Use of a dual active ingredient drug for the preparation of a drug against double hit lymphoma

Abstract

This invention relates to the application of a dual-active-component drug in the preparation of a drug for treating double-hit lymphoma. The active ingredients in the dual-active-component drug consist of a first active component and a second active component. The first active component is selected from homoharringtonine or a pharmaceutically acceptable salt thereof; the second active component is selected from veneclade or a pharmaceutically acceptable salt thereof. This invention demonstrates that homoharringtonine combined with veneclade can significantly inhibit the proliferation of DHL cell lines and induce apoptosis in DHL cell lines. Using a CDX mouse model, it was demonstrated that homoharringtonine combined with veneclade can inhibit the tumorigenesis process of mouse DHL. The combined use of the two active components not only reduces the dosage of each drug and improves drug safety, but also has a more significant effect on improving or treating double-hit lymphoma (DHL) than a single active component, exhibiting a synergistic effect.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to a novel combination drug intervention strategy for double-hit lymphoma, specifically the application of a dual-active-component drug in the preparation of drugs for anti-double-hit lymphoma. Background Technology

[0002] Double-hit lymphoma (DHL) is a highly aggressive non-Hodgkin's lymphoma, defined as a high-grade B-cell lymphoma with MYC and BCL2 and / or BCL6 rearrangements. It has an extremely poor clinical prognosis and is difficult to cure with conventional chemotherapy. This subtype accounts for approximately 5%-10% of diffuse large B-cell lymphomas, and is more common in middle-aged and elderly patients. Clinically, it often presents with rapid onset, rapid progression, significantly elevated LDH levels, high bone marrow and central nervous system involvement, and a high International Prognostic Index (IPI) risk.

[0003] For these patients, the traditional standard treatment regimen R-CHOP (rituximab + cyclophosphamide + doxorubicin + vincristine + prednisone) for diffuse large B-cell lymphoma (DLBCL) is ineffective, with a high relapse rate and significantly lower long-term survival than ordinary DLBCL. Even with intensive chemotherapy or hematopoietic stem cell transplantation, the prognosis remains poor. Therefore, identifying novel molecular targets and biomarkers for DHL 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 DHL patients. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this invention is to provide a new combination drug intervention strategy for double-hit lymphoma, specifically the application of a dual-active component drug in the preparation of a drug for double-hit lymphoma.

[0005] To achieve this objective, the present invention adopts the following technical solution:

[0006] In a first aspect, the present invention provides the application of a dual-active-component drug in the preparation of a drug for treating double-hit lymphoma, wherein the active ingredient in the dual-active-component drug is composed of a first active component and a second active component;

[0007] The first active ingredient is selected from homoharringtonine or its pharmaceutically acceptable salt;

[0008] The second active ingredient is selected from Veneclare or a pharmaceutically acceptable salt thereof.

[0009] Venetoclax is a BCL-2 inhibitor that competitively binds to and releases pro-apoptotic proteins (such as BIM) that are "detained" by BCL-2, thereby initiating cytochrome c release and the caspase cascade, restoring the apoptosis program in cancer cells. Although venetoclax has shown some efficacy in the treatment of chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL), its clinical application often suffers from primary or secondary resistance due to compensatory upregulation of MCL-1, greatly limiting the efficacy of monotherapy. Homoharringtonine (HHT) is a ribosome function inhibitor that directly binds to the peptidyl transfer site (PTC) of the ribosome to inhibit protein translation and rapidly downregulate short-cycle oncogenic proteins such as MCL-1 and c-MYC, thereby exerting an anti-tumor effect.

[0010] This invention develops a novel drug strategy for improving or treating double-hit lymphoma (DHL), specifically by combining homoharringtonine or a pharmaceutically acceptable salt thereof with veneclade or a pharmaceutically acceptable salt thereof to intervene in the progression of DHL. This invention demonstrates that homoharringtonine combined with veneclade can significantly inhibit the proliferation of DHL cell lines and induce apoptosis in DHL cell lines; using a CDX mouse model, it is demonstrated that homoharringtonine combined with veneclade can inhibit the tumorigenesis process of DHL in mice.

[0011] This invention reveals that the combined use of the two active components not only reduces the dosage of each drug and improves drug safety, but also significantly improves or treats double-hit lymphoma (DHL) compared to using a single active component, exhibiting a synergistic effect. Specifically, homoharringtonine combined with veneclade downregulates MCL-1 molecules in DHL cell lines, thereby reversing DHL cell resistance to veneclade and closing the escape route for cancer cells. This invention provides an effective drug combination strategy for the improvement or treatment of double-hit lymphoma (DHL), which is of great significance.

[0012] Preferably, the dual-active-component drug further contains pharmaceutically acceptable excipients.

[0013] Preferably, the dual-active-component drug of the present invention 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, excipient, filler, binder, wetting agent, disintegrant, emulsifier, solubilizer, osmotic pressure regulator, surfactant, coating material, colorant, pH adjuster, antioxidant, antibacterial agent or buffer.

[0014] Preferably, the dual-active-component drug is a single compound preparation or a combination of two separate preparations.

[0015] Preferably, the dual-active-component drug is a combination of two separate formulations, which are applied simultaneously or sequentially.

[0016] Preferably, the preparation is any pharmaceutically acceptable dosage form, such as tablets, powders, suspensions, granules, capsules, solutions, enemas, emulsions, etc.

[0017] In this invention, the first active component of the dual-active component drug inhibits the abnormally high expression of C-myc protein and BCL-2 protein in double-hit lymphoma cells.

[0018] In this invention, the first active component of the dual-active-component drug downregulates the MCL-1 molecule level in double-hit lymphoma cells, reversing the drug resistance of double-hit lymphoma cells to the second active component.

[0019] In a second aspect, the present invention provides the application of a dual-active-component drug in the preparation of a dual-hit lymphoma cell proliferation inhibitor, wherein the active ingredient in the dual-active-component drug is composed of a first active component and a second active component.

[0020] The first active ingredient is selected from homoharringtonine or its pharmaceutically acceptable salt;

[0021] The second active ingredient is selected from Veneclare or a pharmaceutically acceptable salt thereof.

[0022] Preferably, the double-hit lymphoma cells include TMD8 cells and / or Toledo cells.

[0023] According to the research results of this invention, the dual-active-component drug has a significant inhibitory effect on the proliferation of double-hit lymphoma cells. Therefore, this result indicates that the dual-active-component drug can be used as an in vitro experimental drug in the field of scientific research, rather than just for disease treatment. For example, it can be used to study the growth of double-hit lymphoma cells and other cellular metabolic mechanisms or behaviors, and to screen drugs for the prevention or treatment of double-hit lymphoma.

[0024] Thirdly, the present invention provides the application of a dual-active-component drug in the preparation of a double-hit lymphoma cell apoptosis promoter, wherein the active ingredient in the dual-active-component drug is composed of a first active component and a second active component.

[0025] The first active ingredient is selected from homoharringtonine or its pharmaceutically acceptable salt;

[0026] The second active ingredient is selected from Veneclare or a pharmaceutically acceptable salt thereof.

[0027] Preferably, the double-hit lymphoma cells include TMD8 cells and / or Toledo cells.

[0028] According to the research results of this invention, the dual-active-component drug has a significant effect on promoting apoptosis of double-hit lymphoma cells. Therefore, this result indicates that the dual-active-component drug can be used as an in vitro experimental drug in the field of scientific research, rather than just for disease treatment. For example, it can be used to study the growth and other cellular metabolic mechanisms or behaviors of double-hit lymphoma cells, and to screen drugs for the prevention or treatment of double-hit lymphoma.

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

[0030] This invention develops a novel drug strategy for improving or treating double-hit lymphoma (DHL), specifically by combining homoharringtonine or a pharmaceutically acceptable salt thereof with veneclade or a pharmaceutically acceptable salt thereof to intervene in the progression of DHL. This invention demonstrates that homoharringtonine combined with veneclade can significantly inhibit the proliferation of DHL cell lines and induce apoptosis in DHL cell lines; using a CDX mouse model, it is demonstrated that homoharringtonine combined with veneclade can inhibit the tumorigenesis process of DHL in mice.

[0031] This invention reveals that the combined use of the two active components not only reduces the dosage of each drug and improves drug safety, but also significantly improves or treats double-hit lymphoma (DHL) compared to using a single active component, exhibiting a synergistic effect. Specifically, homoharringtonine combined with veneclade downregulates MCL-1 molecules in DHL cell lines, thereby reversing DHL cell resistance to veneclade and closing the escape route for cancer cells. This invention provides an effective drug combination strategy for the improvement or treatment of double-hit lymphoma (DHL), which is of great significance. Attached Figure Description

[0032] Figure 1A This is a statistical chart showing the TMD8 cell proliferation inhibition rate results for each treatment group in Example 1;

[0033] Figure 1B This is a statistical chart of the TMD8 cell survival count results for each treatment group in Example 1;

[0034] Figure 1C This is a statistical chart showing the Toledo cell proliferation inhibition rate results for each treatment group in Example 2;

[0035] Figure 1D This is a statistical chart of Toledo cell survival counts for each treatment group in Example 2;

[0036] Figure 2AThis is a graph showing the statistical results of apoptosis rates of TMD8 cells in each group after treatment in Example 3;

[0037] Figure 2B This is a flow cytometry result of apoptosis levels in each group of TMD8 cells after treatment in Example 3;

[0038] Figure 2C This is a graph showing the statistical results of apoptosis rates in each group of Toledo cells treated in Example 4.

[0039] Figure 2D This is a flow cytometry result of the apoptosis level of Toledo cells in each group after treatment in Example 4;

[0040] Figure 3A These are anatomical images of the tumors in each group of mice in Example 5;

[0041] Figure 3B This is a statistical chart of tumor volume in each group of mice in Example 5;

[0042] Figure 3C This is a statistical chart of tumor weight in each group of mice in Example 5;

[0043] Figure 4A This is a Western blot analysis of the abnormally overexpressed protein in TMD8 cells after 24 h of treatment with homoharringtonine in Example 6.

[0044] Figure 4B This is a Western blot analysis of the abnormally overexpressed protein in Toledo cells after treatment with homoharringtonine for 24 h in Example 6;

[0045] Figure 5A This is a Western blot analysis of the MCL-1 molecule in TMD8 cells after 24 h of treatment with homoharringtonine and veneclade in Example 7.

[0046] Figure 5B This is a Western blot analysis of the MCL-1 molecule after Toledo cells were treated with homoharringtonine and veneclade for 24 h in Example 7. Detailed Implementation

[0047] 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.

[0048] The following examples use homoharringtonine (HHT), a drug purchased from MCE; Venetoclax, a drug purchased from MCE; double-hit lymphoma cell lines (including Toledo cells and TMD8 cells) provided by the Institute of Hematology, Medical School, Xiamen University; and BALB / c-Nu mice purchased from and bred by the Experimental Animal Center of Xiamen University.

[0049] Example 1

[0050] Inhibitory effect of homoharringtonine combined with veneclade on the proliferation of DHL cell line (TMD8)

[0051] The operation method is as follows: Take a quantity of 2 × 10 4 Logarithmic growth phase DHL cell line (TMD8 cell line) was seeded in 96-well plates and set up homoharringtonine (HHT) single drug group, venetoclax single drug group, homoharringtonine and venetoclax combination group, and control group;

[0052] The HHT single-drug concentrations in the experimental groups were 5 nM, 10 nM, 20 nM, and 30 nM, respectively. The Venetoclax single-drug concentrations in the experimental groups were 0.25 nM, 0.5 nM, 1 nM, and 1.5 nM, respectively. The HHT and Venetoclax combination groups had concentrations of 5 nM + 0.25 nM, 10 nM + 0.5 nM, 20 nM + 1 nM, and 30 nM + 1.5 nM, respectively (the former being HHT and the latter being Venetoclax). The control group cells were treated with the same volume of DMSO. After gently shaking and mixing the corresponding volumes of drug or DMSO with the cells in the 96-well cell culture plates, the cells were cultured in a cell culture incubator (Thermo) for 24 h. The cell proliferation level was then detected using a CCK8 assay kit (MCE, Shanghai).

[0053] The cell proliferation inhibition rate results for each group are as follows: Figure 1A As shown.

[0054] Take 2×10 5Logarithmic growth phase DHL cell line TMD8 was seeded in 24-well cell culture plates, with the following groups included: homoharringtonine (HHT) monotherapy group, venetoclax monotherapy group, homoharringtonine and veneclax combination group, and control group; the concentrations were the same as above, and 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 24-well cell culture plates, the cells were cultured in a Thermo cell culture incubator for 24 h, stained with 0.4% trypan blue (Biosharp) for 3 min, and then examined and counted under a light microscope.

[0055] Cell count results for each group are as follows Figure 1B As shown.

[0056] pass Figure 1A and Figure 1B The results show that, compared with higher concentrations of homoharringtonine monotherapy or higher concentrations of veneclade monotherapy, the combination of homoharringtonine and veneclade can significantly reduce the dosage of both drugs while improving the level of DHL cell line proliferation inhibition, thus ensuring both low drug toxicity and excellent anti-double-hit lymphoma efficacy.

[0057] Example 2

[0058] Inhibitory effect of homoharringtonine combined with veneclade on the proliferation of DHL cell line (Toledo)

[0059] The operation method is as follows: Take a quantity of 2 × 10 4 Logarithmic growth phase DHL cell line (Toledo cell line) was seeded in 96-well plates and set up a homoharringtonine (HHT) single drug group, a venetoclax single drug group, a homoharringtonine and venetoclax combination group, and a control group;

[0060] The HHT single-drug concentrations in the experimental groups were 10 nM, 20 nM, 30 nM, and 40 nM, respectively. The Venetoclax single-drug concentrations in the experimental groups were 1.5 nM, 2 nM, 2.5 nM, and 3 nM, respectively. The HHT and Venetoclax combination groups had concentrations of 10 nM + 1.5 nM, 20 nM + 2 nM, 30 nM + 2.5 nM, and 40 nM + 3 nM, respectively (the former being HHT and the latter being Venetoclax). The control group cells were treated with the same volume of DMSO. After gently shaking and mixing the corresponding volumes of drug or DMSO with the cells in the 96-well cell culture plates, the cells were cultured in a cell culture incubator (Thermo) for 24 h. The cell proliferation level was then detected using a CCK8 assay kit (MCE, Shanghai).

[0061] The cell proliferation inhibition rate results for each group are as follows: Figure 1C As shown.

[0062] Take 2×10 5 Toledo DHL cell line in logarithmic growth phase was seeded in 24-well cell culture plates. Four treatment groups were established: a homoharringtonine (HHT) monotherapy group, a venetoclax monotherapy group, a homoharringtonine and venetoclax combination group, and a control group. Cells in the control group were treated with the same concentration as above, and the same volume of DMSO was used. After gently shaking to mix the corresponding volume of drug or DMSO with the cells in the 24-well cell culture plates, the cells were cultured in a Thermo incubator for 24 h. After staining with 0.4% trypan blue (Biosharp) for 3 min, the cells were examined and counted under a light microscope.

[0063] Cell count results for each group are as follows Figure 1D As shown.

[0064] pass Figure 1C and Figure 1D The results show that, compared with higher concentrations of homoharringtonine monotherapy or higher concentrations of veneclade monotherapy, the combination of homoharringtonine and veneclade can significantly reduce the dosage of both drugs while improving the level of DHL cell line proliferation inhibition, thus ensuring both low drug toxicity and excellent anti-double-hit lymphoma efficacy.

[0065] Example 3

[0066] The promoting effect of homoharringtonine combined with veneclade on apoptosis in DHL cell line (TMD8)

[0067] The operation method is as follows: Take a quantity of 2 × 10 5 Logarithmic growth phase DHL cell line (TMD8 cell line) was seeded in 24-well plates and set up homoharringtonine (HHT) single drug group, venetoclax single drug group, homoharringtonine and venetoclax combination group, and control group;

[0068] The HHT single-drug concentrations in the experimental groups were 5 nM, 10 nM, 20 nM, and 30 nM, respectively. The Venetoclax single-drug concentrations in the experimental groups were 0.25 nM, 0.5 nM, 1 nM, and 1.5 nM, respectively. The HHT and Venetoclax combination concentrations were 5 nM + 0.25 nM, 10 nM + 0.5 nM, 20 nM + 1 nM, and 30 nM + 1.5 nM, respectively (the former being HHT and the latter being Venetoclax). 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. The cells were then collected by centrifugation at 300g for 5 min at 4°C. After washing once with PBS, the apoptosis level was detected and the apoptosis rate was calculated using Annexin V / PI (Thermofisher, USA) flow cytometry.

[0069] Figure 2A and Figure 2B The figures show the statistical results of apoptosis rate and the flow cytometry results of apoptosis level in each group of TMD8 cells after 24 h of treatment.

[0070] pass Figure 2A and Figure 2B The results show that, compared with higher concentrations of homoharringtonine monotherapy or higher concentrations of veneclade monotherapy, the combination of homoharringtonine and veneclade can significantly reduce the dosage of both drugs while increasing the apoptosis-promoting level of DHL cell lines, thus ensuring both low drug toxicity and excellent anti-double-hit lymphoma efficacy.

[0071] Example 4

[0072] The promoting effect of homoharringtonine combined with veneclade on apoptosis in DHL cell line (Toledo).

[0073] The operation method is as follows: Take a quantity of 2 × 10 5 Logarithmic growth phase DHL cell line (Toledo cell line) was seeded in 24-well plates and set up a homoharringtonine (HHT) single drug group, a venetoclax single drug group, a homoharringtonine and venetoclax combination group, and a control group;

[0074] The HHT single-drug concentrations in the experimental groups were 10 nM, 20 nM, 30 nM, and 40 nM, respectively. The Venetoclax single-drug concentrations in the experimental groups were 1.5 nM, 2 nM, 2.5 nM, and 3 nM, respectively. The HHT and Venetoclax combination groups had concentrations of 10 nM + 1.5 nM, 20 nM + 2 nM, 30 nM + 2.5 nM, and 40 nM + 3 nM, respectively (the former being HHT and the latter being Venetoclax). 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. The cells were then collected by centrifugation at 300g for 5 min at 4°C. After washing once with PBS, the apoptosis level was detected and the apoptosis rate was calculated using Annexin V / PI (Thermofisher, USA) flow cytometry.

[0075] Figure 2C and Figure 2D The figures show the statistical results of apoptosis rate and the flow cytometry results of apoptosis level in each group of Toledo cells after 24 h of treatment.

[0076] pass Figure 2C and Figure 2D The results show that, compared with higher concentrations of homoharringtonine monotherapy or higher concentrations of veneclade monotherapy, the combination of homoharringtonine and veneclade can significantly reduce the dosage of both drugs while increasing the apoptosis-promoting level of DHL cell lines, thus ensuring both low drug toxicity and excellent anti-double-hit lymphoma efficacy.

[0077] Example 5

[0078] Effects of homoharringtonine combined with veneclade on the tumorigenesis process of DHL

[0079] The specific operating method is as follows:

[0080] (1) Set up a homoharringtonine (HHT) monotherapy group, a venetoclax monotherapy group, a homoharringtonine and venetoclax combination group, and a control group; among them, venetoclax was dissolved in ethanol and then prepared into a solution according to the ratio of PHOSAL® 50 PG:PEG 400=2:1; among them, homoharringtonine was dissolved in PBS for use.

[0081] (2) Constructing a CDX mouse model

[0082] Take 2×10 7TMD8 cells in the logarithmic growth phase were subcutaneously injected into BALB / c-Nu mice to induce tumor formation and establish a CDX mouse model.

[0083] (3) Four days after the injection, the administration began. The dose of Venecra was 25 mg / kg / day, and the dose of Homoharringtonine was 1 mg / kg / day. Venecra was administered by gavage, and Homoharringtonine was administered by intraperitoneal injection. The administration was continued for 10 days. The tumor was then sacrificed, photographed, weighed, and the volume and weight of the tumor were recorded.

[0084] The anatomical appearance of tumors in each group of mice is as follows: Figure 3A As shown, the tumor volume statistics are as follows: Figure 3B As shown in Figure 3C, the tumor weight statistics are as follows. The above experimental results indicate that, compared with the single-drug group, homoharringtonine (HHT) combined with venetoclax is more effective in inhibiting tumorigenesis in the CDX model.

[0085] Example 6

[0086] Homoharringtonine monotherapy downregulates abnormally high expression of C-myc and BCL-2 proteins in DHL.

[0087] The specific operating method is as follows:

[0088] Take 1×10 6 Logarithmically growing DHL cell lines TMD8 and Toledo were seeded in 6-well cell culture plates. Homoharringtonine monotherapy gradients were established, with homoharringtonine concentrations of 0 nM, 5 nM, 10 nM, 20 nM, 30 nM, and 40 nM for TMD8 and Toledo cells, respectively. Control cells were treated with the same volume of DMSO. After gently shaking and mixing the corresponding volumes of drug or DMSO with the cells in the 6-well cell culture plates, the cells were cultured in a cell culture incubator for 24 h. Cells were then collected by centrifugation at 300 g for 5 min at 4°C, washed once with PBS, and lysed using RIPA (Solarbio) lysis buffer. The RIPA lysis buffer contained pre-added protease inhibitor (Solarbio), phosphatase inhibitor (Solarbio), and PMSF (Beyotime) to protect the target protein and its phosphorylation modification.

[0089] The protein was disrupted and lysed using an ice-water bath with ultrasonication at 25% intensity for 5 seconds, followed by a 4-second interval, for a total of ten cycles. After ultrasonication, the sample was centrifuged at 12,000 rpm at 4°C for 10 minutes. The supernatant was carefully transferred to a clean EP tube, and the sample was subjected to BCA protein concentration determination using a BCA assay kit (Yeason). After the determination, the corresponding volume of 5×SDS was added, mixed well, and boiled for 10 minutes. Western blot analysis was then performed to detect C-myc and BCL-2 proteins.

[0090] The results are as follows Figure 4A and Figure 4B As shown, homoharringtonine monotherapy can inhibit the abnormally high expression of C-myc and BCL-2 proteins in DHL cell lines TMD8 and Toledo.

[0091] Example 7

[0092] Homoharringtonine combined with Veneclare downregulates MCL-1 molecules

[0093] The specific operating method is as follows:

[0094] Take 1×10 6 Logarithmically growing DHL cell lines TMD8 and Toledo were seeded in 6-well cell culture plates. Control, homoharringtonine monotherapy, veneclade monotherapy, and dual-drug combination groups were established. In the TMD8 experimental group, homoharringtonine concentration was 10 nM and veneclade concentration was 0.5 nM; control cells were treated with the same volume of DMSO. In the Toledo experimental group, homoharringtonine concentration was 20 nM and veneclade concentration was 2 nM; control cells were treated with the same volume of DMSO.

[0095] After gently shaking and mixing the corresponding volume of drug or DMSO with the cells in the above 6-well cell culture plate, the cells were cultured in a cell culture incubator for 24 h, centrifuged at 300 g for 5 min at 4 °C to collect the cells, washed once with PBS, and lysed with RIPA lysis buffer. The RIPA lysis buffer (Solarbio) was pre-added with protease inhibitor (Solarbio), phosphatase inhibitor (Solarbio), and PMSF (Beyotime) to protect the target protein and its phosphorylation modification.

[0096] The protein was disrupted and lysed using an ice-water bath with sonication at 25% intensity for 5 seconds, followed by 4-second intervals, for a total of ten cycles. After sonication, the sample was centrifuged at 12,000 rpm at 4°C for 10 minutes. The supernatant was carefully transferred to a clean EP tube, and the sample was subjected to BCA protein concentration determination using a BCA assay kit (Yeason). After the determination, the corresponding volume of 5×SDS was added, mixed well, and boiled for 10 minutes. Western blot analysis was then performed to detect MCL-1 protein.

[0097] The results are as follows Figure 5A and Figure 5B As shown, the combination of homoharringtonine and veneclade can downregulate MCL-1 molecules in DHL cell lines TMD8 and Toledo cells, thereby reversing resistance to veneclade and closing the escape route of cancer cells.

[0098] 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.

[0099] 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.

[0100] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. The application of a dual-active component drug in the preparation of a drug for treating double-hit lymphoma, characterized in that, The active ingredient in the dual-active-component drug consists of a first active component and a second active component; The first active ingredient is selected from homoharringtonine or its pharmaceutically acceptable salt; The second active ingredient is selected from Veneclare or a pharmaceutically acceptable salt thereof.

2. The application according to claim 1, characterized in that, The dual-active-component drug also contains pharmaceutically acceptable excipients.

3. The application according to claim 2, characterized in that, The pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carriers, diluents, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH adjusters, antioxidants, antibacterial agents, or buffers.

4. The application according to any one of claims 1-3, characterized in that, The dual-active-component drug is a single compound preparation or a combination of two separate preparations; Preferably, the dual-active-component drug is a combination of two separate formulations, which are applied simultaneously or sequentially. Preferably, the formulation is any pharmaceutically acceptable dosage form.

5. The application according to any one of claims 1-4, characterized in that, The first active component of the dual-active-component drug inhibits the abnormally high expression of C-myc and BCL-2 proteins in double-hit lymphoma cells.

6. The application according to any one of claims 1-5, characterized in that, The first active component of the dual-active-component drug downregulates the MCL-1 molecule level in double-hit lymphoma cells, reversing the drug resistance of double-hit lymphoma cells to the second active component.

7. The application of a dual-active-component drug in the preparation of a double-hit lymphoma cell proliferation inhibitor, characterized in that, The active ingredient in the dual-active-component drug consists of a first active component and a second active component; The first active ingredient is selected from homoharringtonine or its pharmaceutically acceptable salt; The second active ingredient is selected from Veneclare or a pharmaceutically acceptable salt thereof.

8. The application according to claim 7, characterized in that, The double-hit lymphoma cells include TMD8 cells and / or Toledo cells.

9. The application of a dual-active component drug in the preparation of a double-hit lymphoma cell apoptosis promoter, characterized in that, The active ingredient in the dual-active-component drug consists of a first active component and a second active component; The first active ingredient is selected from homoharringtonine or its pharmaceutically acceptable salt; The second active ingredient is selected from Veneclare or a pharmaceutically acceptable salt thereof.

10. The application according to claim 9, characterized in that, The double-hit lymphoma cells include TMD8 cells and / or Toledo cells.

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