Application of combination of Venoclara and regorafenib in treatment of acute myelogenous leukemia

Abstract

The invention discloses an application of a combination of Venoclam and regorafenib in treatment of acute myelogenous leukemia, provides a pharmaceutical composition and a kit for treating leukemia, and also provides a method for inhibiting leukemia cells or tissues in vitro. The invention provides a brand new combined scheme for treating acute myelogenous leukemia, and provides a new scheme for solving the problems of drug resistance and relapse of AML clinically.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and particularly relates to the application of Venetoclax combined with Regorafenib in the treatment of acute myeloid leukemia. Background Art

[0002] Acute Myeloid Leukemia (AML) is a highly heterogeneous hematological malignancy that can affect people of all ages, most commonly the elderly. The therapeutic effect of traditional chemotherapy drugs is limited, the clinical outcomes of AML patients are still not ideal, the recurrence rate is relatively high, and the sensitivity to chemotherapy drugs is lost after recurrence, resulting in patients dying from drug resistance and various complications, which prompts people to search for additional synergistic treatment principles.

[0003] Venetoclax (ABT-199) is a highly selective Bcl-2 inhibitor. Although Venetoclax is combined with DNA hypomethylating agents or low-dose cytarabine, the initial response rate of AML patients is high. However, the heterogeneity of AML cells and complex anti-apoptotic mechanisms make patients develop resistance to ABT-199, or the monotherapy effect is limited, and the condition of most patients will relapse, develop drug resistance and eventually die. Therefore, it is necessary to further study the drug resistance mechanism of Venetoclax to determine the optimal dose of Venetoclax for AML treatment, search for new combination treatment strategies, enhance the anti-AML activity of Venetoclax, and improve the prognosis of AML patients. Summary of the Invention

[0004] In the prior art, some AML patients show natural resistance or acquired resistance to Venetoclax, which is a huge obstacle to the clinical application of Venetoclax in the treatment of AML. The present invention intends to use a novel small molecule inhibitor to enhance the activity of Venetoclax to achieve the goal of treating AML and prolonging the life of patients.

[0005] In order to solve the technical problems existing in the prior art, the present invention provides the following technical solutions.

[0006] The present invention provides the application of a Bcl-2 inhibitor combined with a tyrosine kinase inhibitor in the preparation of a pharmaceutical composition for preventing and treating leukemia.

[0007] Further, the tyrosine kinase inhibitor is in the form of an oral small molecule multi-target.

[0008] Further, the tyrosine kinase inhibitor includes Imatinib, Dasatinib, Nilotinib, Sunitinib, Regorafenib, Bosutinib, Regorafenib or a pharmaceutically acceptable salt thereof.

[0009] Further, the Bcl-2 inhibitor includes venetoclax, orelabrutinib, Lisaftoclax, APG-2575, Tazemetostat or a pharmaceutically acceptable salt thereof.

[0010] Further, the tyrosine kinase inhibitor is regorafenib or a pharmaceutically acceptable salt thereof.

[0011] Further, the Bcl-2 inhibitor is venetoclax or a pharmaceutically acceptable salt thereof.

[0012] Further, the leukemia includes acute leukemia, chronic leukemia, hairy cell leukemia, and prolymphocytic leukemia.

[0013] Further, the acute leukemia includes acute lymphoblastic leukemia L1, acute lymphoblastic leukemia L2, acute lymphoblastic leukemia L3, and acute myeloid leukemia.

[0014] Further, the acute myeloid leukemia includes granulocytic leukemia, erythroleukemia, and megakaryocytic leukemia.

[0015] Further, the chronic leukemia includes chronic lymphocytic leukemia and chronic myeloid leukemia.

[0016] Further, the mass ratio of the Bcl-2 inhibitor to the tyrosine kinase inhibitor is 8:1 - 2:1.

[0017] Further, the concentration range of the Bcl-2 inhibitor is 1 - 2 μM.

[0018] Further, the concentration range of the tyrosine kinase inhibitor is 0.125 - 0.5 μM.

[0019] In some embodiments, the concentration of the Bcl-2 inhibitor is 1 μM, 1.1 μM, 1.2 μM, 1.3 μM, 1.4 μM, 1.5 μM, 1.6 μM, 1.7 μM, 1.8 μM, 1.9 μM, 2 μM. In some embodiments, the concentration of the tyrosine kinase inhibitor is 0.125 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.375 μM, 0.4 μM, 0.45 μM, 0.5 μM. In some embodiments, the mass ratio of the Bcl-2 inhibitor to the tyrosine kinase inhibitor is 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1.

[0020] In the present invention, the "μM" used represents micromolar concentration. μ: the Greek letter "mu", representing the SI prefix "micro-", i.e., 10⁻ 6(parts per million), M: represents molar concentration (mol / L), that is, the amount of substance of the solute in each liter of solution. Therefore, 1 μM = 10⁻ 6 mol / L, that is, each liter of solution contains 1 micromole of solute.

[0021] In some embodiments, venetoclax is combined with regorafenib in a manner that enhances the activity of regorafenib, thereby enhancing the therapeutic effect on leukemia. In some embodiments, the combination of venetoclax and regorafenib produces a synergistic effect, significantly enhancing the efficacy of the combination of the drugs.

[0022] As used herein, the phrase "pharmaceutically acceptable salt" refers to those salts of a compound of interest that are safe and effective upon topical application in mammals and have the desired biological activity. Pharmaceutically acceptable salts include salts of acidic or basic groups present in the designated compound. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, mesylate, esylate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). Certain compounds used in the present invention can form pharmaceutically acceptable salts with various amino acids. Suitable base salts include, but are not limited to: aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, and diethanolamine salts. A review of pharmaceutically acceptable salts can be found in the literature listed: BERGE et al., 66 J. PHARM. SCI. 1-19 (1977).

[0023] The present invention provides a pharmaceutical composition for preventing or treating leukemia, which comprises a Bcl-2 inhibitor and a tyrosine kinase inhibitor as described above, used in combination in a prophylactically effective amount or a therapeutically effective amount.

[0024] Furthermore, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0025] Furthermore, the excipient includes a carrier, adjuvant, excipient, diluent, or other liquid solvent, dispersion aid, suspension aid, surfactant, isotonic agent, thickening agent, emulsifying agent, preservative, solid binder, and lubricant.

[0026] In some embodiments, the pharmaceutically acceptable excipients include any and all solvents, diluents, buffers (e.g., neutral buffered saline, or optionally Tris-HCl, acetate or phosphate buffers), solubilizers (e.g., polysorbate 80), colloids, dispersion media, vehicles, fillers, chelating agents (e.g., EDTA or glutathione), amino acids (e.g., glycine), proteins, disintegrants, binders, lubricants, wetting agents, stabilizers, emulsifiers, sweeteners, colorants, flavoring agents, aromatisers, thickeners, substances for achieving storage effects, coating agents, antifungal agents, preservatives (e.g., ThimerosalTM, benzyl alcohol), antioxidants (e.g., ascorbic acid, sodium metabisulfite), tonicity control agents, absorption delaying agents, adjuvants, bulking agents (e.g., lactose, mannitol), etc. The use of such media and reagents for formulating pharmaceutical compositions is well known in the art. The use of any conventional media or reagents is contemplated in the pharmaceutical composition unless it is incompatible with the active ingredient.

[0027] In some embodiments, the pharmaceutical composition further comprises other drugs that can assist in the treatment of leukemia, specifically including anti-apoptotic agents, mitotic inhibitors (anti-microtubule drugs, representative drugs: vincristine (VCR), paclitaxel (PTX), which block cell division by interfering with tubulin function), immunomodulators (including glucocorticoids (such as prednisone) and monoclonal antibodies (such as rituximab), which enhance the anti-leukemia effect by regulating the immune response or targeting antigens such as CD20), alkylating agents (representative drugs: cyclophosphamide (CTX), busulfan, which inhibit the proliferation of leukemia cells by disrupting the DNA structure and are used in various leukemia chemotherapy regimens), anti-metabolites (representative drugs: methotrexate (MTX), cytarabine (Ara-C), which interfere with nucleic acid metabolism and are widely used in the induction and consolidation treatment of acute leukemia), topoisomerase inhibitors (representative drugs: etoposide (VP-16), irinotecan, which kill leukemia cells by inhibiting the activity of DNA replication-related enzymes), antibiotic drugs (anthracyclines, representative drugs: doxorubicin (ADM), daunorubicin (DNR), which intercalate into the DNA double helix to prevent replication and are the core drugs for acute leukemia chemotherapy), platinum compounds (such as carboplatin, oxaliplatin, which exert anti-leukemia effects by forming DNA cross-linking damage), hormonal drugs (glucocorticoids (dexamethasone) are used in the induction treatment of ALL, which can enhance the chemotherapy sensitivity and inhibit lymphocyte proliferation), targeted drugs (non-TKIs, such as FLT3 inhibitors (midostaurin), CD33 monoclonal antibody (gemtuzumab), which treat relapsed / refractory AML by targeting specific molecular targets), chemotherapy protectants (such as calcium folinate (an antidote for methotrexate), dexrazoxane (which reduces the cardiotoxicity of anthracyclines), which are used to reduce the side effects of chemotherapy).

[0028] In the present invention, a therapeutically effective amount refers to the lowest dose or dose range of a drug that can produce the expected therapeutic effect in a specific patient or population, while ensuring its safety (i.e., the toxicity or side effects are acceptable). This concept runs through drug research and development, clinical trials, and clinical practice, and the following core elements need to be considered comprehensively: 1. Balance between efficacy and toxicity: The core of a therapeutically effective amount lies in balancing the efficacy of the drug and its toxic and side effects. For example, the dose of the anti-tumor drug cytarabine (Ara-C) needs to be adjusted according to the type of leukemia: high doses are used for induction remission of acute myeloid leukemia (AML), but may cause myelosuppression, and the dose needs to be optimized through blood drug concentration monitoring. Similarly, immunomodulators such as interferon γ need to find a balance between enhancing the immune response and avoiding excessive inflammation. 2. Consideration of individual differences: The age, weight, metabolic capacity, genotype, and comorbidities of patients all affect the therapeutically effective amount. For example, elderly AML patients have lower tolerance to anthracyclines (such as daunorubicin), and the dose needs to be reduced to avoid cardiotoxicity. The dose of targeted drugs such as tyrosine kinase inhibitors (imatinib) needs to be adjusted according to the BCR-ABL gene mutation status to overcome drug resistance. 3. Drug mechanism of action and pharmacokinetics: The target and metabolic pathway of the drug directly affect the effective dose. For example: Alkylating agents (such as cyclophosphamide): Kill tumor cells by destroying the DNA structure, and the therapeutically effective amount needs to balance cytotoxicity and the repair ability of normal tissues. Nucleic acid drugs (such as cytarabine): As antimetabolites, they need to be continuously infused to maintain blood drug concentration and inhibit DNA synthesis. Immune checkpoint inhibitors (such as PD-1 antibodies): The dose needs to be adjusted according to the degree of immune cell infiltration in the tumor microenvironment to activate T cells while avoiding autoimmune reactions.

[0029] The present invention provides the use of a Bcl-2 inhibitor in combination with a tyrosine kinase inhibitor in the preparation of a product for inhibiting the proliferation of leukemia cells and promoting the apoptosis of leukemia cells.

[0030] The present invention provides a composition for inhibiting the proliferation of leukemia cells and promoting the apoptosis of leukemia cells, and the composition comprises the following components or is composed of the following component combinations:

[0031] 1) A first active ingredient, and the first active ingredient comprises a Bcl-2 inhibitor;

[0032] 2) A second active ingredient, and the second active ingredient comprises a tyrosine kinase inhibitor.

[0033] In the present invention, the interactions of drugs are classified into additive effect, synergistic effect, and antagonistic effect according to the effects when drugs are used in combination. Synergistic effect means that the effect when drugs used in combination is many times greater than that when used alone. Additive effect means that the effect when drugs used in combination is equivalent to that when used alone. Antagonistic effect means that the effect when drugs used in combination is smaller than that when used alone. In the combination use of the present invention, it is first discovered that the combination of the first preparation and the second preparation has a synergistic effect.

[0034] In some embodiments, "treatment" refers to the improvement, prevention, or reversal of a disease or disorder or at least one distinguishable symptom thereof. In certain specific embodiments, "treatment" refers to the improvement, prevention, or reversal of at least one measurable physiological parameter related to the disease or disorder to be treated, and the parameter is not necessarily recognizable in or recognized by a mammal. In another embodiment, "treatment" refers to inhibiting or slowing down the progression of a disease or disorder, which can be physical, such as stabilizing distinguishable symptoms, or physiological, such as stabilizing physiological parameters, or both. In another embodiment, "treatment" refers to delaying the onset of a disease or disorder.

[0035] In certain specific embodiments, an interested compound is used as a preventive measure. As used herein, "prevention" refers to reducing the risk of acquiring a certain disease or disorder.

[0036] In the present invention, imatinib refers to Imatinib, with action targets: BCR-ABL, KIT, PDGFR. Indications: first-line treatment of CML and gastrointestinal stromal tumor (GIST). Dasatinib refers to Dasatinib, with action targets: BCR-ABL, SRC family kinases. Indications: CML patients resistant or intolerant to imatinib. Nilotinib refers to Nilotinib, with action target: BCR-ABL. Indications: newly diagnosed CML or imatinib-resistant patients. Sunitinib refers to Sunitinib, with action targets: VEGFR, PDGFR, KIT. Indications: renal cell carcinoma, GIST resistant to imatinib. Bosutinib refers to Bosutinib, for CML patients resistant to multiple TKIs. Regorafenib refers to Regorafenib, for GIST patients resistant to imatinib and sunitinib.

[0037] In the present invention, Venetoclax refers to Venetoclax, developed jointly by AbbVie and Roche. Indications: relapsed or refractory chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL); in combination with Obinutuzumab for treatment-naive CLL patients; first-line treatment in combination with chemotherapy drugs for acute myeloid leukemia (AML). ICP-248 (Mesutoclax), developed by Innovent Biologics. Research progress: Approved in February 2025 for a Phase III clinical trial in combination with the BTK inhibitor Orelabrutinib for first-line treatment of CLL / SLL. Lisaftoclax (APG-2575), developed by Ascentage Pharma. Research progress: Conducting clinical trials for relapsed and refractory AML, MDS (myelodysplastic syndrome), etc., initially showing controllable safety and significant efficacy; trials in combination with low-dose chemotherapy drugs (such as homoharringtonine, azacitidine) are ongoing. APG-2575: Structurally similar to Venetoclax, but with reduced risk of drug resistance through optimized design, currently in the global multi-center clinical stage; Tazemetostat: Although it is an EZH1 / 2 inhibitor, studies have shown that it can synergistically enhance the anti-tumor effect with CAR-T therapy by enhancing the immunogenicity of cancer cells and indirectly affecting the Bcl-2 related pathway.

[0038] The present invention provides a method for inhibiting the proliferation of leukemia cells in vitro and promoting the apoptosis of leukemia cells in vitro for non-therapeutic purposes, the method comprising the step of using the composition described above.

[0039] The present invention provides a kit, the kit comprising:

[0040] (1) A first preparation containing a Bcl-2 inhibitor;

[0041] (2) A second preparation containing a tyrosine kinase inhibitor;

[0042] (3) Instructions;

[0043] The first preparation is selected from: Venetoclax, Orelabrutinib, Lisaftoclax, APG-2575, Tazemetostat or a pharmaceutically acceptable salt thereof;

[0044] The second preparation is selected from: Imatinib, Dasatinib, Nilotinib, Sunitinib, Regorafenib, Bosutinib, Regorafenib or a pharmaceutically acceptable salt thereof.

[0045] Furthermore, the instructions indicate that the first preparation and the second preparation are used in combination to treat leukemia, or inhibit the proliferation of leukemia tissues or cells, or promote the apoptosis of leukemia cells or tissues.

[0046] Furthermore, the tyrosine kinase inhibitor is in the form of an orally administered small molecule multi-target.

[0047] Furthermore, the tyrosine kinase inhibitor includes imatinib, dasatinib, nilotinib, sunitinib, regorafenib, bosutinib, regorafenib or a pharmaceutically acceptable salt thereof.

[0048] Furthermore, the Bcl-2 inhibitor includes venetoclax, obutinib, Lisaftoclax, APG-2575, Tazemetostat or a pharmaceutically acceptable salt thereof.

[0049] Furthermore, the tyrosine kinase inhibitor is regorafenib or a pharmaceutically acceptable salt thereof.

[0050] Furthermore, the Bcl-2 inhibitor is venetoclax or a pharmaceutically acceptable salt thereof.

[0051] Furthermore, the leukemia includes acute leukemia, chronic leukemia, hairy cell leukemia, and prolymphocytic leukemia.

[0052] Furthermore, the acute leukemia includes acute lymphoblastic leukemia type L1, acute lymphoblastic leukemia type L2, acute lymphoblastic leukemia type L3, and acute myeloid leukemia.

[0053] Furthermore, the acute myeloid leukemia includes granulocytic leukemia, erythroleukemia, and megakaryocytic leukemia.

[0054] Furthermore, the chronic leukemia includes chronic lymphocytic leukemia and chronic myeloid leukemia.

[0055] Regorafenib (REGB) used in the present invention is an orally administered small molecule multi-target tyrosine kinase inhibitor, which can target the FLT3 pathway and block the downstream signal cascade caused by FLT3-ITD mutation (such as the PI3K / AKT and RAS / RAF / MEK / ERK pathways), thereby inhibiting tumor cell proliferation and inducing apoptosis.

[0056] Advantages and beneficial effects of the present invention:

[0057] The present invention discovers that the combination of REGB and ABT-199 shows stronger tumor inhibitory effects in in vitro experimental models, suggesting that REGB can enhance the anti-AML activity of ABT-199 in acute myeloid leukemia. Description of the Drawings

[0058] Figure 1 It is a diagram showing the results of AnnexinV-FITC / PI double staining and flow cytometry detection of single drug and dual drug combination treatment of AML cell lines.

[0059] Figure 2 It is the result diagram of MTS method for detecting single drug and dual drug combination treatment of AML cell lines. Detailed implementation manners

[0060] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by any ordinary person skilled in the art to which this invention belongs. Alternatively, some of the terms used herein have the meanings listed in the specification. All patents, published patent applications and publications cited herein are incorporated herein by reference as if set forth in full herein. It must be noted that the singular forms "a", "an" and "the" used herein and in the appended claims include plural forms unless the context clearly indicates otherwise.

[0061] Example 1

[0062] 1. Experimental materials

[0063] AML cell lines, AML clinical sample cells.

[0064] 2. Experimental methods

[0065] AnnexinV / PI staining and flow cytometry were used to detect cell apoptosis as follows:

[0066] After treating AML cells (1×10 6 ) with drugs, the cells were collected, centrifuged at 2000 rpm for 5 minutes, the supernatant was discarded, the cell pellet was washed with 1 mL of pre-cooled PBS, transferred to a 1.5 mL EP tube, centrifuged at 2000 rpm for 5 minutes, and the supernatant was discarded as much as possible. 50 μL of binding buffer was added to the 1.5 mL EP tube, then 5 μL of Annexin-FITC was added and mixed well, and stained at room temperature in the dark for 15 minutes. 10 μL of propidium iodide (PI) was added and mixed well, and stained at room temperature in the dark for 5 minutes. 450 μL of binding buffer was added to terminate the reaction. The sample was filtered through a double-layer 200-mesh nylon net, detected by a flow cytometer within 1 hour, the data was saved to make an image, and the data was processed with GraphPad Prism software.

[0067] 3. Experimental results

[0068] AML cell line molm-13 cells were treated with different concentrations of the multi-kinase inhibitor Regorafenib (REGB) and venetoclax (ABT-199) alone or in combination for 48 h. AnnexinV-FITC / PI double staining and flow cytometry were used to detect cell death, and the combination index (CI, CI < 1 indicates a synergistic effect of the two drugs in combination) was calculated using calcusyn software. The results are as Figure 1As shown, where (A) is a flow cytometry representative diagram. (B) is a statistical chart of flow cytometry for detecting apoptosis. **** represents p < 0.0001. It can be seen that the combined application of Regorafenib (REGB) and Venetoclax (ABT-199) significantly improves the therapeutic efficacy and has a synergistic effect.

[0069] Example 2

[0070] 1. Experimental materials

[0071] AML cell line.

[0072] 2. Experimental methods

[0073] The combined effect of the two drugs was determined by the MTS method: According to the solubility of the drugs, the two drugs were serially diluted with DMSO or other solvents so that each drug gradient was 10 times the final concentration. 10 μL of different drugs were added to the corresponding wells of a 96-well plate respectively. The single-drug wells were supplemented with 10 μL of the corresponding solvent, and the control wells were supplemented with 20 μL of the corresponding solvent. 80 μL of cell suspension was added to each well, and the other steps were exactly the same as above. Finally, a standard isobologram was made using the IC50 of the drugs to evaluate the combined effect of the two drugs.

[0074] 3. Experimental results

[0075] The AML cell line molm-13 cells were treated with different concentrations of the multi-kinase inhibitor Regorafenib (REGB) and Venetoclax (ABT-199) alone or in combination for 72 h. The proportion of viable cells was detected by the MTS method, and the IC50 value was calculated using GraphPad Prism and a standard isobologram was made. In the figure, the "points" falling below the diagonal line represent a synergistic effect of the combination of the two drugs, falling on the diagonal line represents an additive effect of the combination of the two drugs, and falling outside the diagonal line represents an antagonistic effect of the combination of the two drugs. The results are as Figure 2 shown, and all the landing points are located below the diagonal line, indicating an additive effect of the combination of the two drugs.

Claims

1. Use of a Bcl-2 inhibitor combined with a tyrosine kinase inhibitor in the preparation of a pharmaceutical composition for preventing and treating leukemia.

2. The use according to claim 1, wherein the tyrosine kinase inhibitor is in the form of an oral small molecule with multiple targets; Preferably, the tyrosine kinase inhibitor comprises imatinib, dasatinib, nilotinib, sunitinib, regorafenib, bosutinib, regorafenib or a pharmaceutically acceptable salt thereof; Preferably, the Bcl-2 inhibitor includes venetoclax, orelabrutinib, lisaftoclax, APG-2575, tazemetostat or a pharmaceutically acceptable salt thereof; Preferably, the tyrosine kinase inhibitor is regorafenib or a pharmaceutically acceptable salt thereof; Preferably, the Bcl-2 inhibitor is venetoclax or a pharmaceutically acceptable salt thereof; Preferably, the leukemia includes acute leukemia, chronic leukemia, hairy cell leukemia, and prolymphocytic leukemia; Preferably, the acute leukemia includes L1 acute lymphoblastic leukemia, L2 acute lymphoblastic leukemia, L3 acute lymphoblastic leukemia, and acute myeloid leukemia; Preferably, the acute myeloid leukemia includes granulocytic leukemia, erythroleukemia, and megakaryocytic leukemia; Preferably, the chronic leukemia includes chronic lymphocytic leukemia and chronic myeloid leukemia.

3. The use according to claim 1, wherein the mass ratio of the Bcl-2 inhibitor to the tyrosine kinase inhibitor is 8:1-2:1; Preferably, the Bcl-2 inhibitor is used at a concentration range of 1-2 μM; Preferably, the tyrosine kinase inhibitor is used at a concentration ranging from 0.125 to 0.5 μM.

4. A pharmaceutical composition for preventing or treating leukemia, comprising a combined use of a preventively effective amount or a therapeutically effective amount of the Bcl-2 inhibitor according to any one of claims 1 to 3 and a tyrosine kinase inhibitor.

5. The pharmaceutical composition according to claim 4, further comprising a pharmaceutically acceptable excipient; Preferably, the auxiliary materials include carriers, adjuvants, excipients, diluents or other liquid solvents, dispersing aids, suspending aids, surfactants, isotonic agents, thickeners, emulsifiers, preservatives, solid adhesives, and lubricants.

6. Application of Bcl-2 inhibitors combined with tyrosine kinase inhibitors in the preparation of products that inhibit leukemia cell proliferation and promote leukemia cell apoptosis.

7. A composition for inhibiting leukemia cell proliferation and promoting leukemia cell apoptosis, the composition comprising the following components or a combination of the following components: 1) A first active ingredient, wherein the first active ingredient comprises a Bcl-2 inhibitor; 2) A second active ingredient, wherein the second active ingredient comprises a tyrosine kinase inhibitor.

8. A method for inhibiting the proliferation of leukemia cells in vitro and promoting apoptosis of leukemia cells in vitro for non-therapeutic purposes, the method comprising the step of using the composition of claim 7.

9. A medicine box, comprising: (1) A first preparation containing a Bcl-2 inhibitor; (2) a second agent containing a tyrosine kinase inhibitor; (3) Instructions; The first agent is selected from: venetoclax, orelabrutinib, lisaftoclax, APG-2575, tazemetostat or a pharmaceutically acceptable salt thereof; The second agent is selected from: imatinib, dasatinib, nilotinib, sunitinib, regorafenib, bosutinib, regorafenib or a pharmaceutically acceptable salt thereof.

10. The drug kit according to claim 9, wherein the instructions indicate that the first agent and the second agent are used in combination to treat leukemia, or inhibit the proliferation of leukemia tissues or cells, or promote apoptosis of leukemia cells or tissues.

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