Application of combination of decitabine and anti-HER2 antibody in preparation of medicine for treating tumors

Abstract

The invention relates to application of a combination of decitabine and an anti-HER2 antibody in preparation of a medicine for treating tumors. The invention provides application of decitabine or an analogue thereof in preparation of a medicine for treating tumors in combination with an anti-HER2 (Human Epidermal Growth Factor Receptor 2) antibody. The invention also provides application of decitabine or an analogue thereof, an anti-HER2 antibody and one, more or all of a cell aging induction medicine, a ZBP1 agonist and an SETDB1 inhibitor in preparation of medicines for treating tumors. The present invention also provides pharmaceutical compositions and kits comprising decitabine or an analog thereof and an anti-HER2 antibody.

Description

Technical Field

[0001] This invention belongs to the field of medicine, and more specifically, this invention relates to the use of decitabine combined with anti-HER2 antibody in the preparation of drugs for treating tumors. Background Technology

[0002] Immune escape is a key behavioral characteristic of tumor cells, one mechanism of which is the overexpression of immunosuppressive signals. HER2 is highly expressed in 20%-30% of breast cancers and other types of cancer, and is closely associated with increased disease invasiveness, recurrence rate, and poor prognosis. Trastuzumab (Herceptin) is a humanized monoclonal antibody targeting HER2, and its effectiveness has been demonstrated in animal and human studies. However, the use of anti-HER2 antibodies alone has limitations, such as high cost, long-term high-dose use being financially unaffordable for some patients, and the limited anti-tumor efficacy of anti-HER2 antibodies alone, which easily leads to drug resistance.

[0003] To address the limitations of using anti-HER2 antibodies alone, researchers have attempted to combine various drugs with anti-HER2 antibodies for anti-tumor therapy. However, the efficacy of different drugs combined with anti-HER2 antibodies for tumor treatment varies, and some drug combinations can cause more severe adverse reactions and may lead to drug resistance in tumor cells, further hindering tumor treatment.

[0004] Therefore, there is an urgent need in this field to explore a drug that can be effectively combined with anti-HER2 antibodies for anti-tumor purposes, so as to enhance the anti-tumor efficacy of anti-HER2 antibodies and reduce the occurrence of drug resistance. Summary of the Invention

[0005] The purpose of this invention is to provide the use of decitabine in combination with anti-HER2 antibody in the preparation of a medicament for treating tumors.

[0006] A first aspect of the invention provides the use of decitabine or an analogue thereof in combination with an anti-HER2 antibody in the preparation of a medicament for treating tumors.

[0007] A second aspect of the invention provides the use of decitabine or an analogue thereof, an anti-HER2 antibody, and one, more or all of a selection of cell senescence-inducing drugs, ZBP1 agonists and SETDB1 inhibitors in the preparation of a medicament for treating tumors.

[0008] In one or more embodiments, the decitabine has a structural formula as shown in Formula I:

[0009]

[0010] In one or more embodiments, the decitabine analogues include decitabine hydrates, solvates, enantiomers, diastereomers, pharmaceutically acceptable salts, prodrugs, and complexes.

[0011] In one or more embodiments, the decitabine analogues include 5-azacytidine, 5-fluoro-2-deoxycytidine, and zebularine.

[0012] In one or more embodiments, the anti-HER2 antibody is an antibody or antigen-binding fragment thereof that binds to the epidermal growth factor 2 (HER2) antigen.

[0013] In one or more embodiments, the antibody includes: monoclonal antibody, polyclonal antibody, multispecific antibody, humanized full-length human antibody, chimeric antibody and camel-derived single-domain antibody; the antigen-binding fragment includes: Fab, Fab', F(ab')2, Fv, double-chain antibody, linear antibody, single-chain antibody, nanobody.

[0014] In one or more embodiments, the anti-HER2 antibody includes: trastuzumab, pertuzumab, inetuzumab, and maggotuximab.

[0015] In one or more specific embodiments, the anti-HER2 antibody is antibody 7.16.4.

[0016] In one or more embodiments, the cell senescence-inducing drug is a drug capable of inducing cell senescence and / or inducing necrosis and apoptosis.

[0017] In one or more embodiments, the cell senescence-inducing drug includes: chemotherapeutic drugs, RAS / RAF / MEK inhibitors, CDK4 / 6 inhibitors, telomerase inhibitors, DNA methyltransferase inhibitors, histone deacetylase inhibitors, and MDM2 inhibitors.

[0018] In one or more embodiments, the chemotherapeutic agents include doxorubicin, etoposide, camptothecin, cisplatin, carboplatin, oxaliplatin, Nutlin 3, and RG7112; the RAS / RAF / MEK inhibitors include sorafenib and lysyl The inhibitors include oxidase, Selumetinib (AZD6244), Trametinib, and Pimasertib; CDK4 / 6 inhibitors include Palbociclib, Ribociclib, Abemaciclib, and Dalpiciclib; telomerase inhibitors include Imetelstat (GRN163L, Rytelo), BIBR1532, and THIO (6-thio-dG, 6-thio-2'-deoxyguanosine); DNA methyltransferase inhibitors include azacitidine, zabulolin, RG108, GSK3685032, and SGI-110; histone deacetylase inhibitors include vorinostat, romidesin, GSK-LSD1, Entinostat, Valproic acid, Tubacin, and ACY-1215; and MDM2 inhibitors include Nutlin-3a, APG-115, MI-773, AMG-232, and Idasanutlin.

[0019] In one or more embodiments, the ZBP1 agonist is any substance that can enhance the activity of ZBP1, increase the stability of ZBP1 or its encoding gene, upregulate the expression of ZBP1, increase the effective duration of ZBP1, promote the transcription and translation of the ZBP1 gene, or promote ZBP1-mediated cell necrosis and apoptosis.

[0020] In one or more embodiments, the ZBP1 agonist includes CBL0137, hydrogen peroxide, arsenite, cisplatin, and ADAR1 inhibitors.

[0021] In one or more embodiments, the SETDB1 inhibitor is a substance that can reduce the activity of SETDB1, reduce the stability of SETDB1 or its encoding gene, downregulate the expression of SETDB1, reduce the effective duration of SETDB1, inhibit the transcription and translation of the SETDB1 gene, or promote necrotic apoptosis by inhibiting SETDB1.

[0022] In one or more embodiments, the SETDB1 inhibitor includes Mithramycin A, MithralogEC-8042, 3'-deazaneplanocin A (DZNep), Paclitaxel, (R,R)-59, and SETDB1-TTD-IN-1.

[0023] In one or more embodiments, the treatment of tumors includes: inhibiting methylation of tumor cells, promoting tumor cell senescence, promoting tumor cell necrosis and apoptosis, inhibiting tumor cell growth, and / or reducing tumor cell drug resistance.

[0024] In one or more embodiments, the promotion of tumor cell senescence includes inducing SETDB1 inactivation; the promotion of tumor cell necrosis and apoptosis includes activating ZBP1.

[0025] In one or more embodiments, the tumor cells are HER2-positive tumor cells, preferably HER2-positive breast cancer cells.

[0026] In one or more embodiments, the tumor is a tumor containing or expressing the HER2 antigen, preferably including primary or metastatic cancer.

[0027] In one or more embodiments, the tumor is a HER2-positive tumor; more preferably, the tumor includes breast cancer, gastric cancer, colon cancer, liver cancer, small cell lung cancer, non-small cell lung cancer, melanoma, pancreatic cancer, skin cancer, uterine cancer, ovarian cancer, Kaposi's sarcoma, squamous cell carcinoma, or a combination thereof.

[0028] A third aspect of the present invention provides a pharmaceutical composition comprising an anti-HER2 antibody and decitabine or an analogue thereof.

[0029] In one or more embodiments, the decitabine or its analogues are as described in any embodiment of the present invention.

[0030] In one or more embodiments, the anti-HER2 antibody is as described in any embodiment of the present invention.

[0031] In one or more embodiments, the pharmaceutical composition further includes a pharmaceutically acceptable carrier and / or one or more additional active ingredients.

[0032] In one or more embodiments, the additional active ingredient is selected from one, more, or all of the following: cell senescence-inducing drugs, ZBP1 agonists, and SETDB1 inhibitors.

[0033] In one or more embodiments, the cell senescence-inducing drug is as described in any embodiment of the present invention.

[0034] In one or more embodiments, the ZBP1 agonist is as described in any embodiment of the present invention.

[0035] In one or more embodiments, the SETDB1 inhibitor is as described in any embodiment of the present invention.

[0036] In one or more embodiments, the pharmaceutical composition is a pharmaceutical composition for treating tumors.

[0037] In one or more embodiments, the treatment of the tumor is as described in any embodiment of the present invention, and / or the tumor is as described in any embodiment of the present invention.

[0038] A fourth aspect of the present invention provides a medicine box comprising a pharmaceutical composition as described in any embodiment of the present invention.

[0039] Other aspects of the invention will be apparent to those skilled in the art from the disclosure herein. Attached Figure Description

[0040] Figure 1 Figure 1 shows the tumor volume detection results in mice treated with a combination of decitabine and anti-HER2 antibody. Figure 1 A is a schematic diagram of the drug administration time points for each group of mice during the experiment of Example 1. Figure 1 B represents the tumor growth curves of mice in each group during the experiment. Figure 1 C is a representative graph showing the tumor size of mice in each group on day 16 after tumor inoculation.

[0041] Figure 2 The antitumor effect of decitabine combined with anti-HER2 antibody depends on the ZBP1 protein. Figure 2 A shows the activation status of the necrosis-apoptosis signaling pathway in ZBP1-MLKL cells after combined drug administration. Figure 2 B shows the effect of combined drug administration on the activation of the cell necrosis-apoptosis signaling pathway after ZBP1 gene knockout. Figure 2 C depicts the effect of combined drug administration on tumor curves in each group of mice with ZBP1 gene knockout.

[0042] Figure 3 The antitumor effect of decitabine combined with anti-HER2 antibody is partly dependent on cellular senescence. Figure 3 A shows the staining results of galactosidase after combined drug administration, scale bar = 40 μm. Figure 3 B and Figure 3 C shows the P21 levels of mice in each group after combined drug administration. Figure 3 B) and Ki-67( Figure 3 C) Staining condition, scale bar = 50 μm. Figure 3 D depicts the changes in the expression levels of key proteins in the necrosis-apoptosis signaling pathway of ZBP1-MLKL cells after drug clearance of senescent cells and combined drug administration.

[0043] Figure 4Decitabine combined with anti-HER2 antibody induces cellular senescence and promotes loss of SETDB1 function. Figure 4 AB showed the expression of SETDB1 protein after combination therapy. Figure 4 C depicts the changes in SETDB1 protein expression levels after drug clearance of senescent cells and combined drug administration. Figure 4 D depicts the changes in expression levels of key proteins in the signaling pathway of cell necrosis and apoptosis in SETDB1 knockout cells after drug clearance of senescent cells and combined drug administration. Detailed Implementation

[0044] Through in-depth research, the inventors discovered that the combination of decitabine and anti-HER2 antibodies can produce a significant synergistic anti-tumor effect. This synergistic effect induces SETDB1 inactivation through cellular senescence, effectively activating ZBP1-mediated necrotizing apoptosis, directly inhibiting tumor cell growth, and exerting an anti-tumor effect.

[0045] the term

[0046] The term "ZBP1" refers to a cell necrosis and apoptosis-related protein. The amino acid sequence of human ZBP1 can be found at NCBI locus number: NP_001153889.1.

[0047] The term "SETDB1" refers to SET domain bifurcated histone lysine methyltransferase 1. The amino acid sequence of human SETDB1 can be found at NCBI locus number: NP_001138887.1.

[0048] The “effective amount” of a drug refers to the amount necessary to induce physiological changes in the cells or tissues to which it is administered.

[0049] A "therapeuticly effective amount" of a pharmaceutical agent, such as a pharmaceutical composition, refers to the amount that effectively achieves the desired therapeutic or preventative outcome at the necessary dosage and time period. A therapeutically effective amount of a pharmaceutical agent may eliminate, reduce, delay, minimize, or prevent adverse effects of disease.

[0050] The “individual” or “subject” is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). Preferably, the individual or subject is a human.

[0051] The term "pharmaceutical composition" refers to a formulation in which the biological activity of the active ingredient contained therein is effective and which does not contain any other ingredients that would have unacceptable toxicity to a subject who would receive the composition.

[0052] "Pharmaceutically acceptable carriers" refer to components in a drug composition that are non-toxic to the subjects, other than the active ingredient. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0053] The term "tumor" refers to a wide variety of diseases characterized by the uncontrolled growth of abnormal cells in the body. For example, unregulated cell division, growth, and proliferation lead to the formation of malignant tumors, which can invade adjacent tissues and can also metastasize to distant parts of the body via the lymphatic system or bloodstream.

[0054] The term "treatment / prevention" (and its grammatical variations) refers to an attempt to alter the natural course of disease in an individual being treated, and can be a clinical intervention performed for prevention or during the course of clinicopathological processes. The desired effects of treatment include, but are not limited to, preventing the onset or recurrence of disease, relieving symptoms, reducing any direct or indirect pathological consequences of the disease, preventing metastasis, slowing the rate of disease progression, improving or alleviating the disease state, reducing drug resistance, and eliminating or improving prognosis.

[0055] Combined use of decitabine and anti-HER2 antibodies

[0056] Specific embodiments of the present invention target HER2 + In breast cancer, it has been found that the combination therapy of decitabine and anti-HER2 antibody can produce a significant synergistic effect by inhibiting SETDB1 expression, increasing sensitivity to immunotherapy and resulting in better treatment outcomes.

[0057] Further research revealed that the combination therapy of decitabine and anti-HER2 antibodies first inhibits SETDB1 expression through epigenetic mechanisms and activates ZBP1-mediated necroptosis, thereby enhancing the tumor's endogenous immunogenicity and increasing its sensitivity to immunotherapy. Furthermore, the cellular senescence induced by the combination therapy also inhibits SETDB1 expression and activates ZBP1-mediated necroptosis, further amplifying the tumor's endogenous immunogenicity and enhancing the efficacy of immunotherapy. The combination therapy of decitabine and anti-HER2 antibodies directly inhibits tumor cell growth, providing a new treatment option for HER2-positive tumor patients and helping to reduce the occurrence of drug resistance.

[0058] Based on the above-mentioned findings of the inventors, this invention provides the use of decitabine in combination with an anti-HER2 antibody in the preparation of a medicament for treating tumors. This invention also provides a method for treating tumors, comprising administering a therapeutically effective amount of decitabine and a therapeutically effective amount of an anti-HER2 antibody to the individual in need.

[0059] In this article, "decitabine", "5AZA", "5-aza-2′-deoxycytidine" or "5-aza-2'-deoxycytidine" all refer to an epigenetic drug with the trade name Dacogen and CAS number 2353-33-5, which exerts its anti-tumor effect by inhibiting DNA methyltransferase.

[0060] The structural formula of decitabine is shown in Formula I:

[0061]

[0062] In this document, "decitabine" also includes its analogues. The term "analyte" generally refers to a substance having a structure similar to decitabine and a function similar to decitabine (e.g., inhibiting DNA methyltransferases, inducing demethylation, or exerting cytotoxic effects). Such analogues include decitabine hydrates, solvates, enantiomers, diastereomers, pharmaceutically acceptable salts, prodrugs, and complexes.

[0063] For example, the decitabine analogue may be 5-azacytidine, 5-fluoro-2'-deoxycytidine, or zebularine.

[0064]

[0065] As used herein, “antibody” refers to any form of antibody or fragment thereof capable of achieving the desired biological or binding activity. Therefore, it is used in the broadest sense, but is not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies, humanized full-length human antibodies, chimeric antibodies, and camel-derived single-domain antibodies. The term “antibody” includes: naturally occurring and non-naturally occurring Abs; monoclonal and polyclonal Abs; chimeric and humanized Abs; human or non-human Abs; fully synthetic Abs; and single-chain Abs. Non-human Abs can be humanized through recombinant methods to reduce their immunogenicity in humans. The term “antibody fragment” or “antigen-binding fragment” refers to an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the specific ability of a full-length antibody to bind to an antigen, such as a fragment retaining one or more CDR regions. Examples of antibody-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; double-chain antibodies; linear antibodies; single-chain antibody molecules; nanobodies; and multispecific antibodies formed from antibody fragments. Furthermore, the term “sequence” as used herein should generally be understood to include both the relevant amino acid sequence and the nucleic acid or nucleotide sequence encoding the sequence, unless a more specific interpretation is required herein.

[0066] In this article, "anti-HER2 antibody" refers to an antibody that binds to the epidermal growth factor 2 (HER2) antigen. In the stated uses or methods, the anti-HER2 antibody blocks the dimerization activation of the human HER2 receptor and / or mediates the killer effect of the immune system (ADCC), thereby activating the immune system, including T cells, and targeting and inhibiting tumor growth. The human HER2 amino acid sequence can be found at NCBI locus number: NP_004439.2. In this invention, the "anti-HER2 antibody" includes, but is not limited to: trastuzumab, pertuzumab, inetuzumab, and maggotuximab. In some specific embodiments, the anti-HER2 antibody is antibody 7.16.4, information of which can be found in Drebin, JA, et al. (1984). "Monoclonal antibodies identify a cell-surface antigen associated with an activated cellular oncogene" Nature 312(5994):545-548.

[0067] In this invention, the tumor is a tumor containing or expressing the HER2 antigen; including but not limited to primary or metastatic cancer. In some embodiments, the tumor is a HER2-positive tumor, preferably including breast cancer, gastric cancer, colon cancer, liver cancer, small cell lung cancer, non-small cell lung cancer, melanoma, pancreatic cancer, skin cancer, uterine cancer, ovarian cancer, Kaposi's sarcoma, squamous cell carcinoma, or combinations thereof. In some specific embodiments, the tumor cells are HER2-positive tumor cells, preferably TUBO HER2+ breast cancer cells.

[0068] When a therapeutically effective dose of decitabine and a therapeutically effective dose of an anti-HER2 antibody are used in combination, they preferably result in inhibition of tumor growth, reduction of tumor volume, increase in the frequency and duration of asymptomatic periods, or reduction of resistance to the anti-HER2 antibody. For example, for the treatment of HER2-related tumors (including, for example, breast cancer, ovarian cancer, endometrial cancer, cervical cancer, etc.), the "therapeutically effective dose" preferably inhibits cell growth or tumor growth by at least about 10%, preferably at least about 20%, more preferably at least about 30%, more preferably at least about 40%, more preferably at least about 50%, more preferably at least about 60%, more preferably at least about 70%, and more preferably at least about 80%, relative to untreated subjects. The ability to inhibit tumor growth can be evaluated in an animal model system that predicts the efficacy of treatment for human tumors. Alternatively, it can also be evaluated by examining the ability to inhibit cell growth, which can be determined in vitro by assay.

[0069] Pharmaceutical Composition

[0070] The present invention also provides a pharmaceutical composition comprising the anti-HER2 antibody described herein and decitabine. The pharmaceutical composition may further comprise various pharmaceutically acceptable carriers in the art.

[0071] Pharmaceutically acceptable carriers are non-toxic to recipients at the doses and concentrations used, and may include, but are not limited to: buffers such as acetates, Tris, phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (such as octadecyl dimethyl benzyl ammonium chloride; chlorhexidine diammonium chloride; benzalkonium chloride, benzyl chloride; phenols, butanol, or benzyl alcohol; hydroxybenzoic acid esters, such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); proteins, such as blood... Albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; tension modifiers, such as trehalose and sodium chloride; sugars, such as sucrose, mannitol, trehalose, or sorbitol; surfactants, such as polysorbate; salt-forming counterions, such as sodium; metal complexes (such as Zn-protein complexes); and / or nonionic surfactants, such as... Or polyethylene glycol (PEG).

[0072] The pharmaceutical compositions of the present invention can be formulated into any conventional dosage form using conventional methods. The dosage form can be diverse, as long as it enables the active ingredient to effectively reach the mammalian body. Examples include: injections, infusions, tablets, capsules, and pills. The active components (anti-HER2 antibody and decitabine) can be present in suitable solid or liquid carriers or diluents. The pharmaceutical compositions of the active components of the present invention can also be stored in sterile instruments suitable for injection or infusion. The effective dose of the active components (anti-HER2 antibody and decitabine) in the composition can vary depending on the administration method and the severity of the disease being treated, based on the experience and advice of the clinician.

[0073] The pharmaceutical compositions of the present invention can be administered orally, as well as intravenously, intramuscularly, or subcutaneously. Injection is preferred. Suitable forms for injection include sterile aqueous solutions or dispersions and sterile powders. Suitable forms for oral administration include, but are not limited to, tablets, powders, capsules, sustained-release formulations, etc. In all cases, these forms must be sterile and must be fluids that can be easily dispensed using a syringe.

[0074] The therapeutically effective dose of the anti-HER2 antibody and decitabine described in this invention can vary depending on the administration method and the severity of the disease to be treated. The selection of the therapeutically effective dose can be determined by those skilled in the art based on various factors (e.g., through clinical trials). These factors include, but are not limited to: the pharmacokinetic parameters of the anti-HER2 antibody and decitabine, such as bioavailability, metabolism, and half-life; the severity of the disease to be treated, the patient's weight, the patient's immune status, and the route of administration.

[0075] In specific embodiments of the present invention, some dosing regimens for animals such as mice are provided. Converting the dosage for animals such as mice to a dosage suitable for humans is readily performed by those skilled in the art, for example, using the Meeh-Rubner formula: Meeh-Rubner formula: A = k′(W² / 3) / 10,000. Where A is the body surface area, expressed in meters. 2 Calculations are as follows: W represents body weight in grams; K is a constant that varies depending on the animal species, generally 9.1 for mice and rats, 9.8 for guinea pigs, 10.1 for rabbits, 9.9 for cats, 11.2 for dogs, 11.8 for monkeys, and 10.6 for humans. It should be understood that dosage conversions can vary depending on the drug and clinical circumstances, and are based on the assessment of an experienced pharmacist.

[0076] The pharmaceutical compositions of the present invention may further comprise other active ingredients, i.e., be used in combination with other drugs. For example, the combination therapy may be the anti-HER2 antibody and decitabine in combination with at least one other antitumor drug.

[0077] Based on the inventors' research, the combined treatment strategy of decitabine and anti-HER2 antibodies first inhibits SETDB1 expression through epigenetic mechanisms and activates ZBP1-mediated necroptosis, thereby enhancing the tumor's endogenous immunogenicity and increasing its sensitivity to immunotherapy. The combined treatment of decitabine and anti-HER2 antibodies also induces cellular senescence, which similarly inhibits SETDB1 expression and activates ZBP1-mediated necroptosis, further amplifying the tumor's endogenous immunogenicity and thus enhancing the efficacy of immunotherapy.

[0078] Therefore, the other active ingredients or drugs may be one, more, or all of the following: cell senescence-inducing drugs, ZBP1 agonists, and SETDB1 inhibitors.

[0079] The term "cell senescence-inducing drugs" generally refers to drugs that can induce cell senescence and / or induce cell necrosis and apoptosis, including but not limited to: chemotherapy drugs (such as doxorubicin, etoposide, camptothecin, cisplatin, carboplatin, oxaliplatin, Nutlin 3, RG7112), and RAS / RAF / MEK inhibitors (such as sorafenib, lysyl...). Oxidase, Selumetinib (AZD6244), Trametinib, Pimasertib), CDK4 / 6 inhibitors (such as Palbociclib, Ribociclib, Abemaciclib, Dalpiciclib), telomerase inhibitors (such as Imetelstat (GRN163L, Rytelo), BIBR1532, THIO (6-thio-dG, 6-thio-2'-deoxyguanosine)), DNA methyltransferase inhibitors (such as azacitidine, decitabine, zabralin, RG108, GSK3685032, SGI-110), histone deacetylase inhibitors (such as vorinostat, romedixin, GSK-LSD1, Entinostat, Valproic acid, Tubacin, ACY-1215), and MDM2 inhibitors (such as Nutlin-3a, APG-115, MI-773, AMG-232, Idasanutlin).

[0080] The term "ZBP1 agonist" refers to any substance that can enhance the activity of ZBP1, increase the stability of ZBP1 or its encoding gene, upregulate ZBP1 expression, increase the effective duration of ZBP1 action, promote the transcription and translation of the ZBP1 gene, or promote ZBP1-mediated cell necrosis and apoptosis. These substances can be used in this invention as ZBP1 agonists, thereby inhibiting tumor growth and / or reducing tumor drug resistance. The ZBP1 agonists include, but are not limited to, CBL0137, hydrogen peroxide, arsenite, cisplatin, and ADAR1 inhibitors.

[0081] The term "SETDB1 inhibitor" refers to any substance that can reduce the activity of SETDB1, decrease the stability of SETDB1 or its encoding gene, downregulate SETDB1 expression, reduce the effective duration of SETDB1 action, inhibit the transcription and translation of the SETDB1 gene, or promote cell necrosis and apoptosis by inhibiting SETDB1. These substances can be used in this invention as SETDB1 inhibitors, thereby inhibiting tumor growth and / or reducing tumor drug resistance. The SETDB1 inhibitors include, but are not limited to, Mithramycin A, Mithralog EC-8042, 3'-deazaneplanocin A (DZNep), Paclitaxel, (R,R)-59, and SETDB1-TTD-IN-1.

[0082] The present invention also provides a kit containing the aforementioned pharmaceutical composition or directly containing the aforementioned decitabine and anti-HER2 antibody. Furthermore, the kit may also include instructions on how to use the medication contained therein.

[0083] Beneficial effects:

[0084] This invention reveals that combining decitabine with an anti-HER2 antibody produces a significant synergistic effect. This combination therapy strategy first inhibits SETDB1 expression through epigenetic mechanisms and activates ZBP1-mediated apoptosis, thereby enhancing the tumor's endogenous immunogenicity and increasing its sensitivity to immunotherapy. Furthermore, the combination therapy-induced cellular senescence also inhibits SETDB1 expression and activates ZBP1-mediated apoptosis, further amplifying the tumor's endogenous immunogenicity and enhancing the efficacy of immunotherapy. Finally, the combination therapy of decitabine and the anti-HER2 antibody directly inhibits tumor cell growth, providing a new treatment option for HER2-positive tumor patients and helping to reduce the occurrence of drug resistance. This combination therapy not only promises to alleviate the economic burden on patients but also improves treatment accessibility and patient compliance.

[0085] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods and reagents in the following embodiments that do not specify specific conditions are, unless otherwise stated, conventional methods and reagents in the art.

[0086] Example 1: The combination of the DNA methyltransferase inhibitor decitabine and anti-HER2 antibody for the treatment of HER2 + Breast cancer

[0087] I. Experimental Materials

[0088] The experimental animals were 6-8 week old SPF-grade female BALB / c mice. The tumor cell line was TUBO HER2. + Breast cancer cells were cultured in DMEM containing 10% fetal bovine serum at 37°C and 5% CO2. The DNA methyltransferase inhibitor decitabine 5AZA was purchased from Sigma-Aldrich (CAS No.: 2353-33-5), and the anti-HER2 antibody was purchased from BioXcell (Lot: BE0277), diluted with phosphate-buffered saline (PBS) before use. The fetal bovine serum was an Excel product, and the DMEM medium was a GIBCO product.

[0089] II. Experimental Methods

[0090] 1. Tumor model creation and grouping

[0091] In the culture of tumor cells in the logarithmic growth phase, the original culture medium was first removed, and the cells were then washed with PBS. Next, the cells were treated with trypsin until they detached from the culture wall. After trypsin treatment, the trypsin was removed, fresh culture medium was added, and the cells were gently pipetted to disperse them. The cells were then transferred to centrifuge tubes and centrifuged at 1000 rpm for 5 minutes to pellet the cells. After centrifugation, the supernatant was removed, and the cells were washed again with PBS to prepare a cell suspension. Finally, the cell suspension was diluted to 0.1 mL per mouse, containing 5 × 10⁶ cells / mL. 5 A concentration of [number] cells was administered subcutaneously to the back of mice. When the tumor volume grew to approximately 100 mm... 3 At that time, the mice were randomly divided into four groups for subsequent experiments:

[0092] (A) Ctrl group;

[0093] (B) Group 5AZA;

[0094] (C) anti-HER2 group;

[0095] (D)5AZA+anti-HER2 group.

[0096] 2. Drug intervention, tumor curve plotting, and calculation of tumor inhibition rate.

[0097] The tumor, inoculated with tumor cells, grew to 100mm. 3 At that time, the patients were randomly assigned to groups before administration. Each group received an intraperitoneal injection of the following doses of anti-HER2 antibody on day 9, and an intratumoral injection of the following doses of 5-AZA on days 9 and 16:

[0098] (A) Solvent control group (Ctrl): Administered an equal volume of solvent (Vehicle);

[0099] (B) 5AZA group: 5AZA 2mg / kg;

[0100] (C) Anti-HER2 group: 50 μg of anti-HER2 antibody per animal;

[0101] (D) 5AZA+anti-HER2 group: 5AZA 2mg / kg + anti-HER2 antibody 50μg / each.

[0102] Tumor volume was recorded twice a week, and a tumor growth curve was plotted. The tumor volume was calculated using the following formula: V = (L × W × H) / 2; where L is the length, W is the width, and H is the height. The tumor inhibition rate was calculated using the following formula: Tumor inhibition rate (%) = [1 - (average tumor volume in the drug intervention group / average tumor volume in the solvent control group)] × 100%.

[0103] 3. Determination of synergistic effects of two drugs used together

[0104] The synergistic effect of the two drugs was determined according to the literature method (Jin Zhengjun, Additive Effect in Combined Drug Use [J]. Chinese Journal of Pharmacology, 1980, 1(2): 70-76). The specific determination method is as follows: Q = E(a+b) / (Ea+Eb-Ea×Eb). In this formula, E(a+b) represents the actual tumor inhibition rate when the two drugs are used in combination, and Ea and E represent the tumor inhibition rates when the two drugs are used alone, respectively. The denominator (Ea+Eb-Ea×Eb) represents the expected combined effect. The Q value is the ratio of the actual effect to the expected effect. When the Q value is between 0.85 and 1.15, it indicates that the combined effect of the two drugs is additive; when the Q value is greater than 1.15, it indicates that there is a synergistic effect; and when the Q value is less than 0.85, it indicates that there is an antagonistic effect.

[0105] 4. Statistical Analysis

[0106] The experimental results were statistically analyzed using Prism software (GraphPad Prism 8.0). Data are expressed as mean ± SEM. One-way ANOVA was used to analyze differences among multiple groups, and the Turkey test was used for post-parameter testing. Significant differences were defined as follows: ** for P < 0.01; **** for P < 0.0001.

[0107] III. Experimental Results

[0108] At the end of the experiment, the number of surviving mice in each group was 5, which met the requirements of the pharmacological experiment. Figure 1 A represents the drug administration time points for each group of mice during the experiment. Figure 1 B represents the tumor growth curves of mice in each group during the experiment. Figure 1C is a representative graph of tumor size in each group of mice on day 16 after tumor inoculation. Table 1 shows the tumor inhibition rate (%) and Q value calculation results for each drug intervention group.

[0109] Table 1. Calculation results of tumor inhibition rate (%) and Q value in the drug intervention group.

[0110]

[0111] Experimental results showed that, compared with the groups treated with 5AZA or anti-HER2 antibodies alone, the combination therapy group exhibited better anti-tumor effects in terms of tumor growth curve and tumor size, with significant differences. The Q value of the combined use of 5AZA and anti-HER2 antibody in anti-tumor therapy was >1.15, indicating a significant synergistic effect. Therefore, decitabine can be combined with anti-HER2 antibodies for anti-tumor treatment to enhance the anti-HER2 antibody efficacy, reduce the development of drug resistance, and lower the economic burden on patients.

[0112] Example 2: The antitumor effect of the combination therapy of the DNA methyltransferase inhibitor decitabine and anti-HER2 antibody depends on ZBP1-mediated cell necrosis and apoptosis.

[0113] I. Experimental Materials

[0114] The experimental animals were 6-8 week old SPF-grade female BALB / c mice. The tumor cell line was CRISPR-Ctrl, ZBP1-gRNATUBO cells, cultured in DMEM containing 10% fetal bovine serum at 37°C and 5% CO2. The DNA methyltransferase inhibitor decitabine 5AZA was purchased from Sigma-Aldrich (CAS No.: 2353-33-5), and the anti-HER2 antibody was purchased from BioXcell (Lot: BE0277), diluted with phosphate-buffered saline (PBS) before use. The fetal bovine serum was an Excel product, and the DMEM medium was a GIBCO product.

[0115] II. Experimental Methods

[0116] 1. Tumor cell grouping, xenograft model creation and grouping

[0117] In the in vitro combined therapy experiment, TUBO cells were seeded into culture plates the night before the experiment. The experiment began on day 1, with TUBO cells treated with a combination of 5-AZA (0.15 μM) and anti-HER2 antibody (1.5 μg / mL). The following day, the 5-AZA was removed, and the cells were cultured again with anti-HER2 antibody (1.5 μg / mL) for 4 days.

[0118] In the culture of tumor cells in the logarithmic growth phase, the original culture medium was first removed, and the cells were then washed with PBS. Next, the cells were treated with trypsin until they detached from the culture wall. After trypsin treatment, the trypsin was removed, fresh culture medium was added, and the cells were gently pipetted to disperse them. The cells were then transferred to centrifuge tubes and centrifuged at 1000 rpm for 5 minutes to pellet the cells. After centrifugation, the supernatant was removed, and the cells were washed again with PBS to prepare a cell suspension. Finally, the cell suspension was diluted to 0.1 mL per mouse, containing 5 × 10⁶ cells / mL. 5 A concentration of [number] cells was administered subcutaneously to the back of mice. When the tumor volume grew to approximately 100 mm... 3 Then, the mice were randomly divided into four groups for subsequent experiments:

[0119] (A) CRISPR-Ctrl group;

[0120] (B) CRISPR-Ctr-5AZA+anti-HER2 group;

[0121] (C)ZBP1-gRNA-Ctrl group;

[0122] (D)ZBP1-gRNA-5AZA+anti-HER2 group.

[0123] 2. Drug intervention and tumor curve plotting

[0124] To assess the impact of this combination therapy on the necrosis-apoptosis pathway, Western blot analysis was used to detect the expression levels of key proteins in the ZBP1-MLKL cell necrosis-apoptosis signaling pathway. Five days after drug administration, total cellular proteins were extracted and subjected to Western blot experiments to observe the specific effects of the combined use of 5AZA and anti-HER2 antibody on the expression of necrosis-apoptosis-related proteins (ZBP1 and MLKL) in TUBO cells, thereby further understanding the potential mechanisms and efficacy of this combination therapy.

[0125] The tumor, inoculated with tumor cells, grew to 100mm. 3 At that time, the patients were randomly assigned to groups before administration. Each group received an intraperitoneal injection of the following doses of anti-HER2 antibody on day 9, and an intratumoral injection of the following doses of 5-AZA on days 9 and 16:

[0126] (A) CRISPR-Ctrl group: equal volume of solvent;

[0127] (B) CRISPR-Ctr-5AZA+anti-HER2 group: 5AZA 2mg / kg + anti-HER2 antibody 50μg / each;

[0128] (C)ZBP1-gRNA-Ctrl group: equal volume of solvent;

[0129] (D)ZBP1-gRNA-5AZA+anti-HER2 group: 5AZA 2mg / kg+anti-HER2 antibody 50μg / each.

[0130] Tumor volume was recorded twice a week, and a tumor growth curve was plotted. The tumor volume was calculated using the following formula: V=(L×W×H) / 2; where L is the length, W is the width, and H is the height.

[0131] 3. Statistical Analysis

[0132] The experimental results were statistically analyzed using Prism software (GraphPad Prism 8.0). Data are expressed as mean ± SEM. One-way ANOVA was used to analyze differences among multiple groups, and the Turkey test was used for post-parameter testing. Significant differences were defined as follows: **** was considered P < 0.0001.

[0133] III. Experimental Results

[0134] At the end of the experiment, five mice survived in each group, a result that meets the standard requirements for pharmacological experiments. Figure 2 A shows the activation status of the necrosis-apoptosis signaling pathway in ZBP1-MLKL cells after combined drug administration, while Figure 2 B shows the effect of combined drug administration on the activation of the cell necrosis-apoptosis signaling pathway after ZBP1 gene knockout. Furthermore, Figure 2 C depicts the effect of combined drug administration on tumor curves in each group of mice with ZBP1 gene knockout.

[0135] Experimental data revealed that 5AZA alone can trigger ZBP1-MLKL-mediated apoptosis. When 5AZA is used in combination with an anti-HER2 antibody, a synergistic effect is produced, more effectively activating the signaling pathway for apoptosis. Further experiments, including in vivo and in vitro studies, confirmed that the antitumor effect of the combination therapy was significantly weakened when ZBP1 protein was knocked out, indicating that the combination of decitabine and anti-HER2 antibody depends on ZBP1-mediated apoptosis.

[0136] Example 3: Combination therapy of decitabine, a DNA methyltransferase inhibitor, and anti-HER2 antibody induces cellular senescence and promotes anti-tumor effects.

[0137] I. Experimental Materials

[0138] The experimental animals were 6-8 week old SPF-grade female BALB / c mice. The tumor cell line was TUBO cells, cultured in DMEM containing 10% fetal bovine serum at 37°C and 5% CO2. The DNA methyltransferase inhibitor decitabine 5AZA was purchased from Sigma-Aldrich (CAS No.: 2353-33-5), the anti-HER2 antibody (7.16.4) from BioXcell (Lot: BE0277), and ABT-263 from Selleck (CAS No.: S1001), diluted with phosphate-buffered saline (PBS) before use. The fetal bovine serum was an Excel product, and the DMEM medium was a GIBCO product.

[0139] II. Experimental Methods

[0140] 1. Tumor cell grouping, xenograft model creation and grouping

[0141] In the in vitro combination therapy experiment, TUBO cells were seeded into culture plates the night before the experiment. The experiment began on day 1, with TUBO cells treated with a combination of 5-AZA (0.15 μM), anti-HER2 antibody (1.5 μg / mL), and ABT-263 (0.25 μM). The following day, 5-AZA was removed, and the cells were cultured for another 4 days with anti-HER2 antibody (1.5 μg / mL) and ABT-263 (0.25 μM).

[0142] In the culture of tumor cells in the logarithmic growth phase, the original culture medium was first removed, and the cells were then washed with PBS. Next, the cells were treated with trypsin until they detached from the culture wall. After trypsin treatment, the trypsin was removed, fresh culture medium was added, and the cells were gently pipetted to disperse them. The cells were then transferred to centrifuge tubes and centrifuged at 1000 rpm for 5 minutes to pellet the cells. After centrifugation, the supernatant was removed, and the cells were washed again with PBS to prepare a cell suspension. Finally, the cell suspension was diluted to 0.1 mL per mouse, containing 5 × 10⁶ cells / mL. 5 A concentration of [number] cells was administered subcutaneously to the back of mice. When the tumor volume grew to approximately 100 mm... 3 Then, the mice were randomly divided into four groups for subsequent experiments:

[0143] (A) Ctrl group;

[0144] (B) Group 5AZA;

[0145] (C) anti-HER2 group;

[0146] (D)5AZA+anti-HER2 group.

[0147] 2. Drug intervention, Ki-67 and P21 immunohistostaining

[0148] To assess whether the combination therapy induced cellular senescence, TUBO cells treated with the drug were stained with galactosidase to observe whether cellular senescence was induced after the combination therapy. To evaluate the important role of cellular senescence in the combination therapy, Western blot was used to detect the expression levels of key proteins in the ZBP1-MLKL cell necrosis-apoptosis signaling pathway after eliminating senescent cells. Five days after drug treatment, total cellular protein was extracted and Western blot was performed to observe the specific effects of cellular senescence on the expression of the necrosis-apoptosis-related protein (ZBP1) after the combined use of 5AZA and anti-HER2 antibody, thereby further understanding the potential mechanism and efficacy of this combination therapy.

[0149] The tumor, inoculated with tumor cells, grew to 100mm. 3 At that time, the patients were randomly assigned to groups before administration. On day 9, each group received an intraperitoneal injection of the following doses of anti-HER2 antibody drug and an intratumoral injection of the following doses of 5AZA:

[0150] (A) Solvent control group: equal volume of solvent;

[0151] (B) 5AZA group: 5AZA 2mg / kg;

[0152] (C) Anti-HER2 group: 50 μg of anti-HER2 antibody per animal;

[0153] (D) 5AZA+anti-HER2 group: 5AZA 2mg / kg + anti-HER2 antibody 50μg / each.

[0154] Animal tissue was harvested on day 16. Tumor tissue from each group was completely removed, prepared into frozen sections, and subjected to Ki-67 and P21 immunohistochemical staining.

[0155] III. Experimental Results

[0156] At the end of the experiment, five mice survived in each group, a result that meets the standard requirements for pharmacological experiments. Figure 3 A shows the staining results of galactosidase after combined drug administration, while Figure 3 B and Figure 3 C shows the staining of P21 and Ki-67 in each group of mice after combined drug administration. Figure 3 D depicts the changes in the expression levels of key proteins in the necrosis-apoptosis signaling pathway of ZBP1-MLKL cells after drug clearance of senescent cells and combined drug administration.

[0157] Experimental results showed that the combined application of 5AZA and anti-HER2 antibody significantly promoted cellular senescence in both in vitro and in vivo experiments. Further research revealed that ZBP1 activation levels were significantly reduced after these senescent cells were cleared using the drug. This indicates that the combined drug strategy, by inducing cellular senescence, can more effectively activate the signaling pathways of necrosis and apoptosis, thereby exerting a powerful anti-tumor effect.

[0158] Example 4: Combination therapy of the DNA methyltransferase inhibitor decitabine and anti-HER2 antibody promotes anti-tumor effects by inducing SETDB1 inactivation through cellular senescence.

[0159] I. Experimental Materials

[0160] The experimental animals were 6-8 week old SPF-grade female BALB / c mice. The tumor cell line was CRISPR-Ctrl, SETDB1-gRNA TUBO cells, cultured in DMEM containing 10% fetal bovine serum at 37°C and 5% CO2. The DNA methyltransferase inhibitor decitabine 5AZA was purchased from Sigma-Aldrich (CAS No.: 2353-33-5), the anti-HER2 antibody from BioXcell (Lot: BE0277), and ABT-263 from Selleck (CAS No.: S1001), diluted with phosphate-buffered saline (PBS) before use. The fetal bovine serum was an Excel product, and the DMEM medium was a GIBCO product.

[0161] II. Experimental Methods

[0162] 1. Tumor cell grouping, xenograft model creation and grouping

[0163] In in vitro combined therapy experiments, TUBO cells or CRISPR-Ctrl, SETDB1-gRNA TUBO cells were seeded into culture plates the night before the experiment. The experiment began on day 1, with TUBO cells treated with a combination of 5-AZA (0.15 μM), anti-HER2 antibody (1.5 μg / mL), and ABT-263 (0.25 μM). The following day, 5-AZA was removed, and the cells were cultured for another 4 days with anti-HER2 antibody (1.5 μg / mL) and ABT-263 (0.25 μM).

[0164] In the culture of tumor cells in the logarithmic growth phase, the original culture medium was first removed, and the cells were then washed with PBS. Next, the cells were treated with trypsin until they detached from the culture wall. After trypsin treatment, the trypsin was removed, fresh culture medium was added, and the cells were gently pipetted to disperse them. The cells were then transferred to centrifuge tubes and centrifuged at 1000 rpm for 5 minutes to pellet the cells. After centrifugation, the supernatant was removed, and the cells were washed again with PBS to prepare a cell suspension. Finally, the cell suspension was diluted to 0.1 mL per mouse, containing 5 × 10⁶ cells / mL. 5 A concentration of [number] cells was administered subcutaneously to the back of mice. When the tumor volume grew to approximately 100 mm... 3 At that time, the mice were randomly divided into four groups for subsequent experiments:

[0165] (A) Ctrl group;

[0166] (B) Group 5AZA;

[0167] (C) anti-HER2 group;

[0168] (D)5AZA+anti-HER2 group.

[0169] 2. Drug intervention, SETDB1 immunohistostaining

[0170] To assess whether the combination therapy affected SETDB1 protein expression, Western blotting was used to examine TUBO cells after drug treatment to observe whether the combination therapy inhibited SETDB1 protein expression. To evaluate whether the combination therapy could induce SETDB1 inactivation through cellular senescence and promote anti-tumor effects, Western blot was used to detect the expression levels of SETDB1 protein after eliminating senescent cells and after combination therapy. Western blot was also used to examine the specific effects of drug elimination of senescent cells and combination therapy on the expression of necrosis-associated protein (ZBP1) in SETDB1 knockout cells.

[0171] The tumor, inoculated with tumor cells, grew to 100mm. 3 At that time, the patients were randomly assigned to groups before administration. On day 9, each group received an intraperitoneal injection of the following doses of anti-HER2 antibody drug and an intratumoral injection of the following doses of 5AZA:

[0172] (A) Solvent control group: equal volume of solvent;

[0173] (B) 5AZA group: 5AZA 2mg / kg;

[0174] (C) Anti-HER2 group: 50 μg of anti-HER2 antibody per animal;

[0175] (D) 5AZA+anti-HER2 group: 5AZA 2mg / kg + anti-HER2 antibody 50μg / each.

[0176] Animal tissue was harvested on day 16. Tumor tissue from each group was completely removed, prepared into frozen sections, and subjected to SETDB1 immunohistoscopy.

[0177] III. Experimental Results

[0178] At the end of the experiment, five mice survived in each group, a result that meets the standard requirements for pharmacological experiments. Figure 4 AB presented the expression of SETDB1 protein after combination therapy. Figure 4 C depicts the changes in SETDB1 protein expression levels after drug clearance of senescent cells and combined drug administration; Figure 4 D depicts the changes in expression levels of key proteins in the signaling pathway of cell necrosis and apoptosis in SETDB1 knockout cells after drug clearance of senescent cells and combined drug administration.

[0179] Experimental results showed that the combined application of 5AZA and anti-HER2 antibody significantly inhibited SETDB1 protein expression in both in vitro and in vivo experiments. Further research revealed that after clearing these senescent cells with the drug, the protein expression level of SETDB1 partially recovered after combined administration. Finally, in SETDB1 knockout cells, activation of the cell necrosis and apoptosis signaling pathway was observed after both drug clearance of senescent cells and combined administration, indicating that the combined treatment of 5AZA and anti-HER2 antibody promotes anti-tumor effects by inducing SETDB1 inactivation through cellular senescence.

[0180] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims. Furthermore, all documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference.

Claims

1. Use of decitabine or its analogues, in combination with anti-HER2 antibodies, in the preparation of medicaments for the treatment of tumors.

2. Use of decitabine or its analogues, anti-HER2 antibodies, and one, more or all of the following selected senescence-inducing drugs, ZBP1 agonists and SETDB1 inhibitors in the preparation of medicaments for treating tumors.

3. The use as described in claim 1 or 2, characterized in that, The decitabine has the structural formula shown in Formula I: And / or, The decitabine analogues include decitabine hydrates, solvates, enantiomers, diastereomers, pharmaceutically acceptable salts, prodrugs, and complexes. And / or, The decitabine analogues include 5-azacytidine, 5-fluoro-2-deoxycytidine, and zebularine.

4. The use as described in any one of claims 1-3, characterized in that, The anti-HER2 antibody is an antibody or its antigen-binding fragment that binds to the epidermal growth factor 2 (HER2) antigen. Preferably, the antibody comprises: monoclonal antibody, polyclonal antibody, multispecific antibody, humanized full-length human antibody, chimeric antibody, and camel-derived single-domain antibody; the antigen-binding fragment comprises: Fab, Fab', F(ab')2, Fv, double-chain antibody, linear antibody, single-chain antibody, and nanobody; More preferably, the anti-HER2 antibody includes: trastuzumab, pertuzumab, inetuzumab, and maggotuximab.

5. The use as described in any one of claims 2-4, characterized in that, The cell senescence-inducing drug is a drug that can induce cell senescence and / or induce cell necrosis and apoptosis; Preferably, the cell senescence-inducing drug includes: chemotherapeutic drugs, RAS / RAF / MEK inhibitors, CDK4 / 6 inhibitors, telomerase inhibitors, DNA methyltransferase inhibitors, histone deacetylase inhibitors, and MDM2 inhibitors; More preferably, the chemotherapeutic drugs include doxorubicin, etoposide, camptothecin, cisplatin, carboplatin, oxaliplatin, Nutlin3, and RG7112; the RAS / RAF / MEK inhibitors include sorafenib and lysyl The inhibitors include oxidase, Selumetinib (AZD6244), Trametinib, and Pimasertib; CDK4 / 6 inhibitors include Palbociclib, Ribociclib, Abemaciclib, and Dalpiciclib; telomerase inhibitors include Imetelstat (GRN163L, Rytelo), BIBR1532, and THIO (6-thio-dG, 6-thio-2'-deoxyguanosine); DNA methyltransferase inhibitors include azacitidine, zabulolin, RG108, GSK3685032, and SGI-110; histone deacetylase inhibitors include vorinostat, romidesin, GSK-LSD1, Entinostat, Valproic acid, Tubacin, and ACY-1215; and MDM2 inhibitors include Nutlin-3a, APG-115, MI-773, AMG-232, and Idasanutlin.

6. The use as described in any one of claims 2-5, characterized in that, The ZBP1 agonist is any substance that can enhance the activity of ZBP1, increase the stability of ZBP1 or its encoding gene, upregulate the expression of ZBP1, increase the effective duration of ZBP1, promote the transcription and translation of the ZBP1 gene, or promote ZBP1-mediated cell necrosis and apoptosis. Preferably, the ZBP1 agonist includes CBL0137, hydrogen peroxide, arsenite, cisplatin, and ADAR1 inhibitor.

7. The use as described in any one of claims 2-6, characterized in that, The SETDB1 inhibitor is a substance that can reduce the activity of SETDB1, reduce the stability of SETDB1 or its encoding gene, downregulate the expression of SETDB1, reduce the effective action time of SETDB1, inhibit the transcription and translation of the SETDB1 gene, or promote cell necrosis and apoptosis by inhibiting SETDB1. Preferably, the SETDB1 inhibitor includes Mithramycin A, Mithralog EC-8042, 3'-deazaneplanocin A (DZNep), Paclitaxel, (R,R)-59, and SETDB1-TTD-IN-1.

8. The use as described in any one of claims 1-7, characterized in that, The tumor treatment includes: inhibiting tumor cell methylation, promoting tumor cell senescence, promoting tumor cell necrosis and apoptosis, inhibiting tumor cell growth, and / or reducing tumor cell drug resistance; preferably, promoting tumor cell senescence includes: inducing SETDB1 inactivation; promoting tumor cell necrosis and apoptosis includes: activating ZBP1; more preferably, the tumor cells are HER2-positive tumor cells, more preferably HER2-positive breast cancer cells; and / or, The tumor is a tumor containing or expressing the HER2 antigen, preferably including primary or metastatic cancer; more preferably, the tumor is a HER2-positive tumor; even more preferably, the tumor includes breast cancer, gastric cancer, colon cancer, liver cancer, small cell lung cancer, non-small cell lung cancer, melanoma, pancreatic cancer, skin cancer, uterine cancer, ovarian cancer, Kaposi's sarcoma, squamous cell carcinoma, or a combination thereof.

9. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises an anti-HER2 antibody and decitabine or an analogue thereof; Preferably, the decitabine or its analogues are as described in claim 3, and / or the anti-HER2 antibody is as described in claim 4; Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or one or more additional active ingredients; more preferably, the additional active ingredients are selected from one, more, or all of the following: a cell senescence-inducing drug, a ZBP1 agonist, and a SETDB1 inhibitor; more preferably, the cell senescence-inducing drug is as described in claim 5, the ZBP1 agonist is as described in claim 6, and / or the SETDB1 inhibitor is as described in claim 7; Preferably, the pharmaceutical composition is a pharmaceutical composition for treating tumors; more preferably, the treatment of tumors is as described in claim 8, and / or the tumor is as described in claim 8.

10. A medicine box, characterized in that, The medicine box includes the pharmaceutical composition as described in claim 9.

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