells by activating the cGAS-STING signaling pathway, our analysis of RNA-seq data from GDC-DLBCL-2018 diffuse large B-cell lymphoma showed that the expression of pyroptosis-related genes was positively correlated with STING1 ( Figure 6 A). Single-cell sequencing results also showed that the subpopulation of DLBCL tumor cells expressing high STING has a high pyroptosis state ( Figure 6 B). In order to clarify whether the STING gene actually mediates the apoptosis and pyroptosis of tumor cells promoted by the BR regimen, we used shRNA technology to knock down STING in lymphoma cells, and the killing effect of the BR regimen on tumor cells was significantly reduced ( Figure 6 CE), while the expression of pyroptosis- and apoptosis-related proteins decreased significantly ( Figure 6 D) These results suggest that the BR regimen kills tumors by activating STING to promote pyroptosis of DLBCL cells.

[0061] Example 2 BR regimen activates cGAS-STING to induce immunogenic death of tumor cells and generate immune response

[0062] (1) BR regimen activates cGAS-STING to induce inflammatory chemotaxis and change the tumor microenvironment

[0063] Experimental procedures: OCI-LY1 and SU-DHL2 cell lines were selected as experimental subjects and divided into control group, BR treatment group, BR+NHS group (normal human serum was added to the culture medium), BR+ICNHS group (human serum with inactivated complement was added to the culture medium). After the cell lines were treated with rituximab (10μM) and bendamustine (200μM) for 12h, the cell survival rate was detected.

[0064] Our in vitro experiments found that the addition of normal human serum can promote the killing of lymphoma cells by the BR regimen, and the killing rate unexpectedly increased by about 30%. However, the killing rate was significantly reduced by the addition of normal human serum with inactivated complement ( Figure 7 A). The experimental results suggest that the BR regimen has achieved good results in the treatment of elderly DLBCL patients, which may be closely related to immune factors. Immunogenic cell death (ICD) is a specific type of cell death. Cells that die in this way can release signal molecules, thereby triggering the activation of the immune system and inflammatory response. Literature reports show that after STING is activated in tumor cells such as colon cancer, glioma and melanoma, highly immunogenic tumor cell fragments or type I interferons can be produced, inducing immunogenic death of tumors, thereby activating the immune system and further killing tumors. RNA sequencing analysis results showed that after BR regimen treatment, tumor cells produced more inflammatory chemokines and related receptor expressions ( Figure 7 B) It may be a phenomenon of immunogenic death. In previous work, we have made it clear that the BR regimen activates the cGAS-STING signaling pathway to directly kill tumor cells, and there is a close relationship between cGAS-STING and immunogenic death.

[0065] We performed weighted gene co-expression network analysis on the GDC-DLBCL-2018 cohort and found that two co-expression modules related to cGAS-STING were enriched in the TNF pathway, cytokine, chemokine, and T cell activation pathways ( Figure 7 CE), among which TNF is a key mediator affecting the inflammatory response in the tumor microenvironment. Single-cell sequencing results of DLBCL showed that tumor cells are the main source of TNF ( Figure 7 F). Our preliminary experiments also confirmed that the BR regimen induced high expression of TNF in lymphoma cells; however, in STING knockout lymphoma cells, TNF expression was significantly decreased after BR treatment ( Figure 7 GI), indicating that the production of TNF depends on STING. Our experimental data also showed that the BR regimen can induce fragmentation of DLBCL and upregulate the type I interferon pathway ( Figure 5 AC, E). Based on the above research results, we propose the hypothesis: BR regimen may induce lymphocytes by activating cGAS-STING Immunogenic death of tumor cells, upregulation of pro-inflammatory factors, formation of immune-hot tumor microenvironment, and promotion of immune cell recruitment( Figure 1 ).

[0066] To verify this hypothesis, we preliminarily studied the relationship between STING expression and immune cell infiltration in DLBCL tumors: We used the TCGA database to analyze DLBCL patients, and found that patients with high STING expression had more abundant T cell infiltration in tumor tissues ( Figure 8 A). Our experiments showed that lymphoma cells treated with BR were more able to attract T lymphocytes to shuttle through the Transwell membrane ( Figure 8 B), further confirmed the above hypothesis.

[0067] (2) BR regimen activates cGAS-STING signaling pathway to induce T cell activation

[0068] Experimental steps: A co-culture system of tumor supernatant, tumor cells and T lymphocytes was established respectively. After treating the cell lines with rituximab (10 μM) and bendamustine (200 μM) for 12 h, the cell survival rate was detected and the differences among the groups were compared.

[0069] WGCNA analysis revealed a co-expression module associated with cGAS-STING enriched in the T cell activation pathway ( Figure 7 C, 7D). Studies have reported that cGAS-STING, as part of the innate immune pathway, can activate adaptive immunity, especially the immune response of T cells. Therefore, we are eager to know whether the anti-tumor function of T cells is enhanced after BR treatment in DLBCL. We cultured normal human T lymphocytes with tumor supernatant and tumor cells in vitro, and found that in the co-culture system, BR-treated tumor cells can better activate T cells and stimulate T cells to secrete more IFN-γ ( Figure 8 C, 8D), suggesting the key role of DLBCL cell surface molecules in regulating T cells. It is known that the direct interaction between T cells and tumor cells includes the mutual recognition and binding of MHC molecules on tumor cells and TCR molecules on T cells. High expression of MHC molecules in tumor cells can significantly affect the recognition and activation of T cells. In addition, studies have reported that the activation of STING-IFN-I signaling can upregulate the expression of MHC molecules on the surface of tumor cells. Therefore, we sequenced and analyzed DLBCL cells and found that tumor cells treated with BR highly expressed MHC molecules ( Figure 8 E). Flow cytometry further confirmed that tumor cells highly expressed MHC class I and MHC class II molecules after BR treatment ( Figure 8 FI).

[0070] Example 3 Therapeutic effect of BR regimen on patients with high tumor burden DLBCL and heart failure symptoms

[0071] Since previous studies have found that the BR regimen involves the cGAS-STING pathway, we speculated that this regimen may be beneficial for symptoms of heart failure, so we used the BR regimen in the treatment of 3 patients with DLBCL with heart failure. All 3 patients with confirmed DLBCL had symptoms of heart failure, decreased ejection fraction and increased BNP levels. Clinical evaluation showed that the patients could not tolerate CHOP chemotherapy (Table 1). The BR regimen (bendamustine 100 mg / m 2 Day 1-2, rituximab 375 mg / m 2 After 4 courses of treatment (21 days for one course), PET-CT showed that all high-metabolism tumor lesions were in an inactivated state, and the primary disease was evaluated as complete remission (Table 1, Figure 1 ), and the patients' cardiac function indicators were significantly improved (Table 2).

[0072] Table 1. Clinical data and characteristics of patients

[0073]

[0074] Table 2 Cardiac function indexes of patients after 4 courses of treatment

[0075]

[0076] The above examples show that bendamustine combined with rituximab can be used in the preparation of anti-tumor drugs for DLBCL patients, especially in the preparation of anti-tumor drugs for DLBCL combined with cardiac function symptoms. While fighting against tumors, it not only does not aggravate the symptoms of cardiac dysfunction, but also has a certain therapeutic effect on the symptoms of cardiac dysfunction.

[0077] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to the above embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. The use of rituximab and bendamustine in the combined preparation of a drug for treating diffuse large B-cell lymphoma symptoms combined with heart failure symptoms, characterized in that: The diffuse large B-cell lymphoma is in a high tumor burden state, the heart failure symptom is a decreased left ventricular ejection fraction accompanied by an increased Pro-BNP level, and the treatment of diffuse large B-cell lymphoma symptoms combined with heart failure symptoms is that the high metabolic tumor lesions are all in an inactivated state and the left ventricular ejection fraction and Pro-BNP return to normal levels.

2. The use according to claim 1, characterized in that: The method for preparing a drug for treating diffuse large B-cell lymphoma symptoms combined with heart failure symptoms is specifically to prepare a drug for treating diffuse large B-cell lymphoma symptoms combined with heart failure symptoms by the following route: Directly induce tumor cell apoptosis and inhibit tumor cell cycle, induce cell pyroptosis, activate the cGAS-STING signaling pathway to kill tumor cells, and activate cGAS-STING to induce immunogenic death of tumor cells.

3. The use according to claim 1-2, characterized in that: The medicament also includes human serum.

4. The use according to claim 1-2, characterized in that: The medicament also includes complement.

5. The use according to any one of claims 1 to 2, characterized in that: The medicaments also include other cytotoxic agents.

6. The use according to any one of claims 1 to 2, characterized in that: The medicament also includes a tumor stem cell differentiation agent.

7. The use according to any one of claims 1 to 2, characterized in that: The drug also includes a pharmaceutically acceptable carrier or excipient.

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