A combined pharmaceutical composition for preventing or treating diffuse large b-cell lymphoma and use thereof

Abstract

The present application relates to a kind of combination drug composition for preventing or treating diffuse large B-cell lymphoma and its application, the active component of the combination drug composition is composed of first active component and second active component;The first active component is selected from selinexor or any one or at least two combinations of its pharmaceutically acceptable salt, isomer, solvate, metabolite;The second active component is selected from ritonavir or any one or at least two combinations of its pharmaceutically acceptable salt, isomer, solvate, metabolite.The combination drug composition has excellent treatment effect of diffuse large B-cell lymphoma, not only can reduce the dosage of selinexor, improve the safety of drug, but also has more significant inhibitory effect on diffuse large B-cell lymphoma than using single selinexor.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to a novel strategy for the prevention, improvement or treatment of diffuse large B-cell lymphoma, specifically to a combination drug composition for the prevention or treatment of diffuse large B-cell lymphoma and its application. Background Technology

[0002] Diffuse large B-cell lymphoma (DLBCL) is an aggressive tumor originating from mature B cells and is the most common type of non-Hodgkin's lymphoma. It exhibits high heterogeneity in morphology, biology, immunophenotype, genetics, and clinical manifestations. Clinically, there is no definitive optimal salvage regimen; some patients relapse within two years after first-line treatment, and some develop refractory disease. For DLBCL patients who fail initial treatment or experience short-term relapse, although high-dose salvage chemotherapy and hematopoietic stem cell transplantation can improve clinical outcomes, most patients still experience disease relapse and may even eventually die, resulting in a significant social and economic burden.

[0003] Therefore, identifying novel molecular targets and biomarkers for DLBCL relapse, drug resistance, and disease progression, and establishing novel targeted therapies and intervention strategies, is of great significance for improving the clinical efficacy and prognosis of DLBCL patients. Selinexor, the first oral selective nuclear export inhibitor, has been approved by the FDA for the treatment of refractory / relapsed diffuse large B-cell lymphoma (DLBCL) after at least two lines of therapy. The SADAL trial showed an overall response rate of 28%, but most patients experienced depression, fatigue, nausea, and decreased appetite.

[0004] Therefore, it is essential to develop a strategy that can reduce the dosage and toxic side effects of celinixol while enhancing its sensitivity to killing diffuse large B-cell lymphoma. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide a novel strategy for the prevention, improvement or treatment of diffuse large B-cell lymphoma, specifically a combination drug composition for the prevention or treatment of diffuse large B-cell lymphoma and its application.

[0006] To achieve this objective, the present invention employs the following technical solution:

[0007] In a first aspect, the present invention provides a combination pharmaceutical composition for the prevention or treatment of diffuse large B-cell lymphoma, wherein the active component of the combination pharmaceutical composition is composed of a first active component and a second active component.

[0008] The first active ingredient is selected from any one or a combination of at least two of celiniso or its pharmaceutically acceptable salts, isomers, solvates, and metabolites;

[0009] The second active component is selected from any one or a combination of at least two of ritanserin or its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

[0010] This invention develops a novel drug combination therapy, which combines the XPO1 inhibitor celiniso (or its pharmaceutically acceptable salts, isomers, solvates, and metabolites) with ritanserin (or its pharmaceutically acceptable salts, isomers, solvates, and metabolites). This combined drug composition exhibits excellent therapeutic effects on diffuse large B-cell lymphoma. The study found that the two active components, when inhibiting diffuse large B-cell lymphoma, not only reduce the dosage of celiniso (or its pharmaceutically acceptable salts, isomers, solvates, and metabolites), improving drug safety, but also have a more significant inhibitory effect on diffuse large B-cell lymphoma than using celiniso (or its pharmaceutically acceptable salts, isomers, solvates, and metabolites) alone, achieving a synergistic effect.

[0011] This invention first demonstrates, using diffuse large B-cell lymphoma cell lines (TMD8 cells and / or Toledo cells), that the combined drug composition can significantly inhibit the proliferation of diffuse large B-cell lymphoma cell lines and induce apoptosis; using a CDX mouse model, it is further demonstrated that the combined drug composition can inhibit tumorigenesis in mice. This invention provides an effective drug combination strategy for the improvement or treatment of diffuse large B-cell lymphoma, and is of great significance.

[0012] In this invention, the combined pharmaceutical composition is a single compound preparation or a combination of two separate preparations.

[0013] Preferably, the combined pharmaceutical composition is a combination of two separate formulations, which are administered simultaneously or sequentially.

[0014] The combined drug composition can be a single compound preparation or a combination of two separate preparations; when it is a combination of two separate preparations, it can be administered simultaneously, alternately, or sequentially.

[0015] In this invention, the formulation is any pharmaceutically acceptable dosage form.

[0016] Preferably, the combined pharmaceutical composition further contains pharmaceutically acceptable excipients.

[0017] Preferably, the combined pharmaceutical composition can be administered alone or in combination with excipients to form an appropriate dosage form for administration. The pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carrier, diluent, binder, wetting agent, disintegrant, emulsifier, solubilizer, osmotic pressure regulator, surfactant, coating material, colorant, pH adjuster, antioxidant, antibacterial agent, or buffer.

[0018] In a second aspect, the present invention provides the use of the combined pharmaceutical composition according to the first aspect in the preparation of a medicament for the prevention, improvement or treatment of diffuse large B-cell lymphoma.

[0019] Preferably, the diffuse large B-cell lymphoma is relapsed or refractory diffuse large B-cell lymphoma.

[0020] This invention develops a novel preventative or therapeutic approach for diffuse large B-cell lymphoma, particularly relapsed or refractory diffuse large B-cell lymphoma. It involves the combined use of the XPO1 inhibitor celinizol (or its pharmaceutically acceptable salts, isomers, solvates, or metabolites) and ritanserin (or its pharmaceutically acceptable salts, isomers, solvates, or metabolites). This combined drug composition exhibits excellent therapeutic efficacy against diffuse large B-cell lymphoma. The invention's research reveals that the two active components, when inhibiting diffuse large B-cell lymphoma, not only reduce the dosage of celinizol (or its pharmaceutically acceptable salts, isomers, solvates, or metabolites), improving drug safety, but also significantly inhibit diffuse large B-cell lymphoma more effectively than using celinizol (or its pharmaceutically acceptable salts, isomers, solvates, or metabolites) alone, achieving a synergistic effect.

[0021] Thirdly, the present invention provides the use of the combined pharmaceutical composition according to the first aspect in the preparation of a diffuse large B-cell lymphoma cell proliferation inhibitor.

[0022] According to the research results of this invention, the combined pharmaceutical composition significantly inhibits the proliferation of diffuse large B-cell lymphoma cells. Therefore, this result indicates that the combined pharmaceutical composition can be used as an in vitro experimental reagent in scientific research, such as studying the growth, apoptosis, and metabolic mechanisms or behaviors of diffuse large B-cell lymphoma cells, and screening therapeutic drugs. The proliferation inhibitor claimed in this invention is not intended to eliminate the cause or lesion; that is, it is an application in the preparation of a diffuse large B-cell lymphoma cell proliferation inhibitor for a non-therapeutic purpose.

[0023] Preferably, the diffuse large B-cell lymphoma cells include TMD8 cells and / or Toledo cells.

[0024] Preferably, the diffuse large B-cell lymphoma cells are tp53 mutant cell lines.

[0025] Fourthly, the present invention provides the use of the combined pharmaceutical composition according to the first aspect in the preparation of an apoptosis promoter for diffuse large B-cell lymphoma cells.

[0026] According to the research results of this invention, the combined pharmaceutical composition significantly induces apoptosis in diffuse large B-cell lymphoma cells. Therefore, this result indicates that the combined pharmaceutical composition can be used as an in vitro experimental reagent in scientific research, such as studying the growth, apoptosis, and metabolic mechanisms or behaviors of diffuse large B-cell lymphoma cells, and screening therapeutic drugs. The apoptosis promoter claimed in this invention is not intended to eliminate the cause or lesion; that is, it is an application in the preparation of an apoptosis promoter for diffuse large B-cell lymphoma cells for a non-therapeutic purpose.

[0027] Preferably, the diffuse large B-cell lymphoma cells include TMD8 cells and / or Toledo cells.

[0028] Preferably, the diffuse large B-cell lymphoma cells are tp53 mutant cell lines.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] This invention develops a novel drug combination therapy and a new preventive or therapeutic approach for diffuse large B-cell lymphoma, especially relapsed or refractory diffuse large B-cell lymphoma. It involves the combined use of the XPO1 inhibitor celinizol (or its pharmaceutically acceptable salts, isomers, solvates, or metabolites) and ritanserin (or its pharmaceutically acceptable salts, isomers, solvates, or metabolites). This combined drug composition exhibits excellent therapeutic efficacy against diffuse large B-cell lymphoma. The invention's research found that the two active components, when inhibiting diffuse large B-cell lymphoma, not only reduce the dosage of celinizol (or its pharmaceutically acceptable salts, isomers, solvates, or metabolites), improving drug safety, but also significantly inhibit diffuse large B-cell lymphoma more effectively than using celinizol (or its pharmaceutically acceptable salts, isomers, solvates, or metabolites) alone, achieving a synergistic effect. Attached Figure Description

[0031] Figure 1A This is a graph showing the cell count results of TMD8 cells after 24 hours of treatment with litanserin and celiniso.

[0032] Figure 1B This is a graph showing the cell count results of Toledo cells after 24 hours of treatment with litanserin and celiniso.

[0033] Figure 2A This is a flow cytometry result of apoptosis level in TMD8 cells after 48 hours of treatment with litanserin and celiniso.

[0034] Figure 2B The graph shows the statistical results of apoptosis rate in TMD8 cells after 48 hours of treatment with litanserin combined with celiniso.

[0035] Figure 2C This is a flow cytometry result of Toledo cells treated with litanserin and celiniso for 24 hours to detect apoptosis levels.

[0036] Figure 2D This is a statistical result of the apoptosis rate of Toledo cells after 24 hours of treatment with litanserin combined with celiniso.

[0037] Figure 3A This is an anatomical view of a CDX mouse model after treatment with litanserine and celiniso.

[0038] Figure 3B This is a tumor growth curve of a CDX mouse model after treatment with litanserin combined with celinisoxol;

[0039] Figure 3C This is a graph showing the statistical results of tumor weight in a CDX mouse model after treatment with litanserin combined with celinisoxol. Detailed Implementation

[0040] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention in any way.

[0041] The processes, conditions, reagents, and experimental methods used in implementing this invention, except as specifically mentioned below, are all common knowledge and general knowledge in the field, and this invention does not have any particular limitations. Experimental methods in the embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer.

[0042] Unless otherwise stated, all technical terms and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. However, in the event of any conflict, the specification containing the definitions shall prevail.

[0043] The drug celinisosole involved in the following examples was purchased from Hanxiang Biotechnology, product number 1393477-72-9; the drug litanserin was purchased from Taoshu Biotechnology, product number 87051-43-2.

[0044] TMD8 cells and Toledo cells were provided by the Institute of Hematology, School of Medicine, Xiamen University.

[0045] CB-17SCID mice were purchased from the Experimental Animal Center of Xiamen University and were bred by the Experimental Animal Center.

[0046] Example 1

[0047] The inhibitory effect of the combined drug combination on the proliferation of diffuse large B-cell lymphoma cell lines:

[0048] The operation method is as follows: take the number of items in the logarithmic growth phase as 2 × 10. 4 Diffuse large B-cell lymphoma cell lines (TMD8 cells and Toledo cells) were seeded in 96-well plates. Control group, celinixol monotherapy group, celinixol combined with 5 μM ritanserin group, and celinixol combined with 7.5 μM ritanserin group were included.

[0049] The experimental group cells were treated with 0.125 μM, 0.25 μM, 0.5 μM and 1 μM of celinixol, respectively, while the control group cells were treated with the same volume of DMSO. After gently shaking and mixing the corresponding volume of drug or DMSO with the cells in the 96-well cell culture plate, the cells were cultured in a cell culture incubator (Thermo) for 24 h and the cell proliferation level was detected by CCK8 kit (ZETA, Shanghai) and flow cytometry (Quanteon / ACEA).

[0050] Experimental results: Cell viability results for each group are as follows Figure 1A (TMD8 cells) Figure 1B (Toledo cells) shown. Through Figure 1A , Figure 1B The results showed that, in both cell lines of diffuse large B-cell lymphoma, the combination of celinixol and ritanserin had a better inhibitory effect on cell proliferation at a lower concentration of celinixol compared to either celinixol monotherapy or ritanserin monotherapy.

[0051] Example 2

[0052] The promoting effect of the combined drug composition on apoptosis in diffuse large B-cell lymphoma cell lines:

[0053] The operation method is as follows: take the number of items in the logarithmic growth phase as 2 × 10. 5 Diffuse large B-cell lymphoma cell lines (TMD8 cells and Toledo cells) were seeded in 24-well plates. Control group, celinixol monotherapy group, and celinixol plus ritanserin combination group were established.

[0054] The concentrations of celinixol alone in the experimental groups were 0.125 μM, 0.25 μM, 0.5 μM, and 1 μM, respectively. For TMD8 cells, the concentrations of celinixol combined with ritanserin were 0.125 μM + 7.5 μM, 0.25 μM + 7.5 μM, 0.5 μM + 7.5 μM, and 1 μM + 7.5 μM, respectively (the former being celinixol and the latter ritanserin). For Toledo cells, the concentrations of celinixol combined with ritanserin were 0.125 μM + 5 μM, 0.25 μM + 5 μM, 0.5 μM + 5 μM, and 1 μM + 5 μM, respectively (the former being celinixol and the latter ritanserin). The control group cells were treated with the same volume of DMSO.

[0055] After gently shaking and mixing the corresponding volume of drug or DMSO with the cells in the 24-well cell culture plate, the cells were cultured in a cell culture incubator for 24 h (Toledo cells) or 48 h (TMD8 cells). The cells were then collected by centrifugation at 300g for 5 min at 4°C, washed once with PBS, and the apoptosis level was detected and the apoptosis rate was calculated using Annexin V / PI (Thermofisher, USA) flow cytometry.

[0056] Experimental results: The flow cytometry results for each group are as follows Figure 2A As shown in (TMD8) and 2C (Toledo), the statistical results for cell apoptosis rate are as follows: Figure 2B (TMD8) Figure 2D (As shown in Toledo). The results show that, in both cell lines of diffuse large B-cell lymphoma, the combination of celiniso and ritanserin showed a better promoting effect on apoptosis at lower drug concentrations compared to either celiniso or ritanserin monotherapy. Furthermore, the apoptosis-promoting effect gradually increased with increasing drug concentration.

[0057] Example 3

[0058] Effects of combined drug combinations on the in vivo tumorigenesis process of diffuse large B-cell lymphoma cells:

[0059] The operation method is as follows:

[0060] (1) Constructing a CDX mouse model

[0061] Take 3.27 × 10 7 TMD8 cells in the logarithmic growth phase were subcutaneously injected into CB-17SCID mice to induce tumor formation and establish a CDX mouse model.

[0062] (2) CDX mice (weighing 18-20g) were divided into four groups of five mice each: a control group, a celinisosole monotherapy group, a ritanserin monotherapy group, and a celinisosole and ritanserin combination group. Celinisosole was administered using a suspension prepared with 100mM celinisosole stock solution and PBS, while ritanserin was administered using a suspension prepared with 50mM ritanserin stock solution and PBS. The control group received a solution prepared with the same volume of DMSO and PBS.

[0063] (3) Four days after modeling, the drug was started. The dose of celinixol was 10 mg / kg / day and the dose of ritanserin was 2.5 mg / kg / day. Both were administered subcutaneously in situ for 5 consecutive days. The tumor size was recorded and the experimental results were statistically analyzed with the start of the drug administration diary as 4 days.

[0064] Experimental results: After the experiment, the tumor anatomy of each group of mice was shown in the following figures. Figure 3A As shown; the tumor growth curves of each group of mice are as follows. Figure 3B As shown; the tumor weight statistics of mice in each group after the experiment are as follows. Figure 3C As shown above, the results indicate that celiniso combined with ritanserin can better inhibit tumorigenesis of diffuse large B-cell lymphoma cells in vivo compared to celiniso alone or ritanserin alone, demonstrating a superior anti-diffuse large B-cell lymphoma effect.

[0065] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0066] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0067] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable way without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. The use of a combination pharmaceutical composition in the preparation of a medicament for improving or treating diffuse large B-cell lymphoma, characterized in that, The active component of the combined pharmaceutical composition consists of a first active component and a second active component; The first active ingredient is selected from celiniso or a pharmaceutically acceptable salt thereof; The second active ingredient is selected from ritanserin or its pharmaceutically acceptable salt.

2. The application according to claim 1, characterized in that, The combined drug composition is a single compound preparation or a combination of two separate preparations.

3. The application according to claim 2, characterized in that, The combined drug composition is a combination of two separate formulations, which are administered simultaneously or sequentially.

4. The application according to claim 2, wherein the formulation is any pharmaceutically acceptable dosage form.

5. The application according to claim 1, characterized in that, The combined pharmaceutical composition also contains pharmaceutically acceptable excipients.

6. The application according to claim 5, characterized in that, The pharmaceutically acceptable excipients are selected from any one or a combination of at least two of the following: carriers, diluents, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, coating materials, colorants, pH adjusters, antioxidants, or antibacterial agents.

7. The application according to claim 1, characterized in that, The diffuse large B-cell lymphoma mentioned is relapsed or refractory diffuse large B-cell lymphoma.

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